Usage of hydrogen peroxide as active substance in the treatment of diabetic retinopathy diseases and the symptoms resulting from these diseases

Abstract

The present invention relates to usage of hydrogen peroxide compounds as active substance in elimination of diabetic retinopathy diseases and the symptoms resulting from these diseases, and pharmaceutical compositions comprising hydrogen peroxide as active substance in the treatment of said diseases.

Description

[0001] USAGE OF HYDROGEN PEROXIDE AS ACTIVE SUBSTANCE IN THE TREATMENT OF DIABETIC RETINOPATHY DISEASES AND THE SYMPTOMS RESULTING FROM THESE DISEASES

[0002] TECHNICAL FIELD

[0003] The present invention relates to usage of hydrogen peroxide compounds as active substance in elimination of diabetic retinopathy diseases and the symptoms resulting from these diseases, and pharmaceutical compositions comprising hydrogen peroxide as active substance in the treatment of said diseases.

[0004] The present invention belongs to pharmacology technical field; and relates to usage of hydrogen peroxide compounds for elimination of the clinical problems created by diabetic retinopathy and the diseases that have similar disease mechanism.

[0005] PRIOR ART

[0006] As known in the art, diabetes disease leads to various diseases like cataract, glaucoma, refractive errors, optic nerve inflammation, cycloplegia and various diseases in the retina. The most frequent reason of blindness that occurs due to diabetes is diabetic retinopathy disease. Diabetic retinopathy can exist in all persons who suffer from diabetes. The biggest factor that triggers diabetic retinopathy disease is the disease duration of diabetes; if the disease duration is longer, the diabetic retinopathy disease risk is higher.

[0007] In diabetic retinopathy, symptoms occur depending on the place where the lesion exists and depending on the intensity of the damage. There is no symptom in early phase of the disease, and no symptom may occur during long durations. The first effect of the disease can be blurry vision in patients. In the following phases, instant bleeding in the eyes can occur and / or the patient can begin not to see. In this perspective, diabetic retinopathy disease has substantially dangerous activities for the persons who suffer from the disease.

[0008] In the related technical field, diabetic retinopathy disease can be treated by surgical operations like laser photocoagulation treatment, intravitreal injections, vitrectomy. In elimination or prevention of said diseases and symptoms, there are substantially no studies related to the presence of active substances that provide technical solution. Since the known surgical treatment methods are tedious for troublesome and patient persons and besides, since said methods have high costs, it has been obligatory to search other treatment methods in the technical field.

[0009] BRIEF DESCRIPTION OF THE INVENTION

[0010] The present invention relates to an active substance that will be used for treatment in elimination of diabetic retinopathy diseases and in elimination of the symptoms resulting from these diseases.

[0011] The present invention relates to obtaining a new active substance in diabetic retinopathy diseases and in elimination of the symptoms resulting from these diseases.

[0012] The present invention relates to usage of active substances that have healing characteristic instead of surgical operations in elimination of diabetic retinopathy diseases and in elimination of the symptoms resulting from these diseases.

[0013] The present invention relates to a pharmaceutical composition which comprises active substance that is suitable in diabetic retinopathy diseases and in elimination of the symptoms resulting from these diseases.

[0014] In order to realize all of these objects, the present invention relates to usage of hydrogen peroxide (H2O2) compound as the treating active substance which provides inhibition of proteasome enzyme(s) in elimination of diabetic retinopathy diseases which lead to diseases like cataract, glaucoma, refractive errors, optic nerve inflammation, cycloplegia and various diseases in the retina, blindness depending on diabetes, and the symptoms resulting from these diseases, and thus which provides increase of GAPDH amount.

[0015] In a possible embodiment of the present invention, said proteasome enzymes are trypsin, chemotrypsin and / or caspase-like enzyme.

[0016] In another perspective, the present invention relates to a pharmaceutical composition comprising hydrogen peroxide (H2O2) compound as the treating active substance which provides inhibition of proteasome enzyme(s) in elimination of diabetic retinopathy diseases which lead to diseases like cataract, glaucoma, refractive errors, optic nerve inflammation, cycloplegia and various diseases in the retina, blindness depending on diabetes, and the symptoms resulting from these diseases, and thus which provides increase of GAPDH amount.

[0017] In a possible embodiment of the present invention, hydrogen peroxide (H2O2) compound exists as active substance at an amount between 0.004 nM and 4 nM in said pharmaceutical composition.

[0018] In a possible embodiment of the present invention, said pharmaceutical composition comprises at least one active substance besides hydrogen peroxide (H2O2) compound.

[0019] In a possible embodiment of the present invention, hydrogen peroxide (H2O2) compound exists as active substance, and it moreover comprises at least one auxiliary component.

[0020] BRIEF DESCRIPTION OF THE FIGURES

[0021] In Figure 1 , the increase in GAPDH amount with respect to duration of existence of the hydrogen peroxide compound in HepG1 cells is shown.

[0022] In Figure 2, the increase in GAPDH amount of hydrogen peroxide compound also in T98G cell that is human glioma cell is shown.

[0023] In Figure 3, the increase in GAPDH amount of hydrogen peroxide compound also in Hek293 cell that is the immortalized kidney epithelial cell is shown.

[0024] In Figure 4, it has been shared that the hydrogen peroxide compound decreases the proteasome chemotrypsin activity in proteasome chemotrypsin activity test.

[0025] In Figure 5, the increase in ubiquitin amount that shows inhibition of proteasome activity with respect to duration of existence of the hydrogen peroxide compound in HepG2 cells is shown

[0026] DETAILED DESCRIPTION OF THE INVENTION

[0027] In this detailed description, the subject matter relates to usage of hydrogen peroxide (H2O2) compounds in elimination of diabetic retinopathy diseases and the symptoms resulting from these diseases, and pharmaceutical compositions comprising hydrogen peroxide as active substance in the treatment of said diseases, and is explained with references to examples without forming any restrictive effect only in order to make the subject more understandable. In the invention, “pharmaceutical component” is used in the meaning of active substance which provides treatment of a disease or elimination of the symptoms resulting from diseases. Pharmaceutical composition is formulations comprising said pharmaceutical component.

[0028] “Diabetic retinopathy” used within the scope of the present invention means a disease which leads to disorders like cataract, glaucoma, refractive errors, optic nerve inflammation, cycloplegia, various diseases of the retina and blindness. In the invention, the term diabetic can also be described as diabetes disease.

[0029] The term “treatment” used within the scope of the present invention describes elimination of findings of diabetic retinopathy within the scope of the present invention.

[0030] The term “elimination of the symptoms resulting from diabetic retinopathy” used within the scope of the present invention describes elimination of at least one of the symptoms and / or diseases formed by said diseases.

[0031] In the invention, “active substance” is described as the component that forms pharmaceutical effect for elimination of the problems resulting from diabetic retinopathy diseases.

[0032] The invention owners have made some studies based on the teachings known in the related art where diabetic retinopathy diseases lead to optic loss problems in patient persons depending on diabetes. In the subject matter, the amount of GAPDH protein, that has an important function in patient persons, is increased. The invention owners recommend the usage of hydrogen peroxide compound as active substance for the patient persons for increasing said GAPDH amount. In patient persons, it decreases chemotrypsin activity of the proteasome system, and as a result of this, it provides reduction of GAPDH protein degradation and it provides increasing of GAPDH amount, it has been detected that hydrogen peroxide compound has a pharmaceutical activity. It is known that there is correlation between the activity of glyceraldehyde-3-phosphate dehydrogenase (it shall be abbreviated as GAPDH) and diabetes retinopathy pathogen. It has been mentioned that the reduction of said GAPDH activity in patient persons - in persons who suffer from diabetes - leads to increase of diabetic retinopathy pathogenesis. Depending on this, the present invention owners have made studies for increasing GAPDH activity in persons who suffer from diabetes and for bringing GAPDH activity to the values that are present in healthy persons. In the invention, “glyceraldehyde 3-phosphate dehydrogenase” is described as an enzyme which is approximately 37 kDa and which catalyzes sixth step of glycolysis and which serves fragmenting of glucose for energy and carbon molecules. In human body, GAPDH catalyzes transformation of glyseraldehyde-3-phosphate into D-glycerate-1 ,3-biphosphate.

[0033] The cysteine residue that exists in GAPDH active region, namely C152, is needed for inducing apoptosis by means of oxidative stress.

[0034] In the subject matter, the amount of expression of GAPDH protein that has an important function in patient persons is increased. The invention owners recommend the usage of hydrogen peroxide compound for patient persons as active substance for increasing said GAPDH expression amount. In patient persons, it decreases the chemotrypsin activity of the proteasome system, and as a result of this, it has been detected that the hydrogen peroxide compound has a pharmaceutical activity in order to provide reduction of GAPDH protein degradation and provide increase of GAPDH amount.

[0035] Moreover, in the invention, this finding, which shows that total ubiquitin amount increases in the cell in the presence of H2O2 and thus which shows the inactivation of the proteasome system, supports chemotrypsin inactivation finding.

[0036] Depending on the increase of GAPDH protein expression amount, the present invention solidifies the correction of the symptoms of diabetic retinopathy.

[0037] Moreover, depending on the increase of GAPDH protein expression amount, the present invention solidifies the correction of the complications of diabetic retinopathy.

[0038] Moreover, depending on the increase of GAPDH protein expression amount, in the present invention, it has been foreseen that AGE products (advanced glycation end products), that play a role in diabetic retinopathy pathogenesis, and protein kinase (PKC) can provide homeostasis in hexosamine pathway.

[0039] In the invention, “hydrogen peroxide compound” is shown by formula 1 . In the art, hydrogen peroxide compound is a compound known to be in substantially light blue color when it is in pure form, and becomes a colorless form when it is diluted. Hydrogen peroxide is a compound which has viscosity that is bigger than the viscosity of water and which shows weak acidic characteristic, and at the same time, it is a strong oxidizer.

[0040] Formula 1 .

[0041] The invention owners have made studies related to increasing of the GAPDH amount of hydrogen peroxide compound and the correlation of said disease and GAPDH activity after the invention owners have detected the information related to the low level of GAPDH activity in diabetic retinopathy disease.

[0042] In the first study realized by the present invention owners, it has been detected that the hydrogen peroxide compound has a surprising effect in increasing of GAPDH amount depending on existence duration of hydrogen peroxide in HepG2 cells. In the tests made, the existence duration of hydrogen peroxide compound in HepG2 cells is between 2 and 4 hours. The realized experiment results are shared as Figure 1. After HepG2, T98G and Hek293 cells have been treated with 0.4 nM H2O2 for 2 hours and 4 hours, protein extraction has been realized with western blot protein extraction lysis buffer (50 mM Tris [pH 8.0], 5 mM EDTA, 150 mM NaCI, 0.5% NP-40, 1 mM PMSF and 0.01 mg of aprotinin / ml), and after the proteins executed in SDS-PAGE are transferred to nitro-cellulose membrane, it has been blotted with GAPDH antibodies and afterwards with secondary antibodies, GAPDH amounts have been determined on the film by means of chemiluminescent substrate.

[0043] Another study made by the invention owners relates to increasing of GAPDH amount also in T98G cell, which is human glioma cell, by the hydrogen peroxide compound. The related test result has been shared as Figure 2.

[0044] Another study made by the invention owners relates to increasing of GAPDH amount also in Hek293 cell, which is immortalized kidney epithelial cell, by the hydrogen peroxide compound. The related test result has been shared as Figure 3.

[0045] In another study, the invention owners have detected that the hydrogen peroxide compound has decreased chemotrypsin activity of proteasome in proteasome chemotrypsin activity test in HepG2 cells. The realized experiment results are shared as Figure 4.

[0046] In another study made by the invention owners, the invention owners have detected that the Ubiquitin amount has increased which is the finding that shows that the proteasome activity is inhibited in HepG2 cells by hydrogen peroxide compound. The realized experiment results are shared as Figure 5.

[0047] As a result of the made tests and the search and development activities, it has been detected that hydrogen peroxide can be used as treating active substance in elimination of diabetic retinopathy diseases which lead to diseases like cataract, glaucoma, refractive errors, optic nerve inflammation, cycloplegia and various diseases in the retina, blindness depending on diabetes, and hydrogen peroxide can be provided in the pharmaceutical compositions that can be used for the treatment of these diseases. In a preferred application, the pharmaceutical composition comprises hydrogen peroxide at a value between 0.004 nM and 4 nM in order to show technical effect in the pharmaceutical composition. In case preferred, the pharmaceutical composition can comprise auxiliary active substance and / or auxiliary components.

[0048] As a result of the studies made within the scope of the present invention, it has been detected that the inhibition of proteasome chemotrypsin enzyme has taken place in the presence of hydrogen peroxide compound, and depending on this, degradation of GAPDH protein decreases and thus, GAPDH protein amount increases in the cell. As a result, it has been detected that the reduction of GAPDH amount, which leads to occurrence frequency of diabetic retinopathy disease in persons who suffer from diabetes, can be prevented by using hydrogen peroxide compound.

[0049] In a perspective, the present invention relates to pharmaceutical compositions comprising hydrogen peroxide compound as active substance in elimination of diabetic retinopathy disease that occurs depending on diabetes and in elimination of symptoms resulting from these diseases. In obtaining said pharmaceutical compositions, more than one active substance can be used besides the hydrogen peroxide compound. Moreover, it is possible to add more than one auxiliary component in obtaining said pharmaceutical compositions. The protection scope of the present invention is the solutions that include hydrogen peroxide compounds as active substance for increasing GAPDH amount in the bodies of patient persons in said pharmaceutical compositions.

[0050] The protection scope of the present invention is set forth in the annexed claims and cannot be restricted to the illustrative disclosures given above, under the detailed description. It is because a person skilled in the relevant art can obviously produce similar embodiments under the light of the foregoing disclosures, without departing from the main principles of the present invention.

Claims

CLAIMS The present invention is usage of hydrogen peroxide (H2O2) compound as the treating active substance which provides inhibition of proteasome enzyme(s) in elimination of diabetic retinopathy diseases which lead to diseases like cataract, glaucoma, refractive errors, optic nerve inflammation, cycloplegia and various diseases in the retina, blindness depending on diabetes, and the symptoms resulting from these diseases, and thus which provides increase of GAPDH amount. The usage according to claim 1 , wherein said proteasome enzymes are trypsin, chemotrypsin and / or caspase-like enzyme. The present invention is a pharmaceutical composition comprising hydrogen peroxide (H2O2) compound as the treating active substance which provides inhibition of proteasome enzyme(s) in elimination of diabetic retinopathy diseases which lead to diseases like cataract, glaucoma, refractive errors, optic nerve inflammation, cycloplegia and various diseases in the retina, blindness depending on diabetes, and the symptoms resulting from these diseases, and thus which provides increase of GAPDH amount. The pharmaceutical composition according to claim 3, wherein hydrogen peroxide (H2O2) compound exists as active substance at an amount between 0.004 nM and 4 nM in said pharmaceutical composition. The pharmaceutical composition according to claims 3-4, wherein said pharmaceutical composition comprises at least one active substance besides hydrogen peroxide (H2O2) compound. The pharmaceutical composition according to claims 3-5, wherein hydrogen peroxide (H2O2) compound exists as active substance, and it moreover comprises at least one auxiliary component.

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