IT202000015583A1 - OPHTHALMIC COMPOSITION - Google Patents

Description

OPHTHALMIC COMPOSITION

DESCRIPTION

Field of invention

The present invention relates to combinations, compositions, formulations and kits comprising at least one plant extract belonging to the Laurus genus and to their use for the prevention and/or treatment of ocular pathologies and/or disorders. State of art

There are many pathologies that can affect the eyes and there are several biochemical and pathological mechanisms that can lead to their development, including oxidative stress. One of the eye diseases that most afflicts the population, especially in the age? senile, ? the cataract. Different types of cataract can be distinguished including nuclear cataract (affecting the center of the lens), cortical cataract (involving the edges of the lens), and posterior subcapsular cataract. The etiopathogenesis of cataract ? mainly due to the oxidation of the crystalline proteins, to the alteration of the cell membranes, to the so-called ?phase separation? and other risk factors briefly discussed below.

Oxidation of lens proteins

As anticipated by Benedek in 1971, the oxidation and aggregation of lens proteins, especially native crystallines, in high molecular weight protein aggregates (HMWP) contributes to the formation of opacities. of the nucleus that characterize cataract, especially the nuclear one <1,19>. Using the "quasi-elastic light scattering" technique, ? been demonstrated that the increase of? intensity? of the light scattered by the nucleus of the human lens correlates linearly with the increase in the concentration of HMWP<10> which, in turn, tends to increase with age<22>. Therefore, the transparency of the crystalline depends on the maintenance of soluble and insoluble proteins in the reduced state and what? requires a balance between oxidizing factors such as the reactive forms of oxygen (ROS) (for example, superoxide, hydroxyl radical, singlet oxygen, hydrogen peroxide) whose concentration increases due to the effect of UVB and UVA radiation, and enzymatic antioxidants (superoxide dismutase, glutathione system, catalase) and non-enzymatic (ascorbic acid, vitamin E-TPGS, xanthophylls) which act as "scavengers" of oxygen free radicals. Spector et al.<1, 19, 37 >report that the concentration of hydrogen peroxide (H2O2) in the aqueous humor in patients with cataract ? more high and even more? in diabetics compared to non-diabetics. The activities enzyme enzymes and the concentration of antioxidants in the body decrease with aging, making proteins more? susceptible to oxidation. In fact, when the tertiary structure of proteins is modified, even by low molecular weight molecules, the protein unfolds making the sulfhydryl groups (-SH) in particular of cysteine vulnerable to oxidants leading to the formation of intermolecular covalent bonds (disulfide bridges -S-S-). Among these low molecular weight molecules there are, for example, simple sugars, cyanate and thiocyanate, glucocorticoids and aldehydes which also derive from the peroxidation of polyunsaturated fatty acids (PUFA) of the cell membrane. Alterations of cell membranes

A second cause of clouding? been identified in the alteration of the membranes of the fibers of the lens, which is found above all in the opacities? cortices (Duncan et al., 1975; <32.>). There? ? to be related to the oxidation of the lipid and protein components of the membranes (Tomba et al., 1985; <39.>) which causes an alteration of the normal permeability? selective to sodium, potassium and calcium. The opacification of the cortex of the lens? in fact characterized by an increase in the content of sodium and calcium ions, by a decrease in potassium and taurine and by an increase in the water content which manifests itself with the formation of lakes and lamellar dissociation and therefore areas of inhomogeneity? optics with spaces of different refractive power. To the lipoperoxidation of the membranes? Also attributed is the formation of fluorescent products which, although present in the normal lens, increase during cataractogenesis and with age. ? a tendency towards an increase in saturated fatty acids with a decrease in unsaturated ones in the membrane phospholipids of the cataract crystalline nuclei was also shown.

Phase separation

Benedek (1997) observed that lowering the culture temperature of a crystalline lens up to the critical value of phase separation of cytoplasmic proteins in regions rich and poor in proteins with different refractive powers creates the reversible phenomenon called "cold cataract? which manifests itself at about 15°C. Various models of experimental cataracts would be due to the raising of the critical temperature which, rising above that of the body, would induce the formation of opacities. This elevation is inhibited by the use of active substances on the phase separation temperature For example, it has been demonstrated in vitro that pantethine is capable of inhibiting the raising of the critical phase separation temperature and therefore the clouding of the lens in experimental models.

UV radiation

UVB (320-290 nm) and UVA (321-399) rays are cytotoxic for the lenticular epithelium and represent the main environmental source of photo-oxidation, the cause of oxidative stress of the lens.

Diabetes mellitus

That diabetes mellitus is a potent risk factor in the genesis of age-related cataract? (quite different from the diabetic cataract proper, snowflake and rapidly evolving, very rare) ? proven for a long time and confirmed in all the most studies? recently, although in the Framingham study the risk is evident only under the age of 65. The association? been found mainly with posterior subcapsular cataract and nuclear cataract. Furthermore, the severity of retinopathy and levels of glycosylated hemoglobin represent additional elements of risk.

Body weight

? It has been shown that calorie restrictions of as little as 21% lead to more energy. low incidence of cataract. In a 1998 study<17>, subjects of age? between 64-74 years of both sexes, for whom weights at birth and at one year of age were available? Well yes ? found that there is a negative association between the disease and weight at one year of age. This singular observation, if confirmed, would suggest that the risk factors for cataracts have a very long incubation period. The proteins of the nucleus of an elderly person date back to fetal life or the first years of life: a disturbance of their synthesis or membrane formation in this period could persist for life and manifest itself in the form of opacities? charged to the nucleus, only many decades pi? late.

Smoke

A positive correlation between smoking and cataracts? been demonstrated (Solberg Y, Rosner M, Belkin M. The association between cigarette smoking and ocular diseases. Surv Ophthalmol.1998;42(6):535?547; Raju P, George R, Ve Ramesh S, Arvind H, Baskaran M , Vijaya L. Influence of tobacco use on cataract development. Br J Ophthalmol.2006;90(11):1374?1377). For what? that concerns the pathogenesis, you can? to believe that smoke contains substances that directly damage the proteins of the lens (cyanate and thiocyanate form protein adducts with the lens, facilitating their oxidation) and substances that block the antioxidant defences. In conclusion, smoking appears to be significantly correlated with nuclear cataract and possibly also with posterior subcapsular cataract. Alcohol

There are epidemiological data that support an increased risk of cataractogenesis from alcohol consumption. One of them (Clayton, 1980) showed a J-shape of the dose-cataract curve, suggesting a protective effect mediated by moderate alcohol consumption. Furthermore, in the Australian Blue Mountains Eye Study<14>, alcohol consumption ? been associated with a reduced prevalence of cortical cataract; only in subjects who are both heavy drinkers and smokers? been observed an increased risk of opacities? nuclear power, higher than that due to smoking alone.

Hypertension

In the Framingham Study, ? a positive association was found with systolic blood pressure; no report instead in the NHANES study (Kahn HA, Leibowitz HM, Ganley JP, et al. The Framingham Eye Study. II. Association of ophthalmic pathology with single variables previously measured in the Framingham Heart Study. Am J Epidemiol. 1977;106(1 ):33?41. The Beaver Dam Study<26 >found association with posterior subcapsular cataract, it is difficult to separate the effect of hypertension, especially if it is severe, from that of hypotensive drugs, particularly thiazide diuretics and furosemide.The Beaver Dam study showed a negative association of thiazide use with nuclear but positive association with posterior subcapsular cataract.In the Blue Montains Eye Study<14>, potassium-sparing diuretics were the only ones to show any positive association with posterior subcapsular cataract, concluding however for the absence of effects, favorable or harmful, by hypotensive drugs, on the transparency of the lens.

Cholesterol synthesis inhibitor drugs

The plasma membranes of the fibers of the lens contain high amounts of cholesterol and cholesterol is likely to be synthesized locally. This summary, what? indispensable for the formation of the membranes intended to wrap the fibers, it plays a critical role in maintaining the transparency of the lens and, for this reason, the attention of pharmacologists? for a long time addressed to the relationship between cataract and cholesterol synthesis inhibitor drugs. For example, triparanol ? was withdrawn from the market in 1962 due to its side effects, including cataracts. HMG-CoA reductase inhibitors such as statins, although having the same efficacy on the enzyme in lens homogenates, have shown some differences: only simvastatin, fluvastatlna and lovastatin have in fact shown a cataractogenic action in dogs at overpharmacological doses. Many epidemiological studies have been conducted in humans from which ? for example, it emerged that lovastatin in treatments of up to two years does not generate opacities? n? increase opacity pre-existing. After five years, ? been reported the appearance of opacities? cortices, but Chylack et al., in 745 patients did not observe an increased incidence of cataract surgery. To the same conclusions? the Blue Mountains Eye Study<14> has arrived.

From the above, it appears evident that oxidative stress, in particular that caused by reactive oxygen species, plays a key role in the pathogenesis of cataract both in experimental animals and in humans (Nita and Grzybowski 2016) in fact, when free radicals accumulate in the lens, polyunsaturated fatty acids are oxidized initiating cataractogenesis. Furthermore, ? It is also known that malondialdehyde (MDA), one of the end products of lipid peroxidation, due to its high ability to of cross-linking towards amino acids, contributes to deforming the spatial structure of proteins, leading to the formation of protein adducts. Therefore, the formation of a Schiff base by reaction of the carbonyl of MDA with the primary amino groups of phospholipids can play an important role in cataractogenesis (Bhuyan KC, et al., Molecular mechanisms of cataractogenesis: IV. Evidence of phospholipid malondialdehyde adduct in human senile cataract. Mech Aging Dev. 1986; 34: 289-96).

As a result, various remedies based on synthetic and/or natural active ingredients with active properties have been proposed in the art. antioxidant, potentially useful in the prevention and/or treatment of eye diseases.

Most of these antioxidants are reducing agents, such as polyphenols, which are capable of interrupting the chain reactions that lead to the release of free radicals.

In addition, several phytochemicals have shown activity preventives in models of oxidative stress. For example, ellagic acid inhibited selenite-induced cataract formation in rats (Sakthivel et al., 2008) and was shown to possess anti-denaturing agent on lens proteins (Sakthivel et al., 2011) and Moringa oleifera - rich in polyphenols - showed the same activity in the selenite cataract model (Sasikala et al., 2010). ? It is also known that flavonoids, phenolic acids, carotenoids, vitamins and lactoferrin are natural antioxidant molecules which, as such, could have anti-cataract (Sunkireddya et al., 2013).

Even a diet rich in fruits, vegetables, fish, legumes and starchy foods can exercise a protective role against cataractogenesis (Theodoropoulou et al., 2014), since? Hypercholesterolemia, hypertriglyceridemia, high levels of LDL cholesterol (low-density lipoprotein cholesterol), saturated fat intake, and elevated fasting blood sugar are known to correlate with the development of cataracts and other degenerative eye diseases (Heydari et al. 2012). So, ? It is possible that changes in diet and/or the intake of antioxidants may reduce the risk of cataracts.

However, while the role of oxidative stress in the pathogenesis of cataract is? now established, the results about the prevention of cataractogenesis through the use of antioxidants are very controversial. In fact, while there is no evidence that supplementation with antioxidants can prevent or slow the progression of cataracts in humans (Chiu and Taylor, 2007; Mathew et al., 2012), experimental evidence demonstrates that the administration of antioxidants in animal models of cataractogenesis is able to prevent or delay the onset of cataract (Soni P. et al., ?Effects of a novel isoflavonoid from the stem bark of Alstonia scholaris against fructose-induced experimental cataract? J Integr Med.2019 , pii: S2095-4964(19)30075-5).

Therefore, c?? still the need? of a product which is effective in the prevention and/or treatment of eye diseases, in particular in the prevention and/or treatment of eye diseases including cataracts.

SUMMARY OF THE INVENTION

Since many natural substances have shown to be effective in therapy and above all safe, bioavailable and well tolerated by patients, the present invention is based on the research and identification of a combination of natural substances which, when used in combination, exert an effective action and enhanced in the prevention and/or treatment of ocular pathologies such as, for example, cataracts.

After extensive experimentation, the inventors have found that the association of at least one plant extract belonging to the Laurus genus, at least one plant extract belonging to the Aloe genus and hyaluronic acid provides a natural solution to the clinical problem of eye pathologies.

Therefore, the present invention refers to the association of at least one plant extract belonging to the Laurus genus, at least one plant extract belonging to the Aloe genus and hyaluronic acid.

The present invention also refers to compositions, preferably for topical use, more preferably in the form of eye drops, comprising the association of at least one plant extract belonging to the Laurus genus, at least one plant extract belonging to the Aloe genus and hyaluronic acid and one or more? carriers suitable cos? such as kits of parts comprising the above separate and independently formulated active ingredients in suitable dosage form for sequential or concurrent administration of the different active ingredients.

Furthermore, the present invention relates to the use of the aforementioned association, compositions and kits in the prevention and/or treatment of ocular pathologies such as, for example, cataract, keratoconus, dry eye, conjunctival allergy, conjunctivitis, blepharitis, keratitis , actinic keratitis and damage from exposure to UV rays and / or non-therapeutic use of the above combinations, compositions and kits as an eye lubricant.

Thanks also to the synergy of its components, the combination object of the present invention allows to simultaneously obtain the reduction of various symptoms associated with ocular pathologies, such as a reduction of the opacity of the eyes. of the lens, conjunctival hyperaemia, dry eyes and foreign body sensation.

Other advantages and features of the present invention will become apparent from the following detailed description.

Glossary

The terms used in the present description are as generally understood by the person skilled in the art, unless otherwise indicated.

With the term ?extract?, in the context of the present description, we mean any product attributable to a vegetable drug including all products deriving from mechanical treatments (pulverization, grinding, mixing and/or other methods) or from extractive treatments (extraction with solvent , distillation, maceration and/or other specific methods) performed on a drug, including mother tinctures and essential oils.

DETAILED DESCRIPTION OF THE INVENTION

As mentioned above, the present invention refers to a combination comprising extract of at least one plant belonging to the genus Laurus, extract of at least one plant belonging to the genus Aloe and also hyaluronic acid.

The main active ingredients of the combination of the invention are described below.

Laurel

The laurel ? an evergreen plant belonging to the Lauraceae family, genus Laurus. It is thought to have originated in the Asia Minor region, where is it from? been distributed throughout the Mediterranean and other parts of Asia. It grows in very warm climates, particularly in southern Europe and around the Mediterranean Sea area (Chmit et al., 2014). The Laurus nobilis extract has high nutritional levels thanks to the content of proteins, free sugars, organic acids, PUFA and tocopherols together with active? antioxidant, scavenging and inhibition of lipid peroxidation (Dias et al., 2014). Laurus nobilis leaves have been shown to improve activity? of insulin in in vitro studies, while in vivo human intake of 1-3 g per day ? been correlated with a significant reduction in fasting serum glucose and LDL cholesterol and triglyceride levels, along with an increase in HDL cholesterol (Khan et al., 2009).

Aloe

Aloe vera? It is a xerophytic plant, belonging to the Liliaceae family, capable of avoiding excessive dehydration and therefore surviving in periods of long drought. It has no trunk, has thick and fleshy leaves, capable of accumulating water. From the leaves of Aloe we obtain a gel whose properties healthy and the uses have been known since ancient times.

The most outside of the leaves contains the aloin which has outstanding properties? laxative, purgative or tonic-digestive, depending on the dosage; the most internal contains a mucilaginous gel that? made up of 98.5% water and the remainder of active substances. Aloe gel? particularly known for its properties? moisturizing, emollient, protective, anti-inflammatory, healing, ? able to maintain or restore the elasticity? skin and, in addition, has a draining and purifying function, as well as antiseptic and bacteriostatic. These properties? medicinal products are mainly mediated by biologically active components present in aloe and its extracts such as, for example, anthraquinones, flavonoids, phenolic acids, antioxidant vitamins, enzymes and other non-nutrient compounds of the plant.

Hyaluronic acid

Hyaluronic acid? an anionic glycosaminoglycan with properties? viscoelastic. It is mainly found in connective tissue, vitreous humor and aqueous humor. It has gained widespread application in lubricants used to treat dry eyes because it effectively binds water, resists dehydration, and exhibits excellent biocompatibility. Previous studies have shown that hyaluronic acid protects corneal epithelial cells, stimulates epithelial migration and improves the quality of the skin. of the retinal image (Nakamura M, Hikida M, Nakano T, et al. Characterization of water retentive properties of hyaluronan. Cornea. 12:433?436, 1993; Johnson ME, Murphy PJ, and Boulton M. Effectiveness of sodium hyaluronate eye drops in the treatment of dry eye. Graefes Arch. Clin. Exp. Ophthalmol.244:109?112, 2006; Montes-Mico R, Cervino A, Ferrer-Blasco T, et al. Optical quality after instillation of eye drops in dry eye syndrome. J. Cataract Refract. Surg.36:935?940, 2010). In eye drops, hyaluronic acid? used to improve the irregularity? of the ocular surface, to stabilize the precorneal tear film and to improve the intensity of dry eye symptoms.

Although to obtain optimal efficacy the presence of the three active ingredients mentioned above is preferred, the present invention also refers to an association comprising one, two or more? of the following: extract of at least one plant belonging to the genus Laurus, extract of at least one plant belonging to the genus Aloe and hyaluronic acid, variously combined. By way of example but in no way limiting, the association can? comprising extract of at least one plant belonging to the genus Laurus and hyaluronic acid or extract of at least one plant belonging to the genus Laurus and extract of at least one plant belonging to the genus Aloe.

The present invention relates in particular to a combination comprising extract of at least one plant belonging to the genus Laurus, extract of at least one plant belonging to the genus Aloe and hyaluronic acid.

With the wording ?at least one plant extract belonging to the genus Laurus? in the context of this description it is understood that the composition can? include at least one or more plant extracts belonging to the genus Laurus. The extract or extracts can be chosen, for example, from Laurus nobilis L. extract (also known as common laurel), Laurus azorica extract, Laurus chinensis extract, Laurus melissifolia extract and mixtures of these, and ? preferably extract of Laurus nobilis L. The extract can? be prepared according to any of the procedures known in the state of the art.

According to one embodiment, the extract can be a mother tincture or an essential oil of laurel (1:3) prepared according to the Official Pharmacopoeia (FU, D.L.S.75/2018).

To obtain the mother tincture or the essential oil? an 80% hydroalcoholic solution in ethyl alcohol was previously prepared. In the hydroalcoholic solution the dry leaves of Laurus nobilis were macerated for 20/25 days, at room temperature, in the ratio of 1:3 by mass, for example 10 g of bay leaves in 30 g of hydroalcoholic solution at 80 % in ethyl alcohol.

Obviously, other mass ratios are also suitable for the purposes of the present invention, by way of example drug:solvent mass ratios can range from 1:1 to 1:20, preferably from 1:3 to 1:10.

It is also understood that if, for the preparation of the mother tincture, not one but more species (for example two, three, four or five species) of plant belonging to the Laurus genus mixed together will be? sufficient that the mass ratio is satisfied for the total mixture.

For the purpose of the present invention, the mother tincture of laurel, obtainable as described above or by any other method known to the expert technician, can be used as such or be subjected to further processing. By way of example, but in no way limiting, pu? be prepared a mixture of mother tincture of laurel and chloroform (CHCl3) in a 1:1 ratio, prepared for example by mixing 25 mL of mother tincture of laurel and 25 mL of chloroform, the resulting mixture ? carried to dryness in a rotavapor at 200 rpm at 37°C and, of the residue obtained, 0.3 mg is added to 5.0 mL of water obtaining a 0.06 mg/mL aqueous solution, which can then be used for the preparation of pharmaceutical forms according to the invention.

According to another embodiment, the laurel extract can be an extract obtainable by hot maceration in water, for example by maceration of 2-10 g, preferably 4 g, of Laurus nobilis leaves in 1 L of water at 100°C for 15 minutes with continuous stirring.

Therefore, the present invention also refers to an association comprising extract of at least one plant belonging to the Laurus genus, extract of at least one plant belonging to the Aloe genus and hyaluronic acid, in which said extract of at least one plant belonging to the Laurus genus? obtainable by hot maceration of fresh vegetable material, preferably leaves, in water, preferably in distilled water, or in which said extract of at least one plant belonging to the genus Laurus ? obtainable by extraction in chloroform of mother tincture or laurel essential oil. How will it turn out? obvious to a person having general knowledge in the art, other extraction procedures known in the art are however suitable.

For the purposes of the present invention, laurel extracts prepared according to the methods described above are particularly suitable, but also commercially available laurel extracts are in any case suitable, for example the essential oil of laurel marketed by ACEF.

The laurel extract, in particular of Laurus nobilis, can be administered for example with a dosage regimen between 0.1 and 0.5 ?g/day, in particular between 0.1 and 0.2 ?g/day, preferably in 1-2 separate administrations (for example 0, 1 ?g x 2 times a day, or 0.2 ?g 1 time a day).

In particular, the inventors have demonstrated that it is possible to successfully use quantities equal to about 0.2 ?g per day of laurel extract, preferably of Laurus nobilis in combination with aloe extract and hyaluronic acid as described herein, obtaining, among other beneficial effects, also a significant regression of the opacity in the treated subjects.

By way of example only, but in no way limiting, 2 drops/day of eye drops comprising 0.1 ?g each of laurel extract, in particular Laurus nobilis, can be administered.

With the wording ?at least one plant extract belonging to the genus Aloe? in the context of this description it is understood that the composition can? include at least one or more plant extracts belonging to the genus Aloe. According to one embodiment, the extract or extracts can be selected from Aloe Vera extract, Aloe Arborescens extract, Aloe Chinensis extract, Aloe Ferox extract, Aloe Humilis extract and mixtures thereof and ? preferably Aloe Vera extract. The extract can be prepared according to the procedures known in the state of the art, for example, could? be a dry or aqueous extract of Aloe vera leaves. The aloe extract can, for example, be prepared by aqueous extraction of fresh leaves, previously washed and shredded, in distilled water at a temperature of 95-98°C for 30 minutes. After cooling, the extract can be subjected to filtration twice and then placed in glass containers for sterilization in an autoclave (120?C for 20 minutes) and in a pressure cooker (100?C for 20 minutes, once and for another 30 minutes once second time). The integrity of the active principles of the extract? preserved (Kodym A, et al. Technology of eye drops containing aloe (Aloe arborescens Mill.--Liliaceae) and eye drops containing both aloe and neomycin sulphate. Acta Pol Pharm.2003;60(1):31?39).

Therefore, the present invention also refers to a combination comprising extract of at least one plant belonging to the Laurus genus, extract of at least one plant belonging to the Aloe genus and hyaluronic acid, in which said extract of at least one plant belonging to the Aloe genus? obtainable by hot extraction of fresh plant material, preferably leaves, in distilled water. How will it turn out? obvious to a person having general knowledge in the art, other extraction procedures known in the art are however suitable. The extract could, for example, be prepared by extracting the leaves in water or in another suitable solvent and optionally freeze-dried. According to a preferred embodiment, the extraction solvent does not comprise alcohol.

For the purposes of the present invention, aqueous extracts of aloe prepared according to the methods described above are particularly suitable. Preferably the aqueous extract has a concentration of 15%-19% preferably 17% by mass but other commercially available aloe extracts are however suitable.

It is understood that, where not one but bens? more extracts (for example two, three, four or five extracts) of plants belonging to the genus Aloe mixed together, will be? sufficient that the concentration of 17% by mass is satisfied for the total mixture of plant species belonging to the genus Aloe.

For ?hyaluronic acid? in the context of the present description it is meant hyaluronic acid or a salt thereof, preferably sodium salt. For the purposes of the present invention it can for example hyaluronic acid can be used, preferably hyaluronic acid sodium salt, having a molecular weight between 100 and 1000 kDa, preferably between 200 KDa and 800 KDa, more? preferably between 300 KDa and 600 KDa. The association can obviously also understand how ?hyaluronic acid? a mixture of hyaluronic acids having different molecular weight, as long as? said molecular weight falls within the aforementioned range.

The sector technician can adjust the quantity? of the extracts used and/or of the active ingredients used in the preparation of the formulations to be administered according to the needs. Will the attending physician know? also identify the optimal dosage for the subject to be treated based on age, gender, weight and general state of health. Therefore, the dosage of the individual components can? be adapted even during the period of taking the combination or composition of the invention according to the results obtained over time.

Although it is not an essential feature of the invention, the inventors have identified the optimal weight ratio between the components of the combination. Preferably, the weight ratio between extract of at least one plant belonging to the genus Laurus, extract of at least one plant belonging to the genus Aloe and hyaluronic acid? between 1:1:1 and 20:175:7, and ? preferably 15:170:2.

The inventors have also observed that the greater efficacy of the association, also in virtue? of the greatest synergy between the various active ingredients, occurs when:

- the plant extract belonging to the genus Laurus ? present in a concentration of between 0.5 mg and 2 mg per 100 mL, preferably 1.2 mg per 100 mL;

- the plant extract belonging to the genus Aloe? present in a concentration between 15% and 20%, preferably 17% by mass, in the aqueous solution to which ? added laurel extract.

- hyaluronic acid? present in a concentration between 0 and l?1%, preferably at 0.2% by mass in the aqueous solution to which ? added laurel extract.

How will it look? evident from the examples, for ?aqueous solution to which ? added laurel extract? means a previously prepared solution which includes Aloe vera gel (17%), hyaluronic acid (0.2%) and buffered water at pH 7.2 with phosphate buffer (82.8%).

According to the example given, the three components are all added in the form of an aqueous solution.

According to one embodiment, the combination of the present invention can include one or more extracts of each of the active ingredients described herein, provided that it comprises at least one plant extract belonging to the Laurus genus, at least one plant extract belonging to the Aloe genus and hyaluronic acid.

The association according to any of the embodiments described up to now can? comprise, as a further active ingredient, an antioxidant preferably selected from the group comprising, but not limited to: Vitamin A, Vitamin C, Vitamin E, d-alpha-tocopheryl poly(ethylene glycol)-1000-succinate (VETPGS), selenium, carotenoids, lycopene , coenzyme Q-10, lipoic acid and mixtures thereof. Therefore, the present invention also refers to a combination comprising, as the main active ingredients, at least one plant extract belonging to the Laurus genus, preferably Laurus nobilis extract, at least one plant extract belonging to the Aloe genus, preferably Aloe vera, hyaluronic acid and optionally, one or more antioxidants as defined above.

According to one embodiment, the combination comprises at least one plant extract belonging to the Laurus genus, preferably Laurus nobilis extract, at least one plant extract belonging to the Aloe genus, preferably Aloe vera and hyaluronic acid as the only active ingredients in the treatment and / or in the prevention of eye diseases, especially cataracts. According to another embodiment, the combination comprises at least one plant extract belonging to the Laurus genus, preferably Laurus nobilis extract, at least one plant extract belonging to the Aloe genus, preferably Aloe vera, hyaluronic acid and one or more? antioxidants as defined above as the only active ingredients in the treatment and/or prevention of ocular pathologies, in particular of cataracts. In the latter case, the association can? include other active ingredients, provided that said other active ingredients are not indicated in the treatment of ocular pathologies, in particular of cataracts.

The present invention also relates to compositions comprising the association according to any of the embodiments described herein and one or more suitable vehicles.

According to one embodiment, the composition comprises, as main active ingredients, at least one plant extract belonging to the genus Laurus, preferably extract of Laurus nobilis, at least one plant extract belonging to the genus Aloe, preferably of Aloe vera, hyaluronic acid and one or more suitable vehicles. The composition can understand as a further active ingredient also one or more? antioxidants as defined above. Suitable vehicles for the purposes of the present invention are those commonly known in the art as suitable for ocular administration, for example, a buffered isotonic solution at pH 7.2.

In a further embodiment, the composition comprises at least one plant extract belonging to the genus Laurus, preferably extract of Laurus nobilis, at least one plant extract belonging to the genus Aloe, preferably of Aloe vera, hyaluronic acid and, optionally, one or more ? antioxidants as defined above, as the only active ingredients in the treatment and/or prevention of ocular pathologies. In the latter case, the composition can? comprise other components, such as for example excipients, carriers, stabilizers, preservatives and the like and/or also other active ingredients, provided that said other active ingredients are not indicated in the treatment of ocular pathologies, in particular of cataracts.

The compositions according to any of the embodiments provided in the present description can be formulated in any form and for any route of administration and associated with any other component, in a variety of forms. of ways.

According to one embodiment, the compositions of the invention are compositions for topical use in liquid, semi-liquid, semi-solid or solid form such as, for example, eye drops, solution, sterile solution, suspension, eye drops, spray, ophthalmic bath, gel, ointment ointment, cream, medicated wipe, occlusive dressing (e.g. in the form of medicated gauze), ophthalmic inserts (e.g. controlled-release medicated hydroxypropylcellulose inserts).

Therefore, the present invention also relates to the combination or composition according to any of the embodiments described above in the form of eye drops, solution, sterile solution, suspension, eye drops, spray, ophthalmic bath, gel, ointment, ointment, cream , medicated wipe, occlusive dressing and ophthalmic inserts. According to one embodiment, the composition for topical use according to the present invention will be? in the form of a buffered isotonic solution at pH 7.2.

According to another embodiment, the compositions of the invention are compositions for oral use in solid form such as, for example, capsules, soft capsules, tablets, pills, lozenges, jellies, powders or granules or in liquid form, such as, for example, emulsions, solutions, prepared or extemporaneous suspensions, syrups and elixirs. Therefore, the present invention also relates to the combination or composition according to any of the embodiments described above in solid form, preferably in the form of capsules, soft capsules, tablets, pills, lozenges, jellies, powders or granules or in liquid, preferably emulsions, solutions, prepared or extemporaneous suspensions, syrups and elixirs. In the case of solid forms, the excipients will preferably be selected from, but not limited to, the following: a) carriers, such as for example sodium citrate and calcium phosphate, b) fillers such as for example starch, lactose, microcrystalline cellulose, sucrose, glucose, mannitol, triglycerides and colloidal silica, c) humectants, such as for example glycerol, d) disintegrating agents, such as alginates, calcium carbonate, starches, derivatives of starch, cellulose and polyvinylpyrrolidone, silicates and sodium carbonate and ) binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, polymeric derivatives of cellulose, starch derivatives f) retarding agents such as paraffin, cellulose polymers, fatty acid esters g) absorption accelerators, such as quaternary ammonium compounds, h) wetting agents and surfactants such as cetyl alcohol and glycerol monostearate, i) adsorbents, such as benthic clays and kaolin, k) lubricants such as talc, calcium stearate, magnesium stearate, polyethylene glycol, sodium lauryl sulfate, sodium stearyl fumarate j) glidants such as talc, colloidal silica.

The solid dosage forms, such as tablets, capsules, soft capsules, jellies, pills and granules, may optionally be coated with enteric, gastric or other types of coatings known in the state of the art possibly allowing the release of the active ingredients only or preferably in a certain section of the intestine, possibly in a delayed manner. Substances which may permit such delayed use include, but are not limited to, polymers and waxes.

In the case of liquid forms, suitable excipients include, but are not limited to, diluents such as water or other solvents, solubilizing agents and emulsifiers selected from ethyl alcohol, polyalcohols, propylene glycol, glycerol, polyethylene glycol and sorbitan esters. These formulations may also contain sweeteners and flavors.

Preferably, the compositions according to the present invention do not contain excipients and/or other additives, but only the active ingredients according to any of the embodiments described herein, dissolved or suspended in a suitable vehicle.

However, where necessary, suitable excipients can be selected from those normally known in the state of the art and include, but are not limited to: surfactants (for example sodium lauryl sulfate and polysorbates), adsorbents (for example silica gel, talc, starch, bentonite, kaolin), glidants and anti-adherents (e.g. talc, colloidal silica, corn starch, silicon dioxide), antioxidants, binders (e.g. gums, starch, gelatin, cellulose derivatives, sucrose, sodium alginate), plasticizers (e.g. ethyl cellulose and other cellulose derivatives, acrylates and methacrylates, glycerol and sorbitol), preservatives (e.g. parabens, sulfur dioxide, polyhexamethylene biguanide or PHMB), disodium edetate (EDTA), thickeners, emulsifiers, humectants, wetting agents, chelators and their blends.

By way of example, the composition may include laurel extract, aloe extract, hyaluronic acid, optionally one or more? oxidants as defined above, EDTA and possibly also PHMB in buffered isotonic solution at pH 7.2.

The compositions of the present invention will be, for example, a medical device, a food supplement, a nutraceutical, dietetic and nutritional composition, a food product, a beverage, a medicament, a medicated food, a food for special medical purposes, or a composition pharmaceutical.

The compositions according to any of the embodiments described here can be used in any subject affected by the above mentioned ocular pathologies, also in the veterinary field.

The combination of the active ingredients mentioned above can be used formulated in a single composition according to the various embodiments described above or in a kit comprising the several separate ingredients, for example, in single compositions formulated in suitable dosage form for topical use as defined above, for sequential administration or simultaneously of the various active ingredients.

Therefore, the present invention also relates to kits of parts comprising the different active ingredients of the combination according to any of the embodiments described herein separated and formulated in suitable dosage form for sequential or simultaneous administration of the different active ingredients. The present invention relates, for example, to a kit of parts comprising a first composition for topical use comprising at least one plant extract belonging to the genus Laurus, a second composition for topical use comprising at least one plant extract belonging to the genus Aloe and a third composition for topical use comprising hyaluronic acid, for use in the prevention and/or treatment of ocular pathologies, by means of the sequential or simultaneous administration of the three compositions. evident to the person skilled in the art from the examples provided below, the present invention constitutes a valid support in the reduction or resolution of the symptoms associated with the aforementioned ocular pathologies, in particular with cataracts, thanks to the combined and diversified action of its components.

The action of the individual components and of the association object of the present invention ? been evaluated by means of in vitro and/or in vivo tests which, moreover, have shown how the composition of the invention? particularly effective in the treatment of eye diseases thanks to the reinforcing (synergistic) action of its components.

Therefore, the present invention also relates to the combination, compositions and kit, according to any of the embodiments described above, for use as a medicament.

Furthermore, the present invention also relates to the combination, to the compositions and to the kit, according to any of the embodiments described herein, for use in the prevention and/or treatment of ocular pathologies such as, for example, cataracts , keratoconus, dry eye, conjunctival allergy, conjunctivitis, blepharitis, keratitis, actinic keratitis and damage from UV exposure, both in humans and animals.

In addition to this, the object of the present invention is also the non-therapeutic use of the combination, composition or kit according to any of the embodiments described above, for example, as an eye lubricant.

The association of the above active principles may? be formulated in a single composition or in a kit according to the various embodiments described above and prepared for example by mixing the selected active ingredients in a suitable vehicle suitable for topical ocular administration with any other active ingredients and/or excipients as known to the person skilled in the art sector.

EXAMPLES

Some non-limiting examples of the compositions according to the present invention are given below. Modifications or variations of the embodiments exemplified herein, obvious to one skilled in the art, are encompassed by the appended claims.

EXAMPLE 1: eye drops, solution

Process for the preparation of laurel mother tincture

To obtain the laurel mother tincture? an 80% hydroalcoholic solution in ethyl alcohol was previously prepared. The dry leaves of Laurus nobilis were macerated in the hydroalcoholic solution for 20/25 days, at room temperature, in the ratio of 1:3 by mass.

Process for the preparation of Laurus extract

The laurel extract? was obtained by mixing 25 mL of laurel mother tincture with 25 mL of chloroform (CHCl3) and dried in a rotavapor at 200 rpm (revolutions per minute) at 37°C. Of the water-soluble residue, 0.3 mg is added to 5 mL of water to obtain a 0.06 mg/mL aqueous solution.

Process for preparing eye drops according to the invention

2 mL are taken from the aqueous solution at a concentration of 0.06 mg/mL and added to 8.0 mL of a previously prepared solution which includes Aloe vera gel (17%), hyaluronic acid (0.2%), disodium EDTA (0.0001%) and buffered water pH 7.2 with phosphate buffer (82.8%) cos? to obtain 10 mL of eye drops according to the present invention. The eye drops ? placed in a special dropper vial.

EXAMPLE 2: eye drops, solution

The eye drops ? was prepared by taking 2 mL of aqueous laurel extract as obtained in example 1 and further adding 1 mL of 0.02% w/v aqueous solution of D-alpha-tocopheryl poly(ethylene glycol)-1000-succinate (VE-TPGS ), obtained from the esterification of vitamin E-succinate with polyethylene glycol 1000. Under these conditions, the solution is? still stable why? VE-TPGS performs its surfactant functions: are micelles formed which keep the compound and the resulting suspension in solution? make up to volume with 7 mL of isotonic buffered solution at pH 7.2. The solution ? been mixed and then placed in a special dropper vial.

EXAMPLE 3: eye drops, solution

The eye drops ? was prepared by taking 2 mL of laurel aqueous extract as obtained in example 1 and further adding 0.5 mL of 0.02% w/v vitamin E solution and 0.5 mL of 0.2% vitamin E solution C and made up to volume with 7 mL of buffered isotonic solution pH 7.2. The solution ? been mixed and then placed in a special dropper vial.

EXAMPLE 4: medicated wipes

Wipes of non-woven fabric were soaked in a sterile environment with the solution according to Examples 1-3 and sealed in aluminum sachets.

Experimental evidence

The authors of the present invention have observed that the topical conjunctival administration of a composition comprising the association of at least one plant extract belonging to the Laurus genus, in particular Laurus nobilis, together with at least one plant extract belonging to the Aloe genus, in particular Aloe vera, and hyaluronic acid is effective in reducing the symptoms of various eye diseases, such as cataracts, thanks also to the synergistic or reinforced action of its components.

Furthermore, ? It was possible to ascertain that the association of at least one plant extract belonging to the Laurus genus with at least one plant extract belonging to the Aloe genus and hyaluronic acid allows to obtain an increase in the lubrication of the ocular surface and a concomitant reduction in the irritative/inflammatory state of the itself significantly higher than what is obtained with the administration of single active ingredients or of only two active ingredients.

The effectiveness of the present invention ? been evaluated as described below, but pu? be evaluated through a variety? of in vitro tests and/or in vivo tests on experimental animals known to the person skilled in the art.

Animal preparation

Twenty albino rabbits of about one year of age were used for the study. and weighing about 1.5-2 kg. The animals were kept on a normal diet, with adequate water and temperature and normal light. The rabbits were divided into 5 groups of 5 animals each (group A, B, C, D and E) as defined below:

- Group A: healthy control group (no treatment and no therapy); - Group B: cataract induction and no therapy;

- Group C: cataract induction and therapy with eye drops based on hyaluronic acid sodium salt 0.2%, Aloe Vera gel 0.1% in buffered isotonic solution at pH 7.2, instilled twice a day for 5 days before ?cataract induction and for 30 days after cataract induction. - Group D: cataract induction and therapy with eye drops of example 1, i.e. comprising hyaluronic acid sodium salt, Aloe Vera gel and Laurus nobilis extract in isotonic buffered solution at pH 7, instilled twice a day for 5 days before the cataract cataract induction and for 30 days after cataract induction.

- Group E: cataract induction and therapy with eye drops comprising only laurel extract (without hyaluronic acid and without aloe extract) in isotonic buffered solution at pH 7.2, instilled twice a day for 5 days before cataract induction and for 30 days after cataract induction. Cataract induction

The cataract ? was induced by subcutaneous injection of 1 mL sodium selenite solution (0.1% w/v in saline) on the neck in groups B, C, D, and E on day zero of the study (see KABIR F, et al< 34>).

For cataract induction, animals in groups B, C, D and E were anesthetized by intramuscular injection of ketamine (50 mg/ml) at a dose of 50 mg/kg and intramuscular injection of diazepam (10 mg/2 ml). at a dose of 2 mg/kg. Were all procedures performed in accordance with the with guidelines no.

86/609/EEC stipulated by the Committee and guidelines of the European Union for animal experimentation. At the end of the experimentation the animals - after being anesthetized - were sacrificed by gaseous embolism, the eyes were immediately enucleated and the lenses removed under optical microscopy.

Clinical monitoring

For the clinical monitoring of cataractogenesis the following parameters were analysed:

- Opacity? of the lens

The morphological examination of rabbit lenses? was performed similarly to the procedure described in Carey et al. (Carey JW, Pinarci EY, Penugonda S, Karacal H, Ercal N. In vivo inhibition of l-buthionine-(S,R)-sulfoximine-induced cataracts by a novel antioxidant, N-acetylcysteine amide. Free Radic Biol Med.2011 ;50:722?9). An hour before the exam? one drop of 2.5% phenylephrine and 1% tropicamide was instilled in each eye to initiate mydriasis. The cataract ? been classified according to the following scale: grade 0 = transparent lens; grade 1 = lens with slight opacity; grade 2 = lens with partial opacity? nuclear; grade 3 = lens with dense opacity? nuclear and grade 4 = opacity? (Muranov K, Poliansky N, Winkler R, Rieger G, Schmut O, Horwath-Winter J. Iodide protection of lens from selenite-induced cataract. Graefes Arch Clin Exp Ophthalmol. 2004;242:146-151).

- Conjunctival hyperemia

? the conjunctiva of both eyes was inspected in order to detect the presence or absence of redness (Macdonald M, The examination of the eye. In: Munro J. and Edwards C. Macleod?s Clinical Examination.10th. ed. Churchill Livingstone. Edinburgh, 2000, 257-271).

- Measurement of eye pressure (IOP)

The IOP? was measured with a Schiotz tonometer. because the Schiotz tonometer does not measure pressure directly, readings were compared to conversion tables to convert scale readings into intraocular pressure estimates. Schiotz tonometry? a form of indentation tonometry. An indentation tonometer measures eye pressure by measuring depth of deformity? caused by a small metal plunger resting on the cornea (Moses RA, Intraocular pressure. In: Moses RA and Hart WM, Alder's Physiology of the eye Clinical Application. 9th ed. Mosby Company. St. Louis, 1997, 223 -245).

Biochemical monitoring

Preparation of crystallines

Each lens, after being removed ? been washed with physiological saline solution; each sample ? was homogenized for 6 seconds in equal volume of 50 mM phosphate buffer (pH 7.2) and centrifuged at 12000 rpm for 15 minutes at 4°C. The supernatant obtained? was used for subsequent analyses.

For biochemical monitoring of cataractogenesis, the following parameters were analysed:

- TBARS dosage

Colorimetric method based on the reaction of 2-thiobarbituric acid in which the quantity is determined? of TBAR (thiobarbituric acid reactive substances) present in the crystalline homogenate with the use of a spectrophotometer. The sample ? was mixed with 2 volumes of 10% w/v trichloroacetic acid (TCA) to precipitate the proteins. The precipitated pellet ? been removed and an aliquot of the supernatant ? was reacted with an equal volume of 0.67% w/v thiobarbituric acid for 10 minutes. The absorbance of TBARS ? was detected at 532 nm and the results were expressed in ?mol/g wet weight.

- TAC essay

The capacity? total antioxidant (TAC) of the lens ? was determined according to the method of Re et al. (1999). As standard ? Has Trolox been used<? > (6-hydroxy-2,5,7,8-tetramethylcroman-2-carbosyl acid) and the ability? champions total antioxidant ? been defined as the concentration of activity? Trolox equivalent to wet weight in ?mol/g.

Dosage of alpha-tocopherol and retinol

Alpha-tocopherol and retinol were determined using the procedure of Zhao et al. (2004). Samples were analyzed by an HPLC system (Kontron Instruments, Milan, Italy). The quantification? was performed using the Geminyx system (version 1.91). The results were expressed as ?mol/mg wet weight.

Results

For the whole duration of the experiments the physical conditions of the animals were satisfactory. There were no changes in ocular tone in any animal studied.

What type of cataract do you have? developed in treated animals (groups B, C, D and E) was of the posterior subcapsular type (SCP), so? how it appeared at the biomicroscopic examination.

- In group A (control) the lenses remained transparent (grade 0) for the whole duration of the experiment (30 days).

- In group B (selenite) after cataract induction the mean grade was 2.5 ? 1.5 to 7? post-induction day and 4 ? 0,00 to each of the 15 days? and 30? of the follow-up.

- In group C (selenite and laurel-free eye drops) after cataract induction the mean score was 2.3 ? 1.7 to 7? day post-induction and 3.8 ? 0,2 at each of 15 days? and 30? of the follow-up.

- In the animals that had received the laurel eye drops (group D) the mean score was 0.5 ? 0.1 to 7? day, which worsened to 1.5 ? 0.5 on 15 days to remain stable at 1.5? 0.2 up to 30? day.

- In the animals that had received eye drops with laurel but without hyaluronic acid and aloe (group E) the average score was 0.6? 0.1 to 7? day, which worsened to 1.6 ? 0.5 to 15? day to remain stable at 1.5 ? 0.2 up to 30? day, i.e. results comparable to those obtained for group D. However, ? been ascertained that the in vivo administration of this eye drops containing only laurel causes conjunctival hyperemia and inflammation from the very first applications and ? generally poorly tolerated by patients.

In other words, these results demonstrate that although both eye drops comprising laurel extract are effective in the prevention and treatment of cataracts, only the eye drops comprising the combination of the present invention, i.e. laurel extract together with aloe extract and acid hyaluronic, is suitable for in vivo administration. In fact, without wanting to be tied to any theory, the inventors have noticed that the administration of laurel extract alone (group E) can have some side effects including conjunctival hyperaemia, redness and irritation.

Biochemical monitoring of the lens

TAC levels were significantly reduced in animals in groups B and C compared with those in groups A (controls) and D-E (laurel-treated). TBARS values were lower in the bay group than in the control group.

The levels of alpha-tocopherol and retinol were significantly reduced in animals belonging to groups B and C. Conversely, the administration of eye drops containing laurel increased the levels of alpha-tocopherol and retinol, demonstrating a protective effect against induced oxidation from selenite (group D and group E).

Claims (12)

1. Association of at least one plant extract belonging to the Laurus genus, at least one plant extract belonging to the Aloe genus and hyaluronic acid. 2. Association according to claim 1, wherein the plant extract belonging to the genus Laurus ? chosen from Laurus nobilis L. extract, Laurus azorica extract, Laurus chinensis extract, Laurus melissifolia extract and mixtures of these, and ? preferably extract of Laurus nobilis L. 3. Association according to claim 1 or 2, wherein the plant extract belonging to the genus Aloe? chosen from Aloe Vera extract, Aloe Arborescens extract, Aloe Chinensis extract, Aloe Ferox extract, Aloe Humilis extract and blends of these, and ? preferably Aloe Vera extract. 4. Association according to one or more? of claims 1 to 3, wherein said hyaluronic acid is hyaluronic acid sodium salt, preferably with a molecular weight between 100 and 1000 kDa, more? preferably between 200 KDa and 800 KDa, even more? preferably between 300 KDa and 600 KDa. 5. Combination according to any one of claims 1 to 4 which further comprises an antioxidant, preferably selected from the group consisting of: Vitamin A, Vitamin C, Vitamin E, D-alpha-tocopheryl poly(ethylene glycol)-1000-succinate (VE- TPGS), selenium, carotenoids, lycopene, coenzyme Q-10, lipoic acid and mixtures thereof. 6. Composition comprising the combination according to any one of claims 1 to 5 and a suitable vehicle and/or excipient, preferably wherein said vehicle is? isotonic buffered solution at pH 7.2. 7. Composition according to claim 6 for topical use, preferably in a form selected from: eye drops, solution, suspension, eye drops, spray, ophthalmic bath, gel, ointment, ointment, cream, medicated wipe, occlusive dressing and ophthalmic inserts. 8. Composition according to claim 6 for oral use, preferably in a form selected from: capsules, soft capsules, tablets, pills, pastilles, jellies, powders, granules, emulsions, solutions, suspensions, syrups and elixirs. 9. Association according to any one of claims 1 to 5 or composition according to any one of claims 6 to 8 for use as a medicament, preferably for use in the prevention and/or treatment of ocular pathologies. 10. Association or composition for use according to claim 9, wherein said eye pathologies are cataract, keratoconus, dry eye, conjunctival allergy, conjunctivitis, blepharitis, keratitis, actinic keratitis and damage from exposure to UV rays. 11. A kit of parts comprising a first composition for topical or oral use comprising at least one plant extract belonging to the genus Laurus, a second composition for topical or oral use comprising at least one plant extract belonging to the genus Aloe and a third composition for topical or oral use comprising hyaluronic acid, for use in the prevention and/or treatment of ocular pathologies, by means of the sequential or simultaneous administration of said first, second and third compositions. 12. Non-therapeutic use of the combination, composition or kit according to any of the preceding claims as an ocular lubricant.