WO2007046083A2

WO2007046083A2 - Compositions for treatment of eye diseases

Info

Publication number: WO2007046083A2
Application number: PCT/IL2006/001168
Authority: WO
Inventors: Yoav Sharoni; Joseph Levy; Yoram Sela; Zohar Nir
Original assignee: Lycored Ltd.
Priority date: 2005-10-16
Filing date: 2006-10-05
Publication date: 2007-04-26
Also published as: JP2009515821A; WO2007046083A3; EP1937240A2; AU2006305546A1; RU2008119234A; MX2008004981A; IL190779A0

Abstract

The present invention relates to pharmaceutical compositions comprising a mixture of carotenoids and adjuvants, which provides synergistic therapeutic potential. The compositions of the present invention, which can be administered orally or topically to the eye, are useful in preventing, attenuating or treating eye diseases such as age-related macular degeneration (AMD), glaucoma, diabetic retinopathy and cataracts.

Description

COMPOSITIONS FOR TREATMENT OF EYE DISEASES

FIELD OF THE INVENTION

The present invention relates to the field of ophthalmology, in particular to compositions useful in treating age-related macular degeneration (AMD) and other eye diseases. The novel formulations comprise a mixture of carotenoids and their biologically active adjuvants, and provide synergistic therapeutic potential.

BACKGROUND OF THE INVENTION Eve Pathologies

Eye diseases, including age-related macular degeneration (AMD), cataracts, diabetic retinopathy and glaucoma, are a leading cause of blindness affecting millions of people worldwide. As the global population ages, the number of individuals affected is expected to increase substantially. Macular degeneration, or age-related macular degeneration (AMD), affects the central part of the retina and is the leading cause of blindness in people over age 65 in the United States. AMD affects 13 million people and causes impairment in about 1.2 million. The prevalence of AMD increases with age from 16.8% in patients 55-64 to 25.6% in patients 65-74 and up to 42% in patients over 75. There are two types of AMD: dry and wet. Dry AMD accounts for about 90 percent of all cases and is sometimes referred to as atrophic, nonexudative, or drusenoid macular degeneration. In dry AMD, deposits called drusen accumulate in the retinal pigment epithelium (RPE) tissue beneath the macula. These deposits are thought to interfere with the function of photoreceptors in the macula, causing progressive degeneration of these cells. Vision loss from dry AMD occurs very gradually over the course of many years. Dry AMD often progresses into wet AMD. In wet AMD, abnormal blood vessels grow beneath the macula. These vessels leak blood and fluid into the macula, thereby damaging photoreceptor cells. Wet AMD progresses rapidly and can cause severe damage to central vision. If wet AMD is diagnosed early, laser surgery can prevent extensive central vision loss by destroying the leaky blood vessels. No nonsurgical treatments are available.

Diabetic Retinopathy (DR) is a complication of diabetes that is caused by changes in the blood vessels of the retina. Damaged blood vessels in the retina may leak blood and grow fragile, brush-like branches and scar tissue. This can blur or distort the vision images that the retina sends to the brain. DR is a leading cause of blindness in the United States and untreated diabetics are more at risk for blindness than the general population.

Cataracts are the leading cause of blindness in the world, and the leading cause of reversible visual loss for people over age 65. Cataracts are a loss of the transparency of the lens of the eye causing blurred vision, glare, sensitivity to light, poor night vision, halos around lights and color distortion. About 4 million people have cataracts in the United States, and 40,000 develop cataracts each year. With the life expectancy over 76 years, the incidence of cataracts is expected to double in the next 12 years. Cataract surgery is the most common surgical procedure for the elderly. Glaucoma is the leading cause of irreversible blindness in the world, the second most common cause of irreversible blindness in the United States, and the most common cause of blindness among blacks. An estimated 2.5 million persons in the U.S. have glaucoma. Worldwide, it is estimated that about 66.8 million people have visual impairment from glaucoma, with 6.7 million suffering from blindness. Glaucoma is not a single disease, but rather a group of disorders that damage the optic nerve. This pattern usually occurs in the presence of high intraocular pressure, but can also occur with normal or even below-normal eye pressure.

Dietary supplements have been shown to be effective in reducing the risk of and in treating certain eye diseases. The Age-Related Eye Disease Study (AREDS), a major clinical trial performed to evaluate the effect of high oral doses of antioxidants and zinc on the progression of AMD and cataract, showed that the consumption of high levels of antioxidants and zinc reduce the risk of developing advanced AMD by about 25 percent. The AREDS composition comprises vitamin A, vitamin C, vitamin E and the minerals zinc and copper. Intake of this composition was shown to be associated with side effects, including genitourinary problems, anemia, skin yellowing and an increased risk of lung cancer in smokers.

Another study, the Lutein Antioxidant Supplement Trial (LAST)₅ evaluated the effects of orally administered lutein and lutein combined with antioxidants on atrophic (dry) AMD. Both treatments significantly improved some measures of visual function, including glare recovery, contrast sensitivity, and visual acuity. The long-term safety of dietary lutein supplementation has not yet been determined.

Dietary Anti-oxidants

Carotenoids are naturally occurring pigments found in certain plants and algae. For example, the red color of tomatoes and yellow color of corn derive from carotenoids. Mammals are incapable of synthesizing carotenoids in situ and must obtain them through their diet. High dietary intake of carotenoids has been shown to be associated with reduced incidence of certain types of cancers, cardiovascular disease and hypertension. A dermal photoprotective effect is also associated with a high dietary intake of carotenoids. Epidemiological studies have shown that dietary anti-oxidants such as carotenoids delay the progression of eye disease, including the progression of cataracts and AMD. Carotenoids such as lycopene, the main tomato carotenoid, became the subject of more intensive investigation. Epidemiological studies have established that a low serum concentration of carotenoids, especially lycopene, is associated with risk of AMD. Other carotenoids, including lutein and zeaxanthin, have been shown to have a protective role in the development of cataract and macular degeneration (Mares-Perlman, 2002).

Lycopene

Lycopene is the major carotenoid present in the diet and provides the familiar red color of tomato products. More than 80% of dietary intake of lycopene is derived from tomato sources, such as ketchup, tomato juice, spaghetti sauce, tomato soup and pizza sauce. Lycopene can be prepared synthetically, or can be obtained as a natural tomato extract. For example, lycopene is obtainable under the name Lyc-O-Mato® (LycoRed, Israel) as an all-natural antioxidant formula containing tomato lycopene, tocopherols, beta- carotene, phytoene, phytofluene, tomato oil, phospholipids, and other important bioactive phytochemicals naturally occurring in tomato oleoresin.

Lycopene has been shown to provide eye protection and endogenous protection of UV induced skin damage when administered orally. Several studies have shown an anti- cataract effect of dietary lycopene in animals (Pollack, et al.). Lycopene prevents galactose- induced morphological changes in cultured human lens cells, suggesting its use as an anti- cataract agent (Mohanty, L, et al). Without wishing to be bound to theory, it is believed that lycopene has a protective effect on lutein and zeaxanthin.

Lutein and Zeaxanthin Lutein and its stereoisomer zeaxanthin belong to the xanthophyll family of carotenoids and are the only two known carotenoid specifically accumulated from plasma and deposited in the lens and macula lutea. The macular pigment comprises lutein and zeaxanthin, and functions as a filter to protect the light-sensitive photoreceptor cells that are also known to scavenge reactive oxygen species (ROS). Dietary intake of these carotenoids results in increased macular pigment and improved vision in patients with AMD and other ocular diseases (reviewed in Alves-Rodrigues and Shao).

Green leafy vegetables are the best dietary source of lutein, with spinach, kale and parsley providing high levels. Purified crystalline lutein has been classified generally recognized as safe (GRAS) and can be added to food and beverages. Pure lutein may also be isolated from certain plants. For example, US Patent No. 5,382,714 describes a process for isolation, purification, and recrystallization of lutein from saponified marigold oleoresin, and US Patent No. 5,648,564 teaches a process for the formation, isolation and purification of comestible xanthophyll crystals from plants, with lutein a preferred product.

Seddon (Seddon et al, 1994) reported the first direct relationship between carotenoid intake and AMD risk. Lutein and zeaxanthin were most strongly associated with a decreased AMD risk. An oral dose of 6% wt/wt per day was shown to translate into a 57% lower risk of disease. Carnosic Acid. Phytoene and Phytofluene

Carnosic acid is an antioxidant extracted from rosemary (Rosemarinus spp) and other herbs, which has been shown to inhibit LDL oxidation in a synergistic manner with lycopene. Phytoene and phytofluene are carotenoids found in tomatoes, and may be found in tomato oleoresin.

Phase II Enzymes

The biochemical mechanisms involved in the protective effects of fruits and vegetables in general and of tomatoes in particular are not completely understood. In recent years, evidence has accumulated indicating that the beneficial action is due, at least in part, to the induction of phase II detoxification enzymes (Talalay, 2000). These enzymes detoxify many harmful substances by converting them to hydrophilic metabolites that can be excreted readily from the body.

The coordinated induction of phase II enzymes, such as NAD(P)H: quinone oxidoreductase (NQOl) and γ-glutamylcysteine synthetase (GCS) is mediated through cis- regulatory DNA sequences located in the promoter or enhancer region, which are known as antioxidant responsive elements (ARE). Stimulation of the ARE transcription system is an established mechanism for the mobilization of the body's defense system against carcinogens and other harmful compounds. The major ARE activating transcription factor Nrf2 (nuclear factor E₂- related factor 2) plays a central role in the induction of antioxidant and detoxifying genes. Under basal conditions, Nrf2 is located in the cytoplasm and is bound to an inhibitory protein, Keapl. Upon challenge with inducing agents, it is released from Keapl and translocates to the nucleus. It has recently been shown by some of the applicants of the present invention that in transiently transfected cancer cells, lycopene is capable of transactivating the expression of reporter genes fused with ARE sequences (Ben Dor et al, 2005). A mixture of two other tomato carotenoids, phytoene and phytofluene, was also effective in activation of ARE. By activating this system, tomato carotenoids induce the production of phase II detoxification enzymes. Carotenoid Formulations

Certain formulations comprising carotenoids have been disclosed in the art. US

Patent Nos. 6,261,598 and 6,509,029 (hereinafter '598 and '029, respectively) disclose oil containing dispersion and an emulsion or dry powder produced therefrom, respectively, comprising a mixture of β-carotene, lycopene and lutein. US Patent Application Publication

No. 2003/0031706 discloses carotenoid formulations comprising specific mixtures of β- carotene, lycopene and lutein. The '598 and '029 patents teach away from an oral composition comprising Lyc-O-Mato^®, in particular a high concentration carotenoid- containing gelatin capsule due to the high phospholipid content and viscosity of the oily dispersion and the exemplified formulations comprise pure crystalline carotenoids.

US Patent No. 6,103,756 teaches oral compositions comprising effective amounts of vitamins, minerals, and plant extracts. The essential ingredients include, inter alia, vitamins A, C and E, and bilberry, lutein and lycopene extracts.

US Patent No. 6,573,299 teaches compositions for topical application to the outer eyelid comprising at least one permeation enhancer and at least one alpha hydroxy acid.

Those compositions, which penetrate the skin and permeate the underlying tissues, are said to improve disorders associated with the aging eye including age-related changes to the eyelids such as wrinkles and dry skin and diseases such as cataracts and AMD.

US Patent Application Publication No. 2003/0050283 relates to an ophthalmic composition for the treatment of pathological retinal disorders comprising as an essential element at least one negatively charged phospholipid, other than cardiolipin. That composition may further comprise at least one carotenoid.

US Patent No. 6,291,519 teaches a method for preventing damage to mammalian eye tissues by applying to the eye a composition comprising an amount of vitamin A and an amount of vitamin E effective to absorb UV-irradiation, destroy ozone, or both.

US Patent No. 5,527,533 teaches a method of improving the vision of an individual suffering from retinal damage or disease, comprising administering a therapeutically effective amount of astaxanthin, a carotenoid found in aquatic animals. Parenteral, oral and topical compositions are disclosed, however the invention is exemplified only by parenteral and oral compositions.

US Patent Application No. 2004/0258769 teaches compositions for intraocular injection comprising anecortave acetate in conjunction with a daily regimen of ocular vitamins.

There remains an unmet need in the art compositions that are effective in preventing and treating eye diseases such as AMD, cataract, diabetic retinopathy and glaucoma.

SUMMARY OF THE INVENTION The present invention relates to compositions effective in treating eye diseases. The compositions comprise a mixture of phytochemicals, which produce a synergistic effect in attenuating and treating eye diseases including glaucoma, cataracts, diabetic retinopathy (DR) and age-related macular degeneration (AMD). Specifically, the compositions are effective in preventing the progression of dry AMD (non-neovascular) to wet AMD (neo vascular).

Certain carotenoids and other phytochemicals are known to be effective in reducing the risk of certain eye diseases and may even improve visual function when ingested in high concentrations. The compositions of the present invention offer a significant advantage over known formulations, as they include a mixture of carotenoids and plant derived adjuvants that act synergistically and afford unexpected results at low concentrations of lycopene. As such, the concentration of lycopene which provides a therapeutic effect is lower (e.g., 10- fold lower) as compared with a corresponding formulation not containing these adjuvants.

In one aspect, the present invention provides a composition comprising lutein, zeaxanthin, lycopene, phytoene, phytofluene and carnosic acid, and a pharmaceutically acceptable carrier or excipient.

The components of the composition are provided as percent weight per total weight (% w/w). In one exemplary embodiment, lutein is present in the composition in a concentration range of about 0.025% w/w to about 5% w/w. A range of about 0.25% w/w to about 2.5% w/w is preferred, and a concentration of about 2% w/w is more preferred. In some embodiments lutein is synthetically prepared. In other embodiments lutein is isolated and purified from a natural source.

The present composition further comprises zeaxanthin, a stereoisomer of lutein. In one exemplary embodiment, zeaxanthin is present in the composition in a concentration range of about 0.001% w/w to about 2% w/w. A range of about 0.025% w/w to about 1% w/w is preferred, and a concentration of about 0.5% w/w is more preferred. In some embodiments zeaxanthin is synthetically prepared. In other embodiments zeaxanthin is isolated and purified from a natural source.

In other embodiments, zeaxanthin is contained in the lutein fraction. In accordance with these embodiments, the total concentration of lutein and zeaxanthin is about 0.035% to about 7% w/w, preferably in the range of about 1.5% to about 2.5% w/w, more preferably about 2% to about 2.2% w/w total lutein and zeaxanthin.

Lycopene is preferably provided in the composition as a natural compound. In some embodiments, lycopene is provided as an extract, example, as an extract of tomato oleoresin, such as Lyc-O-Mato®. In some embodiments the lycopene extract further comprises phytoene, phytofluene, lutein and zeaxanthin. According to one exemplary embodiment, lycopene is provided in the present composition in a concentration range of about 0.025% w/w to about 5% w/w. A range of about 0.25% w/w to about 2.5% w/w is preferred, and a concentration of about 1.5% w/w is more preferred.

Phytoene and phytofluene are provided as adjuvants in the present compositions. In an exemplary embodiment, phytoene and phytofluene are present at about 10% of the concentration of lycopene. In another exemplary embodiment, each of phytoene and phytofluene are provided in a concentration range of about 0.0025% w/w to about 1.25% w/w. A range of about 0.025% w/w to about 0.25% w/w of each phytochemical is preferred, and a concentration of 0.15% w/w total of both phytoene and phytofluene, corresponding to about 10% of the lycopene content, is more preferred. Carnosic acid or derivatives thereof, (e.g., carnosol, 6,7-dehydrocarnosic acid or 7- ketocarnosic acid) is also provided as an adjuvant in the present compositions. Carnosic acid or its derivatives is preferably provided in a concentration range of about 0.025% w/w to about 2.5% w/w. A range of about 0.25% w/w to about 2% w/w is preferred, and a concentration of 1% w/w is more preferred

In another currently preferred embodiment, the composition further comprises tomato oleoresin.

According to one embodiment the composition of the present invention comprises from about 0.025% w/w to about 5% w/w lutein; from about 0.001% w/w to about 2% w/w zeaxanthin; from about 0.025% w/w to about 5% w/w mg lycopene; from about 0.0025% w/w to about 1.25% w/wmg phytoene; from about 0.0025% w/w to about 1.25% w/wphytofluene; from about 0.025% w/w to about 2.5% w/w carnosic acid or derivative thereof; and a pharmaceutically acceptable carrier or excipient.

In a currently preferred embodiment, the composition of the present invention comprises; about 1.5% w/w lycopene; about 2% w/w total of lutein and zeaxanthin, about

0.15% w/w total of phytoene and phytofluene; about 1% w/w carnosic acid or a derivative thereof; and a pharmaceutically acceptable carrier or excipient. In one embodiment, the composition further comprises tomato oleoresin.

Additional therapeutic agents, including vitamins, minerals, other carotenoids, and micronutrients can optionally be incorporated in the compositions of the present invention.

In some embodiments the composition further comprises a source of zinc, which can be zinc oxide or a zinc salt. Any zinc salt that is compatible with the eye is acceptable. Examples of zinc salts include but are not limited to zinc chloride, zinc acetate, zinc gluconate, zinc carbonate, zinc sulfate, zinc borate, zinc nitrate and zinc silicate. In some embodiments a concentration of about 0.25% to about 15% w/w zinc is provided. In preferred embodiments a concentration of about 7.5% w/w zinc is provided; preferably in the form of zinc gluconate. In certain embodiments a source of copper is provided in the present composition. In some embodiments cupric oxide is preferred. In some embodiments a copper is provided at a concentration of about 0.01% to about 5% w/w; preferably at a concentration of about 0.25% w/w. Certain preferred vitamins include vitamin A, vitamin C and vitamin E. The vitamins can be provided as natural or synthetically produced compounds. Derivatives, including provitamins of the preferred vitamins are acceptable. In some preferred embodiments vitamin A is provided as provitamin A, specifically β-carotene. In some embodiments, combinations of the above vitamins are included in the composition. The addition of vitamin E, vitamin C and β-carotene is preferred. Vitamin E and vitamin C can each be included in the present composition at a concentration of about 0.25% to about 25% w/w. Preferable concentrations of vitamin E and vitamin C are about 12.5%. β-carotene can be included in the present composition at a concentration of about 0.025% to 2.5% w/w. Preferable concentrations of β-carotene are about 0.75% w/w. According to some embodiments, the composition of the present invention comprises lutein, zeaxanthin, lycopene, phytoene, phytofluene, carnosic acid or a derivative thereof, vitamin E or a derivative thereof, vitamin A or a derivative thereof (e.g. β- carotene), vitamin C, zinc and copper; and a pharmaceutically acceptable carrier or excipient. In one preferred embodiment, the composition comprises: from about 0.025% w/w to about 5% w/w lutein; from about 0.001% w/w to about 2% w/w zeaxanthin; from about 0.025% w/w to about 5% w/w mg lycopene; from about 0.0025% w/w to about 1.25% w/wmg phytoene; from about 0.0025% w/w to about 1.25% w/w phytofluene; from about 0.025% w/w to about 2.5% w/w carnosic acid or derivative thereof; from about 0.25% w/w to about 25% w/w vitamin E; from about 0.25% w/w to about 25% w/w vitamin C; from about 0.025% to about 2.5% w/w β-carotene; from about 0.25% w/w to about 15% w/w mg zinc; from about 0.01% w/wto about 5% w/w copper; and a pharmaceutically acceptable carrier or excipient.

In another currently preferred embodiment, the composition comprises: about 1.5% w/w lycopene; about 2% w/w total of lutein and zeaxanthin, about 0.15% w/w total of phytoene and phytofluene; about 1% w/wcarnosic acid or a derivative thereof; about 12.5% w/w vitamin E; about 12.5% w/w vitamin C; about 0.75% w/w β-carotene; about 7.5% w/w zinc; about 0.25% w/w copper; and a pharmaceutically acceptable carrier or excipient.

The present invention relates to pharmaceutical compositions for oral use or for topical administration to the eye. In one embodiment the composition is formulated for oral use in a form selected from a tablet, caplet, capsule, microcapsule, pellet, pill, powder, syrup, gel, slurry, granule, suspension, dispersion, emulsion, liquid, solution, dragee, bead and beadlet and the like. A beadlet is a polysaccharide complex, in the shape of a bead, in which small droplets containing the active material are embedded. In certain preferred embodiments the composition is formulated as a soft gelatin capsule or as a hard gelatin capsule. In other embodiments the composition is formulated as a beadlet based on alginates, gelatin or other natural or synthetic polymers.

The present invention also relates to a composition formulated for application to the eye. The composition can be formulated as a liquid, cream, paste, ointment, emulsion including submicron emulsion, gel, thermogel, or suspension.

The composition can also be dispensed as dry formulation, for example as powder, granules, microcapsules or capsules, for reconstitution as a liquid, dispersion, emulsion or suspension.

In some embodiments the composition is provided in an aqueous or non-aqueous medium, and is preferably sterile (microbe-free).

In another aspect, the present invention provides a method of preventing, attenuating or treating an eye disease comprising the step of administering an effective amount of a composition comprising lutein, lycopene, phytoene, phytofluene and carnosic acid; and a pharmaceutically acceptable carrier or excipient, in an amount effective to prevent attenuate or treat the eye disease. Non-limiting examples of eye diseases include AMD (both wet and dry), glaucoma, diabetic retinopathy and cataract. Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based on the surprising discovery that a composition comprising a mixture of carotenoids and other phytochemicals is effective in treating eye diseases. Without wishing to be bound to any particular theory or mechanism, it is believed that lycopene and the adjuvants carnosic acid, phytoene and phytofluene work synergistically in the formulations of the invention. This synergistic effect allows for the inclusion of lower amounts of lycopene e.g., 10 times lower than the amount of lycopene in known formulations, i.e., formulations which do not contain carnosic acid, phytoene and phytofluene, while still achieving a beneficial therapeutic effect. Lycopene, in turn, has a protective effect on lutein and zeaxanthin.

Accordingly the present invention provides a pharmaceutical composition comprising lutein, zeaxanthin, lycopene, phytoene, phytofluene, carnosic acid; and a pharmaceutically acceptable carrier or excipient.

In preferred embodiments lycopene is provided as a natural tomato extract, which can be extracted from tomato oleoresin.

In specific embodiments, the composition of the present invention comprises from about 0.025% w/w to about 5% w/w lutein; from about 0.001% w/w to about 2% w/w zeaxanthin; from about 0.025% w/w to about 5% w/w mg lycopene; from about 0.0025% w/w to about 1.25% w/wmg phytoene; from about 0.0025% w/w to about 1.25% w/w phytofluene; from about 0.025% w/w to about 2.5% w/w carnosic acid or derivative thereof; and a pharmaceutically acceptable carrier or excipient.

In a currently preferred embodiment, the composition of the present invention comprises; about 1.5% w/w lycopene; about 2% w/w total of lutein and zeaxanthin, about 0.15% w/w total of phytoene and phytofluene; about 1% w/w carnosic acid or a derivative thereof; and a pharmaceutically acceptable carrier or excipient.

In another currently preferred embodiment, the composition further comprises tomato oleoresin. The tomato oleoresin can be obtained from tomato or tomato products in accordance with methods well known to a person of skill in the art.

Additional therapeutic agents can be added to the composition. Therapeutic agents include anti-inflammatory, antipruritic, pain relievers, antibiotics, wetting agents, moisturizing agents, vitamins, minerals, micronutrients and other phytochemicals, including carotenoids.

Certain preferred vitamins include vitamin A, vitamin C and vitamin E, and derivatives thereof. The vitamins may be provided as natural or synthetically produced compounds. In some preferred embodiments vitamin A is provided as provitamin A, specifically as β-carotene. Other acceptable forms of vitamin A include retinal, esters of retinol, retinoic acid, and the like. Vitamin E is meant to include α-tocopherol and isomers and racemates of α-tocopherol and derivatives thereof (e.g., tocopherol acetate). Vitamin C can be provided per se or as a derivative of ascorbic acid. For example, ascorbic palmitate, a fat-soluble ester, is stable and acceptable in the present compositions.

Vitamin E can be included in the composition at a concentration range of about

0.25% w/w to about 25% w/w, preferably at a concentration range of about 0.1% to about

15%, more preferably at a concentration of about 12.5% w/w. Without wishing to be bound to any particular mechanism or theory, it is believed that vitamin E increases the stability and cellular uptake of lycopene.

Vitamin A or derivatives thereof, for example β-carotene, can be included in the composition at a concentration range of about 0.025% w/w to about 2.5% w/w; preferably at a concentration of about 0.75% w/w. Vitamin C and derivatives thereof, including ascorbic acid are optionally included in the composition of the present invention. In certain embodiments vitamin C is included in a range of about 0.25% w/w to about 25% w/w, preferably at a concentration range of about 0.1% to about 15%, more preferably at a concentration of about 12.5% w/w. Minerals can also be added to the formulations of the present invention. Preferred minerals include zinc and copper, which are preferably added together to reduce the risk of zinc-related anemia. Zinc, in the form of a zinc salt or a zinc oxide, is preferably included in the composition at a range of about 0.25% w/w to about 15% w/w, preferably about 7.5% w/w zinc. Preferred zinc salts include but are not limited to zinc chloride, zinc acetate, zinc gluconate, zinc carbonate, zinc sulfate, zinc borate, zinc nitrate and zinc silicate. In specific embodiments, the composition comprises zinc gluconate. A source of copper can be included at a concentration range of about 0.01% w/w to about 5% w/w. In preferred embodiments cupric oxide is included at a concentration of about 0.25% w/w.

According to some embodiments, the composition of the present invention comprises lutein, zeaxanthin, lycopene, phytoene, phytofluene, carnosic acid or a derivative thereof, vitamin E or a derivative thereof, vitamin A or a derivative thereof (e.g. β- carotene), vitamin C, zinc and copper; and a pharmaceutically acceptable carrier or excipient. In one preferred embodiment, the composition comprises: from about 0.025% w/w to about 5% w/w lutein; from about 0.001% w/w to about 2% w/w zeaxanthin; from about 0.025% w/w to about 5% w/w mg lycopene; from about 0.0025% w/w to about 1.25% w/wmg phytoene; from about 0.0025% w/w to about 1.25% w/w phytofluene; from about 0.025% w/w to about 2.5% w/w carnosic acid or derivative thereof; from about 0.25% w/w to about 25% w/w vitamin E; from about 0.25% w/w to about 25% w/w vitamin C; from about 0.025% to about 2.5% w/w β-carotene; from about 0.25% w/w to about 15% w/w mg zinc; from about 0.01% w/w to about 5% w/w copper; and a pharmaceutically acceptable carrier or excipient. In another currently preferred embodiment, the composition comprises: about 1.5% w/w lycopene; about 2% w/w total of lutein and zeaxanthin, about 0.15% w/w total of phytoene and phytofluene; about 1% w/wcarnosic acid or a derivative thereof; about 12.5% w/w vitamin E; about 12.5% w/w vitamin C; about 0.75% w/w β-carotene; about 7.5% w/w zinc; about 0.25% w/w copper; and a pharmaceutically acceptable carrier or excipient.

Mechanism of Action

Without wishing to be bound by any particular mechanism and theory, it believed that the lycopene may exert its therapeutic effects by inducing antioxidant response element (ARE), which in turn induces the expression of phase II detoxification enzymes. These enzymes detoxify many harmful substances by converting them into hydrophilic metabolites that can be excreted readily from the body. Examples of phase II enzymes, are NAD(P)H:quinone oxidoreductase (NQOl) and g glutamylcysteine synthetase (GCS). The major ARE-activating transcription factor Nrf2 plays a central role in the induction of antioxidant and detoxifying genes.

In one non-limiting embodiment described herein for purpose of illustration and not for limitation, lycopene increases the activity of ARE by forming Keapl adducts. This may cause in turn its dissociation from Nrf2 and the activation of the latter. Keapl is cysteine- rich cytoplasmic protein that negatively regulates the activation of Nrf2. The central domain of Keapl is the most cysteine-rich domain and is required for cytoplasmic sequestration and inhibition of Nrf2. Dissociation of Nrf2 from Keapl represents a regulatory step that allows Nrf2 to translocate to the nucleus and activate transcription of ARE-dependent genes. The dissociation of Nrf2 from Keapl -Nrf2 may be regulated by the redox status of these specific cysteine residues. Adducts of specific aldehydes were observed in LC-MS-MS analyses of purified human Keapl. The formation of these adducts was coincident with. Nrf2 stabilization, nuclear Nrf2 translocation and ARE dependent gene activation.

As contemplated herein, the lycopene and the adjuvants (i.e., carnosic acid, phytoene and phytofluene) in the formulations of the present invention act synergistically and afford unexpected results at low concentrations of lycopene. As such, the concentration of lycopene which provides a therapeutic effect is lower (e.g., 2 fold, 5 fold or even 10-fold lower) as compared with a corresponding formulation not containing these adjuvants. This provides a significant advantage over known formulations.

Formulations

As disclosed herein, the compositions of the present invention comprise lutein, lycopene, zeaxanthin phytoene, phytofluene and carnosic acid, in combination with chemical components such as physiologically suitable carriers and excipients. Pharmaceutical compositions for use in accordance with the present invention can be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the active compounds into preparations which can be used pharmaceutically. The carotenoids are formulated as pharmaceutical compositions and administered to a mammalian subject, such as a human patient in a variety of forms such as liquid, solid, and semisolid. The pharmaceutical compositions can be administered to a subject by any method known to a person skilled in the art, such as orally, topically, parenterally, paracancerally, transmucosally, transdermally, intramuscularly, intravenously, intradermally, subcutaneously, intraperitonealy, intraventricularly, intracranially or intratumorally. For oral administration, the compounds can be formulated by combining the active compounds with pharmaceutically acceptable carriers known in the art. The compositions can be formulated in any solid or liquid dosage form known in the art, including but not limited to, tablet, caplet , capsule, microcapsule, pellet, pill, powder, syrup, gel, slurry, granule, suspension, dispersion, emulsion, liquid, solution, dragee, bead and beadlet. The oral compositions can be formulated as immediate release formulations, or as controlled or sustained release formulations allowing for extended release of the active ingredient(s) over a predetermined time period.

Suitable excipients for solid formulations include but are not limited to fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; starch based excipients such as maize starch, wheat starch, rice starch, potato starch and the like, gelatin, gum tragacanth, cellulose based excipients as microcrystalline cellulose, carboxymethylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, methylhydroxypropylcellulose, hydroxypropylcellulose and the like. Polymers such as polyvinylpyrrolidone (PVP) and cross-lined PVP can also be used. In addition, the compositions may further comprise binders (e.g. acacia, cornstarch, gelatin, carbomer, ethyl cellulose, guar gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, povidone), disintegrating agents (e.g. cornstarch, potato starch, alginic acid, silicon dioxide, croscarmelose sodium, crospovidone, guar gum, sodium starch glycolate), surfactants (e.g. sodium lauryl sulfate), and lubricants (e.g. stearic acid, magnesium stearate, polyethylene glycol, sodium lauryl sulfate).

For liquid formulations, pharmaceutically acceptable carriers may be aqueous or non-aqueous solutions, suspensions, emulsions or oils. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters. Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media. Examples of oils include but are not limited to petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, mineral oil, olive oil, sunflower oil, and fish-liver oil.

Preferred oral pharmaceutical compositions include capsules made of gelatin as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. In certain preferred embodiments the capsules exclude components of animal origin and are acceptable for vegetarians and vegans. Soft gelatin capsules and methods of preparing them are known in the art. Non- limiting examples can be found in US Patent Nos. 6,217,902; 6,258,380; 5,916,591, and 4,891,229, all of which are incorporated herein by reference. In another embodiment, the present invention further relates to a composition formulated for application to the eye. The composition can be formulated as drops, solution (aqueous and non-aqueous), cream, paste, ointment, gel, emulsions, suspension, and the like

For ophthalmic administration, the composition can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as liquids, gels, emulsions, thermogels, slurries, suspensions, and the like, for ophthalmic use by a patient. Alternatively the composition can be formulated as a solid, for resuspension. In other embodiments, pharmaceutical compositions for ophthalmic administration include aqueous solutions of the active ingredients in water-soluble form.

Other acceptable excipients and additives known to the person with skill in the art may be included in the compositions of the present invention, for example stabilizers, solubilizers, tonicity enhancing agents, buffer substances, preservatives, thickeners, complexing agents and other excipients, as well as additional therapeutic agents. A solubilizer can be for example, tyloxapol, fatty acid glycerol polyethylene glycol esters, fatty acid polyethylene glycol esters, polyethylene glycols, glycerol ethers or mixtures of those compounds. A specific example of a solubilizer well tolerated by the eye is a polyoxyethylated castor oil for example, the commercial products Cremophor ^®or Cremophor^® RH40. Another example of a solubilizer is tyloxapol. The concentration used depends especially on the concentration of the active ingredient. The amount added is typically sufficient to solubilize the active ingredient. For example, the concentration of the solubilizer is from 0.1 to 5000 times the concentration of the active ingredient.

Examples of buffer substances are acetate, ascorbate, borate, hydrogen carbonate/carbonate, citrate, gluconate, lactate, phosphate, propionate and TRIS (tromethamine) buffers. The amount of buffer substance added is, for example, that necessary to ensure and maintain a physiologically tolerable pH range. The pH range is typically in the range of from 5 to 9, preferably from 5.2 to 8.5.

Tonicity enhancing agents are selected from ionic and non-ionic agents. For example, iom^'c compounds, include alkali metal or alkaline earth metal halides, such as, for example, CaCl₂ KBr, KCl, LiCl, NaI, NaBr or NaCl₅ or boric acid. Non-ionic tonicity enhancing agents are, for example, urea, glycerol, sorbitol, mannitol, propylene glycol, or dextrose. For example, sufficient tonicity enhancing agent is added to impart to the ready- for-use ophthalmic composition an osmolality of approximately from 50 to 1000 mOsmol. Examples of preservatives are quaternary ammonium salts such as benzalkonium chloride, benzoxonium chloride or polymeric quaternary ammonium salts, alkyl-mercury salts of thiosalicylic acid, such as, for example, thiomersal, phenylniercuric nitrate, phenylmercuric acetate or phenylmercuric borate, parabens, such as, for example, methylparaben or propylparaben, alcohols, such as, for example, chlorobutanol, benzyl alcohol or phenyl ethanol, guanidine derivatives, such as, for example, chlorohexidine or polyhexamethylene biguanide, or sorbic acid. Where appropriate, a sufficient amount of preservative is added to the ophthalmic composition to ensure protection against secondary contaminations during use caused by microbes.

The compositions of the present invention may comprise further non-toxic excipients, such as, for example, emulsifiers, wetting agents or fillers, such as, for example, the polyethylene glycols (PEG200, 300, 400 and 600) or Carbowax^® (CarbowaxlOOO,

1500, 4000, 6000 and 10000). Other excipients that may be used if desired are listed below but they are not intended to limit in any way the scope of the possible excipients. They can be complexing agents, such as disodium-EDTA or EDTA, antioxidants, such as ascorbic acid, acetylcysteine, cysteine, sodium hydrogen sulfite, butyl-hydroxyanisole, butyl- hydroxytoluene; stabilizers, such thiourea, thiosorbitol, sodium dioctyl sulfosuccinate or monothioglycerol; or other excipients, such as, for example, lauric acid sorbitol ester,

Methanol amine oleate or palmitic acid ester.

Optionally, the suspension may also contain suitable stabilizers or agents, which increase the solubility of the compounds, to allow for the preparation of concentrated solutions. For example, US Patent No. 5,576,311, the contents of which are incorporated by reference herein, teaches stable aqueous suspension of drugs suitable for therapeutic administration to the eye comprising cyclodextrin type-suspending agents. Other useful formulations include submicron ocular emulsions, for example an ocular drug delivery vehicle as disclosed in US Patent No. 5,496,811, the contents of which are incorporated by reference as if fully set forth herein.

The amount and type of excipient added is in accordance with the particular requirements and is generally in the range of from approximately 0.0001 to approximately 90% by weight.

The amount of a composition to be administered will, of course, depend on many factors including the subject being treated, the severity of the affliction, the manner of administration, and the judgment of the prescribing physician. However, the dose employed will generally depend on a number of factors, including the age and sex of the patient, the precise disorder being treated, and its severity.

Preferably, the preparations are in unit dosage form, intended for oral administration. In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active components. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, for example, tablets, capsules, and powders in vials or ampoules. The unit dosage form can also be a capsule, cachet, or tablet itself or it can be the appropriate number of any of these in packaged form.

The dosing schedule of the compositions of the present invention can vary according to the particular application and the potency of the active ingredients. Determination of the proper dosage is within the skill of the art. For convenience, a single daily dose is preferred. Alternatively, the total daily dosage may be divided and administered in portions during the day such as twice daily, thrice daily and the like. Biweekly, weekly, bimonthly and monthly administration are also contemplated.

Therapeutic Use In another aspect, the present invention provides a method of preventing, attenuating or treating an eye disease comprising the step of administering an effective amount of a composition comprising lutein, lycopene, phytoene, phytofluene and carnosic acid; and a pharmaceutically acceptable carrier or excipient. Non-limiting examples of eye diseases include AMD (both wet and dry), glaucoma, diabetic retinopathy and cataract. Thus, in one embodiment, the present invention provides a method of preventing, attenuating or treating AMD comprising the step of administering an effective amount of a composition comprising lutein, lycopene, phytoene, phytofluene and carnosic acid; and a pharmaceutically acceptable carrier or excipient, in an amount effective to prevent, treat or attenuate AMD. As used herein, the term "AMD" includes both dry and wet AMD, as well as the transition from dry AMD to wet AMD.

In another embodiment, the present invention provides a method of preventing, attenuating or treating glaucoma comprising the step of administering an effective amount of a composition comprising lutein, lycopene, phytoene, phytofluene and carnosic acid; and a pharmaceutically acceptable carrier or excipient, in an amount effective to prevent, treat or attenuate glaucoma.

In another embodiment, the present invention provides a method of preventing, attenuating or treating diabetic retinopathy comprising the step of administering an effective amount of a composition comprising lutein, lycopene, phytoene, phytofluene and carnosic acid; and a pharmaceutically acceptable carrier or excipient, in an amount effective to prevent, treat or attenuate diabetic retinopathy.

In another embodiment, the present invention provides a method of preventing, attenuating or treating cataract, comprising the step of administering an effective amount of a composition comprising lutein, lycopene, phytoene, phytofluene and carnosic acid; and a pharmaceutically acceptable carrier or excipient, in an amount effective to prevent, treat or attenuate cataract.

As used herein, the term "attenuating" means to soothe, calm or ease the symptoms of any of the aforementioned eye diseases. As used herein, the term "treating" includes preventative as well as disorder remittive treatment. As used herein, the term "administering" refers to bringing a subject in contact with the formulation of the present invention. In one embodiment, the present invention encompasses administering the formulations of the present invention to a human subject.

The following examples are presented in order to more fully illustrate some embodiments of the invention. They should, in no way be construed, however, as limiting the broad scope of the invention. One skilled in the art can readily devise many variations and modifications of the principles disclosed herein without departing from the scope of the invention.

EXAMPLES

Example 1 : Oral Compositions

Preparations were prepared, as follows:

Preparation 1: about 6 mg lycopene (about 1.5% w/w); about 8 mg lutein and zeaxanthin (combined - about 2% w/w); about 0.6 mg of phytoene and phytofluene (combined - about 0.15%); and about 4 mg carnosic acid (about 1% w/w).

Preparation 2:

Preparation 2 contains all of the ingredients of preparation 1 and in addition: about 3 mg β-carotene (about 0.75%); about 30 mg zinc (about 7.5% w/w); about 1 mg copper (about 0.25% w/w); about 50mg vitamin E (about 12.5% w/w); and about 50 mg vitamin C (about 12.5% w/w).

Preparation 3:

Preparation 3 comprises the following ingredients

Lyc-O-Mato® ² Carnosic Acid

The above compositions are mixed with mineral oil and or vegetable oil and are prepared as soft gelatin capsules containing about 400 mg total weight.

Alternatively the above compositions are admixed with fillers and excipients and are prepared as solid oral dosage forms containing about 400 mg total weight.

Example 2 - Ophthalmic Compositions

Each of preparations 1 to 3 above were mixed with an ophthalmically acceptable carrier and formulated into ophthalmic formulations.

While certain embodiments of the invention have been illustrated and described, it will be clear that the invention is not limited to the embodiments described herein.

Numerous modifications, changes, variations, substitutions and equivalents will be apparent to those skilled in the art without departing from the spirit and scope of the present invention as described by the claims, which follow. REFERENCES

Alves-Rodrigues, A. and Shao, A. "The science behind lutein." Toxicology Letters, 150:57-83, 2004.

Ben-Dor A, Steiner M, Gheber L, Danilenko M, Dubi N, Linnewiel K, Zick A, Sharoni Y, and Levy J. Carotenoids activate the antioxidant response element transcription system. MoL Cancer Therp 4(1): 177-186. 2005

Mares-Perlman JA, Millen AE, Ficek TL, Hankinson SE. "The body of evidence to support a protective role for lutein and zeaxanthin in delaying chronic disease. Overview." J Nutr. 2002 Mar;132(3):518S-524S. Mohanty, I, Joshi, S, Trivedi, D, Srivastava, S., and Gupta, S.K. " Lycopene prevents sugar-induced morphological changes and modulates antioxidant statues of human lens epithelium." British J Nutrition, 88:347-354, 2002.

Pollack A, Madar Z, Eisner Z, Nyska A, and Oren, P. Inhibitory effect of lycopene on cataract development in galactosemic rats. Metab Pediatr Syst Ophthalmol. 1996- 1997;19-20:31-6.

Seddon JM, Ajani UA, Sperduto RD, Hiller R, Blair N, Burton TC, Farber MD, Gragoudas ES, Haller J, Miller DT, et al. Dietary carotenoids, vitamins A, C, and E, and advanced age-related macular degeneration. Eye Disease Case-Control Study Group. JAMA. 272(18):1413-20, 1994 Talalay P. Chemoprotection against cancer by induction of phase 2 enzymes.

Biofactors; 12: 5-11. 2000

Claims

1. A pharmaceutical composition comprising: lutein, zeaxanthin, lycopene, phytoene, phytofluene, carnosic acid; and a pharmaceutically acceptable carrier or excipient.

2. The composition according to claim 1, wherein the lycopene is provided as a natural tomato extract.

3. The composition according to claim 2, wherein the lycopene is extracted from tomato oleoresin.

4. The composition according to claim 1, wherein the lycopene, phytoene, phytofluene and carnosic acid act synergistically to achieve a therapeutic effect of lycopene at a lower concentration as compared with a corresponding formulation not containing phytoene, phytofluene and carnosic acid.

5. The composition according to claim 1, comprising from about 0.025% w/w to about 5% w/w lutein; from about 0.001% w/w to about 2% w/w zeaxanthin; from about 0.025% w/w to about 5% w/w mg lycopene; from about 0.0025% w/w to about 1.25% w/wmg phytoene; from about 0.0025% w/w to about 1.25% w/w phytofluene; from about 0.025% w/w to about 2.5% w/w carnosic acid or derivative thereof; and a pharmaceutically acceptable carrier or excipient.

6. The composition according to claim 5, comprising: about 2% w/w total of lutein and zeaxanthin; about 1.5% w/w lycopene; about 0.15% w/w total of phytoene and phytofluene; about 1% w/w carnosic acid or a derivative thereof; and a pharmaceutically acceptable carrier or excipient.

7. The composition according to claim 1, further comprising at least one mineral.

8. The composition according to claim 7, wherein the mineral is zinc.

9. The composition according to claim 8, wherein the zinc is provided as zinc oxide.

10. The composition according to claim 7, wherein the mineral is copper.

11. The composition according to claim 10, wherein the copper is provided as cupric oxide.

12. The composition according to claim I₅ further comprising at least one vitamin.

13. The composition according to claim 12, wherein the at least one vitamin is selected from the group consisting of vitamin A, vitamin C, vitamin E, derivatives thereof and combinations thereof.

14. The composition according to claim 1, comprising lutein, zeaxanthin, lycopene, phytoene, phytofluene, carnosic acid, β- carotene, vitamin E, vitamin C, zinc, copper, and a pharmaceutically acceptable carrier or excipient.

15. The composition according to claim 14, comprising: from about 0.025% w/w to about 5% w/w lutein; from about 0.001% w/w to about 2% w/w zeaxanthin; from about 0.025% w/w to about 5% w/w mg lycopene; from about 0.0025% w/w to about 1.25% w/wmg phytoene; from about 0.0025% w/w to about 1.25% w/w phytofluene; from about 0.025% w/w to about 2.5% w/w carnosic acid or derivative thereof; from about 0.25% w/w to about 25% w/w vitamin E; from about 0.25% w/wto about 25% w/w vitamin C; from about 0.025% to about 2.5% w/w β-carotene; from about 0.25% w/wto about 15% w/w mg zinc; from about 0.01% w/wto about 5% w/w copper; and a pharmaceutically acceptable carrier or excipient.

16. The composition according to claim 14, comprising about 2% w/w total of lutein and zeaxanthin; about 1.5% w/wlycopene; about 0.15% w/w total of phytoene and phytofluene; about 1% w/w carnosic acid or a derivative thereof; about 12.5% w/w vitamin E; about 12.5% w/w vitamin C; about 0.75% w/w β-carotene; about 7.5% w/w zinc; about 0.25% w/w copper; and a pharmaceutically acceptable carrier or excipient.

17. The composition according to claim 1, wherein the lycopene induces the production of phase II enzymes.

18. The composition according to claim 1, further comprising tomato oleoresin.

19. The composition according to any one of claims 1-18, wherein the composition is formulated for oral administration.

20. The composition according to claim 19, in a form selected from the group consisting of a tablet, caplet , capsule, microcapsule, pellet, pill, powder, syrup, gel, slurry, granule, suspension, dispersion, emulsion, liquid, solution, dragee, bead and beadlet.

21. The composition according to claim 20, wherein the capsule is selected from a soft gelatin capsule and a hard gelatin capsule.

22. The composition according to any one of claims 1-18, wherein the composition is formulated for topical administration to the eye.

23. The composition according to claims 22, in a form selected from the group consisting of a solution, eye drops, liquid, cream, paste, ointment, emulsion, submicron emulsion, gel, thermogel, semi-solid, solid and suspension.

24. A method of preventing, attenuating or treating an eye disease comprising administering to a subject in need thereof an effective amount of a composition comprising lutein, lycopene, phytoene, phytofluene zeaxanthin and carnosic acid; and a pharmaceutically acceptable carrier or excipient, in an amount effective to prevent, attenuate or treat the eye disease.

25. The method according to claim 24, wherein the formulation is administered orally.

26. The method according to claim 24, wherein the formulation is administered topically to the eye.

27. The method according to claim 24, wherein the eye disease is selected from the group consisting of glaucoma, cataracts, diabetic retinopathy (DR) and age related macular degeneration (AMD).

28. The method according to claim 27, wherein the eye disease is dry

AMD.

29. The method according to claim 27, wherein the eye disease is wet

AMD.

30. A pharmaceutical composition comprising: about 6 mg lycopene (about 1.5% w/w); about 8 mg lutein and zeaxanthin (about 2% w/w); about 0.6 mg of phytoene and phytofluene (about 0.15%); about 4 mg carnosic acid (about 1% w/w); about 50mg vitamin E (about 12.5% w/w); about 50mg vitamin C (about 12.5% w/w); about 3 mg β-carotene (about 0.75%); about 30 mg zinc (about 7.5% w/w); about 1 mg copper (about 0.25% w/w); and and a pharmaceutically acceptable carrier or excipient.

31. The composition according to claim 30, further comprising tomato oleoresin.

32. The composition according to claim 30, in a form selected from a soft gelatin capsule and a hard gelatin capsule.

33. The composition according to claim 30, wherein the zinc is provided as zinc oxide.

34. The composition according to claim 30, wherein the copper is provided as cupric oxide.