KR20050011350A - Therapeutic containing human integrin binding protein or peptide for treating ophthalmopathy - Google Patents

Abstract

PURPOSE: Provided is a therapeutic for ophthalmopathy which containing recombinant disintegrin protein, saxatilin and human integrin binding protein which has a similar sequence to the saxatilin, as active ingredient. Therefore, it is useful for treatment of ophthalmopathy associated with abnormal angiogenesis. CONSTITUTION: The therapeutic for treatment of angiogenesis related ophthalmopathy is characterized by containing, as an active ingredient, human integrin binding proteins which has a protein sequence of saxatilin represented by the SEQ ID NO: 1 or a similar sequence thereto. Its effective amount is 100ng-1mg/100kg of body weight.

Description

Therapeutic containing human integrin binding protein or peptide for treating ophthalmopathy}

The present invention relates to an ophthalmic disease therapeutic agent comprising a recombinant disintegrin polypeptide. More specifically, the present invention relates to a disintegrin-based protein containing an Arg-Gly-Asp (RGD) sequence, which is dose-dependently acting on abnormal angiogenesis and used as a therapeutic agent for disease.

Angiogenesis refers to the process by which sprouts are formed from existing microvascular vessels, and these sprouts multiply to form new capillaries. This is a very important normal process in fetal growth, uterine saline, placental proliferation, corpus luteum formation and wound healing in the human body (Gunther G. et al ., Oncology 54: 177-184 (1997)). Solid cancer is typically a disease with many neovascularizations, but the neovascularization is not secondary to the etiology of cancer and is a secondary feature that is only produced for the supply of nutrients and oxygen to rapidly growing cancer tissues. In cancer, the treatment of neovascularization can be suppressed to some extent, but the underlying treatment is impossible. On the contrary, there are diseases in which neovascularization itself is a etiology caused by abnormally increased neovascularization or failure to regulate normally. Representative diseases include neovascular diseases, rheumatoid arthritis, psoriasis, and pyogenic granulomas. These diseases can be treated by eliminating neovascularization causes as the disease progresses. As in chemotherapy, there are methods that use drugs that directly inhibit abnormal neovascular growth that has a structure that easily damages blood vessels, and neutralize or eliminate blood vessel-inducing cytokines. By strengthening the vascular structure of the fundamentally eliminate the causes of blood vessel induction is a method of treatment. Strengthening the structure of existing vessels does not cause secondary ischemic state due to vascular destruction and neovascularization does not occur.

In the eye, neovascularization must be turned off, but when the neovascularization is turned on due to a false signal, it causes serious eye disease, which is a major cause of acquired blindness (Lois E. H.et al.,Nat Med.5: 1390-1395 (1999). Representative neovascular diseases include diabetic retinopathy, blood vessels in the retina, retinopathy of prematurity, and age-related macular degeneration in which the blood vessels form in the choroid (Amal). AEet al.,Retina11: 244-249 (1991); Constantin J. P.et al.,Ophthalmology97: 1329-1333 (1990); Jin-Hong C.et al.,Current opinion in Ophthalmology12: 242-249 (2001); Peter A. C.,J of Cellular Physiology184: 301-310 (2000). Retinopathy of prematurity (ROP) occurs in two stages as the leading cause of infant blindness. At birth, retinal vessels are incomplete, especially in premature babies with ongoing ROP (Flynn J.T.et al.,Arch ophthalmol95: 217-223 (1977). Thus, the blood vessels form an avascular retina, which is the state of the hypoxic peripheral retina. (ROP stage 1) The non-perfusion level of the ROP stage 1 retinal detachment and blindness by neovascularization, which is the next stage 2 Determine the degree of destrutive stage (Penn JS)et al.,Invest Ophthalmol Vis Sci35: 3429-435 (1994). If blood vessels form normally in premature infants, the secondary neovascularization of ROP will not occur. Premature infants found that high levels of oxygen use were associated with the disease. This suggests that an oxygen-regulated factor is included. Essential for normal blood vessel development VEGF, known as an oxygen regulator, is expected to play a significant role in ROP, and several studies have shown that it plays an important role in the first and second phases of ROP (Pierce E.A.et al., Arch Ophthalmol114: 1219-1228 (1996). In particular, ROP animal model (high supplement oxygen) has been shown to affect vascular growth by inhibiting VEGF expression in the first stage. Diabetic retinopathy is one of the most important complications of microvascular complications in which hyperglycemia is a major cause and is the primary cause of acquired blindness in adults (Brownlee M.,Nature414: 813-820 (2001). Severe visual impairment associated with diabetic retinopathy may result in retinal neovascularization (Battegay E.J.,J Mol Med73: 333-346 (1995)) and its resulting vitreous hemorrhage and four tonic retinal detachment (Cai J., Boulton M.,Eye16: 242-260 (2002)). Pathophysiological changes in diabetic retina include loss of pericapillary cells, basal membrane thickening, impaired retinal vascular control, capillary circulation, microvascular flow, and IRMA (intraretinal microvascular abnormalities). Generates a Ryu Area (Lip PLet al., Invest Ophthalmol Vis Sci41: 2115-2119 (2000); Hammes H.P.et al., Diabetes51: 3107-3112 (2002)). These changes continue to lead to increased vascular permeability, chronic retinal hypoxia and retinal ischemia, which eventually lead to macular edema or neovascularization leading to proliferative diabetic retinopathy (Aiello L.P.et al., Diabetes Care21: 143-156 (1998). Diabetics have high levels of VEGF, and this factor appears to cause retinopathy by destroying the vascular retinal barrier. Age-related macular degeneration is the leading cause of blindness in people over 50. The cause of severe vision loss is caused by neovascularization from choroidal capillaries (Ferris F.L. 3rdet al., Arch Ophthalmol102: 1640-1642 (1984)). AMD is divided into two different forms: wet and dry. Wetness usually occurs after dry outbreaks. Dryness refers to the case of macular degeneration due to loss of pigmentation and retinal pigment epithelium (RPE). Wetness is a change in dryness. Subretinal neovascularization (subretinal scar), subretinal hemorrhage, and retinal pigment epithelial cell detachment occur. Subretinal neovascularization is actually scar tissue that is enlarged for the treatment of space caused by diseased retinal pigment epithelial cells. New blood vessels grow, leaking plasma and fibrin and causing small retinal detachment (Mousa S.A. et al., J Cell Biochem 74: 135-43 (1999)). In addition, scarring of the subretinal scar also contributes to decreased vision.

Current treatments for these eye diseases include laser therapy, photocoagulation, cryocoagulation, and photodynamic therapy (Visudyne) (Edwin EB et al ., Ophthalmology 88: 101-107 (1981)). All of these treatments are surgical treatments, and medical treatments are still in development. Surgical treatment not only has a big limitation that it can not be applied to all patients, but also has a low success rate and a very high cost, which causes a great social and economic burden. Most patients who can't operate have blindness in the absence of special treatment. As human lifespan is prolonged, these diseases continue to increase, and the development of appropriate therapeutics is urgent. Therefore, research and development of angiogenesis inhibitors are currently being actively conducted, and most of the eye disease treatment agents under development include steroids, MMP inhibitors, and antibodies against angiogenic growth factors (Jeremy G. et al. , Am J Pathology 160: 1097-1103 (2002)).

Angiogenesis is achieved by vascular cell adhesions in smooth muscle and endothelial cells. Adhesion requires binding of matrix proteins to specific cell surface receptors, including the integrin family (Brian P. Eliceiri, Circ Res . 89: 1104-1110 (2001)). Angiogenesis has been reported to exist in two pathways by the vascular cell integrins αvβ3 and αvβ5 (Lin L. et al ., Exp. Eye Res . 70: 537-546 (2000)). These two integrins are expressed in angiogenic vascular cells and are known to play an important role in bFGF- and VEGF-induced angiogenesis (Guoquan Gao et al ., FEBS Lett. 489: 270-276). 2001); Joan W. Millers et al ., Ophthalmology 100: 9-14 (1993); Napoleone Ferrara, Curr Opin Biotech. 11: 617-624 (2000); Martin Friedlander et al ., Proc. Natl. Acad. Sci 93: 9764-9769 (1996)). In general, disintegrin is known as a potent antagonist of αvβ3 integrin expressed by bFGF, and the disintegrin used in the present invention is a polypeptide derived from snake venom (Sung-Yu H. et al ., Thrombosis Res) ., 105: 79-86 (2002)). Most disintegrins contain Arg-Gly-Asp (RGD) or Lys-Gly-Asp (KGD) sequences, which are structural motifs of fibrinogen. Reported studies have shown that disintegrins containing RGD sequences inhibit the formation of new blood vessels by preventing cells from adhering to the ECM (Victor IR and Michael SG Prostate 39: 108-118 (1999); Yohei M. et al ., J of Biological chemistry 276: 31959-31968 (2001).

It has been reported that the neovascularization pathway expressed by VEGF is more important in the eye, and we are the first to see whether disintegrin, which actually contains the RGD sequence, can also inhibit intraocular neovascularization mainly involving VEGF but not bFGF. Clear results were obtained for.

Therefore, saxatrin has been confirmed to be effective in the treatment of neovascularization of intraocular diseases such as diabetic retinopathy, prematurity retinopathy, age-related macular degeneration, etc., by dose-dependently regulating intraocular neovascularization.

The present invention solves the above problems, and the object of the present invention is to provide a therapeutic agent for ocular diseases caused by neovascularization by inducing a dose-dependent enhancement of normal angiogenesis.

FIG. 1A is a surgical micrograph of saxatrin inhibiting corneal neovascularization in VEGF-induced mice upon intraperitoneal injection (dosage) administration (5 mg / kg / day).

FIG. 1B is a surgical micrograph showing that saxatrin stimulates corneal neovascularization in VEGF-induced mice upon intraperitoneal dosage administration (10 ng −1 ug / kg / day).

Figure 2a is a photograph of fluorescence microscopy observed after treatment by concentration (0, 12.5, 25, 100 ug / ml) to confirm the basic toxicity of saxatrin in primary cultured retinal cells.

Figure 2b is a graph of the lactate dehydrogenase enzyme released in the medium after treatment by concentration (0, 12.5, 25, 50, 100 ug / ml) to confirm the basic toxicity of saxatrin in primary cultured retinal cells.

Figure 3a is a photograph of the observation of inhibiting abnormal neovascularization and normal development of vascular maturation of high concentration saxatrin injected intraperitoneally in an animal model that lowered to normal oxygen partial pressure after hyperbaric oxygen treatment (75%) to induce retinal neovascularization.

FIG. 3b is a photograph showing observation that the dose-dependent improvement of normal blood vessels and abnormal blood vessels are suppressed by low-dose saxatrine injected intraperitoneally in an animal model that induces retinal neovascularization by reducing to normal oxygen partial pressure after hyperbaric oxygen treatment (75%). admit.

Figure 4 is a graph of the level of abnormal retinal vascular inhibition by saxatrin dose-dependently intraperitoneally injected in the animal model to lower the normal oxygen partial pressure after hyperbaric oxygen treatment (75%) to induce retinal neovascularization.

FIG. 5 shows that the blood vessel leakage is reduced by low-dose saxatrine injected intraperitoneally in an animal model that induces retinal neovascularization by reducing to normal oxygen partial pressure after hyperbaric oxygen treatment (75%) using the fluorescent substance FITC-dextran. Observation pictures.

Figure 6a shows how the vascular structure is formed after treatment of cells with a recombinant saxatrin at a concentration of 1 ug / ml or less using a BCE cell (bovine capillary endothelial cell) microtubule system, a research system for vascular structure formation. Photomicrograph.

Figure 6b is a microscopic picture showing the migration of cells after the migration experiments using BCE cells in a study on the movement of cells treated with recombinant saxatrin at a concentration of 1 ug / ml or less.

FIG. 7A shows angiography of blood vessels in the choroid of the experimental group not treated with saxatrin in the CNV animal model using a laser and a group administered 10 ug / kg every 2 days for 3 weeks in a CNV animal model. Photos.

FIG. 7B is angiography photographs comparing blood vessels generated in the choroid of the saxatin-treated experimental group in the CNV animal model and the group administered with 0.1 ug / kg and 10 ug / kg every 2 days for 3 weeks.

Figure 7c is a micrograph taken after H-E staining tissue sections of the eye group of the group treated with no saxatrin in the CNV animal model and the group administered 10 ug / kg every 2 days for 3 weeks.

In order to achieve the above object, the present invention provides a therapeutic agent for neovascular diseases related to human integrin binding protein having a saxatrine protein sequence of SEQ ID NO: 1 or the like sequence as an active ingredient.

The effective amount for the vascular maturation and stabilization of the therapeutic agent is 100 ng-1 mg / kg body weight / day dose in the case of angioplasty to provide a neovascular related eye disease treatment. Above this dose affects normal vascular growth, and below, vascular maturation efficacy is not complete.

Eye diseases that can be applied in the present invention are prematurity retinopathy, diabetic retinopathy and age-related macular degeneration.

The invention is briefly described below.

In the present invention, it is possible to treat neovascularization-related ocular disease with a different mechanism of action than a therapeutic agent having angiogenesis inhibitor efficacy. Its contents are based on disintegrin, which is effective in promoting vascular maturation, thereby stabilizing vascular structure, thereby eliminating angiogenesis and causing angiogenesis with an incomplete vascular structure that is easily damaged. A drug for eye diseases has been developed.

In the present invention, it has been found that there is a new function of vascular maturation as well as a known integrin inhibitor of disintegrin. The disintegrin function differed according to the dose (1-10 mg / kg body weight: high dose) and below (100 ng-1 mg / kg body weight: low dose) used as a cancer treatment through the animal model.

At high doses, abnormal neovascularization was inhibited as expected, and normal retinal blood vessel growth was inhibited during development. In addition, some of the developing blood vessels did not show a complete therapeutic effect due to pathological incomplete structure. At low doses, however, vascular structures were observed to enhance the formation of blood vessels similar to the retinal vessels under normal conditions, thereby reducing the ischemic environment and suppressing abnormal neovascularization induced by ischemia.

The effects of purely purified purified saxatrin on intraocular neovascularization under in vivo conditions were analyzed and the following novel phenomena were discovered. First, saxatrin acts dose-dependently on angiogenesis expressed by hypoxia in the eye, inhibiting integrin or inducing normal angiogenesis. Second, the low concentration saxatrin treatment at the developmental stage through the oxygen partial pressure animal model does not inhibit angiogenesis that is normally generated at all, but rather helps to form normal blood vessels and stabilizes the structure of the blood vessels. Blood leakage in the blood vessels of the phenomenon has been reduced. Therefore, the use of the present invention is preferred for diabetic retinopathy and age-related macular degeneration, which are diseases related to abnormal neovascularization induced by prematurity retinopathy and destruction of normal vascular structure, which are diseases caused by the normal development of blood vessels in eye diseases. Do.

Hereinafter, the present invention will be described in more detail with reference to non-limiting examples.

Example 1 Effects of Saxatrin with Dose on VEGF-induced Neovascularization in Vascular Corneal Tissues.

To investigate the effects of saxatrin's intraocular neovascularization, a micropocket was created in the cornea of the mouse eye and a pellet containing 300 ng of VEGF was inserted to induce angiogenesis. At this time, in order to see the efficacy of saxatrine, saxatrine was intraperitoneally injected. Five days later, the eyes of the mice were examined by anatomic microscope to determine whether neovascularization was generated. As a result, it was confirmed that the high dose (5 mg / kg body weight / day) effectively inhibits the growth of VEGF-induced ocular vascular endothelial cells during the saxtrin peritoneal injection, low dose (10 ng-1 ug / kg body weight / day) Abdominal injection of saxatrin induced angiogenesis rather than growth inhibition of neovascularization. (See FIG. 1A, FIG. 1B)

Inhibition of VEGF-induced neovascularization in ocular avascular tissues by high dose saxatrin can be explained by VEGF-induced integrin and saxatrin blocking it in blood vessels of ocular tissue. In the case of low-dose saxatrine, it is the first fact that disintegrin-based saxatrin does not block integrin and shows angiogenesis efficacy. On the other hand, no side effects such as corneal haze by saxatrin were observed in the mice used in the experiment.

Example 2 Basic Toxicity Test of Saxatrin in Primary Cultured Retinal Cells

The eye is part of the central nervous system. In particular, the retinal tissue is the optic nerve (optic nerve), various interneurons (retinal ganglion cells), and photoreceptors (photoreceptors) are the majority. These cells are so sensitive that apoptosis can occur even with small stimuli, and once killed, they do not regenerate. Therefore, it is very important to investigate whether saxatrin is toxic to these neurons. To verify these, primary cultures of the retina from the eyes of two-day-old ICR mice were treated with saxatrin concentrations (0, 12.5, 25, 50, 100 ug / ml), and cytotoxicity was determined by LDH assay and Hoechst-PI staining. Investigate. (See Figures 2A, 2B)

As a result, it was confirmed that the toxicity of saxatrin retinal cells in accordance with the concentration compared to the control group was little at the experimental concentration. Thus, when saxatrin is used for the treatment of eye diseases, at least there is no toxicity problem in the nervous system.

Example 3 Effect of Saxatrin in Retinal Neovascularization-induced Mouse Model Using Oxygen Partial Pressure Change

Artificial retinal neovascularization caused by oxygen partial pressure difference is similar to that of prematurity and diabetic retinopathy in humans. The mice exposed to the high oxygen environment (75%) at the beginning of birth were tested using the principle of spontaneously producing abnormal neovascularization when returning to normal oxygen partial pressure (20%) (Higgins RD. Et al ., Curr. Eye Res . 18: 20-27 (1999); Bhart N. et al. , Pediatric Res. 46: 184-188 (1999); Gebarowska D. et al., Am. J. Pathol. 160: 307-313 ( 2002)). To this end, after 7 days of birth in a device capable of adjusting the oxygen partial pressure, it is left for 5 days in a high-pressure oxygen environment maintaining a 75% oxygen partial pressure, and then left for 5 days again at a normal oxygen partial pressure of 20% oxygen. At this time, saxatrin was administered to the mouse intraperitoneally once a day for 5 days to observe the neovascularization of the retina. In order to observe the blood vessels, a solution of 50 mg of FITC-dextran having a molecular weight of 2 × 10 ⁶ in 1 ml of saline was injected through the left ventricle. Immediately after injection, the mice's eyes were removed. The extracted eye was washed with physiological saline and fixed with 4% paraformaldehyde for 4 to 24 hours, and then the lens was removed from the eye, the retina was flattened on the glass slider, sutured with glycerin-gelatin, and observed using a fluorescence microscope. .

Based on the dose of 1 mg / kg / day that saxatrine has anticancer efficacy in the existing rat animal experiment, the high dose treatment group 1 mg, 5 mg, 10 mg / kg / day and 1 mg / kg / day Experiments were divided into low dose treatment groups of 0.1 ug, 1 ug, 10 ug, and 100 ug / kg / day. As a result, it was found that neovascularization was severely formed in the periphery of the retina of the mice treated with physiological saline only after hyperbaric oxygen treatment, but the neovascularization was effectively inhibited in the high-dose saxatrin-treated group. The neovascular suppression pattern was dose-dependent. These results confirm that saxatrin acts as an antagonist of integrin expressed by VEGF from the existing results of VEGF-induced and neovascularization in the retina in this oxygen partial pressure animal model. (Note: FIG. 3A)

On the other hand, the young mice left in hyperbaric oxygen did not develop vascular tissues of the normal retina at the developmental stage as compared with the well-grown mice. This was the case even when saxatrin was treated at high doses. However, the doses of 100 μg, 10 μg, 1 μg, 100 ng, and 10 ng / kg / day were observed to determine the effect of saxatrin at low doses below 1 mg / kg / day. 100 ng-100 μg / At kg / day no morphological features of abnormal neovascularization were observed, and well-formed blood vessels that were dose-dependently normal were observed (see FIG. 3B). After comprehensively treating the saxatilin concentration, the retinopathy scoring system (Higgins RD. Et al ., Curr. Eye Res . 18: 20-27 (1999)) was used to measure the degree of retinopathy (see Fig. 4). In the low dose saxatrin dose range from 100 ng / kg / day to 1 mg / kg / day, retinopathy was actually very low. This shows that saxatrin can be used in the treatment of eye diseases as a very interesting result in that it plays a role in supporting normal blood vessel growth in addition to the advantage that it has no effect on normal blood vessels. Disintegrin-based drugs have been shown for the first time to improve the formation of these normal blood vessels in addition to the effects of neovascularization depending on the treatment concentration. It is inferred that soluble polypeptides such as saxatrin are attached to integrins and carry signaling associated with cell growth. Induction of renal neovascularization induced by the low dose treatment in the retinal neovascularization-inducing mouse model using this oxygen partial pressure induced normal blood vessel development, which shows pathological inhibitory effect by reducing hypoxia and eventually eliminating the cause of neovascularization. It may be used to treat prematurity retinopathy. In preterm retinopathy, animal models showed that induction of normal angiogenesis through low-dose treatment was more effective than high-dose saxatrin treatment to prevent abnormal neovascularization. In addition, it was observed through the FITC-dextran fluorescence leak test that low-dose saxatrin treatment stabilized the structure of the blood vessels, so that no blood leakage occurred (see FIG. 5). Retinal vessels have blood-retina-barrier (BRB), such as the blood-brain-barrier (BBB) of the cerebrovascular system, so large molecules do not easily exit the blood vessels. The leakage of relatively high polymers such as FITC-dextran into the retina indicates significant damage to the microstructure of the retinal vessels and has been demonstrated experimentally that saxatrin prevented the damage. Therefore, Saxatrin maintains the vascular structure even when the disease occurs due to abnormalities such as blood leakage of blood vessels in the early stages of the disease, such as diabetic retinopathy and age-related macular degeneration. It may be used as an initial treatment for.

Example 4 Induction of Angiogenesis of Low Dose Saxatrin

The following experiments were performed to determine the function of low-dose saxatrin that is different from the neovascularization inhibitory function of high-dose saxatrin. Low dose saxatrine treatment in animal models tended to help normal blood vessel formation rather than neovascular suppression. Therefore, microtubule and migration experiments were performed to investigate the function of low-dose saxatrin administration. First, microtubule experiments were performed to determine how low-dose saxatrine affects angiogenesis. BCE (bovine capillary endothelial cell) cells activated with VEGF were observed for the formation of tubules using ECM625, a Chemicon company. As a result, it was observed that tubule formation was well at low saxatrin concentration of less than 1 ug / ml. (Note: FIG. 6A)

The migration experiments were performed to investigate the effect of saxatrin on cell migration according to the observation that the angiogenesis was good at low concentrations of saxatrin. After culturing the activated BCE cell with VEGF, a portion of the cell was removed and the cell movement was observed. As a result, it was confirmed that the movement of cells in the low concentration saxatrin of 1 ug / ml or less. (Note: Figure 6b)

As a result, low concentration of saxatrin was found to affect vascular structure by affecting cell migration and tubule formation, unlike saxatrin causing cell death at high concentration.

Example 5: Saxatrin Efficacy in CNV Animal Model Using Laser

CNV is an angiogenesis in the choroid. Currently, the animal model of this disease is an animal model induced by laser retinal pigment epithelial (RPE) destruction (Takahashi K. et al., Invest.Ophthal. Vis. Sci. 44: 409-415 (2003). Lai C.-C. et al. Invest.Ophthal.Vis.Sci. 42: 2401-2407 (2001)). In this way, the laser treatment was performed on colored rabbits, followed by intramuscular injection of saxatrin once every two days. Angiography was performed to observe neovascularization. As a result, when 10 ug / kg injection of saxatrin for 3 weeks after the procedure did not show any blood vessels in the laser sopt (reference: Figure 7a). In one experiment with a lower dose, some vessels were observed when compared to 10 ug / kg (see FIG. 7B). This shows that saxatrin acts dose-dependently. As a result of HE staining of the ocular tissue, many macrophages were observed in the retinal layer and many ECMs were observed in the laser spot of the drug-free group. In addition, the retinal pigment epithelium completely disappeared (see FIG. 7C). On the other hand, there was less ECM expression and monolayer covering of retinal pigment epithelial cells in the laser spot of saxatrin treated group. This result shows that saxatrin reduces the negative effect of retinal pigment epithelium caused by laser burn and is effective in scarless wound healing. As a result, saxatrin effectively reduces the blood vessels generated in the choroid at low doses and is a treatment for age-related macular degeneration. Prove that it can be used.

The present invention proposes a new treatment method through a therapeutic drug in addition to the neovascular disease-related ophthalmic disease treatment method that was only dependent on the existing surgical operation. Surgical operation is expensive and has limitations that cannot be applied to all patients, but the present invention is a breakthrough in the treatment of neovascular diseases related to ocular diseases, and prevents blindness. Administration of an appropriate dose of saxatrin has no effect on the formation of existing normal blood vessels and normal neovascularization of the newly developing stage. Rather, it helps to produce normal blood vessels in the developmental stage, which is a great advantage for patients in developmental stages such as prematurity retinopathy. If you suppress all neovascularization, it cannot be applied to prematurity retinopathy. Therefore, the value of saxatrin as a therapeutic agent for retinopathy of prematurity is very high. In the case of diabetic retinopathy, angiogenesis can be effectively suppressed by administering a dose (low dose) that can protect the structure of blood vessels at an early stage, and administering a high dose of saxatrine during the formation of a large amount of neovascularization. Seems to be. Therefore, the present invention has a new content that can maximize the efficacy of treatment by administering in an appropriate dose according to the characteristics and timing of the ophthalmic disease. In addition, saxatrin may inhibit abnormal blood vessel growth by helping normalize vascular structure in age-related macular degeneration.

<110> EYEGENE CO., LTD. <120> Therapeutic containing human integrin binding protein or peptide          for treating ophthalmopathy <160> 1 <170> KopatentIn 1.71 <210> 1 <211> 73 <212> PRT Glosius saxatilis <400> 1 Glu Ala Gly Glu Glu Cys Asp Cys Gly Ala Pro Ala Asn Pro Cys Cys   1 5 10 15 Asp Ala Ala Thr Cys Lys Leu Arg Pro Gly Ala Gln Cys Ala Glu Gly              20 25 30 Leu Cys Cys Asp Gln Cys Arg Phe Met Lys Glu Gly Thr Ile Cys Arg          35 40 45 Met Ala Arg Gly Asp Asp Met Asp Asp Tyr Cys Asn Gly Ile Ser Ala      50 55 60 Gly Cys Pro Arg Asn Pro Phe His Ala  65 70

Claims (3)

A therapeutic agent for neovascular diseases related to a human integrin binding protein having a saxatrin protein sequence of SEQ ID NO: 1 or a similar sequence thereof as an active ingredient. The method of claim 1, The ocular disease is an ocular disease treatment, characterized in that prematurity retinopathy, diabetic retinopathy, or age-related macular degeneration. The method of claim 1, The effective amount for the vascular maturation and stabilization of the therapeutic agent is an intravascular disease therapeutic agent, characterized in that the range of 100 ng ~ 1 mg per Kg of body weight when vascular injection.