CN101218253A

CN101218253A - Cardiovascular disease therapies

Info

Publication number: CN101218253A
Application number: CNA2006800244832A
Authority: CN
Inventors: I·朗赛特默帕罗伯克; C·T·雅各布; D·Y·刘
Original assignee: Fibrogen Inc
Current assignee: Fibrogen Inc
Priority date: 2005-05-05
Filing date: 2006-05-05
Publication date: 2008-07-09
Also published as: CN101213213A; ZA200710200B; ZA200710198B

Abstract

The present invention relates to methods and agents for treating impaired vascular and cardiac function. Methods and agents for treating various physiological and pathological features associated with vascular dysfunction and cardiac dysfunction are also provided.

Description

Cardiovascular disease therapies

The series number that the application requires on May 5th, 2005 to submit to is 60/678, the series number that 495 U.S. Provisional Application, on June 15th, 2005 submit to is 60/690, the series number that 957 U.S. Provisional Application and on November 7th, 2005 submit to is 60/734,433 U.S. Provisional Application No., each application is introduced the application as a reference in full.

Technical field

The present invention relates to be used for the treatment of the method and the medicament of impaired blood vessel and heart function.The present invention also is provided for treating the various physiology relevant with dysfunction of blood vessel and cardiac dysfunction and the method and the medicament of pathology performance.

Background technology

Diabetic vascular complications has destructiveness, can influence each major organs.The diabetic subject has higher atherosclerosis, cardiovascular, peripheral blood vessel and cerebrovascular disease sickness rate.Vascular complication and vascular disease are the major causes of most of glycosuria patients' death and morbidity.

Several factors causes vascular disease takes place in the diabetic subject, comprises hyperglycemia, hypertension, obesity and hyperlipemia.For example, the glucose level that increases (being hyperglycemia) is considered to the microvascular complication major cause of (comprising retinopathy, ephrosis and neuropathy), also is considered to relevant with the great vessels complication (as cardiovascular disorder and peripheral vascular disease) of accelerated development too early.Glucose level that increases and oxidative stress all can quicken the formation of advanced glycation end products (AGE) as the feature of diabetes microenvironment.Hyperglycemia and AGE can cause the variation (as damage) of the normal configuration and the function of endothelium, and can cause endothelial function disturbance.In addition, hyperglycemia is relevant with various vascular complications (comprising the damage to the capillary blood vessel system and the great vessels system of human body) with AGE.Moreover hyperglycemia is relevant with inner membrance middle level thickness with hyperlipemia, and inner membrance middle level thickness is the mark of the cardiovascular and cerebrovascular disease of generally acknowledging.

The diabetes cardiovascular complication is serious, and is the major cause of this disease M ﹠ M.These complication comprise coronary heart disease (CHD), congestive heart failure, apoplexy, peripheral arterial disease, myocardosis, ephrosis, retinopathy and neuropathy.

Vascular disease can take place in the lump with other complication (for example diabetic nephropathy) in the diabetic subject, and the glycosuria ephrosis is a kind of common and common very serious illness.Cardiovascular disorder is to suffer from the diabetic subject's of end stagerenaldisease (ESRD) underlying cause of death (McMillan et al. (1990) BMJ301:540-544; Hirschl et al. (1992) Am J KidneyDis.20:564-568 and Rischen-Vos et al. (1992) Nephrol Dial Transplant 7:433-437.).

The diabetic subject has serious congestive heart failure risk.Some factors are facilitated the high incidence of diabetic cardiomyopathy, comprise long-term hypertension, chronic hyperglycemia, serious coronary atherosclerosis etc.The diabetic subject compares with the patient of non-diabetic, and the mortality ratio that apoplexy causes increases near 3 times (Stamler et al. (1993) Diabetes Care 16:434-444.).In addition, diabetes can increase the possibility (Folsom et al. (1994) Stroke 16:434-444. and O ' Leary et al. (1992) Stroke 25:66-73.) of serious carotid atherosclerosis.

The sickness rate that the diabetic subject suffers from peripheral vascular disease (PVD) than high about 4 times of the patient of non-diabetic (referring to, H.E.Lebovitz for example, ed. Therapy for diabetes mellitus(3 ^RdEdition), the Gibbons among the pp.290-302. (1998) Peripheral vascular disease).Peripheral vascular disease is because atherosclerosis causes leg arteries, the illness that causes the arm artery to narrow down sometimes.It causes the ulcer of lower limb, poor wound healing and resists the ability reduction of infection.Its reason comprises and postpones or stop oxygen (local asphyxia), nutrient substance and microbiotic are transported to infected zone, and immunne response impaired (referring to aforementioned Gibbons).The illness that other is relevant with diabetes can further increase the chance that PVD takes place the diabetic subject as hypertension, obesity and hyperlipemia.

At present with treatment or reduce the progress of diabetic vascular complications and seriousness or used several different methods, comprise and optimize glycemic control (by changing diet and/or insulinize), hypertension control (comprise and bestow Zinc metallopeptidase Zace1 (ACE) inhibitor and angiotensin receptor blocker (ARB)), cholesterol lowering therapeutic etc. to reduce hypertension with the therapeutic strategy that occurs as target that prevents described complication.But described therapy is not general success, and is often reversing vascular complication relevant with diabetes and symptom (for example, damaging) or be invalid in the function aspects of improvement influenced blood vessel, organ and tissue.Moreover, because for example aging population, and fat and sedentary life factor such as popular increase in the U.S. and global population, global diabetes popularity is all in increase.Therefore, diabetes are global major risk factors of vascular disease.Therefore, the demand of method to the treatment diabetic vascular complications is arranged in described technical field, the demand to the method for the progress that reduces these vascular complications and seriousness is also arranged, and to the demand of the method for the generation that prevents these complication.In addition, method and medicament to effective reverse vascular complication relevant with diabetes in described technical field also have demand, comprise and reverse symptom and, and improve the function that is subjected to blood vessel, organ and tissue that the vascular complication relevant with diabetes influence the damage of blood vessel.

The present invention confirms first: in diabetes animal model, the specificity of CTGF is suppressed to alleviate dysfunction of blood vessel can improve vascular function significantly again, thereby can satisfy above-mentioned needs.Especially, the present invention confirms to use that anti-CTGF antibody capable alleviates blood vessel hardness, vascular permeability effectively, exosmose (for example oedema) and angiosteosis in diabetes animal model.Method of the present invention and medicament also can reverse the symptom relevant with diabetes and effectively to the damage of blood vessel.

The present invention also confirms the specificity of CTGF is suppressed to improve heart function.Especially, the present invention confirms that using anti-CTGF antibody can significantly improve heart function, and this improvement by following parameter is confirmed: ventricular diastole, ventricular systole, diastole end pressure, diastole end volume, ejection fraction, arterial elasticity, stroke output and cardiac output.

Description of drawings

Fig. 1 has shown that method of the present invention and medicament improve cardiac ejection fraction effectively in mammalian object.

Fig. 2 has shown that method of the present invention and medicament reduce left ventricular end diastolic effectively and press in mammalian object.

Fig. 3 has shown that method of the present invention and medicament improve myocardial contraction effectively in mammalian object.

Fig. 4 has shown that method of the present invention and medicament improve ventricular diastole effectively in mammalian object.

Fig. 5 has shown that method of the present invention and medicament alleviate carotid artery effectively and axially harden in mammalian object.

Fig. 6 has shown that method of the present invention and medicament alleviate carotid artery effectively and circumferentially harden in mammalian object.

Fig. 7 has shown that method of the present invention and medicament alleviate effectively with the reverse carotid artery relevant with diabetes and has axially hardened in mammalian object.

Fig. 8 has shown that method of the present invention and medicament alleviate effectively with the reverse carotid artery relevant with diabetes and has circumferentially hardened in mammalian object.

Fig. 9 has shown that method of the present invention and medicament reverse the reduction of axial tension rate in the body effectively in mammalian object.

Figure 10 has shown that method of the present invention and medicament reverse the increase of Aorta ring open-angle effectively in mammalian object.

Figure 11 has shown that method of the present invention and medicament alleviate oedema outside the blood vessel effectively in mammalian object.

Figure 12 has shown that method of the present invention and medicament reduce vascular permeability effectively in mammalian object.

Figure 13 has shown that method of the present invention and medicament reduce oedema outside vascular permeability relevant with diabetes with reverse and the blood vessel effectively in mammalian object.

Figure 14 has shown that method of the present invention and medicament alleviate angiosteosis effectively in mammalian object.

Figure 15 has shown that method of the present invention and medicament reduce cardiac weight effectively in diabetes animal model.

Figure 16 has shown that method of the present invention and medicament reduce the ratio of cardiac weight and body weight effectively in mammalian object.

Figure 17 has shown that method of the present invention and medicament reduce low-density lipoprotein in the blood (LDL) level effectively in mammalian object.

Figure 18 has shown that method of the present invention and medicament reduce HbA1c level in the blood effectively in mammalian object.

Figure 19 has shown that method of the present invention and medicament reduce HbA1c level in the blood effectively in mammalian object.

Figure 20 has shown that anti--CTGF antibodies angiotensin receptor blocker (ARB) therapy has increased blood middle-high density lipoprotein (HDL) level in mammalian object.

Summary of the invention

The invention provides a kind of method that is used to alleviate the dysfunction of blood vessel of object or improves vascular function, described method comprises the anti-CTGF agent of using significant quantity to described object, thereby alleviates the dysfunction of blood vessel of described object or improve vascular function.

In each embodiment, described to as if the object suffering from diabetes or have the risk of suffering from diabetes.

In specific embodiment, vascular function can be capillary blood vessel function or great vessels function, perhaps relevant with peripheral blood vessel vascular function.

In some embodiments, method of the present invention be particularly useful for reducing endothelial function disturbance, treat or alleviate arteriosclerosis particularly axially, radially or circumferentially arteriosclerosis, reduce vascular permeability, reduce and exosmose, reduce the inner membrance middle level and thicken or reduce arteria carotis communis inner membrance middle level and thicken.Especially, method of the present invention can be used for alleviating oedema or tissue edema.Similarly, method of the present invention also can be used for alleviating damage or the dysfunction of blood vessel to vascular function; Or alleviate angiosteosis.

Method of the present invention can be used in particular for treating the vascular complication relevant with diabetes.

The object of described method can be a Mammals.Especially, described object can be the people.

The anti-CTGF agent that is used for method of the present invention can for example be polypeptide, polynucleotide or small molecules.Especially, described anti-CTGF agent can be and CTGF or its fragment bonded antibody, antisense molecule, siRNA or micromolecular compound.In specific embodiment, described anti-CTGF agent is at CTGF or its segmental monoclonal antibody.In other specific embodiment, described anti-CTGF agent is CLN-1 or its fragment of describing among the WO 2004/108764.

Detailed Description Of The Invention

They should be appreciated that the specific method, scheme, clone, mensuration and the reagent that the invention is not restricted to described in the application, because can change.Also should understand the used term of the application and be intended to describe specific implementations of the present invention, rather than in order to limit the scope of the present invention shown in claims.

Must be noted that unless context has clear indication, otherwise used singulative " a kind of " and " described " comprises plural form in the application and claims.Therefore, for example " a kind of fragment " comprised numerous these class fragments, and a kind of " antibody " refers to the Equivalent of one or more antibody and this antibody well known by persons skilled in the art, or the like.

Except as otherwise noted, employed all technology and scientific terminology have the identical meanings of one skilled in the art of the present invention institute common sense among the application.Although all can implement and test among the present invention and use with any method and the material of the method for describing among the application and materials similar or equivalence, what describe in this application be preferable methods, device and material.All publications that the application quotes are introduced the application as a reference in full with it, its objective is describe and public publication in report can be used for method of the present invention, reagent and means.Any content among the application should not be regarded as all admitting that the present invention is not prior to this open according to what formerly invent.

Unless specialize, enforcement of the present invention will be adopted the ordinary method in chemistry, biological chemistry, molecular biology, cytobiology, genetics, immunology and the area of pharmacology.These technology are fully explained in the literature.Referring to, Gennaro for example, A.R.ed. (1990) Remington ' sPharmaceutical Sciences, 18th ed.Mack Publishing Co.Colowick, S.et al.eds.Methods In Enzymology, Academic Press, Inc.Handbook ofExperimental Immunology, Vols.I-IV (D.M.Weir and C.C.Blackwell, eds.1986, Blackwell Scientific Publications); Maniatis, T.et al.eds. (1989) Molecular Cloning:A Laboratory Manual, 2nd edition, Vols.I-III, Cold SpringHarbor Laboratory Press; Ausubel, F.M.et al.eds. (1999) Short Protocols inMolecular Biology, 4th edition, John Wiley ﹠amp; Sons; Ream et al.eds. (1998) Molecular Biology Techniques:An Intensive Laboratory Course, AcademicPress); PCR (Introduction to Biotechniques Series), 2nd ed. (Newton ﹠amp; Grahameds.1997, Springer Verlag).

The present invention partly relates to a discovery, and described discovery refers to Connective Tissue Growth Factor (CTGF) in cardiovascular disorder, comprises the particular aspects of vascular function impaired (being dysfunction of blood vessel) and heart function impaired (being cardiac dysfunction), the performance keying action.

The present invention's part has unexpected benefit based on a plurality of concrete aspect that is suppressed at treatment dysfunction of blood vessel and cardiac dysfunction of finding CTGF.Before having reduced, data acknowledgement provided by the invention, the inhibition of CTGF think symptom with the irrelevant various cardiovascular disordeies of CTGF.In some aspects, evidence provided by the invention shows, provides a kind of in order to treatment or prevention dysfunction of blood vessel and the specific physiology of cardiac dysfunction and the methods of treatment of pathological condition by suppressing CTGF.

The invention provides the method and the medicament that are used for reducing, improve or reverse the object complication relevant with multiple different pathologic process, described multiple different pathologic process and impaired vascular function and impaired heart function are relevant.In some embodiments, described optimally is Mammals to liking animal, is the people optimization.

The present invention also is provided for the medicament of method of the present invention.Described medicament can comprise micromolecular compound, peptide and protein (comprising antibody or its functionally active fragment), reach polynucleotide (comprising small interference ribonucleic acid (siRNA), little RNA (miRNA), ribozyme and antisense sequences).(referring to, Zeng (2003) Proc Natl Acad Sci USA 100:9779-9784 for example; And Kurreck (2003) Eur JBiochem 270:1628-1644.)

Impaired vascular function

Impaired vascular function (for example, dysfunction of blood vessel) can be caused by various unusual institutes, comprises that the structure of vascular system for example and/or function are interfered or damage.The various pathologies of impaired vascular function and blood vessel are relevant with damage, cause deleterious variation, and for example cause arteriosclerosis, blood vessel is penetrating and angiosteosis.

Arterial stiffness comprises for example characteristics such as blood vessel swelling property, conformability and Young's modulus, and has been proved to be and can predicts well that coronary heart disease and cardiovascular disorder mortality ratio are (referring to, O ' Rourke et al (2002) Am J Hypertens 15:426-444 for example; Boutouyrie et al (2002) Hypertension39:10-15; Blacher et al (1999) Circulation 99:2434-2439).Generally speaking, the arterial stiffness increase can cause systolic pressure increase, ventricular mass to increase and coronary perfusion reduction diastole.The arterial stiffness increase also reduces relevant (referring to, Suzuki et al (2001) Diabetes Care 24:2107-2114 for example) with the artery of lower extremity flow.

It is relevant with the numerous disease state that comprises diabetes and chronic nephropathy that arterial stiffness increases (being arteriosclerosis), also is the sign of aging course simultaneously.Arteriosclerosis is paid close attention in diabetes or metabolism syndrome patient especially, and in these diseases, arteriosclerosis occurs in that all age groups are consistent.Arteriosclerosis also is to suffer from the mark that risk of cardiovascular diseases increases, and described cardiovascular disorder comprises for example myocardial infarction, heart failure, general mortality rate, apoplexy, dementia and ephrosis.

Arteriosclerosis is relevant with deleterious effect to the 26S Proteasome Structure and Function of blood vessel and heart.In artery, arteriosclerosis influences the variation of mechanicalness vascular stimulation, causes endothelial function disturbance and vascular disease.In heart, arteriosclerosis influences the perfusion of ventricle working load, cardiac ejection efficient and heart itself.

Arteriosclerotic clinical alternative terminal point mark comprises that isolated systolic hypertension and pulse pressure increase.Pulse pressure difference is poor between systolic pressure and the diastolic pressure.Arteriosclerosis increases relevant with pulse pressure.Pulse pressure increases to and is higher than normal value indication and has arteriosclerosis.Pulse wave velocity (PWV) is measured and also can be assessed arterial stiffness (referring to, Lehman et al (1992) Diabetic Med 9:114-119. for example).Pulse wave velocity is that the Ve pressure wave leaves heart along artery progressive speed.Pulse wave is decided according to the hardness of this artery along the artery rate of advance.Measuring pulse wave velocity carries out between carotid artery and femoral artery usually.Slow wave speed expression artery has plasticity-, and fast velocity of wave is represented arteriosclerosis; Therefore, pulse wave velocity is high more, the rigidity of vessel wall and hardness are high more and swelling property is low more (referring to, Nichols andO ' Rourke in McDonald ' s blood flow in arteries.Theoretical for example, experimental andclinical principles.Fourth Edition, London, Sydney, Auckland:Arnold are E.1998.).The method that these methods and other are used to measure arterial stiffness is well-known in the present technique field and is that those skilled in the art are available at any time.

Angiosteosis (comprising coronary artery calcification, valvular calcification, calcific aortic and arteriosteogenesis) has a direct impact vascular function and heart function.Angiosteosis is relevant with expansion with the sclerosis of the wall of great vessels (for example aorta and arteria carotis communis), and influences vascular function by contraction and the diastole that weakens blood vessel.Angiosteosis is the fixed index of coronary artery disease and vascular disease, in the generation of cardiovascular disorder (comprising that the impaired and heart function of vascular function is impaired) and pathogenesis, account for the key player, thereby with the myocardial infarction of high risk and dead relevant (referring to, Doherty et al (2004) Endocr Rev 25:629-627. for example).Angiosteosis all can take place in endarterium and tunica media of artery.

The invention provides the method and the medicament that can be used for treating impaired vascular function.In each embodiment, method of the present invention and medicament can be used for treating impaired vascular function, and wherein said impaired vascular function is arteriosclerosis, vascular permeability, exosmoses and angiosteosis.

In some aspects, the present invention relates to be used for alleviate dysfunction of blood vessel and improve the method for vascular function at the object of suffering from diabetes or having a risk of suffering from diabetes.In addition, the present invention relates to be used for suffering from diabetes or having the method for the object reverse of the risk of suffering from diabetes vascular complication, symptom or the damage relevant with the glucose level of diabetes or rising.In the preferred implementation of each method of following explanation, preferably to as if the people.

The invention provides a kind of method that is used for alleviating at the object of suffering from diabetes or having a risk of suffering from diabetes dysfunction of blood vessel, described method comprises the anti-CTGF agent of using significant quantity to described object, thereby alleviates the dysfunction of blood vessel of object.Term " anti-CTGF agent " is meant expression or the active medicament of any inhibition CTGF in this application.The present invention also provides a kind of to be suffered from diabetes or is having the method for improving vascular function in the object of the risk of suffering from diabetes, and described method comprises the anti-CTGF agent of using significant quantity to described object, thereby improves vascular function.

The invention provides a kind of method that is used for reversing vascular complication, symptom or damage at the object of the risk of suffering from the vascular complication relevant or have trouble the vascular complication relevant with diabetes with diabetes, described method comprises the anti-CTGF agent of using significant quantity to described object, thereby reverses vascular complication, symptom or the damage of described object.

Because described some method of the application relates to the object of suffering from diabetes or having the risk of suffering from diabetes, when judging whether described object suffers from diabetes or have the risk of suffering from diabetes, can use this area skill to state any method that personnel accept and use.For example, the people (by for example fasting plasma glucose test, oral glucose tolerance test or random blood sugar test determine) of glucose level more than about 200mg/dL can be regarded as suffering from the object of diabetes.Therefore, say in a sense that the people of glucose level more than about 200mg/dL is suitable for the object for the treatment of with the use of method provided by the present invention or medicine.Can identify object (people who for example has the risk of suffering from diabetes) by assessing one or more known factor relevant with the increase of diabetes occurrence risk with risk of suffering from diabetes, described factor comprises family history, some ethnic group or racial group, gestational diabetes medical history, obesity (especially high-caliber internal organ or stomach fat), sedentary life, age, hypertension and the schizophrenia etc. that diabetes are arranged in the family, comprises that also glucose metabolism changes (comprising that glucose tolerance reduces (IGT) or pre-diabetes).

In each embodiment of method of the present invention, described dysfunction of blood vessel is capillary blood vessel dysfunction or great vessels dysfunction.In some embodiments, described dysfunction is a cardiovascular functional disorder, or the cerebrovascular function obstacle, or the dysfunction of blood vessel relevant with ephrosis, retinopathy or neuropathy.In a specific embodiment, described dysfunction is a ventricular dysfunction, preferred left ventricular dysfunction.In one embodiment, described dysfunction is the peripheral blood vessel dysfunction.

The present invention also comprises the method for improving vascular function.On the one hand, the invention provides the method that is used for improving at the object of suffering from diabetes or having a risk of suffering from diabetes vascular function, described method comprises the anti-CTGF agent of using significant quantity to described object, thereby improves the vascular function of described object.

In each side of the present invention, described vascular function is capillary blood vessel function or great vessels function.In some embodiments, described vascular function is cardiovascular function or cerebrovascular function, or the vascular function relevant with ephrosis, retinopathy or neuropathy.In described function aspect certain is ventricular function, preferred left ventricular function.On the one hand, described vascular function is relevant with peripheral blood vessel.

The present invention further comprises the method that is used for alleviating at the object of suffering from diabetes or having a risk of suffering from diabetes endothelial function disturbance, and described method comprises the anti-CTGF agent of using significant quantity to described object, thereby alleviates the endothelial function disturbance of described object.The present invention also is provided to be suffered from diabetes or has the method for improving endothelial function in the object of the risk of suffering from diabetes, and described method comprises the anti-CTGF agent of using significant quantity to described object, thereby improves endothelial function.Especially, the present invention relates to reverse the method for endothelial function disturbance.

In one embodiment, the invention provides a kind of method of suffering from diabetes or having vascular complication relevant in the object of the risk of suffering from diabetes that is used for the treatment of with diabetes, described method comprises to described object uses anti-CTGF agent, thereby treats described vascular complication.Anti-CTGF agent is the medicament of the active and/or expression of any inhibition CTGF.The present invention also provides to be suffered from diabetes or is having the method for slowing down the process of vascular complication in the object of the risk of suffering from diabetes or alleviating its seriousness, described method comprises the anti-CTGF agent of using significant quantity to described object, thereby slows down the process of described vascular complication or alleviate its seriousness.The method that the present invention further provides prevention or slow down and suffer from diabetes or have the generation of the vascular complication in the object of the risk of suffering from diabetes, described method comprises the anti-CTGF agent of using significant quantity to described object, thus the generation of the prevention or the vascular complication that slows down.In another embodiment, the invention provides the method that is used for reversing the object of suffering from the vascular complication relevant, symptom or damage or has the object of the risk of suffering from vascular complication, symptom or the damage relevant vascular complication, symptom or the damage relevant with diabetes with diabetes with diabetes, described method comprises the anti-CTGF agent of using significant quantity to described object, thereby reverses vascular complication relevant with diabetes in the described object, symptom or damage.

In some embodiments, described vascular complication is the great vessels complication; Be microvascular complication in other embodiments.In each embodiment, described complication is selected from heart trouble, ephrosis, neuropathy and retinopathy.In one embodiment, described complication is cardiovascular complication or cerebrovascular complication.In another embodiment, described complication is the peripheral blood vessel complication.

The present invention also provides and has been used for the treatment of arteriosclerotic method, and described method comprises to suffering from diabetes or have the anti-CTGF agent that the object of the risk of suffering from diabetes is used significant quantity, thus treatment arteriosclerosis.In every respect, described arteriosclerosis be selected from axially, radially and circumferential arteriosclerosis.The present invention includes and reduce vascular permeability, reduce and exosmose (for example oedema or tissue edema), alleviate the inner membrance middle level and thicken and alleviate the method that arteria carotis communis inner membrance middle level thickens.In every kind of situation, described method comprises to suffering from diabetes or having the anti-CTGF agent that the object of the risk of suffering from diabetes is used significant quantity, thereby reach described desired result, for example be respectively and reduce vascular permeability, reduce and exosmose (for example oedema or tissue edema), alleviate the inner membrance middle level and thicken and alleviate arteria carotis communis inner membrance middle level and thicken.

On the one hand, the present invention relates to be used for alleviating blood vessel injury or the handicapped method of suffering from diabetes or having the object of the risk of suffering from diabetes, described method comprises to the object of suffering from diabetes or having a risk of suffering from diabetes uses the anti-CTGF agent of significant quantity, thereby alleviates the blood vessel injury or the dysfunction of described object.In every respect, described blood vessel may be the blood vessel in the great vessels system, for example main blood vessel such as aorta, coronary artery, carotid artery, cerebral artery vessel, the Renal artery, iliac artery, femoral artery He popliteal artery etc.; Maybe may be the blood vessel in the capillary blood vessel system, the little blood vessel of retina arteriole, renal glomerulus arteriole, vasa nervorum, heart arteriole and capillary bed relevant with peripheral nervous system etc. for example with eye, kidney, heart and maincenter.

On the one hand, the invention provides the method for the angiosteosis that is used for reducing object, described method comprises the anti-CTGF agent of using significant quantity to described object, thereby reduces the angiosteosis of described object.In some sense, as in the preceding method, described to as if the object suffering from diabetes or have the risk of suffering from diabetes.

One preferred embodiment in, described to as if Mammals, in most preferred embodiments, described to as if the people.In some embodiments, described to as if the object suffering from diabetes or have the risk of suffering from diabetes.

Heart function is impaired

Heart function impaired (for example cardiac dysfunction) and heart trouble can be from various unusual, comprise that the contraction of for example heart and cardiac muscle and/or diastole are interfered or damage.From clinical angle, the impaired unusual state of any heart function that can be considered of heart function, described state cause the speed of cardiac pumping not match with anabolic requirement and/or must it be matched with unusual higher ventricular diastole volume.Heart function impaired (for example cardiac dysfunction, heart failure) may be cardiac systolic function obstacle or cardiac diastolic function obstacle (being called systolic heart failure and diastolic heart failure again).

In the cardiac systolic function obstacle, main is unusually ventricle can not normal contraction and is discharged competent blood.The cardiac systolic function obstacle is impaired relevant with myocardial contraction, and the impaired heart contraction that causes of myocardial contraction weakens, thereby causes stroke output and cardiac output minimizing, ventricle emptying deficiency, cardiac dilatation and common ventricular diastole voltage rise height.Adult cardiac systolic function obstacle is defined as left ventricular ejection fraction and is lower than 45%.In the cardiac diastolic function obstacle, main is normally diastole and/or full of ventricle unusually.Ventricular diastole relevant with diastolic heart failure and full impaired meeting cause all raising in any diastolic volume ventricular diastole pressure.The ventricular systole of many heart function impaired subjects and ventricular diastole all present unusually.

There have few techniques can allow those skilled in the art be able to evaluation in object, diagnosis or evaluate cardiac function to be impaired.In general, these technology comprise for example electrocardiogram(ECG, ultrasonic cardiography, cardiac catheterization, vasography and heart nuclear imaging technology (comprising the radioisotope angiography art).

In some cases, when described object is in static and VEDP is normal or when raising, may present cardiac output under cardiac dysfunction or the situation in heart failure and stroke output reduces existing.In many cases, heart function may enough made the time spent deficiency when static.

Usually by measuring ejection fraction evaluate cardiac function, promptly measure the mark of the process cardiac pump blood of at every turn beating.Ejection fraction is the ratio of stroke output and EDD, method of calculation be the blood volume that penetrates from ventricle (be called stroke output, SV) divided by the blood volume behind the ventricular filling (EDD, EDV).The normal value of adult's object ejection fraction is 50-70%.Heart and injury can damage the ability that heart is effectively penetrated blood, thereby reduces ejection fraction.Ejection fraction will be hanged down clinically can show as cardiac dysfunction or heart failure.Ejection fraction is one of most important prediction index of prognosis relevant with cardiac dysfunction; Typically, the object prognosis mala of ejection fraction reduction.

Ejection fraction is measured with ultrasonic cardiography usually, wherein measures the volume of the chambers of the heart (for example ventricle) in cardiac cycle.Ejection fraction also can be measured with cardiac MRI, rapid scanning axial cardiac tomoscan (CT) imaging, ventriculography, gate SPECT and radiocontrast vasography or RAG.

Normal left ventricle is whenever fought and is penetrated the 50-80% of its EDD, and promptly its ejection fraction is 0.50 to 0.80.In the grownup, the left ventricular volume normal value is as follows: EDD is 72 ± 15mL/m ²(mean+SD), end-systolic volume are 20 ± 8mL/m ²

The present invention also comprises other method, comprises improving heart function respectively, improving ventricular diastole, increase ventricular systole power, increase the diastole end and press, increase EDD, improve ejection fraction, improve arterial elasticity, increase stroke output and increase kinemic method.In these methods each all comprises to suffering from diabetes or having the anti-CTGF agent that the object of the risk of suffering from diabetes is used significant quantity, thereby reach desired effect, for example in described object, improve heart function respectively, improve ventricular diastole, increase ventricular systole power, increase diastole end pressures, the phase EDD, improve ejection fraction, improve arterial elasticity, increase stroke output and increase cardiac output.

In one embodiment, the invention provides a kind of method that is used for the treatment of or prevents cardiac complication, described method comprises to object uses anti-CTGF agent, thus treatment or prevention cardiac complication.In some embodiments, described to as if the object suffering from diabetes or have the risk of suffering from diabetes.Described method is particularly useful for treatment or prevention is selected from cardiac hypertrophy, congestive heart failure and myocardiac cardiac complication.Cardiac complication be treated or be prevented to method of the present invention also can by LDL level in the reduction blood.

Anti-CTGF agent

In any method as described above, described anti-CTGF agent can be polypeptide, polynucleotide or small molecules especially, for example with CTGF bonded antibody, antisense molecule, siRNA and micromolecular compound etc.Especially, suppressing CTGF can realize by expression and the active method of any well-known adjusting CTGF in the present technique field.Preferable methods is to use anti-CTGF agent, and for example at the human monoclonal antibodies of CTGF, the method for the generation of the method for the expression of gene of CTGF, inhibition CTGF or the active method of inhibition CTGF also can be used for the present invention yet any inhibition is encoded.

The exemplary antibodies of using in the method for the invention is at for example US5, and 408,040, description is arranged among WO99/07407, WO99/33878 and the WO00/35936.An exemplary antibodies of using in the method for the invention has description in WO2004/10874, incorporate the document into the application in full by reference.Described antibody or its fragment can be used by the various means that those skilled in the art are familiar with.For example often be to come injection of antibodies by intravenous injection, peritoneal injection or subcutaneous injection.

The present invention has also described CTGF and has expressed and/or active micromolecular inhibitor; For example WO96/38172 confirms that for example the cAMP instrumentality of Toxins,exo-, cholera and 8Br-cAMP and so on is the inhibitor that CTGF expresses.Therefore, be confirmed as prostaglandin(PG) for example and/or prostacyclin analogs such as Iloprost compound (referring to, for example WO 00/02450; Ricupero et al. (1999) AmJ Physiol 277:L1165-1171; Also referring to Ertl et al. (1992) Am Rev Respir Dis 145:A19) and the potential phosphodiesterase IV inhibitors (referring to, Kohyama et al. (2002) Am J Respir Cell Mol Biol 26:694-701 for example), all can be used for regulating the expression of CTGF.In addition, serine/threonine mitogen-activated protein kinase inhibitor (especially p38), cyclin dependent kinase (for example CDK2), glycogen synthase kinase (GSK)-3 also may be relevant with the reduction that CTGF expresses (referring to for example, Matsuoka et al. (2002) Am J Physiol Lung Cell MolPhysiol 283:L103-L112; Yosimichi et al. (2001) Eur J Biochem 268:6058-6065; WO 01/38532 and WO 03/092584).These compounds can and be used by blas preparation in the present technique field.

In addition, polynucleotide comprise small interference ribonucleic acid (siRNA), micro ribonucleic acid (miRNA), ribozyme and antisense sequences, can be used in the method for the present invention in order to suppress expression and/or the generation (referring to for example Kondo et al. (2000) Biochem Biophys Res Commun278:119-124.) of CTGF.These technology are well-known in various equivalent modifications.The antisense constructs that target CTGF expresses has been described and has been used to reduce that CTGF expresses (referring to for example WO 96/38172 in the dissimilar cells; WO 00/27868; WO 00/35936; WO 03/053340; Kothapalli et al. (1997) Cell Growth Differ 8 (1): 61-68; Shimo et al. (1998) JBiochem (Tokyo) 124 (1): 130-140; And Uchio et al. (2004) Wound Repair Regen12:60-66.).This antisense constructs can be used for reducing the expression of CTGF, thereby alleviates or prevent CTGF inductive pathologic process.This construct can and be expressed instrumentality with appropriate carriers and is designed for cell-specific or tissue specific expression and composing type or inducible expression.This genetic constructs can and be used by blas preparation in the present technique field.

Therefore, in some embodiment of the present invention, described anti-CTGF agent is a CTGF antibody.In a preferred implementation, described antibody is the CTGF monoclonal antibody.In another preferred implementation, described antibody is people or humanization CTGF antibody.In a specific embodiment, described antibody is the CLN-1 that is put down in writing as in WO 2004/108764.In another embodiment, described medicament is a small molecules.In another embodiment, described medicament is a nucleic acid.In further embodiment, described nucleic acid is selected from ring nucleus thuja acid, oligonucleotide or polynucleotide.In specific embodiment, described medicament is antisense oligonucleotide or siRNA.

The present invention relates to the application of method of the present invention in conjunction with other therapies.In one embodiment, described method and another therapy are united use, for example in order to the curative effect of further enhancing to some pathological state.Two kinds of treatments can be implemented simultaneously or implement in succession, for example during a treatment time-histories, or after sb.'s illness took a turn for the worse and alleviate.In another embodiment, described method is united use with the methods of treatment (for example ACE inhibitor, ARB, statin, advanced glycosylation end product (AGE) inhibitor etc.) that other has similar or different binding modes.Those skilled in the art are familiar with treatment vascular complication and vascular disease prior treatment method, comprise for example ACE inhibitor, angiotensin receptor blocker, statin, advanced glycosylation end product inhibitor, calcium channel blocker etc.Especially, above-mentioned any therapeutical agent and the method for the invention are united use.

Proved that bradykinin can regulate CTGF and express, and the bradykinin b 1 receptor antagonist reduces the vascular permeability of diabetic animal (referring to for example Ricupero et al. (2000) J Biol Chem275:12475-12480; Lawson et al. (2005) Regul Pept 124:221-224; Tan et al. (2005) Am J Physiol Renal Physiol 288:F483-F492; Wilkinson-Berka andFletcher (2004) Curr Pharm Des 10:3313-3330.).Therefore, in some sense, the invention provides be used to regulate kallikrein-kassinin kinin (method of kallikrien-kinin system and medicament, to regulate the vascular complication of CTGF mediation, vascular permeability and exosmosing for example.Especially, use medicament and the method that kassinin kinin is put enzyme-kinin system of regulating separately, or unite use with the medicament and the method that suppress CTGF.

Pharmaceutical formulation and route of administration

Be familiar with as the present technique field, composition of the present invention can directly be used or use in the pharmaceutical composition that contains vehicle.Methods of treatment of the present invention can comprise the compound of the present invention of using significant quantity to the object of suffering from dysfunction of blood vessel or cardiac dysfunction (being that vascular function is impaired or heart function is impaired) or having a risk of suffering from dysfunction of blood vessel or cardiac dysfunction.One preferred embodiment in, described to as if Mammals, in most preferred embodiments, described to as if the people.

By routine test can be easily to determine compound or effective amount of drug (that is dosage), effectively and easily route of administration and proper formula.Provide in the prior art various prescriptions and delivery system (referring to for example above-mentioned Gennaro, ed. (2000) Remington ' s PharmaceuticalSciences; And above-mentioned Hardman, Limbird, and Gilman, eds. (2001) ThePharmacological Basis of Therapeutics.).

Suitable route of administration for example can comprise in oral cavity, rectum, part, nose, lung, eye, intestines and administered parenterally.Main administered parenterally approach comprises intravenous injection, intramuscular injection and subcutaneous injection.Accessory route of administration comprises in intraperitoneal, intra-arterial, intraarticular, intracardiac, the brain pond, in the intracutaneous, intralesional, intraocular, pleura, in the sheath, administration in intrauterine and the ventricle.Formulation types to be used and route of administration and whether preferred topical or whole body administration, according to indication to be treated with and physics, the chemistry and biology characteristic of medicine decide.

The pharmaceutical dosage form of The compounds of this invention can provide with systems such as abrupt release, controlled release, slowly-releasing or target administrations.Formulation commonly used for example comprises solution and suspension, (little) emulsion, ointment, gel and patch, liposome, tablet, coated tablet, soft or hard capsule, suppository, ovule (ovule), implants, amorphous or crystalline powder, aerosol and freeze-dried preparation.According to used route of administration, may need to use described medicine, for example syringe and syringe needle, sucker, pump, injection pen, medicator or special-purpose flask with particular device.Pharmaceutical dosage form often is made up of medicine, one or more vehicle and container/closed system.One or more vehicle (being also referred to as inert fraction) can be added in the compound of the present invention, make described medicine production, stability, administration and security be improved or promote, and the means that obtain required drug release pattern are provided.Therefore, can be depending on multiple factor to be added to the type of the vehicle in the medicine, for example the physics of medicine and chemical property, route of administration and manufacturing processed.In the art, be included in each pharmacopeia, provide acceptable vehicle on the pharmacopedics (referring to for example USP, JP, EP and BP, the FDA webpage ( Www.fda.gov), Inactive IngredientGuide 1996 and Handbook of Pharmaceutical Additives, ed.Ash; SynapseInformation Resources, Inc.2002.).

The pharmaceutical dosage form of The compounds of this invention can any in the art method of knowing be made, for example traditional mixing, screening, dissolving, fusing, granulation, make that coated tablet, compressing tablet, suspension, extruding, spraying drying, grinding, emulsification, (receiving/little) are encapsulated, embedding or freeze-drying method.As mentioned above, composition of the present invention can comprise acceptable inert fraction on one or more physiology, and described inert fraction can promote the activist to form the process of medicinal preparations.

Proper formula depends on required medicine-feeding way.For example when intravenous injection, described composition can be formulated in the aqueous solution, compatible buffers on the available if necessary physiology (comprising for example phosphoric acid salt, Histidine or citrate buffer) reaches tonicity agent (for example sodium-chlor or glucose) to adjust the pH value of prescription.Through mucous membrane or intranasal administration the time, preferred semi-solid state formulation, liquid formulation or patch, and may contain penetration enhancers.Such penetration agent is widely known by the people in the present technique field.In oral administration, described compound can be mixed with the liquid or solid formulation, makes abrupt release formulation, controlled release form or slow release formulation.Suitable oral dosage form comprises tablet, pill, coated tablet, soft capsule and hard capsule, liquid, gel, syrup, paste, suspension and emulsion.Described compound also can be formulated as composition for rectal administration, for example contains the suppository or the enema,retention of conventional suppository bases such as theobroma oil or other glyceride type.

Solid oral dosage form can be made with vehicle, and described vehicle can comprise that filler, disintegrating agent, (do, wet) tackiness agent, dissolving delay agent, lubricant, glidant, antitack agent, Zeo-karb, wetting agent, antioxidant, sanitas, tinting material and seasonings.These vehicle can be synthetic or natural origin.The example of described vehicle comprises derivatived cellulose, citric acid, Lin Suanergai, gelatin, magnesiumcarbonate, Stepanol MG/sodium, N.F,USP MANNITOL, polyoxyethylene glycol, polyvinylpyrrolidone, silicate, silicon-dioxide, Sodium Benzoate, sorbyl alcohol, starch, stearic acid or its salt, sugar (being glucose, sucrose, lactose etc.), talcum, tragacanth mucilage, (hydrogenation) vegetables oil and wax.The second alcohol and water can be used as the granulation co-adjuvant.In some cases, need carry out dressing to tablet with for example taste masking film, anti-hydrochloric acid in gastric juice film or extended release film.Natural and synthetic polymer through being usually used in that tablet is carried out dressing, is made coated tablet in conjunction with pigment, sugar and organic solvent or water.When capsule was more preferred than tablet, the powder of medicine, suspension or its solution can place compatible hard or soft capsule.

In one embodiment, but compound topical application of the present invention, as by skin patch, semisolid or liquid preparation example gel, (little) emulsion, ointment, solution, (receiving/little) suspension or whipping agent.The process that medicine penetrates in skin and the deep tissues can be regulated by the following method: for example use penetration enhancers; Suitable selection and combination by lipotropy, wetting ability or amphipathic vehicle (comprising water, organic solvent, wax, oils, synthetic and natural polymer, tensio-active agent and emulsifying agent); Regulate by the pH value; And use complexing agent.Other technology is iontophoresis for example, can be used for regulating the skin penetration of The compounds of this invention.For example wishing the MIN general exposure of medicine and carrying out in the situation of local application, preferably through skin or local application.

Use inhalation or during via nasal administration; convenient carry the form that is used for compound of the present invention that solution, suspension, emulsion are arranged or from the semi-solid state aerosol of compression wrap or atomizer, described compression wrap or atomizer can use propelling agent (for example halogenation charcoal of being given birth to by methane and ethane, carbonic acid gas or any other suitable gas Xing) usually.For local aerosol, can use hydrocarbon polymer for example butane, iso-butylene and pentane.When using the pressurized spray agent, can measure suitable dose unit by using valve to carry through the amount of measuring.Can prepare for example gelatine capsule or the cartridge case that are used for sucker or insufflator.Typically, they can contain the powder mixes of described compound and suitable powder matrix (as lactose or starch).

Normally aseptic also can the providing by unit dosage of composition of carrying out administered parenterally by injection is provided, and for example in ampoule, syringe, injection pen or in multi-dose container, the latter is contained sanitas usually.Described composition can be suspension, solution or the emulsion in oiliness or aqueous carrier, and can contain prescription medicament (formulatory agents) as damping fluid, tonicity agent, viscosity intensifier, tensio-active agent, suspension agent and dispersion agent, antioxidant, biocompatible polymer, sequestrant and sanitas.According to the injection site, described carrier may contain water, synthetic oil or vegetables oil and/or organic cosolvent.In some cases, for example under the situation that is freeze-dried products or enriched material, the preparation of administered parenterally needs reduction or dilution before administration.Provide the prolonged action preparation of controlled release or slowly-releasing The compounds of this invention to comprise to receive/microparticle or receive/little or non-micronization crystalline injectable suspensions.Except controlled release/sustained-release matrix of being familiar with in the art, also can use polymkeric substance for example poly(lactic acid), polyglycolic acid or its multipolymer as controlled release/sustained-release matrix.Other long-acting delivery system can need the implants of otch and the form of pump to provide.

The suitable carriers that is used for intravenous injection molecule of the present invention, known in the art, it comprises that group water solution for example contains the damping fluid of phosphoric acid salt or Histidine, this group water solution contain matrix (for example sodium hydroxide) with form the ionizing compound, as the sucrose or the sodium-chlor of tonicity agent.Can add cosolvent, for example polyoxyethylene glycol.These water based systems can be dissolved compound of the present invention effectively, and produce low toxicity when systemic administration.Can change each components in proportions in the solution system considerably, and can not damage solubleness and toxic characteristic.In addition, the characteristic of composition also can change.For example, can use hypotoxic tensio-active agent,, also can add biocompatible polymer such as polyvinylpyrrolidone with polyoxyethylene glycol or other cosolvent as polysorbate or poloxamer, and other sugar and polyvalent alcohol can replace glucose.

To can be used for the composition of methods of treatment of the present invention, can treat effective dose according to a preliminary estimate by the various technology of being familiar with for the people in the art.The predose that uses in the zooscopy can be decided according to the effective concentration of establishing in the cell culture test.For example can use and judge the dosage range that is fit to human subjects from the data of zooscopy and cell culture test acquisition.

The treatment effective dose or the amount of compound of the present invention, medicament or medicine are meant dosage or amount that described compound, medicament or medicine cause the doing well,improving in the object or prolong lifetime.The toxicity of described molecule and result of treatment can be determined in cell culture or laboratory animal by the standard pharmacy procedure, for example measure LD50 (mld) and ED50 (median effective dose).Toxicity is exactly therapeutic index with the dosage ratio of curative effect, and this index can be represented with the ratio of LD50/ED50.With the medicament with high therapeutic index serves as preferred.

Significant quantity or treatment have the treatment amount to be meant to cause the amount of the biology of researcher, animal doctor, doctor or other clinicist expectation or medical response (for example vascular function improves and heart function improvement etc.) in tissue, system, animal or human.

Dosage is good with the circulation composition scope that comprises ED50, and with rare or not have toxicity be good.Dosage can be in this scope according to used formulation and/or medicine-feeding way and different.Consider the detail of the object patient's condition, should select definite prescription, medicine-feeding way, dosage and spacing of doses according to method known in the art.

Dosage and spacing of doses can be adjusted individually, with activeconstituents level, i.e. minimum effective concentration (MEC) in the blood plasma that be enough to produce a desired effect (for example vascular function improvement, heart function improvement etc.) are provided.MEC can depend on each compound and be different, but can estimate according to for example vitro data and experimentation on animals.Reach the necessary dosage of MEC and depend on personal feature and medicine-feeding way.Under the situation of topical or selectivity absorption, effective partial concn of described medicine may be uncorrelated with blood substance concentration.

The medicament of using or the amount of composition can be depending on multiple factor, comprise sex, age and body weight, severity of disease, administering mode and the prescriber's of treatment target judgement.

If desired, composition of the present invention can provide in the packing of one or more unit dosage that contains activeconstituents or dispensation apparatus.Described packing or device can comprise for example tinsel or plastic foil (for example blister), or glass and rubber plug (for example in bottle).Described packing or dispensation apparatus can attach the specification sheets of administration.Also can be placed in the proper container, and stick treatment indication label comprising in the pharmaceutical carrier that is formulated in a consistency of composition of The compounds of this invention.

These embodiments and other embodiments of the present invention are that one skilled in the art of the present invention have considered that the application can easily expect after disclosing.

Embodiment

The present invention further understands with reference to following embodiment, and these embodiment only are examples of the present invention.These embodiment only are for the present invention will be described.Scope of the present invention is not limited to only be used to illustrate illustrative embodiments in a certain respect of the present invention.The method of any function equivalence all within the scope of the present invention.With reference to foregoing description and accompanying drawing, various modifications and the disclosed content of the application done at description of the invention will be obvious for those skilled in the art.These modify the scope that also falls into claims protection.

Embodiment 1: anti-CTGF treatment improves hemodynamic parameter and heart function

As follows, with method of the present invention be used for confirming its at animal model to the wide spectrum curative effect aspect some of the vascular complication relevant with diabetes.(65mg/kg) bring out diabetes for Sprague Dawley rat single intravenous injection potion U-9889 (STZ).The diabetes that STZ brings out in the rat cause vascular permeability to increase (Lawson et al. (2005) Regul Pept 124:221-224) and decreased cardiac function (Machackova et al (2004) Mol Cell Biochem 261:271-278).Experimental rat was accepted the intravenous injection of 0.1M citric acid buffered U-9889 (pH value 4.1), dosage 65mg/kg (65mg/ml) at the 0th day.Whether STZ successfully brings out diabetes with the rising of glucose level on an empty stomach in the 2nd day (＞250mg/dl) confirmation.

Diabetic animal is assigned to each treatment group (carrier, 3mg/kg, 5mg/kg; Or the anti-CTGF antibody of 10mg/kg CLN-1).Treated animal and accepted peritoneal injection carrier or anti-CTGF antibody, 6 weeks by a definite date weekly for 3 times.With Animal Anesthesia, the little sharp conductivity cell that pressure transmitter (Millar instruments) will be housed then was placed on left ventricle and measured heart function when finishing 6 weeks.With computer data acquisition system (ADI Instruments) record P-V ring.Computer Analysis P-V ring (PVAN system, Millar Instruments) is to measure hemodynamic parameter.Perhaps, animal is imposed slight anesthesia, measure heart function with the heart ultrasonic technique then.This technology is familiar with in those skilled in the art.

Determine that according to each Hemodynamics and heart function parameter diabetic animal presents decreased cardiac function.Compare with untreated diabetic animal, present heart function with the diabetic animal of anti-CTGF Antybody therapy and strengthen.Specifically, the ventricular diastole, ventricular systole, diastole end pressure, EDD, ejection fraction, arterial elasticity, stroke output and the cardiac output that present improvement with the diabetic animal of anti-CTGF Antybody therapy.To sum up, these results show that inhibition CTGF provides the methods of treatment of the treatment vascular complication relevant with diabetes, for example improves heart and vascular function.

Determine that according to each Hemodynamics and heart function parameter diabetic animal presents decreased cardiac function.Detected the influence of anti-CTGF treatment to ejection fraction (stroke output is divided by cubic capacity).As shown in Figure 1, the contraction of diabetic animal and diastolic function are all impaired or weaken.Especially, compare with the control animals of health, the ejection fraction of diabetic animal (EF) per-cent reduces.Compare with untreated control group diabetic animal, the diabetic animal ejection fraction per-cent of using behind the anti-CTGF antibody presents increase (referring to Fig. 1).Use (from the 6th thoughtful the 12nd week) after the anti-CTGF antibody, the ejection fraction per-cent of diabetic animal is similar or suitable to observed ejection fraction per-cent in the healthy non-diabetic control animals.These data show that using anti-CTGF antibody has stoped the ejection fraction reduction.

Also detected the influence of anti-CTGF treatment to left ventricular end diastolic presssure (LVEDP).Diabetic animal presents left ventricular end diastolic presssure to be increased.As shown in Figure 2, use anti-CTGF antibody and stoped visible left ventricular end diastolic presssure increase in untreated control group diabetic animal.Use after the anti-CTGF antibody, the left ventricular end diastolic presssure of diabetic animal is similar to viewed result in the non-diabetic control animals of health.These results show and use the increase that anti-CTGF antibody has stoped left ventricular end diastolic presssure.

Also detected the influence of anti-CTGF treatment to cardiac contractility and diastole.As shown in Figure 3, compare with the non-diabetic control animals of health, the maximum dP/dt of diabetic animal descends.In addition, use anti-CTGF antibody to diabetic animal and stoped the minimum inotropic decline of observed dP/dt (referring to Fig. 4) in untreated diabetic animal.Maximum dP/dt descends and minimum dP/dt descends minimizing indication cardiac contractility and diastole make moderate progress.

Generally speaking, these results show that anti-CTGF treatment is effective improving on the heart function.

Embodiment 2: anti-CTGF treatment reduces arterial stiffness

Measure the influence (arterial stiffness be vascular function, especially the standard of measurement of great vessels function) of anti-CTGF treatment with the diabetes animal model of the foregoing description 1 to arterial stiffness.Obtain the various measuring results of arterial stiffness, comprising from carotid passive and active pressure-volume data.In order to measure, the PE-50 conduit that is connecting three-way stop-cock is inserted the distal portions of left carotid artery.To improve the Krebs damping fluid through described intubate is filled in the carotid artery.Block carotid near-end with blood vessel blocking device.Pressure transmitter is connected to described three-way stop-cock, measures the power that produces in the process of perfusion buffered soln.The end of described PE pipe also is connected to equidistant force transducer, to measure the equidistant axial force in the pressure process.Weigh the change of the segmental outside radius of described artery with digital image analysis system.With no Ca ²⁺Damping fluid pours into described artery fragment to measure the passive hardness of artery.In perfusion liquid, add vasodilator and vasoconstrictor (for example norepinephrine) and measure the active hardness of arterial wall.In some experiments, from animal excision artery fragment, and with body in-draw length with the analogous instrument assessment of exsomatizing.

The carotid artery hardness of diabetes rat increases.As shown in Figure 5, the isolating carotid power-line of buckling demonstrates carotid artery axial stiffness increase in diabetic animal.Carotid artery axial stiffness with the diabetic animal of the anti-CTGF antibody (CLN-1) of 3mg/kg or 10mg/kg treatment is similar with the control animals of health.These results show anti-CTGF Antybody therapy prevention or (have for example reduced the carotid axial stiffness of diabetic animal, prevention or reduction dysfunction of blood vessel, especially prevent or reduction great vessels dysfunction), and the anti-CTGF Antybody therapy order axial stiffness relevant with diabetes belongs to non-diabetic control group normal level.

In addition, the pressure radius tracing analysis shows circumferentially (promptly radially) hardness increase of carotid artery of diabetic animal.The circumferential hardness of carotid artery similar to the non-diabetic animal (referring to Fig. 6) through the diabetic animal of anti-CTGF Antybody therapy.

To sum up, these data show, suppressing CTGF provides reduction or the prevention artery relevant with diabetes and the method for blood vessel hardness increase.Therefore, suppress the methods of treatment that CTGF provides the treatment vascular complication relevant with diabetes and improved vascular function.

Except that the above experiment, method of the present invention and medicament are used to prove it in animal model, and the others of the vascular complication relevant with diabetes are also had the wide spectrum curative effect.In a series of experiments, measure the influence (arterial stiffness be vascular function, especially the standard of measurement of great vessels function) of anti-CTGF treatment to arterial stiffness with the foregoing description 1 described diabetes animal model.(65mg/kg) bring out diabetes (type i diabetes) for Sprague Dawley rat single intravenous injection 0.1M citric acid buffered (pH4.1) U-9889 (STZ).STZ handles whether successfully bring out diabetes with the rising of glucose level on an empty stomach in the 2nd day (＞250mg/dl) confirmation.The diabetes that STZ brings out in the rat cause vascular permeability to increase (Lawson et al. (2005) Regul Pept 124:221-224).

In the experiment of another series, behind the animal injection STZ, the diabetes of leaving and disease progression 6 weeks relevant with diabetes.After 6 weeks, diabetic animal is divided into following each treatment group: human IgG control group (10mg/kg, peritoneal injection, on every Wendesdays time, 6 weeks by a definite date), anti-CTGF antibody group (CLN-1,10mg/kg, peritoneal injection, on every Wendesdays time, 6 weeks by a definite date), Captopril group is (75mg/kg/ days, oral, in tap water).

The required axial force of axial tension in the body is kept in Fig. 7 demonstration.Data are represented with mean+SD among Fig. 7; The NS=there was no significant difference; The data of replicate measurement are analyzed (Tukey post hoc test) by two-way ANOVA.

Diabetes rat carotid artery hardness increases.Specifically, diabetes make axial passive hardness significantly increase.As shown in Figure 7, each the carotid power-line of buckling confirms to bring out 6 weeks after the diabetes, and the carotid artery axial stiffness increases (referring to " 6 weeks of STZ " among Fig. 7) in the diabetic animal.As shown in Figure 7, compare with the axial passive hardness of non-diabetic control animals, (untreated) diabetic animal in the 6th week or the 12nd week (from the 6th thoughtful the 12nd week through the treatment of control group human IgG) observed axial passive hardness all has increase diabetic animal.Compare with the 6th all viewed axial passive hardness, from the animal of the 6th thoughtful the 12nd week through the treatment of control group human IgG, the 12nd all driven axles increase to hardness after accepting the STZ processing and diabetes take place.These results show that the diabetic animal arterial stiffness increases, and this increase is from the lasting and deterioration of the 6th weeks to the 12 weekly assembly.

Compare with the diabetic animal of having used the human IgG contrast, the diabetic animal of having used anti-CTGF antibody presents axial passive hardness reduction or has the axial passive hardness (referring to Fig. 7) of reduction.The back takes place and began in 6 weeks to use anti-CTGF antibody to diabetic animal in injection STZ and diabetes, and the axial stiffness of animal is compared with the non-diabetic animal and presented increase at this moment.Use during the 6th thoughtful the 12nd week after the anti-CTGF antibody, the axial passive hardness measurement value of animal is lower than viewed value when the 6th week.These data show that using anti-CTGF antibody has stoped the axial passive hardened progress of carotid artery.These data also show uses the axial sclerosis that anti-CTGF antibody can effectively reverse or reduction is relevant with diabetes.

These results show with anti-CTGF Antybody therapy diabetic animal, can (for example prevent and reduce carotid axial stiffness, prevention and alleviate dysfunction of blood vessel, especially prevent and alleviate the great vessels dysfunction), and anti-CTGF Antybody therapy can effectively reverse the axial stiffness relevant with diabetes.

To sum up, these data show, suppressing CTGF provides and be used to reduce or the method for the increase of artery that prevention is relevant with diabetes and blood vessel hardness.Therefore, suppress CTGF the methods of treatment that is used for the treatment of the vascular complication relevant with diabetes and improves vascular function is provided.

In addition, the pressure radius tracing analysis of these animals shows circumferentially (promptly radially) hardness increase of diabetic animal carotid artery.Among Fig. 8 when elevated pressures the pressure radius curve move to left, the expression arterial stiffness increases.Data show, compare with the control animals of health, and the diabetic animal in the 6th week or the diabetic animal for the treatment of with the control group human IgG from the 6th thoughtful the 12nd week all present circumferentially (promptly radially) arterial stiffness increase (referring to Fig. 8) of carotid artery.In Fig. 8, data are represented with mean+SD; The NS=there was no significant difference; The data of replicate measurement are carried out the ANOVA variance analysis by LSD post hoc tests.

After STZ injection and diabetes take place, compare with the animal for the treatment of with the control group human IgG, the circumferential hardness of carotid artery through the diabetic animal of anti-CTGF Antybody therapy during the 6th thoughtful the 12nd week presents remarkable reduction.These results show, use anti-CTGF antibody capable and effectively reduce the circumferential arterial stiffness relevant with diabetes.In addition, these results show that anti-CTGF Antybody therapy can effectively reverse the circumferential passive arterial stiffness relevant with diabetes with improving (for example reducing circumferential arterial stiffness).

To sum up, these data show, suppress CTGF and provide the method for the increase of minimizing, prevention or reverse artery relevant with diabetes and blood vessel hardness (for example alleviate or reverse dysfunction of blood vessel, particularly alleviate or reverse the great vessels dysfunction).Therefore, suppress CTGF the effective methods of treatment that is used for the treatment of the vascular complication relevant with diabetes and improves vascular function is provided.In addition, these results show, the great vessels complication that the method among the present invention can treat effectively, prevent, reduces with compound or reverse is relevant with diabetes, thereby help to improve the great vessels function.

The measurement of non-loaded and zero stress state

With method of the present invention and medicament be used for confirming its at animal model to the wide spectrum curative effect aspect some of the vascular complication relevant with diabetes.In a series of experiments, the following diabetes animal model that utilizes is measured the influence (arterial stiffness be vascular function, especially the standard of measurement of great vessels function) of anti-CTGF treatment to arterial stiffness.Give Sprague Dawley rat single intravenous injection 0.1M citric acid buffered (pH4.1) U-9889 (STZ) (65mg/kg), bring out (I type) diabetes.Whether STZ successfully brings out diabetes with the on an empty stomach rising of glucose level in the 2nd day (＞250mg/dl) confirmation that is as the criterion in animal.

Behind the injection STZ, the diabetes of leaving and progression of disease 6 weeks relevant with diabetes.After 6 weeks, diabetic animal is divided into following different treatment group: the human IgG control group (10mg/kg, peritoneal injection, on every Wendesdays time, 6 weeks by a definite date); Anti-CTGF antibody group (CLN-1,10mg/kg, peritoneal injection, on every Wendesdays time, 6 weeks by a definite date); Captopril group (75mg/kg/ days, oral, in tap water); Losartan group (20mg/kg/ days, in tap water); Anti-CTGF antibody (CLN-1,10mg/kg, peritoneal injection, on every Wendesdays time, 6 weeks by a definite date)+captopril (75mg/kg/day, oral, in tap water) group; Anti-CTGF antibody (CLN-1,10mg/kg, peritoneal injection, on every Wendesdays time, 6 weeks by a definite date)+losartan (20mg/kg/ days, in tap water) group.

The artery Mechanics Performance Testing of describing among the animals received embodiment 1 in this experiment.Before Mechanics Performance Testing, in-vivo measurement the segmental length of artery.20 minutes measure sample length behind the mechanical test is to obtain non-loaded and the zero stress state data.This length is compared with fragment length in the body of extracing blood vessel measurement before.The ratio of gained is registered as axial tension rate in the body.As shown in Figure 9, reduced axial tension rate in the body to the 6th all artificial diabeteses.To the 12nd week, anti-CTGF Antybody therapy has stoped the interior axial tension rate of body further to drop, and descends and the axial tension rate is lasting in the body in the animal of vehicle treatment.The decline indication artery of body in-draw rate just hardens.Anti-CTGF Antybody therapy has prevented arteriosclerosis.

The reverse that Aorta ring open-angle increases

From isolating carotid artery fragment middle part excision Aorta ring and be placed on the Krebs solution.Take the digital photos in the cross section of described ring under no-load condition.Follow and radially cut described ring, it is fan-shaped to make it be opened into.Take digital photos after 20 minutes, allow it that viscoelastic creep takes place.Open-angle is defined as formed angle between the line that (difference) connect internal surface mid point and fan-shaped (two) interior end points, and it is the feature of zero stress state.As shown in figure 10, increased open-angle to the 6th all artificial diabeteses.In the 12nd week, the open-angle that anti-CTGF Antybody therapy order increases is replied normal, is tantamount to the level of normal healthy controls group, and open-angle then continues to rise in vehicle treated animals.Open-angle is the indication that unrelieved stress is arranged in the arterial wall.

These results show the prevention of anti-CTGF Antybody therapy diabetic animal and have reversed the pathological change of carotid wall and (for example prevented and alleviated dysfunction of blood vessel, especially prevent and alleviated the great vessels dysfunction), and anti-CTGF Antybody therapy can effectively reverse the hardness relevant with diabetes.To sum up, these data show, suppress CTGF and provide the method for the increase of minimizing, prevention or reverse artery relevant with diabetes and blood vessel hardness (for example alleviate or reverse dysfunction of blood vessel, particularly alleviate or reverse the great vessels dysfunction).Therefore, suppress effective methods of treatment that CTGF provides the treatment vascular complication relevant with diabetes and improved vascular function.In addition, these results show, the great vessels complication that method of the present invention can treat effectively, prevent, reduces with compound or reverse is relevant with diabetes, thereby help to improve the great vessels function.

Embodiment 3: anti-CTGF treatment reduces vascular permeability

Utilize the foregoing description 1 described diabetes animal model to measure the influence (vascular permeability be vascular function, especially the standard of measurement of capillary blood vessel function) of anti-CTGF treatment to vascular permeability.After bringing out diabetes, in the increase of different time according to the vascular permeability (VP) of following test rat.Give rat intravenous injection Evans Blue (EB) dyestuff (20mg/kg) without anesthesia.After 20 minutes, put to death animal, and extract heart with excessive anesthesia.Excision and weighing are from the skin fragment of the later trunk of shoulder.In this detected, the feature that vascular permeability increases was that dyestuff exosmoses and enters skin.Skin is immersed in 24 ℃ methane amide (4ml/g weight in wet base) 24 hours.Utilize plate reader to be extracted into the absorbancy of the EB dyestuff in the methane amide in the 620nm measurement by spectrophotometry.In this detected, the concentration of EB dyestuff was directly proportional with the vascular permeability degree.In addition, obtain the skin fragment with 6 millimeters biopsy punches.These segmental weight of weighing are then in 60 ℃ of dried overnight.The weight of weighing dry sample, and the ratio of definite weight in wet base and dry weight.Wet/dried ratio increases indication and has tissue edema.

As Figure 11 and shown in Figure 12, wet/dried ratio that Evans Blue exosmoses to be increased and organize increases, and proves that the vascular permeability at the 3rd all diabetic animals increases with tissue edema (comparing with control group non-diabetic animal).Diabetic animal is compared with untreated diabetic animal behind anti-CTGF Antybody therapy, and Evans Blue exosmoses and reduces and wet/dried ratio reduction.

These results show, with anti-CTGF Antybody therapy diabetic animal, have reduced the vascular permeability relevant with diabetes and have alleviated tissue edema (for example alleviating dysfunction of blood vessel, especially the capillary blood vessel dysfunction).To sum up, these results show, the microvascular complication that method of the present invention can be treated effectively, prevent with compound or minimizing is relevant with diabetes, thereby help to improve the capillary blood vessel function.

As shown in figure 13, the Evans Blue of described tissue exosmoses to be increased and wet/dried ratio increase, proof is after STZ injection and diabetes generation, the diabetic animal in the 6th week and used the diabetic animal of human IgG contrast during the 6th thoughtful the 12nd week, vascular permeability increases with tissue edema (comparing with control group non-diabetic animal).(data are analyzed by ANOVA earlier among Figure 13, analyze by LSD post hoc tests then).(from 12 weeks of the 6th week to the) with the diabetic animal of anti-CTGF Antybody therapy, are compared with untreated diabetic animal, and Evans Blue exosmoses and reduces and wet/dried ratio reduction.In addition,, compare, demonstrate vascular permeability reduction and tissue edema and reduce with the control animals of using the human IgG contrast from the animal of 12 weeks of the 6th week to the using anti-CTGF antibody.These results show vascular permeability and the tissue edema that anti-CTGF Antybody therapy can effectively improve, minimizing is relevant with diabetes with reverse.

These results show, diabetic animal is accepted behind the anti-CTGF Antybody therapy can minimizing relevant with diabetes with reverse vascular permeability and tissue edema (for example alleviate and reverse dysfunction of blood vessel, particularly alleviate and reverse the capillary blood vessel dysfunction).To sum up, these results show, the microvascular complication that method of the present invention can treat effectively, prevent, reduces with compound or reverse is relevant with diabetes, thereby help to improve the capillary blood vessel function.

Embodiment 4: anti-CTGF treatment reduces angiosteosis

As described below, detect the influence of anti-CTGF treatment to angiosteosis.Bring out 6 weeks after the diabetes with STZ, to do the Von Kossa dyeing of calcium mineral deposition by the carotid artery fragment that obtains in the animal.The arteria carotis communis fragment of two cm long was placed 10% zinc formalin (0.1M) 24 hours,, use paraffin embedding subsequently with the ethanolic soln dehydration of serial dilution.Do the section of 3 micron thickness from described carotid artery tissue, remove deparaffnize, aquation in distilled water is used 5%AgNO then ₃Handled 60 minutes.Then with distilled water rinsing carotid artery tissue sample and use 5%Na ₂S ₂O ₃Handled two minutes.Then with the rinsing in distilled water and 95% ethanol of described sample order.Then described sample is redyed with eosin, and be placed on inspection under the opticmicroscope, mark to angiolithic degeneration by the furvous/brown colouring that presents along the vessel wall theca interna.Described vessel wall is measured angiosteosis to exist or not exist under observation by light microscope.

The result of this research as shown in Figure 14.Bring out 6 weeks after the diabetes, the von Kossa dyeing of the calcium mineral deposition relevant with described blood vessel makes the black/brown in color pigmented section that occurs strong positive along the carotid artery intima layer of diabetic animal; The carotid artery that obtains from the non-diabetic animal then occurs seldom or does not have calcium dyeing.In addition, from the carotid artery that the diabetic animal (n=6) through human IgG contrast treatment obtains, there is half to be strong positive angiosteosis dyeing.Similarly, the 6th week from the carotid artery that the diabetic animal (n=8) through captopril treatment obtains, have half to present strong calcium dyeing.(CLN-1, the carotid artery that diabetic animal 3mg/kg) obtains present the dyeing of calcium mineral deposition to be reduced from using anti-CTGF antibody.Particularly, the carotid artery that the diabetic animal for the treatment of from the anti-CTGF antibody (CLN-1) of accepting 3mg/kg obtains, fewer than half (43%) is presenting positive calcium dyeing the 6th week.(CLN-1 10mg/kg) can prevent all calcium depositions from the carotid artery section that diabetic animal (n=6) obtains to use the anti-CTGF antibody of higher dosage.These results show with anti-CTGF Antybody therapy diabetic animal can prevent or reduce angiosteosis.These data show that anti-CTGF treatment helps prevention or reduces angiosteosis.

Embodiment 5: anti-CTGF treatment has the treatment curative effect to the cardiovascular complication in the diabetes animal model

Use 8 all male db/db mouse in age (C57BLKS/J-leprdb/leprdb) and their the littermate mouse of age-matched non-diabetic db/+ (C57BLKS/J-leprdb/+) (Jackson Labs, Bar Harbor, ME).The about 40g of db/db mouse initial weight.The about 20g of the initial body weight of non-diabetic db/+ mouse.In some experimental group, tap water adds the losartan of 200mg/L, and this concentration guarantees that the transfer rate of losartan is about 2mg/ days.

(St.Louis Mo.), and passes through the repurity of albumin A chromatography to many donors human IgG of purifying available from Sigma Chemical Corporation.Neutralization immediately contains the acid elutriant of human IgG, and thoroughly dialyses with PBS, and sterile filtration then detects determining purity and no intracellular toxin, and be stored in 4 ℃ up to use (lot number CML052803 or similar, 4.67mg/mL).

In case 100% db/db mouse become hyperglycemia (with glucose level than db/+ control group mice high 2 times or more than be as the criterion), come into effect anti-CTGF monoclonal antibody (CLN-1) treatment ages in 8 weeks.After being as the criterion diabetes definite and taking place with hyperglycemia, the db/db mouse is divided into 10 every group at random, and according to following treatment.One group of db/db mouse is accepted the anti-CTGF antibody of peritoneal injection (3mg/kg) treatment, another winding is subjected to anti-CTGF antibody (CLN-1,10mg/kg) treatment, another winding is subjected to isotype pairing, incoherent human IgG (cIgG, 10mg/kg) treatment, another winding is subjected to losartan (2mg/ days) treatment in the tap water, and another group is accepted losartan (2mg/ days) and the peritoneal injection cIgG (10mg/kg) in the tap water simultaneously.Antibody is through the intraperitoneal dispenser, and initial single injection amount is 300 micrograms, is thereafter 3 times weekly, the 3mg/kg in totally 8 weeks or 10mg/kg dosage (the about 0.5ml of volumetric injection).

When putting to death animal, extract non-empty stomach blood sample from the eye socket venous plexus with the heparinization kapillary.Carry out HbA1c and lipid level analysis with unsegregated heparinized blood.Serum sample is left in-80 ℃ until analysis.Extract left kidney and right kidney, liver and heart from each animal, weigh then.

The following lipid determination that carries out: by commercially available detection kit (PTS Panels, PolymerTechnology Systems, Inc.) level of mensuration total cholesterol (TC), HDL and triglyceride level (TG).Calculate the level of LDL by following formula:

LDL＝TC-HDL-(TG/5)

With the glycated hemoglobin level (per-cent with glycated hemoglobin HbA1c is represented) of measuring fresh whole blood through the commercially available HbA1c detection kit of authorization.

As shown in figure 15, use the diabetic animal of anti-CTGF antibody and compare with the diabetic animal of using the IgG contrast, cardiac weight reduces.These results show that anti-CTGF treatment helps treating the cardiovascular disorder relevant with diabetes, comprises cardiac hypertrophy, congestive heart failure and myocardosis.Figure 16 shows, in the diabetes rat model that as described in example 1 above STZ brings out, uses the ratio that the CTGF antibody capable reduces cardiac weight and body weight.

As shown in Figure 17, use anti-CTGF antibody capable to diabetic animal and reduce blood LDL level.The increase of LDL level is the risk factors that cardiovascular disorder and obstacle take place.Therefore, these results show that anti-CTGF treatment can help reducing the risk of cardiovascular disorder generation or helping preventing its generation by reducing the LDL level.

Figure 18 is shown to diabetic animal and uses anti-CTGF antibody capable reduction HbA1c (glycated hemoglobin) level.Figure 19 shows, in the diabetes rat model that brings out as embodiment 1 described STZ, uses anti-CTGF antibody capable and reduces blood HbA1c level.The glycated hemoglobin horizontal survey can provide the accurate index of average blood sugar concentration in nearest 2 to 3 months of the human body.Human normal (non-diabetic) glycated hemoglobin level is between 4 to 6%.In diabetics research, DCCT finds, compares with the diabetics with higher HbA1c level, the HbA1c level is reduced to or remains on average 7.2%, can make cardiovascular disorder reduce 35%.Therefore, these results show, anti-CTGF treatment can reduce the glycated hemoglobin level, thereby reduce risk that cardiovascular disorder takes place or the generation that prevents cardiovascular disorder.

Figure 20 shows that anti-CTGF antibody and ARB combined treatment can effectively improve blood HDL level.

With reference to foregoing description, the various modifications of making at description of the invention will be tangible for those skilled in the art.These revise the scope that all falls into claims protection.

All reference that the application quotes are introduced the application as a reference in full with it.

Below each reference introduce the application in full as a reference with it.

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Claims

1. method that alleviates the dysfunction of blood vessel of object or improve the vascular function of object, described method comprises the anti-CTGF agent of using significant quantity to described object, thereby alleviates the dysfunction of blood vessel of object or improve the vascular function of object.

2. the process of claim 1 wherein described to as if the object suffering from diabetes or have the risk of suffering from diabetes.

3. the method for aforementioned each claim, wherein said vascular function is capillary blood vessel function or great vessels function.

4. the method for claim 1 or claim 2, wherein said vascular function is relevant with peripheral blood vessel.

5. the method for claim 1 or claim 2, wherein said method is used to alleviate endothelial function disturbance.

6. the method for claim 1 or claim 2, wherein said method are used for the treatment of or reduce arterial stiffness.

7. the method for claim 6, wherein said arterial stiffness is selected from axially, radially reaches circumferential arterial stiffness.

8. the method for claim 1 or claim 2, wherein said method be used to reduce vascular permeability, reduce and exosmose, reduce the inner membrance middle level and thicken or reduce arteria carotis communis inner membrance middle level and thicken.

9. the method for claim 8, wherein said method is used to reduce oedema or tissue edema.

10. the method for claim 1 or claim 2, wherein said method is used to alleviate damage or the dysfunction of blood vessel to blood vessel.

11. the method for claim 1 or claim 2, wherein said method is used to reduce angiosteosis.

12. the method for aforementioned each claim, wherein said method is used for the treatment of the vascular complication relevant with diabetes.

13. the method for aforementioned each claim is wherein said to liking Mammals.

14. the method for aforementioned each claim is wherein said to liking the people.

15. the method for aforementioned each claim, wherein said anti-CTGF agent is polypeptide, polynucleotide or small molecules.

16. the method for claim 15, wherein said anti-CTGF agent is and CTGF bonded antibody or its fragment, antisense molecule, siRNA or micromolecular compound.

17. the method for claim 16, wherein said anti-CTGF agent are monoclonal antibody or its fragments at CTGF.

18. the method for claim 16, wherein said anti-CTGF agent are CLN-1 or its fragment that WO 2004/108764 describes.