COMPOSITIONS BASED ON SAFFRON FOR THE PREVENTION AND/OR TREATMENT OF CORNEAL DYSTROPHIES

Abstract

The present invention relates to a pharmaceutical, dietary and/or food composition, comprising saffron for use in the prevention and/or treatment of corneal dystrophies. The present invention also relates to a combination comprising saffron and at least one antioxidant and to a pharmaceutical dietary and/or food composition comprising said combination for use in the prevention and/or treatment of corneal dystrophies.

Claims

1. Pharmaceutical, dietary and/or food composition, preferably a food supplement, containing effective amounts of saffron, for use in the treatment and/or prevention of corneal dystrophies.

2. Composition for use according to claim 1, characterised in that in said saffron the amount of trans-crocin-4-gentiobiose-gentiobiose is present in an amount equal to or higher than 16.9% by weight with respect to the total weight of the saffron.

3. Composition for use according to claim 1, characterised in that in said saffron the amount of trans-crocin-3-gentiobiose-glucose is preferably present in an amount equal to or higher than 8% by weight with respect to the total weight of the saffron.

4. Composition for use according to any one of the preceding claims, characterised in that the corneal dystrophies are selected from the group comprising epithelial corneal dystrophies, corneal dystrophies of Bowman's layer, stromal corneal dystrophies, and posterior or endothelial corneal dystrophies.

5. Composition for use according to claim 4, characterised in that said corneal dystrophies are epithelial corneal dystrophies.

6. Composition for use according to claim 5, characterised in that said corneal dystrophy is epithelial basement membrane dystrophy.

7. Composition for use according to any one of the preceding claims, characterised in that said saffron is administered at a daily dose comprised between 5 and 50 mg/day, preferably comprised between 10 and 40 mg/day, and still more preferably 20 mg/day or 30 mg/day.

8. Composition for use according to any one of the preceding claims, characterised in that it further comprises effective amounts of at least one antioxidant.

9. Composition for use according to claim 8, characterised in that said at least one antioxidant is a polyphenol.

10. Composition for use according to claim 9, characterised in that said polyphenol is selected from the group comprising quercetin, curcumin and resveratrol.

11. Composition for use according to any of claims 8 to 10, characterised in that said saffron is administered at a daily dose comprised between 5 and 50 mg/day, preferably comprised between 10 and 40 mg/day, and still more preferably 20 mg/day or 30 mg/day and said antioxidant is administered at a daily dose of 50 or 250 mg/day, more preferably at a daily dose of 100 mg/day or 200 mg/day.

12. Composition for use according to any one of the preceding claims, characterised in that it is administered to mammals, in particular to a human.

13. Composition for use according to any one of the preceding claims, further comprising at least one physiologically acceptable excipient.

14. Composition for use according to any one of the preceding claims, in the form of a table, granulate, dragee or capsule, preferably a table.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0049] Additional features and advantages of the invention will become more clearly apparent by the following description of some preferred embodiments thereof, given hereinbelow by way of illustration and not of limitation, with reference to the attached drawings. In such drawings:

[0050] FIG. 1 is a graph illustrating the size of the damaged area (mm.sup.2), in the four experimental groups (A-D), over the course of the seven days following surgery;

[0051] FIG. 2 is a graph illustrating the degree of opacity in the four experimental groups (A-D), over the course of the seven days following surgery.

[0052] The following examples are intended to better understand the invention, without in any way limiting it.

Experimental Part EXAMPLE 1

[0053] It has been demonstrated that administration of a composition comprising effective doses of saffron to a patient suffering from epithelial basement membrane dystrophy was effective in the treatment of such disease.

[0054] The epithelial basement membrane dystrophy had been diagnosed based on clinical manifestations of recurrent corneal erosions and infections that the patient had suffered for several years prior to start drug-based treatment.

[0055] The patient complained of acute pain in the eyes, above all during the latter stages of sleep and upon waking. The patient had initially been treated with steroids and doxycycline (100 mg/day) for 10 months, with concomitant use of tear substitutes.

[0056] The treatment proved effective and the symptoms subsided.

[0057] Nevertheless, the discontinuation of the doxycycline resulted in the return of the symptoms within a few days and likewise the acknowledged side effects. The patient discontinued treatment with doxycycline and subsequently started treatment with two tablets per day of a saffron composition containing:

TABLE-US-00001 saffron 10 mg curcumin 50 mg

[0058] The patient immediately noted the absence of symptoms related to recurrent corneal erosions.

[0059] Given the tolerability of the composition, treatment with saffron was continued and today, approximately two years after starting treatment, the patient is no longer suffering from any symptoms.

EXAMPLE 2

[0060] A patient suffering from epithelial basement membrane dystrophy and the same clinical history as described in Example 1 was treated with two tablets per day of a saffron composition containing:

TABLE-US-00002 saffron 15 mg quercetin 50 mg resveratrol 50 mg

[0061] The patient immediately noted the absence of symptoms related to recurrent corneal erosions.

[0062] The treatment with saffron was continued and today, approximately two months after starting treatment, the patient is no longer suffering from any symptoms. It has therefore been demonstrated that administration of a composition comprising effective doses of saffron, quercetin and resveratrol to a patient suffering from epithelial basement membrane dystrophy was effective in the treatment of such disease.

EXAMPLE 3

[0063] A patient suffering from epithelial basement membrane dystrophy and the same clinical history as described in Example 1 was treated with two tablets per day of a saffron composition containing:

TABLE-US-00003 saffron 10 mg

[0064] The patient immediately noted the absence of symptoms related to recurrent corneal erosions.

[0065] During treatment with saffron, the patient stopped taking steroids and doxycycline and only used the tear substitutes occasionally.

[0066] Today, approximately two years after starting treatment, the patient is no longer suffering from any symptoms.

EXAMPLE 4

(Evaluation of the Effects of Orally Administered Saffron Solution on the Corneal Wound Healing Process on a Murine Model of Surgical Corneal Lesion)

[0067] For the experiment, 40 animals (mice) of the MUS MUSCULUS species were used, all male, healthy, and aged three months. The saffron used had an amount of trans-crocin-4-gentobiosio-gentobiosio amounting to 16.9% and of trans-crocin-3-gentobiosio-glucose amounting to 8%. Both eyes of each mouse were subjected to PRK (photorefractive keratectomy), which consists of a surgery on the central cornea, with a 2 mm ablation area, 45 microns of depth (reaching the epithelium), using an excimer laser.

[0068] The corneal wound healing process was monitored using a stereoscopic microscope, immediately after surgery and at 1, 2, 3 and 7 days thereafter. With a colorimetric test with fluorescein (Alcon Cusí, Barcelona, Spain) the degree of damage to the corneal epithelium was evaluated. This is because the fluorescein accumulates in the areas where the epithelium is damaged; with the wound healing evolution, the marked (coloured) area decreases. The level of opacity of the cornea was evaluated according to the method of Fante et al. (1990) which involves four levels of opacity, ranging from 0 to 4, where 0=completely clear cornea, 4=severe opacity. All clinical evaluations were performed separately, by two operators.

[0069] The animals were divided into four experimental groups:

[0070] Group A: mice with corneal injury, not treated with saffron (drinking water only);

[0071] Group 13: mice with corneal injury, treated with aqueous saffron solution;

[0072] C and D are used to show the groups of animals used as an internal control, which were treated, respectively, with plasma rich in PRGF and Cacicol growth factors (CACICOL-RGTA 20; Thea Laboratoires). The saffron treatment was orally administered (in the diet), while in the two control groups, the treatment was administered topically.

[0073] The ocular features were studied daily just before administration of treatments by microscopic analysis. Each group was analysed at four times: 1, 2, 3 and 7 days of treatment. These time points were selected because they include important events in the wound healing process.

[0074] In the following Table I the experimental schedule is summarised:

TABLE-US-00004 TABLE I Treatment Left eye Right eye N. of Group injured injured Doses Sacrifice time animals A water water ad libitum 7 days 10 B Saffron Saffron 5 mg/kg 7 days 10 solution solution C PRGF PRGF 2.5 μL/eye 7 days 10 D Cacicol Cacicol 2.5 μL/eye 7 days 10 * “Sacrifice time” it is meant the number of days from the surgery through to the time at which the animal is sacrificed.

[0075] The aqueous saffron solution was administered daily by syringe at a dose of 5mg/kg/day. The volume of solution administered was 300 μl/day. All the treatments (saffron, PRGF, and Cacicol) began seven days prior to surgery and were ended seven days later, with the death of the animal. The ocular features were evaluated daily by microscopic analysis. After the sacrifice, the eyes were enucleated and duly processed for immune histological analysis.

[0076] The results are shown in FIGS. 1 and 2, wherein the data is expressed, respectively, in units of damaged surface area (mm.sup.2) and degree of opacity during the seven days following surgery.

[0077] With reference to FIG. 1, 1 day after surgery, the greater re-epithelialisation efficiency corresponds to treatment with the saffron solution according to the invention—group B (0.22±0.02 mm). This value is lower than that obtained with the Cacicol—group D (0.43±0.06 mm), although the difference between the two groups was not statistically significant. Significant differences were observed between the untreated control group (A; 0.65±0.1 1 mm) and the saffron group (B), whereas treatment with Cacicol (group D) showed no significant differences from the mice drinking only water (group A). Treatment with PRGF (group C) was found to be the least efficient in terms of reduction of the injured area (1.19±0.15 mm) with marked differences with respect to the other groups. On the second day, there were no statistically significant differences between the groups, although the smallest average injured area observed was that of the saffron group, in accordance with the invention, i.e. group B (0.02±0.01 mm). Also on the third day, no significant differences between the groups were observed. 7 days after surgery, most of the eyes had completely repaired at the level of the epithelium, while the group treated with drinking water (A) presented epithelial ulcers in 12.5% of cases and the PRGF group (C) in 10% of cases. A 100% success rate was observed in the animals treated with Cacicol (group D) and with saffron (group B).

[0078] With reference to FIG. 2, following surgery on the cornea, the tissue became very opaque during the first 24 hours, due to inflammatory processes and oedema. All the groups observed one day after surgery showed a degree of corneal opacity greater than or equal to 3 according to the Fantes' scale.

[0079] Two days after treatment, the degree of opacity decreased slightly with respect to level 3 in all groups, except the untreated group (group A), even though statistically significant differences between groups can be observed. On the third day of analysis, there was a marked difference between the groups. Treatment with Cacicol (group D), PRGF (group C), and saffron (group B) significantly improved corneal transparency quality. The Cacicol and PRGF treatments showed opacity levels below level 2, while the saffron group showed an average opacity value of (2.15±0.12). Statistically significant differences between the saffron and Cacicol/PRGF groups were observed on the third day. The seventh day after surgery was particularly interesting because that was when the positive controls (Cacicol and PRGF) showed an optimal degree of corneal transparency. After seven days, the saffron group, according to the invention (group B), showed a significant reduction in corneal opacity, which settled at an average value of (1.77±0.14).

[0080] In terms of speed of epithelial healing, treatment with the aqueous solution of saffron for a duration of 14 days (7 prior to surgery and 7 days after) was as efficient as the Cacicol (which is considered the best treatment in the healing process) and was significantly better than treatment with PRGF. The saffron also significantly reduced the level of opacity compared with the untreated mice (drinking water only), although it was less effective than the other substances such Cacicol or PRGF. Regarding this, it is important to note that the administration route differed for each substance: the Cacicol and the PRGF were applied directly to the cornea, while the saffron solution was administered systemically. The concentration of an active ingredient administered by oral route which acts in the cornea healing process cannot be determined with respect to a topical treatment (drops), but this shows that, although administered systemically, saffron can reduce the level of opacity.

[0081] Therefore it has been demonstrated that the orally treatment with saffron, according to the invention, serves in the re-epithelisation of the cornea.