FOLATE COMPOSITIONS

Abstract

A composition is described comprising at least one folate which is useful in the treatment of ocular diseases linked to elevated retinal venous pressure.

Claims

1. A preparation for treatment of systemic and ocular diseases wherein the disease is linked to elevated retinal venous pressure, the preparation comprising at least one folate.

2. The preparation according to claim 1, wherein the ocular disease is selected from the group consisting of diabetic retinopathy, macular degeneration and glaucoma.

3. The preparation according to claim 2, wherein the glaucoma is either a primary open angle glaucoma, a primary angle closure glaucoma or a normal tension glaucoma.

4. The preparation according to claim 1, wherein the preparation further comprises a sulfur donor compound.

5. The preparation according to claim 1, wherein the preparation further comprises at least one vitamin of the B complex.

6. The preparation according to claim 1, wherein the preparation further comprises arginine or arginine esters.

7. The preparation according to claim 1, wherein the preparation further comprises a choline donor.

8. The preparation according to claim 1, wherein the preparation further comprises acetylcholine.

9. The preparation according to claim 1, wherein the preparation further comprises glucosamine.

10. The preparation according to claim 1, wherein the preparation further comprises vitamin D.

11. The preparation according to claim 1, wherein the folate comprises a folate salt comprising the folate and a cation selected from the group consisting of arginine, choline, acetylcholine, 1,1-dimethylbiguanidine, phenylethylbiguanidine, glucosamine and dimethylaminoethanol.

12. The preparation according to claim 1, wherein the folate comprises a folate salt having an anion selected from the group consisting of 5-formyl-(6S)-tetrahydrofolic acid, 5-formyl-(6RS)-tetrahydrofolic acid, 10-formyl-(6R)-tetrahydrofolic acid, 5-methyl-(6S)-tetrahydrofolic acid, 5-methyl-(6RS)-tetrahydrofolic acid, (6S)-tetrahydrofolic acid, 5,10-methylene-(6R)-tetrahydrofolic acid, 5,10-methenyl-(6R)-tetrahydrofolic acid, 5,10-diformyl-(6S)-tetrahydrofolic acid, 5-methyl-10-formyl-(6S)-tetrahydrofolic acid.

13. The preparation according to claim 4, wherein the sulfur donor compound is N-acetylcysteine.

14. The preparation according to claim 5, wherein the at least one vitamin of the B complex is selected from the group consisting of vitamin B.sub.1, vitamin B.sub.2, vitamin B.sub.6, and vitamin B.sub.12.

15. The preparation according to claim 7, wherein the choline donor is betaine.

16. The preparation according to claim 10, wherein the vitamin D is vitamin D.sub.3.

17. A method for treating systemic and ocular diseases wherein the disease is linked to elevated retinal venous pressure in an individual, comprising the steps of administering a preparation to the individual comprising at least one folate.

18. The method according to claim 17, wherein the ocular disease is selected from the group consisting of diabetic retinopathy, macular degeneration and glaucoma.

19. The method according to claim 18, wherein the glaucoma is either a primary open angle glaucoma, a primary angle closure glaucoma or a normal tension glaucoma.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0031] FIG. 1 shows development of RVP before and during supplementation with Ocufolin® forte as discussed in the Examples.

DETAILED DESCRIPTION

[0032] As a normal level of retinal venous pressure (RVP) is considered a pressure to be equal or a few mm Hg above intraocular pressure (IOP). To measure the RVP, intraocular pressure (IOP) is increased by an ophthalmodynamometer (e.g. Löw contact lens dynamometer) or by IOPstim system (by Imedos) until the vein of interest pulsates. This applied force is also called ophthalmodynamometric force (ODF). That is to say, if a spontaneous venous pulsation is present (ODF=0), RVP equals IOP. If no spontaneous venous pulsation is present, the ODF necessary to provoke a spontaneous venous pulsation is measured and RVP is calculated as RVP=ODF+IOP.

[0033] Retinal venous pressure may be measured as describe for instance in Mozaffarieh M. et al., Graefes Arch Clin Exp Ophthalmol, 2014, 252, 1569-1571, by ophthalmodynamometry. The ophthalmodynamometric force may be measured as described in Mustur D. et al., The EPMA Journal, 2017, 8, 339-344.

[0034] Of course, retinal venous pressure can also be increased in a clinically healthy eye. But nevertheless, it can be a strong sign of a systemic disorder (e.g. autoimmune diseases).

[0035] A preparation according to the present invention for use in the treatment of ocular diseases comprises at least one folate or a salt thereof. The ocular disease is linked to an elevated retinal venous pressure.

[0036] Ocular diseases linked to, associated with or caused by elevated retinal venous pressure are diabetic retinopathy, macular degeneration (MD), glaucoma such as primary open angle glaucoma (POAG), primary angle closure glaucoma (PACG) and normal tension glaucoma (NTG).

[0037] Folates are reduced forms of folic acid which may also employed in preparations according to the present invention.

[0038] Folic acid is the oxidized form and the parent compound of biological folate. Because of its stability folic acid is used for supplements and food fortification. Folic acid is however not metabolically active and requires reduction and one carbon substitution before it is converted to 5-methyl tetrahydrofolate by several enzymatically catalyzed steps. While the enzymatic conversion of folic acid itself may be incomplete, disrupted or reduced at several points of its pathway, the effect of its deficiency may be multiplied as the folate metabolism is linked to other metabolic cycles which means, that a malfunction in one cycle may induce malfunctions in other metabolic cycles. A further form is 5-formyl tetrahydrofolate, wherein instead of the methyl moiety a formyl moiety is present.

[0039] In a further embodiment the preparation further comprises a sulfur donor compound, preferably N-acetyl cysteine.

[0040] In another embodiment the preparation additionally comprises at least one compound of the B vitamin complex. These B vitamins are preferably selected from the group consisting of vitamin B.sub.1, vitamin B.sub.2, vitamin B.sub.6, and vitamin B.sub.12. Vitamin B.sub.12 is particularly preferred.

[0041] In yet another embodiment the preparation further comprises arginine or an arginine ester.

[0042] In addition, the preparation may comprise a choline donor. A preferred choline donor is betaine.

[0043] The composition may also comprise the compound acetylcholine.

[0044] Additionally, the preparation may comprise glucosamine.

[0045] In a further embodiment, the composition comprises vitamin D, wherein vitamin D.sub.3 is particularly preferred.

[0046] In a preferred embodiment, the folate salt consists of a folate and a cation as counter ion wherein the cation is selected from the group consisting of arginine, choline, acetylcholine, 1,1-dimethylbiguanidine, phenylethylbiguanidine, glucosamine and dimethylaminoethanol.

[0047] The following folates are preferred, 5-formyl-(6S)-tetrahydrofolic acid, 5-formyl-(6RS)-tetrahydrofolic acid, 10-formyl-(6R)-tetrahydrofolic acid, 5-methyl-(6S)-tetrahydrofolic acid, 5-methyl-(6RS)-tetrahydrofolic acid, (6S)-tetrahydrofolic acid, 5,10-methylene-(6R)-tetrahydrofolic acid, 5,10-methenyl-(6R)-tetrahydrofolic acid, 5,10-diformyl-(6S)-tetrahydrofolic acid, 5-methyl formyl-(6S)-tetrahydrofolic acid. That means the anion of the folate salt is selected from the group consisting of the afore-mentioned folates.

[0048] The preparation may be formulated in different forms, such as a liquid preparation, a cream or in a capsule.

[0049] A liquid form of the preparation is for instance the following, the compositions according to the instant invention comprise the calcium salt, magnesium salt, sodium or zinc salt of levoleucovorin, and one or more of the compounds sodium gluconate, potassium gluconate, sodium lactate, potassium lactate, glycerophosphate disodium salt or glycerophosphate dipotassium salt. Further cations in the salt of levoleucovorin such as arginine may be used.

[0050] The composition may or may not contain additional excipients. Preferably the compositions are free of benzyl alcohol, tromethamine or monothioglycerol. Excipients such as mannitol for acceptable cake formation during the freeze-drying process, or sodium chloride and dextrose to adjust for osmolarity may be added to the compositions. The pH of the solutions is typically in the range of 6.5 to 8.5, and can be adjusted during drug product manufacturing with e.g. small amount of hydrochloric acid or sodium hydroxide. The solution may contain antioxidants to prevent oxidative degradation.

[0051] In the composition at least one additional compound, like the calcium salt, sodium salt, magnesium salt or zinc salt of leucovorin, (6R,S)-tetrahydrofolic acid, (6S)-tetrahydrofolic acid, 5,10-methylene-(6R,S)-tetrahydrofolate, 5,10-methylene-(6R)-tetrahydrofolate, 5-methyl-(6R,S)-tetrahydrofolate or 5-methyl-(6S)-tetrahydrofolate or a mixture of 2, 3 or more of said compounds can be used.

[0052] Preferred compositions according to the instant invention comprise one or more of the compounds sodium gluconate, potassium gluconate, sodium lactate, potassium lactate, glycerophosphate dipotassium salt or glycerophosphate disodium salt.

[0053] Regarding the amounts of the compounds in the compositions the following ratios are preferred. The compositions preferably contain for one mole of the calcium salts, sodium salts, magnesium salts or zinc salts of levoleucovorin, 0.8 to 6.0 moles, advantageously 1.0 to 4.0 moles, of sodium gluconate, potassium gluconate, sodium lactate or potassium lactate. In a practical embodiment the compositions preferably contain for one mole of the calcium salts, magnesium salts or zinc salts of levoleucovorin, 1.5 to 3.0 moles of sodium gluconate, or potassium gluconate. Such a composition may contain for one mole of the salt a minimum of 0.8, preferred of 1.0 moles, advantageous 1.5 moles, and a maximum of 6.0 moles, preferably of 4.0 moles, advantageous 3.0 moles, of sodium gluconate, potassium gluconate, sodium lactate or potassium lactate.

[0054] Other compositions preferably contain for one mole of the calcium salts, sodium salts, magnesium salts or zinc salts of levoleucovorin, 0.4 to 4.0 moles, preferably 0.5 to 3.0 moles, advantageously 0.7 to 2.0, of glycerophosphate disodium salt or glycerophosphate dipotassium salt. Such a composition may contain for one mole of the salt a minimum of 0.4, preferred of 0.5 moles, advantageously 0.7 moles and a maximum of 4.0 moles, preferably of 3.0 moles, advantageously of 2.0 moles of glycerophosphate disodium salt or glycerophosphate dipotassium salt.

[0055] Another exemplary composition or preparation which is suitable to be contained in a capsule is as follows, for example, the preparation contains the components calcium salt of folate, such as L-5-methyl-tetrahydrofolate or L-5-formyl-tetrahydrofolate, a sulfur donor compound such as N-acetylcysteine or its salt, a selene comprising compound such as L-selenomethionine, cholecalciferol, calcium D-panthothenate, vitamin B.sub.12, such as methylcobalamin, vitamin B.sub.6, such as pyridoxal-5′-phosphate, vitamin B.sub.2, such as riboflavin, vitamin B.sub.1, such as thiamine mononitrate, zeaxanthin, lutein, vitamin E, namely D-α-tocopherol, vitamin C, such as calcium ascorbate, a gluconate such as copper gluconate and zinc comprising compound, such as zinc oxide or zinc acetate.

[0056] For example, the preparation contains the components calcium salt of L-5-methyl-tetrahydrofolate in an amount of 0.2 mg to 2.7 mg, N-acetylcysteine or its salt in an amount of 40 mg to 540 mg, L-selenomethionine in an amount of 0.005 mg to 0.06 mg, cholecalciferol in an amount from 0.009 mg to 0.1 mg, calcium D-panthothenate in an amount from 1 mg to 15 mg, methylcobalamin in an amount from 0.003 mg to 0.1.5 mg, pyridoxal-5′-phosphate in an amount of 1 mg to 9 mg, riboflavin in an amount of 2 mg to 30 mg, thiamine mononitrate in an amount of 0.2 mg to 4.5 mg, zeaxanthin in an amount from 1 mg to 3 mg, lutein in an amount from 4 mg to 15 mg, D-α-tocopherol in an amount from 1 mg to 16 mg, calcium ascorbate in an amount from 20 mg to 100 mg, copper gluconate in an amount from 0.1 to 2 mg and zinc oxide in an amount from 7 mg to 80 mg.

[0057] The component N-acetylcysteine can be used both in the form of the free acid and in the form of one of its salts, for example as a sodium or calcium salt. N-acetylcysteine may also be present in the form of N-acetylcysteine-amide. A further sulfur donor compound is lipoic acid.

[0058] The composition further preferably contains one or more auxiliary substances selected from the group consisting of refined soybean oil, type NGM, soybean oil, partially hydrogenated 31°−37° C., soybean oil, partially hydrogenated 36° C.-42° C., rapeseed oil refined, glyceryl monosterate, sunflower or soy lecithin and beeswax. The rapeseed oil or soya oils provide protection of the active components against light and moisture.

[0059] In another preferred version, the composition contains one or more auxiliaries selected from the group consisting of refined soybean oil, type NGM, soybean oil, partially hydrogenated 31°-37° C., soybean oil, partially hydrogenated 36° C.-42° C., rapeseed oil refined, glyceryl monosterate, sunflower or soy lecithin, beeswax, triglycerides of medium or longer chain length (medium chain length: C.sub.6 to C.sub.12, longer chain length: C.sub.13 to C.sub.24), phosphoglycerides with at least one organic phosphoric acid ester, and ester compounds from alcohols with a chain length of C.sub.20 to C.sub.40 and fatty acids with medium or long chain length.

[0060] In another version, the composition is contained in a caspule, for instance a soft gelatine capsule, a vegan soft shell capsule or a hard gelatin capsule. The soft gelatine capsule preferably contains gelatine, glycerol 98%, glycerol 86%, sorbitol 70%, titanium dioxide, sodium copper chlorophylline and water.

[0061] An exemplary formulation for a cream comprises thoroughly mixed phases. Phase A comprises an oil. Phase B comprises glycerol and an emulsifier. The phase C comprises an aqueous folate composition comprising a physiologically effective amount of a folate salt, and optionally at least one compound selected from the group consisting of sodium gluconate, potassium gluconate, glycerophosphate disodium salt and glycerophosphate dipotassium salt. It may optionally comprise further compounds such as a pharmaceutically acceptable buffer, e.g. tris(hydroxymethyl)-aminomethan (TRIS), and a pharmaceutically acceptable antioxidant, e.g. glutathione. Optional phase D comprises one or more matting agents. Titanium dioxide may be used as a white dye. Alternatively, silica dioxide can be used.

[0062] A folate preparation according to the present invention preferably comprises in phase C 0.1 mg to 1000 mg at least one folate salt selected of the group consisting of calcium, magnesium, sodium, zinc, arginine, choline, acetylcholine, 1,1-dimethylbiguanidine, phenylethylbiguanidine, and dimethylaminoethanol salt of the folate per milliliter of a polar solvent. Polar solvents are for instance water, methanol, ethanol, n-propanol, isopropanol, glycerine, dimethylsulfoxide. Mixtures of such polar solvents may also be used. Arginine, choline, acetylcholine, 1,1-dimethylbiguanidine, phenylethylbiguanidine, glucosamine and dimethylaminoethanol salt of the folate are preferred.

[0063] In another embodiment the phase C comprises for one mole of the folate salt 0.8 to 10 molar sodium gluconate or potassium gluconate. If phase C comprises glycerophosphate disodium salt or glycerophosphate dipotassium salt, these salts are preferably present in a concentration of 0.4 to 5 molar.

[0064] Preferably, preparations according to the present invention comprise in phase A an oil consisting of a medium-chain triglycerol. Fatty acid of such medium-chain triglycerols have a chain-length in the range of C.sub.6 to C.sub.12. Most preferred is the oil of caprylic/capric acid triglycerol. Further, it may optionally comprise a dicarbonic acid alcohol. The dicarbonic acid has a chain length in the range of C.sub.2 to C.sub.10 and the alcohol is selected of the group consisting of methyl, ethyl, isopropyl, propyl, butyl, pentyl alcohol.

[0065] Emulsifiers used in phase B of preparations preferably have HLB value equal or greater than 5. The hydrophilic-lipophilic balance of a surfactant is a measure of the degree to which it is hydrophilic or lipophilic, determined by calculating values for the different regions of the molecule, as described by Griffin in 1954. Griffin's method for non-ionic surfactants as described in 1954 works as follows: HLB=20×Mh/M, wherein Mh is the molecular weight of the hydrophilic portion of the compound and M is the total molecular weight of the compound. An HLB value of 0 corresponds to a completely lipophilic/hydrophobic molecule, and a value of 20 corresponds to a completely hydrophilic/lipophobic molecule. An emulsifier having HLB value in the range of 8 to 16 is suitable for stabilizing oil in water (o/w) emulsions.

[0066] A preferred emulsifier is a sucrose ester, wherein the fatty acids have chain length in the range of C.sub.14 to C.sub.20. Such sucrose esters must have a HLB value equal or greater than 5. Most preferred is the emulator sucrose stearate.

Example 1

[0067] In a preliminary study a patient was treated with Ocufolin Forte®, a folate preparation as described above, wherein the preparation is comprised in a capsule. The treatment was performed for ten days. No side effects of the treatment were reported. The retinal venous pressure of the patient was measured as indicated before the treatment cycle and thereafter in both eyes. Before the treatment the RVP was measured as 56 mmHg (right eye) and 59 mmHg (left eye). The intraocular pressure (IOP) was determined to be 18 mmHg (both eyes). After the treatment the RVP was determined at 32 mmHg (right eye) and 16 mmHg (left eye). The IOP was measured as 16 mmHg (both eyes). The results show a massive decrease of the RVP.

[0068] Hcy was measured from patient's blood serum according to known methods. The IOP was measured using CorvisST® device. ODF measurements were performed using an ophtalmodynamometer according to Low. Retinal venous pressure (in mm Hg) is calculated according to the equation RVP=IOP+ODF.

[0069] Further measurement with glaucoma patients gave the following results:

TABLE-US-00001 TABLE 1 Retinal Venous Pressure Right eye Left eye Patient Date Hcy mm Hg mm Hg 1 21 Nov. 2019 12.89 16 Jan. 2020 25.5 74.1 13 Feb. 2020 16.5 58 2 4 Nov. 2019 12.63 35.90 39.9 24 Jan. 2020 21.90 27 31 Jan. 2020 14.00 18 3 26 Nov. 2019 18.63 28.8 71 6 Feb. 2020 18 40 4 21 Nov. 2019 15.41 9 Jan. 2020 32.90 34.1 23 Jan. 2020 15.40 17.3
Patients had supplementation with Ocufolin Forte® (one capsule per day).

Example 2

[0070] In continuation of the pilot study, the effect of vitamin supplementation comprising L-Methylfolate on retinal venous pressure and homocysteine plasma level in patients with glaucoma was further assessed.

[0071] The subjects included were all patients all suffered from glaucoma and/or ocular vascular diseases, such as normal tension glaucoma, primary angle closure glaucoma, and primary open angle glaucoma. The ophtalmodynamometric force initially measured was 15 mm Hg in any vein in or at the optic nerve head (ONH). The serum homocysteine (HCy) levels of the patients was higher than 12 μm/L.

[0072] The data was collected at a total of six time points. At visit 1 baseline measurements of IOP, RVP and serum levels of Hcy were performed (time point pre 1). At the second visit the baseline measurements were repeated and followed by starting the vitamin supplementation (time point pre 2). As a supplement Ocufolin® forte was used at the dosage of one capsule a day for a period of three months. No side effects from supplementation with Ocufolin® forte were reported.

[0073] During the treatment the IOP and RVP were measured after six weeks and after three months. For some of the patients, measurements were done more often, e.g. after two and four weeks (time points post 1, post 2, post 3, and post 4). Hcy was measured from patient's blood serum according to known methods. The IOP was measured using CorvisST® device. ODF measurements were performed using an ophtalmodynamometer according to Low. Retinal venous pressure (in mm Hg) is calculated according to the equation RVP=IOP+ODF.

TABLE-US-00002 TABLE 2 Mean (Standard Deviation) All pre 1 pre 2 post 1 post 2 post 3 post 4 N = 572 N = 106 N = 119 N = 136 N = 116 N = 52 N = 37 N RVP 29.3 34.5 32.7 26.9 26.4 29.5 23.9 539 (14.2) (16.9) (15.0) (12.5) (13.6) (11.7) (8.32) IOP 12.2 12.1 12.6 11.9 12.0 13.0 11.5 572 (2.76) (3.22) (2.81) (2.69) (2.46) (2.32) (2.74) ODF 17.2 22.3 20.0 15.1 14.3 16.5 12.5 539 (13.5) (16.3) (13.9) (11.7) (13.0) (11.5) (8.91) N = total number of veins measured

[0074] FIG. 1 shows the development of RVP before and during supplementation with Ocufolin® forte.

[0075] In addition to the significant reduction of RVP in all patients also mean homocysteine (Hcy) serum levels were statistically significantly lower after Ocufolin® forte supplementation than before.