AU749673B2 - Use of glycosaminoglycans for producing pharmaceutical preparations for treating diabetes-associated diseases of the eye - Google Patents

Description

i 1, The use of glycosaminoglycans for producing pharmaceutical preparations for treating eye disorders associated with diabetes The invention relates to the use of glycosaminoglycans and specifically low molecular weight heparins for producing pharmaceutical preparations for the prevention or treatment of eye disorders associated with diabetes mellitus wherein said low molecular weight heparin has an average molecular weight of greater than 2000 Dalton. The invention further relates to pharmaceutical preparations suitable for treating eye disorders associated with diabetes.

The use of glycosaminoglycans and specifically of low molecular weight heparins and heparinoids for producing pharmaceutical preparations for treating perfusion disorders is well known.

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. Americ. Soc. Nephrol. 8 (1997) 456 462).

15 US 5,032,679 claims the use of glycosaminoglycans for inhibiting the proliferation of S.i: smooth muscle cells and the diseases associated therewith.

US 4,966,894 claims polysulfated heparins for treating diseases caused by retroviruses.

Retinopathy is the commonest and most serious complication of the 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 S and duration of the diabetes. The current findings on the retina also play an important part in the 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 25 the study: A: under 30years 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).

The development of retinopathy was observed and recorded 0480/01179 2 photographically. The patients in the study were checked regularly during the follow-up period and, if necessary, treated by the laser coagulation method.

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 3.2% were blind.

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 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.

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 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 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 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 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.

.Q 45 The pharmaceutical preparations produced for the use according to the invention may contain the abovementioned compounds as free 0480/01179 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 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.

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 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 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 as example thereof. Examples of natural glycosaminoglycans are heparin-,-heparan sulfate, keratan sulfate, dermatan sulfate,chondroit-inor chondfoitin-sulfate. Heparan sulfate corresponds to heparin but has-fewer N- and O-suTrate groups and more N-acety- groups--The-glycosaminoglycans or heparinoids advantageous for the use according to the invention or for the pharmaceutical preparations are selected from the group of dermatan sulfate, heparan sulfate, dextran sulfate (US 5,541, r66~), xylan sulfate (such--as pentosan polysulfate, EP-A-0 184 480, FR -35170), heparin or derivatives of these 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 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 removed by extraction with an organic solvent. Finally, the proteins are removed by proteolytic digestion and thus the 0480/01179

A

4 glycosaminoglycans are obtained. Charles et al. (Biochem. J. (1936) 1927 1933) and Coyne, E. in Chemistry and Biology of Heparin (Elsevier Publishers, North Holland, Lunblad, R.L., eds., 1981) describe methods for isolating heparin, for example.

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 sulfur content of from 6 to 15% by weight, preferably from 13 to 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.

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 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.

Low molecular weight glycosaminoglycans, for example low molecular weight heparins and/or dermatan sulfates, can be prepared by a nunber 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 (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 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 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 Sinhibitors of the renin-angiotensin system or their combination.

0480/01179 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.

Examples of drugs which lower blood glucose are insulin, agents which lower blood glucose for oral use, such as sulfonylureas, or a-glucosidase inhibitors.

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.

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 the invention can be administered in a conventional way orally or parenterally (subcutaneously, intravenously, intramuscularly, intraperitoneally), and oral or intravenous administrations are preferred.

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 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.

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 mg/kg of bodyweight/day on parenteral administration or in a 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 those mentioned above, are in principle all pharmaceutical forms Swhich can be used for oral or parenteral administration, whether 0480/01179 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 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 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 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, 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 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 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 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 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.

The pharmaceutical preparations may moreover comprise preservatives, solubilizers, stabilizers, wetting agents, 0480/01179 7 emulsifiers, sweeteners, colorants, flavorings, salts to alter the osmotic pressure, buffers, coating agents and/or antioxidants.

Examples Retinopathy study a) Preclinical studies Animal experimental studies were carried out to investigate the development of diabetic retinopathy on treatment with various glycosaminoglycans (especially Reviparin®) by comparison with placebo.

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 3 cells of the pancreatic islets of Langerhans. After 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 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 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.

Material and methods Species: rat Type: Sprague Dawley, Janvier, France Sex: male Weight: 220 to 240 g animals were used in the study. All the animals received a single intraveneous 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 Langerhans. After a few days, the blood glucose levels in these rats rose 25 mmol/l in all animals) significantly by 0480/01179 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 blood.

In the s.c. test subcutaneous), the animals received 1 mg/kg Reviparin® administered subcutaneously in a volume of 1 ml/kg. The corresponding control group received the corresponding amount of placebo isotonic saline).

In the p.o. test oral), the animals received 5 mg/kg Reviparin® administered by gavage. The volume was 5 ml/kg.

The corresponding control group received the corresponding amount of placebo isotonic saline).

Number of Group animas Diabetes induction Treatment animals Placebo S 1 10 60 mg/kg streptozotozin Placebo s.c. on days 8 28 2 60 mg/kg streptootoin 1 mg/kg Reviparin® s.c. on days 8 28 3 10 60 mg/kg streptozotozin laceb p.o. on days 8 28 4 10 60 mg/kg streptozotozin 5 mg/kg Reviparin Sp.o. on days 8 28 The animals were treated on workdays. No treatment of the animals took place on Saturday and Sunday.

Protocol Day 0: Induction of diabetes with streptozotozin 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) Scoring system: 0: No emergence of FDex 1: Emergence from <2 leakage points and/or slight increase in the background fluorescence 2: Emergence from 2 leakage points and/or large N increase in the background fluorescence 0480/01179 9 3: Extensive emergence from all vessels within minutes after administration of FDex Results: Of the total of 40 animals used in the study, 5 animals died during the experimental period (group i: 1 animal, group 3: 2 animals, group 4: 2 animals).

Administration of Reviparin® resulted, after subcutaneous administration for 3 weeks, in a significant reduction in the exudation of fluorescence-labeled macromolecule (see Figure Figure 1 illustrates the effect of Reviparin® (1 mg/kg subcutaneously) on vessel permeability in STZ 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.

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® 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 specific changes in the fundus of the eye during treatment with Reviparin.

The study inclusion and exclusion criteria were defined as follows: 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 3 months previously, panfundus laser coagulation performed previously (all causes), clinical need for laser coagulation because of severe non-proliferative or proliferative diabetic 0480/01179 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 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 reliable contraception (hormonal contraception or intrauterine pessary), HbAlc greater than uncontrolled arterial hypertension.

From a population of 36 patients eventually 11 patients were 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 (n time since diagnosis 21.73 10.53 (3 42) years, height 167 11.3 (150 186) cm, weight 79.3 11.3 (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 (54 89) beats per minute.

Concomitant disorders: The following secondary and concomitant disorders were found in the patients: arterial hypertension (n coronary heart disease (n arterial occlusive disease (n previous stroke (n peripheral diabetic neuropathy (n diabetic 3 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 lichen ruber planus (n 1).

There were no allergies in any of the patients. 4 patients were smokers, and 6 patients had hyperlipidemia.

Concurrent medication: The patients received the long-term medication listed in N Table I. No new medicines were prescribed or discontinued during the treatment period. The insulin dose for patients 0480/01179 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.

Physical examination: The clinical examinations for inclusion of the patients revealed no pathological findings which would have excluded 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.

Investigations of safety parameters The following parameters were and will be investigated before, during and after the treatment period (after the 12th 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 phosphatase, gGT [sic], ASAT, ALAT, bilirubin), cholesterol, triglycerides, serum glucose, HbAlc, total protein, albumin, blood picture (leukocytes, erythrocytes, hemoglobin, platelets), coagulation parameters (ATIII, aPTT, HEP test) In the urine: creatinine, total protein, albumin, creatinine clearance As a further parameter, the blood pressure was investigated 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 ,Aq improvement in specific changes in the fundus associated with diabetic retinopathy. For this purpose, before Reviparin® 0480/01179 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 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 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 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 according to the following assessment scale distinctly better, slightly better, the same, slightly worse, distinctly worse.

Interim result after treatment for 12 weeks 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 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 /1]m [sic] another patient developed leukopenia with a minimum of 3.31 [x 109/1]. 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.

0480/01179 13 There were likewise no alterations in blood pressure regulation (see Table IV).

Efficacy variables: of the total of 20 eyes to be examined (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 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 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.

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, 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 weeks show a tendency for the retinopathy to improve.

However, final evaluation of all the eyes has yet to be performed.

0480/01179 14 Table I: Long-term medicinal treatment of the patients recruited for the Reviparin® treatment study Pre- Substance class Individual substances scriptions Platelet aggregation inhibitors 3 acetylsalicylic acid (max. daily dose 100 mg) ACE inhibitors 6 captopril, enalapril, fosinopril, ramipril Diuretics 7 hydrochlorothiazide, furosemide, torasemide Beta-blockers 2 metoprolol Other antihypertensives 2 moxonidine, nisoldipine Cardiac agents 5 digitoxin, isosorbide dinitrate, molsidomine In s 17 normal insulin, NPH insulin, combination Insulins 17 insulin Antilipemics 3 fluvastatin, lovastatin, sivastatin Antidepressants 1 amitriptyline Antiepileptics 2 carbamazepine, lamotrigine Transquilizers 1 lorazepam Gastrointestinal agents 1 omeprazole Mineral products 1 calcium 0480/0117 9 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 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 FAG X X X Blood picture X X X X X X X X X X X HEP test X -X X X X X X aPPT X AT II 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 S. alb. X X X S. crea. X X X X X X X X Crea. CI. 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 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. trigi. X X X gGT [sic] X -X X ALAT X ASAT X -X X S. bili X X X

S.

U.

HEP test alb.

crea.

Crea. Cl.

aPPT AT III chol.

trigl.

gGT [sic]

ASAT

Serum bili =Bilirubin Urine prot. Protein Heparin test BP Blood pressure Albumin TAG [sic] Fluorescence angiography Creatinine chi. =Chloride Creatinine clearance TP =Total protein Activated partial thromboplasmin [sic] time glu. =Glucose Antithrombin III HbAlc Glycosylated hemoglobin Aic [sic] Cholesterol MAU Microalbuminuria Triglycerides y-Glutamintransferase [sic] Alanine aminotransferase Aspartate aminotransferase 0480/01179 Table III: 16 Laboratory results before start of treatment (TO) and after treatment for 12 weeks (Tl) with Reviparin.

Means and standard deviation (SE) are stated.

TO TI [sic] M ±SE M SE 1. Blood picture Leukocytes [x10 9 7.5 2.5 7.5 2.3 Hemoglobin [g/dl] 14.0 1.8 14.2 1.4 Platelets [x10 9 265 36 252 36 2. Coagulation aPPT 25.4 1.6 AT III 90.1 7.7 Hep test 21.1 2.1 73.2 27.1 3. Kidney function Na [mmol/1] 137 2 139 1 K [mmol/1] 4.5 0.5 4.3 0.3 Cl [mmol/l] 95 4 95 4 S. TP 70.8 5.6 70.0 7.2 S. alb 44.1 5.0 43.2 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 U. TP [mg/d] 1261 2019 1394 2454 4. Metabolism Glucose [mg/dl] 211 83 148 HbAlc 7.3 0.8 7.4 1 chol. [mg/dl] 221 42 218 31 trigl. [mg/dl] 266 255 241 251 Liver function gGT [sic] 12 6 16 11 ALAT 10 4 19 6 ASAT 9 3 10 3 bili [mg/dl] 0.5 0.1 0.5 0.1

Claims (11)

1. The use of low molecular weight heparin for producing pharmaceutical preparations for the prevention or treatment of eye disorders associated with diabetes mellitus wherein said low molecular weight heparin has an average molecular weight of greater than 2000 Dalton.

2. The use of low molecular weight heparin as claimed in claim 1, wherein the low molecular weight heparin are administered orally or parenterally.

3. The use of low molecular weight heparin as claimed in claim 1 or 2, wherein the low molecular weight heparin are administered parenterally in a dose of from 0.1 to 30 mg/kg of bodyweight/day.

4. The use of low molecular weight heparin as claimed in claim 1 or 2, wherein the low molecular weight heparin are administered orally in a dose of from 0.2 to 500 mg/kg of bodyweight/day.

5. The use of low molecular weight heparin as claimed in any of claims 1 to 4, wherein the low molecular weight heparin have a sulfur content of from 6 to 15% by weight.

6. The use of low molecular weight heparin as claimed in any of claims 1 to 5, wherein the low molecular weight heparin have an average molecular weight in the range of greater than 2000 Dalton to less than but including 20,000 Dalton.

7. The use of low molecular weight heparin as claimed in any of claims 1 to 6, wherein the low molecular weight heparin, their derivatives or mixtures are used in the form of the free acids, in the form of their physiologically active salts or their mixtures.

8. A pharmaceutical preparation when used for the treatment of eye disorders associated with diabetes mellitus and comprises one or more low molecular weight heparins, physiologically active salts of these substances or their mixtures. 19

9. A combination of a pharmaceutical preparation as claimed in claim 8 and at least one drug which lowers blood pressure or at least one drug which lowers blood glucose, or their combination.

A combination as claimed in claim 9 comprising ACE inhibitors as a drug which lowers blood pressure.

11. A method of treatment or prophylaxis of diabetes mellitus comprising administering to a patient in need of such treatment or prophylaxis an efficacious amount of a combination as defined in claim 9. DATED this 1st day of October 2001 KNOLL AKTIENGESELLSCHAFT WATERMARK PATENT TRADEMARK ATTORNEYS 290 BURWOOD ROAD HAWTHORN VICTORIA 3122 AUSTRALIA SLCG:AMT:SLBP17203AU eoee LCG:AMT:SLB P17203AU00 a