AU1335599A - Use of glycosaminoglycans for producing pharmaceutical preparations for treating diabetes-associated diseases of the eye - Google Patents

Description

The use of glycosaminoglycans for producing pharmaceutical preparations for treating eye disorders associated with diabetes 5 The invention relates to the use of glycosaminoglycans for producing pharmaceutical preparations for the prevention or treatment of eye disorders associated with diabetes mellitus. The invention further relates to pharmaceutical preparations suitable for treating eye disorders associated with diabetes. 10 The use of glycosaminoglycans and specifically of heparins and heparinoids for producing pharmaceutical preparations for treating perfusion disorders is well known. 15 The use of glycosaminoglycans for a number of other diseases has recently been described. Thus, US 5,236,910 claims the use of glycosaminoglycans for treating diabetic nephropathy and neuropathy. The use of low molecular weight heparins for the same indication is described by van der Pijl et al. (J. Americ. Soc. 20 Nephrol. 8 (1997) 456 - 462). US 5,032,679 claims the use of glycosaminoglycans for inhibiting the proliferation of smooth muscle cells and the diseases associated therewith. 25 US 4,966,894 claims polysulfated heparins for treating diseases caused by retroviruses. Retinopathy is the commonest and most serious complication of the 30 eyes in diabetes mellitus. In industrialized countries it is the principal cause of blindness. The risk of an individual diabetic becoming blind is very difficult to estimate. It depends on many factors such as age, type and duration of the diabetes. The current findings on the retina also play an important part in the 35 assessment of the risk. An epidemiological study was carried out in the 1980s in Wisconsin, USA, on about 2500 people selected from a population of 10,000 diabetics. The patients were divided into 3 groups for 40 the study: A: under 30 years of age at onset of diabetes and treatment with an insulin therapy, B: over 30 years of age at onset of diabetes and treatment with an insulin therapy and C: "older" patients without insulin therapy (see Klein et al., Diabetes/Metabolism Reviews, 5 (1989) 559 - 570). 45 The development of retinopathy was observed and recorded 2 photographically. The patients in the study were checked regularly during the follow-up period and, if necessary, treated by the laser coagulation method. 5 The prevalence of retinopathy was 50.1% in the patients in the study, but only 2.2% had criteria associated with a risk of loss of vision. The prevalence of moderate impairment of vision in the "younger" group was 1.4%. 3.2% were blind. 10 At present, laser coagulation is the only effective treatment of retinopathy. Effective medicines for established retinopathy are not available as yet. A first elaborate method for treating retinopathy with the aid of 15 high molecular weight heparin in combination with a detoxified enzyme obtained from the venom of the Brazilian vipers Bothrops jararaca and Lachesis atrox is described and claimed in US 3,869,548. 20 A disadvantage of this method is that the detoxified enzyme must first be obtained in an elaborate process from the venom and then be detoxified. It was possible to treat retinopathy in an animal experiment on guinea pigs only in combination with high molecular weight heparin. Neither the heparin used nor the venom on its own 25 showed an effect in the treatment of retinopathy. It is an object of the present invention to provide means for treating retinopathy and/or macular degeneration which do not have the abovementioned disadvantages and can be employed 30 reliably, easily and simply for treating retinopathy and macular degeneration. We have found that this object is achieved by using glycosaminoglycans for producing pharmaceutical preparations for 35 the prevention or treatment of eye disorders associated with diabetes. The invention additionally relates to pharmaceutical preparations which are suitable for treating eye disorders associated with 40 diabetes and which comprise one or more substances selected from the group of glycosaminoglycan, glycosaminoglycan derivatives, heparinoids, physiologically active salts of these substances or their mixtures. JO 45 The pharmaceutical preparations produced for the use according to o the invention may contain the abovementioned compounds as free 3 compounds or in the form of their physiologically active salts, their tautomers and/or isomeric forms or in the form of the combination of the free compounds and the various salts. Examples of advantageous physiologically active salts which may be 5 mentioned are the Na, Ca or Mg salts. Salts with organic bases such as diethylamine, triethylamine or triethanolamine are also suitable. The pharmaceutical preparations may advantageously contain at least one free substance or at least one compound in the form of its salt or mixtures thereof. 10 The glycosaminoglycans (= mucopolysaccharides) utilized for the use according to the invention mean negatively charged polysaccharides (= glycans) consisting of variously linked disaccharide units in which, for example, 1 molecule of a uronic 15 acid such as D-glucuronic acid or L-iduronic acid is glycosidically linked to the 3 or 4 position of an aminosaccharide such as glucosamine or galactosamine. At least one of the sugars in the disaccharide has a negatively charged carboxylate or sulfate group which may be linked via an oxygen or 20 nitrogen atom. Glycosaminoglycans show a strongly acidic reaction due to the uronic acids and the sulfuric ester groups. These acidic groups may be naturally present and/or have been introduced synthetically into the compounds, for example by sulfation. The sulfation method of US 5,013,724 may be mentioned 25 as example thereof. Examples of natural glycosaminoglycans are heparin, heparan sulfate, keratan sulfate, dermatan sulfate, chondroitin or chondroitin sulfate. Heparan sulfate corresponds to heparin but has fewer N- and O-sulfate groups and more N-acetyl groups. The glycosaminoglycans or heparinoids 30 advantageous for the use according to the invention or for the pharmaceutical preparations are selected from the group of dermataq sulfate, heparan sulfate, dextran sulfate (US 5,541;16f6} iylan sulfate (su aspentosan polysulfate, EP-A-0 184 480, FR 8351)70),heparin or derivatives of these 35 substances. These substances are administered in an amount effective for treating eye disorders. Glycosaminoglycans can advantageously be isolated from animal tissues, such as intestinal mucosa or from the ears of pigs or 40 cattle. The tissues used for isolating the glycosaminoglycans are, for example, autolyzed and extracted with alkali. It is then possible to coagulate the protein and precipitate it, for example by acidification. After the precipitate has been taken up in a polar nonaquoeus solvent such as ethanol or acetone, the fats are 45 removed by extraction with an organic solvent. Finally, the proteins are removed by proteolytic digestion and thus the 4 glycosaminoglycans are obtained. Charles et al. (Biochem. J. 30 (1936) 1927 - 1933) and Coyne, E. in Chemistry and Biology of Heparin (Elsevier Publishers, North Holland, N.Y., Lunblad, R.L., eds., 1981) describe methods for isolating heparin, for example. 5 These glycosaminoglycans isolated from natural sources may advantageously also undergo a derivatization, for example by polysulfation, as described by way of example in US 5,013,724. This polysulfation results in the glycosaminoglycans having a 10 sulfur content of from 6 to 15% by weight, preferably from 13 to 15% by weight. Derivatives of the substances mean compounds which improve the properties of the glycosaminoglycans on use, in terms of their effect, their stability and their elimination from the body. 15 The glycosaminoglycans advantageously used are heparins and/or dermatan sulfate with an average molecular weight of from 1000 to 20,000 dalton, preferably from 1500 to 9000 dalton, particularly preferably from 2000 to 6000 dalton. Low molecular weight 20 heparins and/or dermatan sulfates, which may be polysulfates, are particularly advantageous, in the form of the free acid or in the form of a salt with physiologically tolerated bases or mixtures of these compounds. 25 Low molecular weight glycosaminoglycans, for example low molecular weight heparins and/or dermatan sulfates, can be prepared by a number of-methods-.The preparation of low molecular weight heparins by depolymerization using nitrous acid is described, for example, in EP-B-0 037 319 or in Biochemistry 15 30 (1976) 3932. Low molecular weight heparins and low molecular weight glycosaminoglycans can also be prepared using enzymes (Biochem. J. 108 (1968) 647), using sulfuric acid and chlorosulfonic acid (FR No. 2,538,404), using periodate or using physical methods such as y radiation (EP-A-0 269 937) or 35 ultrasound (Fuchs et al., Lebensm. Unters. Forsch. 198 (1994) 486 - 490). The invention further relates to combination products consisting of pharmaceutical preparations which comprise glycosaminoglycans 40 such as low molecular weight heparins and/or dermatan sulfates, and at least one drug which lowers blood pressure or at least one drug which lowers blood glucose or their combination. Drugs which lower blood pressure mean, for example, inhibitors of endothelin converting enzyme (ECE), endothelin antagonists or inhibitors of the renin-angiotensin system or their combination.

5 Inhibitors of the renin-angiotensin system are renin inhibitors, angiotensin-II antagonists and, in particular, angiotensin converting enzyme (ACE) inhibitors. The combination of drugs achieves an advantageous lowering of blood pressure. 5 Examples of drugs which lower blood glucose are insulin, agents which lower blood glucose for oral use, such as sulfonylureas, or a-glucosidase inhibitors. 10 The combinations of said drugs (glycosaminoglycans, drugs which lower blood pressure and/or blood glucose) can be administered in a single pharmaceutical form or temporally and spatially separate. 15 Concerning the dosage and mode of administration, the same factors must be taken into account as for the corresponding single substances. The pharmaceutical preparations produced and used according to 20 the invention can be administered in a conventional way orally or parenterally (subcutaneously, intravenously, intramuscularly, intraperitoneally), and oral or intravenous administrations are preferred. 25 The dosage depends on the age, condition and weight of the patient and on the mode of administration. The glycosaminoglycans are advantageously administered in a dose of from 0.1 to 500 mg/kg of bodyweight/day. In the case of 30 parenteral use, the glycosaminoglycans are advantageously administered in a dose of from 0.1 to 30 mg/kg of bodyweight/day, and in the case of oral use they are administered in a dose of from 0.2 to 500 mg/kg of bodyweight/day, it being possible to administer the dose in a single dose or in divided doses. 35 Mixtures of, for example, at least one low molecular weight heparin and/or its polysulfated derivative and/or at least one low molecular weight dermatan sulfate and/or its polysulfated derivative are also administered in a dose of from 0.1 to 30 mg/kg of bodyweight/day on parenteral administration or in a 40 dose of from 0.2 to 500 mg/kg/day on oral administration. Pharmaceutical preparations comprising glycosaminoglycans for treating eye disorders caused by diabetes mellitus, or combinations of these polysaccharides with other drugs among 45 those mentioned above, are in principle all pharmaceutical forms 50/ which can be used for oral or parenteral administration, whether 0 6 solid or liquid, such as uncoated or (film-)coated tablets, capsules, powders, granules, suppositories, solutions or suspensions. These are produced in a conventional way. The drugs can moreover be processed with conventional pharmaceutical aids 5 such as tablet binders, bulking agents, preservatives, tablet disintegrants, flow regulators, plasticizers, wetting agents, dispersants, emulsifiers, solvents, release-slowing agents, antioxidants and/or propellant gases (cf. H. Sucker et al.: Pharmazeutische Technologie, Thieme-Verlag, Stuttgart, 1991). The 10 forms obtained in this way normally contain from 0.1 to 90% by weight of the drug. Owing to the potentiation of the effect of the individual components, the combination of the various classes of effect is 15 an ideal addition. The combinations according to the invention are generally administered orally, for example in the form of uncoated or (lacquer-)coated tablets, hard and soft gelatin capsules, 20 solutions, emulsions or suspensions. However, administration can also take place rectally, for example in the form of suppositories, or parenterally, for example in the form of solutions for injection. The drugs can be administered in the form of products which contain both drugs together, such as 25 tablets or capsules, or separately as ad hoc combination of single substances, which can be administered simultaneously or sequentially. To produce uncoated or (lacquer-)coated tablets and hard gelatin 30 capsules, a combination according to the invention can be processed with pharmaceutically inert, inorganic or organic excipients. Excipients which can be used for uncoated or coated tablets and hard gelatin capsules are lactose, cornstarch or derivatives thereof, talc, stearic acid or its salts. Excipients 35 suitable for soft gelatin capsules are vegetable oils, waxes, fats, semisolid and liquid polyols. Excipients suitable for producing solutions and syrups are, for example, water, polyols, sucrose, invert sugar, glucose and the 40 like. Excipients suitable for solutions for injection are water, alcohols, polyols, glycerol, vegetable oils. Excipients suitable for suppositories are natural or hardened oils, waxes, fats, semiliquid or liquid polyols and the like. 45 The pharmaceutical preparations may moreover comprise preservatives, solubilizers, stabilizers, wetting agents, 7 emulsifiers, sweeteners, colorants, flavorings, salts to alter the osmotic pressure, buffers, coating agents and/or antioxidants. 5 Examples Retinopathy study a) Preclinical studies 10 Animal experimental studies were carried out to investigate the development of diabetic retinopathy on treatment with various glycosaminoglycans (especially Reviparin@) by comparison with placebo. 15 To induce diabetes and the associated sequelae (eg. retinopathy, nephropathy), rats were given a single i.v. dose of 60 mg/kg streptozotozin. This treatment led to destruction of the P cells of the pancreatic islets of Langerhans. After 20 a few days, the blood glucose levels in these rats rose significantly by comparison with animals not treated with streptozotozin, and the animals lost weight, caused by the inability to utilize glucose as a consequence of the deficiency of insulin which is necessary for uptake of 25 glucose into the cells from the blood. Daily administration of Reviparin@ (= low molecular weight heparin, average molecular weight 4000 dalton) in a dose of 1 mg/kg intravenously or 5 mg/kg orally significantly, by 30 comparison with placebo, impeded the development of diabetic retinopathy, which in rats can be measured semiquantitatively with the aid of a score through an increase in vessel permeability as early as possible. 35 Material and methods Species: rat Type: Sprague Dawley, Janvier, France Sex: male 40 Weight: 220 to 240 g 40 animals were used in the study. All the animals received a single intraveneous (= i.v.) administration of streptozotozin (= STZ) in a dose of 60 mg/kg. This treatment led to destruction of the P cells of the pancreatic islets of 45 Langerhans. After a few days, the blood glucose levels in these rats rose (> 25 mmol/l in all animals) significantly by 0 8 comparison with animals not treated with streptozotozin, and the animals lost weight, caused by the inability to utilize glucose as a consequence of the deficiency of insulin which is necessary for uptake of glucose into the cells from the 5 blood. In the s.c. test (s.c. = subcutaneous), the animals received 1 mg/kg Reviparin® administered subcutaneously in a volume of 1 ml/kg. The corresponding control group received the 10 corresponding amount of placebo (= isotonic saline). In the p.o. test (p.o. = oral), the animals received 5 mg/kg Reviparin® administered by gavage. The volume was 5 ml/kg. The corresponding control group received the corresponding 15 amount of placebo (= isotonic saline). Group Number of Diabetes induction Treatment animals 1 10 60 mg/kg streptozotozin Placebo 20 s.c. on days 8 - 28 2 10 60 mg/kg streptozotozin 1 mg/kg Reviparin" s.c. on days 8 - 28 3 10 60 mg/kg streptozotozin Placebo p.o. on days 8 - 28 25 4 10 60 mg/kg streptozotozin 5 mg/kg Reviparin" p.o. on days 8 - 28 The animals were treated on workdays. No treatment of the animals took place on Saturday and Sunday. 30 Protocol Day 0: Induction of diabetes with streptozotozin 35 Day 7: Measurement of the blood glucose level Day 28: Measurement of the vascular permeability after injection of a macromolecular marker (FITC-dextran 150,000, = FDex) 40 Scoring system: 0: No emergence of FDex 1: Emergence from < 2 leakage points and/or slight increase in the background fluorescence A 5 2: Emergence from > 2 leakage points and/or large increase in the background fluorescence 9 3: Extensive emergence from all vessels within 5 minutes after administration of FDex Results: 5 Of the total of 40 animals used in the study, 5 animals died during the experimental period (group 1: 1 animal, group 3: 2 animals, group 4: 2 animals). 10 Administration of Reviparing resulted, after subcutaneous administration for 3 weeks, in a significant reduction in the exudation of fluorescence-labeled macromolecule (see Figure 1). Figure 1 illustrates the effect of Reviparin® (1 mg/kg subcutaneously) on vessel permeability in STZ 15 diabetic rats. It was also possible after oral administration of Reviparin® to reduce the extent of exudation (Figure 2). Figure 2 illustrates the effect of Reviparin® (5 mg/kg orally) on vessel permeability in STZ diabetic rats. 20 b) Clinical studies 11 patients with Type I or Type II diabetes mellitus took part in the study. The intention in the study was to inject the patients subcutaneously with 4300 units of Reviparin® 25 once a day in the morning for 12 weeks. Thereafter, the patients were to be followed up for a period of 12 weeks (total observation period 24 weeks. The primary observation variable was defined as the change in 30 specific changes in the fundus of the eye during treatment with Reviparin. The study inclusion and exclusion criteria were defined as follows: 35 Inclusion criteria: Type I or Type II diabetes mellitus, mild or moderate non-proliferative retinopathy, age 18-80 years 40 Exclusion criteria: Severe non-proliferative diabetic retinopathy, proliferative diabetic retinopathy, focal laser coagulation performed up to 5 3 months previously, panfundus laser coagulation performed previously (all causes), clinical need for laser coagulation because of severe non-proliferative or proliferative diabetic 10 retinopathy, treatment with medicines which are known to interact with hemostasis or platelet function (NSAIDs, prostaglandin synthesis inhibitors, anticoagulants [ASA > 100 mg], dextrans), treatment with steroids, known 5 heparin-induced thrombocytopenia, known allergy to heparin and sulfates, gastrointestinal hemorrhages (patients with a history of gastroinestinal hemorrhage should be treated with H2 blockers or with omeprazole), pregnancy or desired pregnancy, female patients of childbearing age must carry out 10 reliable contraception (hormonal contraception or intrauterine pessary), HbAlc greater than 9%, uncontrolled arterial hypertension. From a population of 36 patients eventually 11 patients were 15 selected according to these criteria. The demographic data for the patients are as follows: 4 women and 7 men, aged 52.45 ± 18.73 (31 - 74) years (M ± SE[range]) with diabetes mellitus type I (n = 6) and type II 20 (n = 5), time since diagnosis 21.73 ± 10.53 (3 - 42) years, height 167 ± 11.3 (150 - 186) cm, weight 79.3 ±E11.3 (57 - 97.7) kg, body mass index 28.8 ± 4.83 (24.1 - 38.2) kg/m 2 , average blood pressure for 24-hour measurement 125/75 ± 13/7 (100/58 - 150/82) mmHg, pulse rate 77.4 ± 12.2 25 (54 - 89) beats per minute. Concomitant disorders: The following secondary and concomitant disorders were found 30 in the patients: arterial hypertension (n = 7), coronary heart disease (n = 5), arterial occlusive disease (n = 2), previous stroke (n = 1), peripheral diabetic neuropathy (n = 4), diabetic 35 nephropathy (n = 5, incipient with microalbuminuria in 4 patients, macroalbuminuria in 1 patient), chronic renal insufficiency (n = 4; including one patient with chronic glomerulonephritis with nephrotic sydrome), genuine epilepsy (n = 1), lichen ruber planus (n = 1). There were no allergies in any of the patients. 4 patients 40 were smokers, and 6 patients had hyperlipidemia. Concurrent medication: The patients received the long-term medication listed in Table I. No new medicines were prescribed or discontinued during the treatment period. The insulin dose for patients 0 11 injecting insulin was kept constant. One patient took an antihistamine as required for allergic rhinitis. Three of the patients were taking long-term treatment with acetylsalicylic acid 100 mg a day. 5 Physical examination: The clinical examinations for inclusion of the patients revealed no pathological findings which would have excluded 10 the patients from taking part in the treatment study (particular findings: 2 cases of chronic foot ulcers, one case of left hemiparesis particularly affecting the arm). The findings in the physical examination correlated with the previously known disorders. 15 Investigations of safety parameters The following parameters were and will be investigated before, during and after the treatment period (after the 12th 20 week) and after the follow-up period (after the 24th week): In the blood: retention values (urea, creatinine), serum electrolytes (sodium, potassium, chloride), liver function test results (alkaline 25 phosphatase, gGT [sic], ASAT, ALAT, bilirubin), cholesterol, triglycerides, serum glucose, HbAlc, total protein, albumin, blood picture (leukocytes, erythrocytes, hemoglobin, platelets), coagulation parameters (ATIII, 30 aPTT, HEP test) In the urine: creatinine, total protein, albumin, creatinine clearance As a further parameter, the blood pressure was investigated 35 by 24-hour blood pressure measurement before and at the end of the treatment period and at the end of the follow-up period. The exact times of the investigation of the invididual parameters are to be found in Table II. Investigations of the efficacy The treatment study was started in order to achieve an 5 improvement in specific changes in the fundus associated with diabetic retinopathy. For this purpose, before Reviparin@ 12 treatment started, a fluorescence angiography was performed with the Heidelberger fluorescence angiograph with a maximum image section of 30* per single film. The disk was fixed as nasal edge mark in order to be able to compare the same image 5 segments with one another in all checkups. The films were each taken with the best possible area coverage, taking optimal image quality into account. In this study, the specific changes, detected by fluorescence angiography, of mild non-proliferative retinopathy (in particular number and 10 extent of intraretinal microaneurysms, and point hemorrhages and hard exudates) were compared on two fluorescence angiography films at the start of the treatment and after treatment for twelve weeks. The fluorescence angiography films for 2 patients at the end of the Reviparin® treatment 15 and for all at the end of the follow-up period have yet to be obtained. The fluorescence angiography was evaluated by qualitative criteria (microaneurysms, intraretinal hemorrhage) and 20 according to the following assessment scale ++ distinctly better, + slightly better, = the same, - slightly worse, -- distinctly worse. Interim result after treatment for 12 weeks 25 Safety variables: during the treatment period there were no changes in the retention parameters or alterations in the serum electrolyte concentration and the liver function test results. The serum glucose levels were subject to large 30 variations because the patients were not usually fasting when the blood samples were taken. One patient developed during the Reviparin® treatment a slight leukocytosis up to a maximum of 13,300 [x 10 9 /l]m [sic] another patient developed leukopenia with a minimum of 3.31 35 [x 109/l]. No other changes in leukocytes were observed. There were no alterations in the erythrocyte concentration, the hemoglobin, the hematocrit or the platelet concentration. Further parameters to be examined were anti-Xa activity and creatinine clearance. No deterioriation was found. For protein excretion, two patients with Type I diabetes showed a decrease in microalbuminuria during the treatment (34.7 mg/day before Reviparin® treatment, 730 mg/day after 12 weeks). The laboratory results are summarized in Table III. 0 13 There were likewise no alterations in blood pressure regulation (see Table IV). Efficacy variables: of the total of 20 eyes to be examined 5 (one female patient discontinued the treatment), 16 eyes were examined and evaluated semiquantitatively by the abovementioned criteria. The fundus of the eye was initially examined only by fundus photography. It was therefore possible to investigate, and compare semiquantitatively, only 10 the extent of the hard exudates with this mode of assessment. Examination of the hard exudates showed no alteration from the previous finding in 10 eyes. In 3 eyes there was an improvement (+), and in 3 further eyes there was a deterioration (-) in the fundus finding. None of the eyes 15 showed a distinct improvement or deterioriation. Two of the improved eyes belonged to a patient with Type II diabetes, and the remaining improved eye belonged to a patient with Type I diabetes. All the eyes with slight deterioriation belonged to patients with Type II diabetes. 20 To evaluate specific changes in the fundus of the eye, fluorescence angiography was performed in this study for the first time. This method allows more detailed examination of changes typical of retinopathy, such as microaneurysms, 25 intraretinal hemorrhages or - in cases of severe non-proliferative retinopathy - intraretinal microvascular anomalies (= IRMA). This diagnostic method is thus the gold standard for detecting pathological changes in fundus vessels. The results with this method after treatment for 12 30 weeks show a tendency for the retinopathy to improve. However, final evaluation of all the eyes has yet to be performed. 35 40 0 14 Table I: Long-term medicinal treatment of the patients recruited for the Reviparin® treatment study 5 Substance class Pre- Individual substances Substnce lassscriptions Platelet aggregation inhibitors 3 acetylsalicylic acid (max. daily dose 100 mg) ACE inhibitors 6 captopril, enalapril, fosinopril, ramipril Diuretics 7 hydrochlorothiazide, furosemide, torasemide 10 Beta-blockers 2 metoprolol Other antihypertensives 2 moxonidine, nisoldipine Cardiac agents 5 digitoxin, isosorbide dinitrate, molsidomine Insulins 17 normal insulin, NPH insulin, combination insulin 15 Antilipemics 3 fluvastatin, lovastatin, sivastatin Antidepressants 1 amitriptyline Antiepileptics 2 carbamazepine, lamotrigine Transquilizers 1 lorazepam 20 Gastrointestinal agents 1 omeprazole Mineral products 1 calcium 25 30 35 40 5 15 Table II: Flowchart (times of evaluation of the safety variables) Day Day Week Day Week Week Week Week Week Week Week 0 3 1 10 2 4 8 12 16 20 24 5 History X X X X X X X X Clinical X X X exam. BP X X X X X X X X 24h BP X X X 10 FAG X X X Blood picture X X X X X X X X X X X HEPtest X X X X X X X aPPT X 1 5 AT III X S.Na X X X X X X X X S.K X X X X X X X X S. chl. X X X X X X X X S. prot. X X X 20 S. alb. X X X S. crea. X X X X X X X X Crea. Cl. X X X X X X X U. alb. X X X X X X X U. TP X X X X X X X 25 S. glu. X X X X X X X X X HbAlc X X X X X X X S. chol. X X X S. trigl. X X X 30 gGT [sic] X X X ALAT X X X ASAT X X X S. bili X X X S. = Serum bili = Bilirubin 35 U. = Urine prot. = Protein HEP test = Heparin test BP = Blood pressure alb. = Albumin TAG [sic] = Fluorescence angiography crea. = Creatinine chl. = Chloride Crea. Cl. = Creatinine clearance TP = Total protein 40 aPPT = Activated partial thromboplasmin [sic] time glu. = Glucose AT III = Antithrombin III HbAlc = Glycosylated hemoglobin Alc [sic] chol. = Cholesterol MAU = Microalbuminuria trigl. = Triglycerides gGT [sic] = y-Glutamintransferase [sic] ALAT = Alanine aminotransferase 0 ASAT = Aspartate aminotransferase 16 Table III: Laboratory results before start of treatment (TO) and after treatment for 12 weeks (Tl) with Reviparin. Means (M) and standard deviation (SE) are stated. 5 TO TI [sic] M SE M ±SE 1. Blood picture Leukocytes [x10 9 /l] 7.5 2.5 7.5 2.3 10 Hemoglobin [g/dlJ 14.0 ± 1.8 14.2 1.4 Platelets [x10 9 /1] 265 ± 36 252 36 2. Coagulation 15 aPPT [s] 25.4 ± 1.6 AT III [%] 90.1 ± 7.7 Hep test [s] 21.1 ± 2.1 73.2 27.1 3. Kidney function 20 Na [mmol/l] 137 t 2 139 1 K [mmol/l] 4.5 0.5 4.3 0.3 Cl [mmol/l] 95 ± 4 95 4 S. TP [g/l] 70.8 ± 5.6 70.0 * 7.2 25 S. alb [g/l] 44.1 ± 5.0 43.2 8.0 S. crea. [mg/dl] 1.5 1.1 1.3 0.9 Crea. Cl. 91 68 95 63 MAU [mg/d] 740 1518 856 * 1971 30 U. TP [mg/d] 1261 2019 1394 2454 4. Metabolism Glucose [mg/dl] 211 t 83 148 55 HbAlc [%9] 7.3 0.8 7.4 1 35 chol. [mg/dl] 221 42 218 31 trigl. [mg/dl] 266 255 241 251 5. Liver function 40 gGT [sic] [U/l] 12 6 16 11 ALAT [U/l] 10 4 19 6 ASAT [U/l] 9 3 10 3 bili [mg/dl] 0.5 0.1 0.5 0.1 5 17 Table IV: Ambulatory 24-hour blood pressure measurement before start of treatment (TO) and after treatment for 12 weeks (TI) with Reviparin. The means for the recording period for the systolic and diastolic blood 5 pressures and for the pulse fate of the individual patients are stated; the mean of all the measurements and the standard deviation are indicated at the bottom. 10 Mean Mean Heart Mean Mean Heart Patient code systolic diastolic rate nz [sic] systolic diastolic rate Time loss TO TO TO TI [sic] T1 T1 2R1 128 80 84 127 76 81 2R2 115 72 82 15 2R3 100 58 54 109 64 86 1RI 131 82 67 124 78 69 1R2 124 81 87 116 76 95 1R3 119 78 89 126 83 85 20 1R4 119 71 89 122 74 83 1R6 151 78 77 129 73 81 1R7 133 66 62 1R9 138 78 89 1R12 122 73 76 25 Mean 125 74 78 122 74 83 SE 13 7 12 7 6 9 The study showed that it was possible to improve the patient's 30 retinopathy by administering Reviparin®. 35 40 45

Claims (12)

1. The use of glycosaminoglycans for producing pharmaceutical 5 preparations for the prevention or treatment of eye disorders associated with diabetes mellitus.

2. The use of glycosaminoglycans as claimed in claim 1, wherein at least one glycosaminoglycan selected from the group of 10 dermatan sulfate, heparan sulfate, dextran sulfate, xylan sulfate, heparin or derivatives of these substances is administered in an effective amount.

3. The use of glycosaminoglycans as claimed in claim 1 or 2, 15 wherein a low molecular weight heparin, heparan sulfate, dermatan sulfate or mixtures thereof are administered.

4. The use of glycosaminoglycans as claimed in any of claims 1 to 3, wherein the glycosaminoglycans are administered orally 20 or parenterally.

5. The use of glycosaminoglycans as claimed in any of claims 1 to 4, wherein the glycosaminoglycans are administered parenterally in a dose of from 0.1 to 30 mg/kg of bodyweight/ 25 day.

6. The use of glycosaminoglycans as claimed in any of claims 1 to 4, wherein the glycosaminoglycans are administered orally in a dose of from 0.2 to 500 mg/kg of bodyweight/day. 30

7. The use of glycosaminoglycans as claimed in any of claims 1 to 6, wherein the glycosaminoglycans have a sulfur content of from 6 to 15% by weight. 35

8. The use of glycosaminoglycans as claimed in any of claims 1 to 7, wherein the glycosaminoglycans have an average molecular weight in the range from 1000 to 20,000 dalton.

9. The use of glycosaminoglycans as claimed in any of claims 1 40 to 8, wherein the glycosaminoglycans, their derivatives or mixtures are used in the form of the free acids, in the form of their physiologically active salts or their mixtures.

10. A pharmaceutical preparation which is suitable for treating 45 eye disorders associated with diabetes and comprises one or more substances selected from the group of glycosaminoglycan, glycosaminoglycan derivatives, physiologically active salts 2 of these substances or their mixtures.

11. A combination of a pharmaceutical preparations [sic] as claimed in claim 10 and at least one drug which lowers blood 5 pressure or at least one drug which lowers blood glucose, or their combination.

12. A combination as claimed in claim 11 comprising ACE inhibitors as drug which lowers blood pressure. 10 15 20 25 30 35 40 45