Ophthalmic compositions for treating visual disorders

Abstract

To provide an ophthalmic composition for the treatment and / or prevention of visual disorders caused by changes in the structure of the lens of the eye associated with changes in the proteins forming the lens, and to provide uses thereof.SOLUTION: An ophthalmic composition consisting of a polyphenol compound in combination with a therapeutically effective amount of a terpenoid or a salt or crystalline form thereof, wherein the composition requires a low dose of the bioactive compound to treat the disease with a reduced number of applications. In particular, the ophthalmic compositions of the present invention can be used for the treatment and reversal of visual impairment caused by changes in lens structure, such as cataracts, by being able to reverse lens opacities caused by disease.SELECTED DRAWING: Figure 5

Description

[Technical Field]

[0001] The present invention relates to healthcare, and more particularly to ophthalmic compositions for the treatment of visual disorders, especially those that affect the normal function of the lens of the eye. [Background technology]

[0002] Visual disorders affecting the normal structure of the eye's lens are conditions that alter or change the proteins that make up the lens, resulting in impaired vision. Changes in the transparency and / or rigidity of the lens, caused by the aggregation of lens proteins (crystallins), often occur. This type of recurrent condition is known as "cataract." The term "cataract" referred to in this invention refers to a disease that causes opacity in the eye's lens due to changes in the structure and / or aggregation of existing proteins, reducing the amount of light that enters, and resulting in impaired vision. Cataract formation can be congenital, juvenile, age-related, or secondary to diseases (especially diabetes, myotonic dystrophy, galactosemia, neurofibromatosis type 2, rubella), trauma, etc. Cataracts can be nuclear, cortical, or posterior subcapsular. However, there are other diseases associated with changes in the lens structure, such as presbyopia, lens sclerosis, retinal degeneration, Refsum's disease, Smith-Lemli-Opitz syndrome (SLOS), drusen, Schneider's corneal dystrophy (SCD), abetalipoproteinemia (ABL), familial hypobetalipoproteinemia (FHBL), aging, macular degeneration, and diabetic retinopathy.

[0003] Treatment of these conditions depends on the underlying condition, so any pathology that may predetermine the formation of these conditions should first be addressed. Some symptoms of these conditions (e.g., early cataracts) can be reduced by the use of prescription lenses, anti-reflective lenses for the sun, or medications (e.g., drops, ointments, and pills).

[0004] Another treatment option is surgery. It should be noted that for surgery to be performed, the cataract must be in the mature or opacified stage, and before that, corrective measures using lenses are intended only to reduce the refractive error resulting from the development of opacity. It is worth mentioning that cataract surgery, which consists of removing the crystalline lens and replacing it with an artificial intraocular lens, presents drawbacks such as high costs and side effects such as posterior capsule rupture and corneal edema.

[0005] Due to the problems caused by surgery, state of the art has explored the possibility of using bioactive compounds in the treatment of visual impairment. Perhaps the most studied bioactive compound to date is terpenoids, especially lanosterol, although its use in cataract reversal has not been conclusively demonstrated. The most successful experiment on this compound was conducted by Zhao et al. (Non-Patent Document 1). They demonstrated that the use of 14 intravitreal injections loaded with lanosterol nanoparticles (final dose 1.4 mg) could reduce the cataract stage of these animals by up to 2 degrees, while providing 50 μL of lanosterol droplets to a group of seven dogs three times a day for six weeks. Unfortunately, the number of injections required to achieve results is too large, so it does not significantly reduce the risks resulting from surgery or eliminate problems if the condition progresses.

[0006] Following the efforts of Zhao et al., other authors have not come close to obtaining similar results. Chen et al. (Non-Patent Document 2) confirmed in their study that lanosterol can have positive in vitro activity. They added 10 μM and 200 μM lanosterol and 10 μM and 200 μM 25-hydroxycholesterol to 2 mg / mL protein aggregates obtained from surgically removed cataracts for 14 days. Macroscopic findings suggested that lanosterol and 25-hydroxycholesterol dramatically removed the initial dark solution in a dose-dependent manner after 6 days of treatment at room temperature. The authors found that the efficacy of these compounds was highly dependent on the degree or severity of cataract. However, this study suggested that the concentration required to reach 50% of the estimated mean maximum developmental effect (EC50) was 1.5- to 4-fold higher in the cortical cataract group compared to the nuclear milk spot group. Furthermore, the amount of released protein decreased with increasing severity. This study, as reported by Zhao et al., demonstrates limitations and confirms the need for high-dose and frequent treatment. In severe nuclear cataracts, compounds may not penetrate the interior of the lens. Therefore, in practice, the effective concentration of a compound in human lenses with mature or dense cataracts may be several hundred times higher than the EC50 value determined ex vivo.

[0007] Daszynski et al. (Non-Patent Document 3) conducted an in vitro cataract reversal study in rat lenses, along with a protein solubilization study in human lenses alone. 15 mM lanosterol in liposomes was unable to reverse opacification or prevent its progression. In contrast, opacification in all lanosterol-treated lenses progressed to a more advanced, mature cataract stage with obvious nuclear involvement. Nagai et al. (Non-Patent Document 4) developed an intravitreal injection formulation containing lanosterol nanoparticles (LAN-NPs) based on the findings of Zhao et al. by using a ball milling method. They evaluated the therapeutic effects of LAN-NPs on lens structure collapse and opacification by using two rat cataract models: SCR-N, rats with slight lens structure collapse, and SCR-C, rats with a combination of significant lens structure collapse and opacification. Unfortunately, LAN-NPs did not restore lens structure; they only delayed the onset of opacification. Lanosterol supplementation using LAN-NPs failed to improve the severe structural collapse of the lens.

[0008] Similar results were found by Shanmugam et al. (Non-Patent Document 5). They studied the effect of lanosterol on the nuclei of human lens with age-related cataracts. They used 40 age-related cataracts removed by manual small-incision cataract surgery, randomly immersing 20 nuclei in a 25 mM lanosterol solution, while the remaining nuclei were stored in the dark for 6 days in a control solution without lanosterol. In conclusion, 25 mM lanosterol solution did not reverse the opacification of age-related human cataracts.

[0009] Patent Document 1 describes a method for treating or preventing visual impairment using lanosterol. This paper proposes a dosage of 0.1 mg to 5 g, e.g., 1 mg to 2.5 g, of the compound to be administered systemically to a human weighing approximately 70 kg. However, to evaluate the effect of lanosterol on reducing cataract formation in lens tissue, naturally cataractous rabbit lenses were isolated and incubated for 6 days in a solution containing 25 mM lanosterol. Subsequently, the transparency of the lenses before and after lanosterol treatment was compared, and a strong trend toward reduced cataract severity was observed, as evidenced by increased lens transparency. The information presented in this patent is consistent with that reported in other references regarding limitations found with the use of lanosterol in the treatment of cataracts, particularly using the same concentration of lanosterol, as reported by Shanmugam et al.

[0010] Patent Document 2 describes the preparation and method of pharmaceutical compositions containing lanosterol compounds for ophthalmic use, particularly lanosterol compounds at concentrations of 5 to 250 mM, and the application of the pharmaceutical compositions for the prevention and treatment of eye diseases. The described composition, in the form of eye drops containing 25 mM lanosterol (11.4 mg / mL), was administered to Canis familiaris L. three times daily: 50 μL of 25 mM lanosterol was applied in the morning, afternoon, and evening for 12 consecutive weeks, with an interval of at least 5 hours between each application. Although no clear data are presented, it has been suggested that this composition may completely cure traumatic cataracts in two weeks; however, the aforementioned subsequent experimental evidence does not support its use, particularly in high-grade, advanced cataracts.

[0011] Chemical modifications to lanosterol have also been proposed to enhance its effects. Patent Document 3 (Patent Document 3) describes compositions of lanosterol and 25-hydroxycholesterol derivatives (including pharmaceutically acceptable salts thereof) used in combination with antioxidants, free radical scavengers, regulators of protein carbonylation, lipid peroxidation, redox enzyme enhancers, or antioxidants. The paper suggests inhibitory concentrations of lanosterol derivatives ranging from 0.010% w / v to approximately 5% w / v. Patients diagnosed with cataracts and exhibiting lens opacity were photographed before treatment. The paper reports that "lens opacity dramatically improved after three weeks and almost completely disappeared after six weeks," but no measurement of cataract grade reduction was reported. Therefore, it is unclear whether the modifications performed could be useful in cases of advanced cataract development.

[0012] Patent Document 4 describes aqueous ophthalmic compositions containing steroids, such as lanosterol, in combination with other agents for treating eye diseases, injuries, or damage. However, it emphasizes the importance of excipients in the ultimate therapeutic effect, using 2-hydroxypropyl-β-cyclodextrin (CD) and hydroxypropylmethylcellulose (HPMC). They specifically conducted a study on the treatment of cataractous lenses in three different species of dogs using a procedure involving the injection of nanoparticles loaded with a steroid formulation (100 μg). These included lanosterol and another cholesterol-derived steroid, in a volume equivalent to 50 μL (1 drop), three times daily for three weeks at approximately 7:00 AM, 1:00 PM, and 4:00 PM. Experimental data show a maximum improvement of 20% in the dogs used in this study.

[0013] Other bioactive compounds that have been tried for the treatment of cataracts are polyphenolic compounds of natural origin, particularly in the case of curcumin. As in the case of Manikandan et al., it has been reported that curcumin can inhibit sodium selenite-induced oxidative stress (Non-Patent Document 6). They tested the antioxidant potential of curcumin against sodium selenate-induced (15 μM / kg body weight) cataracts in rat pups. This group of researchers concluded that treatment with curcumin (75 mg / kg body weight as a single dose) resulted in a significant reduction in lipid peroxidation, enzymatic antioxidant, and non-enzymatic antioxidant levels, which may support the idea that consumption of free curcumin in the diet may help prevent the development of senile cataracts. In another study, Manikandan et al. (Non-Patent Document 7) investigated the expression of αA- and αB-crystallins and heat shock protein 70 (Hsp 70) during curcumin treatment of sodium selenite-induced cataract development in Wistar rat pups. They similarly concluded that curcumin suppressed αA- and αB-crystallin expression induced by selenite and Hsp 70. Therefore, curcumin may inhibit and / or prevent cataract formation in rat pups.

[0014] Regarding the effectiveness of curcumin in prevention and treatment, Yogaraj et al. (Non-Patent Document 8) developed and characterized a nanocurcumin formulation using functionalized polyamidoamine dendrimers (PAMAM) (third generation) with encapsulated curcumin. They tested their formulation in the same model as the Manikandan group in Wistar rat pups by expressing pro-inflammatory genes in vitro using a human lens epithelial cell (HLE-B3) system. Their results showed that nanocurcumin proved as effective as free curcumin in reducing sodium selenite-induced cell death, RNA degradation, and gene expression of iNOS (inducible nitric oxide synthase) levels. They therefore concluded that the encapsulated curcumin formulation using QPAMAM compensates for the main limitations of its free counterpart with its higher solubility, sustained release, and higher bioavailability by preventing the development of oxidant-mediated cataracts.

[0015] Patent Document 5 also describes ophthalmic nanosuspension formulations and preparation methods. In particular, nanosuspension ophthalmic preparations are prepared by combining ophthalmic pharmaceutical adjuvants with curcumin. Ophthalmologically acceptable gel and nanosuspension preparations of curcumin were tested in the same rat model with selenite-induced cataracts. Administration on a 3x / 8-day regimen demonstrated beneficial effects of curcumin ophthalmic nanosuspension. There is evidence of better bioavailability with a larger surface area, improving the extent of drug absorption and thus prolonging the duration of action. However, reversion has not been demonstrated in vivo; rather, enzymatic activity has been characterized ex vivo.

[0016] Patent Document 6 relates to an aqueous ophthalmic composition containing isolated or combined natural curcuminoids, such as curcumin, bisdemethoxycurcumin, demethoxycurcumin, and synthetically derived bis-o-demethylcurcumin and / or other demethylated curcuminoids, together with other ophthalmic excipients, along with suitable surfactants and cosolvents as penetration enhancers, which are useful for treating ophthalmic diseases or disorders. Similar to other curcumin-related papers, the study concluded that instillation of a 98% curcumin ophthalmic formulation into the eyes of Wistar rat pups effectively reduced the effects of selenite-induced cataracts and provided a preventative effect.

[0017] However, experimental evidence regarding naturally occurring polyphenolic compounds such as curcumin has not shown that they can reverse cataract formation at high levels, only prevent their formation.Similarly, the development of compositions based on terpenoid compounds such as lanosterol has achieved minimal reduction in cataracts by using repeated, very aggressive treatments at high doses.Efforts to develop compositions based on these types of bioactive compounds have not achieved significant reduction or elimination of cataracts.

[0018] As a result of the above, we are attempting to solve the problems of currently used ophthalmic compositions for the treatment of visual disorders by developing a composition that requires lower doses and a reduced number of applications to be effective, in addition to allowing the reversal of cataracts at an advanced developmental stage. [Prior art documents] [Patent documents]

[0019] [Patent Document 1] U.S. Patent No. 10,471,076 [Patent Document 2] Chinese Patent No. 106344587 [Patent Document 3] International Publication No. 2019 / 097434 [Patent Document 4] US Patent Application Publication No. 2020 / 016176 [Patent Document 5] Chinese Patent Application Publication No. 106511269 [Patent Document 6] European Patent No. 2346520 [Non-patent literature]

[0020] [Non-Patent Document 1] Zhao et al.Lanosterol Reverses Protein Aggregation in Cataracts.Nature.2015;523:607-611 [Non-patent document 2] Chen et al.Lanosterol and 25-hydroxycholesterol Dissociate Isolated Crystalline Aggregates from the Human Cataract Lens Through Different Mechanisms.Biochemical and Biophysical Research Communications.2018;506(4):868-873 [Non-patent document 3] Daszynski et al. Failure of Oxysterols such as Lanosterol to Restore the Clarity of the Cataract Lens.Scientific Reports.2019; 9:8459 [Non-patent document 4] Nagai et al.Intravitreal Injection of Lanosterol Nanoparticles Rescues Collapse of Lens Structure at an Early Stage in Shumiya Cataract Rats.International Journal of Molecular Sciences.2020;21(3):1048 [Non-patent document 5] Shanmugam et al.Effect of Lanosterol on Human Cataract Nucleus.Indian Journal of Ophthalmology.2015;63(12):888-90 [Non-patent document 6] Manikandan et al.Effect of Curcumin on Selenite-Induced Cataractgenesis in Wistar Rat Pups.Current Eye Research.2010;35:122-129 [Non-Patent Document 7] Manikandan et al.Effect of Curcumin on the Modulation of αA-and αB-crystallin and Heat Shock Protein 70 in Selenium-Induced Cataractgenesis in Wistar Rat Pups.Molecular Vision.2011;17:388-394 [Non-patent document 8] Yogaraj et al.Quaternary Ammonium Dendrimeric Poly(Amidoamine)Encapsulated Nanocurcumin Effectively Prevents Cataracts in Rat Pups by Regulating Pro-Inflammatory Gene Expression.Journal of Drug Delivery Science and Technology.2020;58:1773-2247 Summary of the Invention

[0021] Object of the invention In view of the deficiencies of the prior art, it is an object of the present invention to provide an ophthalmic composition for treating visual disorders which requires low doses of biologically active compounds to reverse and / or prevent conditions that modify or alter the normal structure of the eye's lens, primarily the proteins that form it.

[0022] Another object of the present invention is to provide an ophthalmic composition for treating visual disorders that is effective in treating such disorders and requires a reduced number of applications. These and other objectives are achieved by the ophthalmic compositions for treating visual disorders according to the present invention.

[0023] Summary of the Invention To achieve the objectives of the present invention, an ophthalmic composition for treating visual disorders has been invented, characterized in that it contains a therapeutically effective amount of a polyphenolic compound in combination with a terpenoid, making it possible to reverse and / or prevent the changes in conditions that modify or alter the normal structure of the lens, mainly the proteins that form it.

[0024] Another aspect of the present invention contemplates methods of making the ophthalmic compositions of the present invention and their use in treating ocular diseases caused by protein alteration or aggregation. [Brief explanation of the drawings]

[0025] The novel features believed characteristic of the present invention are set forth with particularity in the appended claims, however, certain embodiments, features, and certain objects and advantages thereof will be better understood from the detailed description when read in connection with the accompanying drawings. [Figure 1] FIG. 1 is a set of photographs of different stages of lens opacification in experimental animals to illustrate different degrees of cataract development. [Figure 2] FIG. 1 is a series of photographs of lenses from healthy and disease control animals that developed grade 4 cataracts. [Figure 3] FIG. 1 is a series of photographs of animal lenses before and after treatment with intravitreally applied curcumin nanoemulsion. [Figure 4] FIG. 1 is a series of photographs of animal lenses before and after treatment with intravitreally applied lanosterol nanoemulsion. [Figure 5] FIG. 1 is a series of photographs of animal lenses before and after treatment with intravitreally applied curcumin nanoemulsion containing lanosterol at a 1:1 ratio. [Figure 6] FIG. 1 is a series of photographs of animal lenses before and after treatment with intravitreally applied curcumin nanoemulsion containing lanosterol at a ratio of 1:3. [Figure 7] FIG. 1 is a series of photographs of animal lenses before and after treatment with intravitreally applied free curcumin. [Figure 8] FIG. 1 is a series of photographs of animal lenses before and after treatment with intravitreally applied free lanosterol. DETAILED DESCRIPTION OF THE INVENTION

[0026] It has been found that in several known visual disorders, it is possible to reverse and / or prevent, by a few applications, the changes in the normal structure of the lens, primarily conditions that modify or alter the proteins that form the lens, and to effectively treat these visual disorders with ophthalmic compositions comprising a therapeutically effective amount of a polyphenolic compound in combination with a terpenoid or their respective pharmaceutically acceptable salts or crystalline forms.

[0027] The new combination of polyphenolic compounds and terpenoids exhibits a synergistic effect that makes it possible to more effectively reverse and / or prevent changes in lens structure, with fewer applications and for shorter periods of time, and at lower doses than those reported in the state of the art for both compounds individually.

[0028] According to the principles of the present invention, the ophthalmic composition may contain any ratio between the polyphenolic compound and the terpenoid. In a preferred embodiment, the ophthalmic composition of the present invention contains the polyphenolic compound to the terpenoid in a ratio of 1:10 to 10:1, preferably the polyphenolic compound to the terpenoid in a ratio of 1:3 to 3:1, and more preferably both compounds are present in the ophthalmic composition in a ratio of 1:1.

[0029] In a preferred embodiment of the present invention, the polyphenolic compound is a natural curcuminoid, preferably selected from curcumin, bis-o-demethylcurcumin, demethoxycurcumin and synthetically derived bis-o-demethylcurcumin, and / or other demethylated curcuminoids, stable isomers of the above, or pharmaceutically acceptable salts or crystalline forms thereof, with naturally occurring curcumin being particularly preferred due to its antioxidant and anti-inflammatory activity. These compounds are well described in the state of the art so that a person skilled in the art can obtain them by using the techniques described above in the background of the present invention.

[0030] Terpenoids include lanosterol and its steroid derivatives, dihydrolanosterol; 4,4-dimethylcholesta-8(9),14,24-trien-3β-ol; 4,4-dimethylcholesta-8,24-dien-3β-ol; 4,4-dimethylcholesta-8-en-3β-ol; 4,4-dimethylcholesta-8(9),14-dien-3β-ol; 14-desmethyllanosterol; lathosterol; Δ7,24-cholestadienol; cholesterol; cholesta-7-enol; cholesterol esters; 7-dehydrocholesterol; desmosterol; 7-dehydrodesmosterol; zymosterol; 27-hybrid The compound is selected from the following: hydroxycholesterol; cholesta-7,24-dien-3-β-ol; cholesta-8(9)-en-3-β-ol; 5α-cholestan-3β-6-one; 5-cholesten-3β,25-diol; 5-cholesten-3β,25-OSO3H (5-cholesten-3β,25-sulfate); 5-cholesten-3β-OSO3H,25-ol (5-cholesten-3β-sulfate,25-ol); 5-cholesten-3β,25-diol; disulfate and / or its ester; the stable isomers described above; or their pharmaceutically acceptable salts or crystalline forms, and lanosterol and / or its derivatives are particularly preferred.Similarly, these compounds are described in the state of the art, so that it is clear to those skilled in the art that they can be obtained by referring to the known techniques in the background of the present invention.

[0031] In a particularly preferred embodiment, the composition comprises curcumin as the polyphenolic compound, which is preferably obtained from the rhizomes of the perennial herb Curcuma longa using techniques described in the state of the art.

[0032] In another particularly preferred embodiment, the composition comprises lanosterol as an amphiphilic tetracyclic terpenoid, synthesized from waxes such as squalene or lanolin, also using techniques described in the state of the art.

[0033] Regarding other ophthalmologically acceptable ingredients for preparing the ophthalmic compositions of the present invention, these are preferably selected appropriately for application of the composition via topical, subconjunctival, retrobulbar, periocular, subretinal, suprachoroidal, intracameral, intravitreal, or intraocular routes, including, but not limited to, pharmaceutical forms selected from ophthalmic solutions, ophthalmic ointments, eye washes, intraocular infusions, anterior chamber washes, oral medications, injections, as part of ocular implants, or as preservatives for extracted corneas. Further, ophthalmologically acceptable ingredients and excipients can be used to formulate the compositions of the present invention, including, but not limited to, water, buffer or sodium chloride solutions, surfactants or cyclodextrins, emulsions, liposomes, ophthalmologically acceptable gels or suspensions, or combinations thereof. A particularly preferred excipient consists of an ophthalmic nanoemulsion.

[0034] The compositions of the present invention are effective in reversing and / or preventing changes in lens structure, primarily changes in lens protein, preferably by at least one application of the composition. As such, they are useful for treating visual impairments, regardless of their cause, including cataracts, or other diseases associated with changes in lens structure, such as presbyopia, phacosclerosis, retinal degeneration, Refsum's disease, Smith-Lemli-Opitz syndrome (SLOS), drusen, Schneider's corneal dystrophy (SCD), abetalipoproteinemia (ABL), familial hypobetalipoproteinemia (FHBL), aging, macular degeneration, and diabetic retinopathy. When the disease is cataracts, the compositions of the present invention can substantially completely reverse the disease.

[0035] Techniques for preparing the compositions of the present invention are known in the state of the art. However, any known method described in the state of the art can be used to incorporate polyphenolic compounds and terpenoids into ophthalmic nanoemulsions, such as the method reported by Agame-Lagunes et al. (Curcumin Nanoemulsions Stabilized with Modified Phosphatidylcholine on Skin Carcinogenesis Protocol. Current Drug Metabolism. 2020, 21, (3): 226-234). This method generally consists of mixing an oil phase containing the compounds with an aqueous phase to obtain a nanoemulsion by methods known in the art, either high-energy or low-energy, although high-energy methods are preferred. These methods allow obtaining nanometer-scale particle sizes in the range of 5 to 999 nm, preferably 20 to 300 nm, from one phase, preferably the oil phase.

[0036] In a particular embodiment, for nanoemulsion formation, a mixture having a ratio of aqueous phase with respect to oil phase of 80:20 to 95:5 is provided, with mixtures having a content of aqueous phase of at least 95% being particularly preferred.

[0037] To prepare a nanoemulsion of the compound of the present invention, the oil phase preferably contains an ophthalmologically acceptable oil, a surfactant, and an organic solvent. Oils are preferred, triacylglyceride oils are more preferred, glycerol derivatives and those having three fatty acids are more preferred, and a mixture of medium-chain triacylglycerides is preferred. The surfactant is also preferably amphiphilic, either synthetic or natural, and is preferably natural, such as phosphatidylcholine, mono- and diacylglycerides, more preferably phosphatidylcholine. The ophthalmologically acceptable organic solvent is preferably an alcohol, more preferably one having 1 to 3 carbon atoms, with ethanol being particularly preferred.

[0038] With respect to the aqueous phase, a preferred embodiment comprises deionized water and glycerol as a stabilizer to form an ophthalmically acceptable vehicle. In a preferred embodiment of the invention, the aqueous phase comprises 5% to 60% glycerol, with glycerol in the range of 10% to 45% being preferred, and glycerol in an amount of 15% to 25% being even more preferred.

[0039] The ophthalmic composition obtained according to the principles of the present invention may be prepared in any pharmaceutical form useful for application to the eye, for its use in the treatment and / or prevention of visual impairment caused by structural changes in the eye lens, in particular those associated with protein alterations, and is preferably adaptable for use in at least one application to achieve near complete reversal of the visual impairment, achieving reversal within a maximum of seven days after treatment.

[0040] The present invention will be better understood by the following examples, which are presented for illustrative purposes only to enable a complete understanding of preferred embodiments of the present invention and to guide their realization, and this does not mean that there are no other embodiments not shown that can be implemented based on the foregoing description. [Example]

[0041] A. Preparation of Ophthalmic Compositions for the State of the Art and the Present Invention To illustrate the novel effects of the compositions of the present invention, six examples of compositions shown in Table 1 were prepared as sterile nanoemulsions for their intravitreal application, using curcumin as the polyphenol and lanosterol as the terpenoid, as are the most studied in the state of the art, with 10 mL of base:

[0042] [Table 1]

[0043] To prepare the nanoemulsions to be tested, an oil phase (dispersed) was obtained using a mixture of 10% phosphatidylcholine, 3 mL ethanol, and 5% medium-chain triacylglycerides along with curcumin and / or lanosterol according to the table. Once the mixture of compounds of the present invention was made, the oil phase was subjected to a water bath to remove excess organic solvent before homogenization.

[0044] The aqueous phase (dispersant) consisted of 60% water and 25% glycerol. Both phases were manually combined to create a coarse emulsion for further processing in an ULTRA-TURRAX Digital T25 rotor-stator homogenizer (IKA Works, Inc., Staufen, Germany), which was then sonicated using a Branson Digital Sonifier S-450D (Branson Ultrasonic Corp., Danbury, Connecticut) to reduce particle size and obtain the relevant nanoemulsion.

[0045] The composition was sterilized using a UV light lamp. For sterilization, the system was poured into a sterile culture plate with a volume of 10 mL and then left uncovered under a UV light lamp for 40 minutes. The system was then collected in a sterile syringe in a glass bottle sealed with parafilm until use in the animal model. All procedures were carried out under sterile conditions.

[0046] In addition, compositions E5 and E6 containing free curcumin (C) and free lanosterol (L) were prepared by dissolving only the compounds in the dispersed phase used. At this time, the mixture of medium-chain triacylglycerides was prepared in a volume of 10 mL at the blending ratio shown in the table.

[0047] B. Cataract induction in Wistar rats To evaluate in vivo cataract regression, 7-week-old albino Wistar rats weighing 120-140 g were randomly assigned to groups (n = 3) and kept in a confined area in a controlled environment of 23 ± 2°C temperature and 40-70% relative humidity, with a 12-hour light / 12-hour dark cycle and free access to commercial diet and water. Animals were housed according to the specifications of Mexican Official Standard NOM-062-ZOO-1999.

[0048] Cataracts were induced in 30 rats by intravitreal administration of sodium selenite. After treatment, the experimental animals developed the disease, but only 26.66% of these animals developed cataracts in both eyes. The remainder developed the disease in only one eye, with the left eye being the most affected.

[0049] For intravitreal treatment, rats were anesthetized by intraperitoneal injection of Zoletil 50 (40 mg / kg body weight) in addition to a local anesthetic (tetracaine), an ocular lubricant (hypromellose), and an antibiotic (ciprofloxacin) to avoid infection.

[0050] To perform the intravitreal injection, a stereoscope was used to allow for greater precision during the procedure. To release intraocular pressure and remove vitreous fluid, a 1 mL insulin syringe with a 27S-gauge needle was used to create a puncture hole through the sclera into the vitreous at a 45° angle. A Hamilton syringe with a 26S-gauge removable needle was inserted into the same hole containing the selenite solution and injected into the chamber, avoiding contact with the lens. The needle was then carefully removed, and one drop of antibiotic was placed in each eye.

[0051] Opacity assessment after intravitreal sodium selenite administration was performed weekly for 4 weeks by slit lamp microscopy, while classification of lens opacification was performed as shown in Table 2 below:

[0052] [Table 2]

[0053] The degree of cataract development is shown in Figure 1, which provides photographic evidence of lens opacification in experimental animals. Figure 1 shows a grade 0 lens (G0), a grade 1 lens (G1), a grade 2 lens (G2), a grade 3 lens (G3), and finally a grade 4 lens (G4).

[0054] C. Application of the Ophthalmic Composition to Rats with Cataracts Eight groups of three albino Wistar rats were formed. A healthy control (negative control) group (E0) was not induced with sodium selenite and did not develop cataracts, while another positive or diseased control group (E7) had cataracts induced as described above but did not receive treatment with the curcumin or lanosterol or their mixtures of the present invention. The remaining groups were treated intravitreally with compositions E1 to E6, respectively, using the same technique as used for sodium selenite application, with two applications (except for the control group) at a 5 μL volume of each ophthalmic composition, one week apart. The results for each rat (R1 to R3) were shown in Table 3.

[0055] [Table 3]

[0056] The extent of cataract development and reduction can be seen in the figures shown in Table 3, whereby the synergistic effect obtained from the compositions of the present invention (E3 and E4) can be observed, which reversed almost all opacification. This also occurred with composition E4, where significantly less curcumin, along with lanosterol, achieved more than two-fold reversal, based on prior art results and results obtained in Examples E5 and E6, for example, which showed better reversal with curcumin than free lanosterol, even considering that one skilled in the art would not be motivated to use more lanosterol than curcumin.

[0057] All of the above is evidenced when analyzing the final opacification after treatment, which was approximately 1 in the group of the composition of the present invention. This is highly statistically significant, as confirmed by Duncan's statistical analysis shown in Table 4, where groups E4 and E3 are in subset 2 and have similar results. In Table 4, the final opacification means of the groups within the homogenous subsets are displayed, and a harmonic mean sample size of 3000 is used.

[0058] In this regard, according to Duncan's statistical analysis, the best treatment corresponds to E3, a group found in subset 1 together with E0, reporting no statistically significant difference from E0 (healthy control), indicating that the E3 composition was able to reverse cataracts in a significant way.

[0059] [Table 4]

[0060] It is also noteworthy that the difference in final opacification means between the minimum and maximum values ​​for statistical group 3 is 1 degree (1.3333-2.3333), while the difference between the minimum and maximum values ​​for group 2 is 0.3333 degrees (1.0000-1.3333). This indicates that treatment with the compositions of the present invention is highly effective in treating visual impairment.

[0061] Thus, it has been demonstrated that, under the principles of the present invention, novel effects superior to those obtained by the individual compounds can be achieved. Furthermore, it will be clear to those skilled in the art that combinations of other polyphenolic compounds with other terpenoids that have already demonstrated in the state of the art a much less significant cataract-reversing effect on their own, comparable to that of Examples E1, E2, E5 or E6, can be made to obtain synergistic effects comparable to those obtained with compositions E3 and E4 described in the preceding examples.

[0062] It will also be clear to those skilled in the art that, according to the state of the art, the compositions of the present invention can be formulated in different pharmaceutical forms for ophthalmic use, since the prior art itself shows the effect of different compounds in several ophthalmic compositions, such as topical, subconjunctival, retrobulbar, periocular, subretinal, suprachoroidal, intracameral, intravitreal or intraocular routes.

[0063] In accordance with the above, it is demonstrated that the ophthalmic compositions for the treatment of visual disorders of the present invention have been prepared to achieve the reversal and / or prevention of such disorders, which are primarily caused by alterations in the structure of the lens, primarily due to alterations in lens proteins. It will also be apparent to those skilled in the art that the embodiments of such compositions as described above and illustrated in the accompanying drawings are merely illustrative of the present invention and not limiting thereof, since considerable variations in their details are possible without departing from the scope of the present invention.

[0064] Accordingly, the present invention should not be viewed as limited except as required by the state of the art and the appended claims.

Claims

1. An ophthalmic composition comprising a therapeutically effective amount of a polyphenolic compound and a terpenoid, or their respective pharmaceutically acceptable salts or crystalline forms.

2. 10. The ophthalmic composition of claim 1, further characterized in that the polyphenolic compound is selected from natural curcuminoids or derivatives thereof.

3. 3. The ophthalmic composition of claim 2, further characterized in that the polyphenolic compound is selected from curcumin, bisdemethoxycurcumin, demethoxycurcumin, and synthetically derived bis-o-demethylcurcumin and / or other demethylated curcuminoids, or stable isomers thereof.

4. 4. The ophthalmic composition of claim 3, further characterized in that the polyphenol compound is natural curcumin.

5. 10. The ophthalmic composition of claim 1, further characterized in that the terpenoid is selected from lanosterol and its steroid derivatives.

6. The terpenoids may be lanosterol; dihydrolanosterol; 4,4-dimethylcholesta-8(9),14,24-trien-3β-ol; 4,4-dimethylcholesta-8,24-dien-3β-ol; 4,4-dimethylcholesta-8-en-3β-ol; 4,4-dimethylcholesta-8(9),14-dien-3β-ol; 14-desmethyllanosterol; lathosterol; Δ7,24-cholestadienol; cholesterol; cholesta-7-enol; cholesterol esters; 7-dehydrocholesterol; desmosterol; 7-dehydrodesmosterol; zymosterol; 6. The ophthalmic composition of claim 5, further characterized in that the ophthalmic composition is selected from 27-hydroxycholesterol; cholesta-7,24-dien-3-β-ol; cholesta-8(9)-en-3-β-ol; 5α-cholestan-3β-6-one; 5-cholesten-3β,25-diol; 5-cholesten-3β,25-OSO3H (5-cholesten-3β,25-sulfate); 5-cholesten-3β-OSO3H,25-ol (5-cholesten-3β-sulfate,25-ol); 5-cholesten-3β,25-diol; disulfates and / or esters thereof; or stable isomers thereof.

7. 7. The ophthalmic composition of claim 6, further characterized in that the terpenoid is lanosterol.

8. 5. The ophthalmic composition of claim 4, further characterized in that the curcumin is derived from the rhizome of the perennial herb Curcuma longa.

9. 8. The ophthalmic composition of claim 7, further characterized in that the terpenoid is lanosterol, an amphiphilic tetracyclic terpenoid synthesized from a wax such as squalene or lanolin.

10. 10. The ophthalmic composition of claim 1, further characterized by comprising an ophthalmologically acceptable excipient for topical, subconjunctival, retrobulbar, periocular, subretinal, suprachoroidal, intracameral, intravitreal, or intraocular application.

11. 11. The ophthalmic composition of claim 10, further characterized in that its pharmaceutical form is selected from an ophthalmic solution, an ophthalmic ointment, an eye wash, an intraocular injection, an anterior chamber wash, an oral medication, an injection, as part of an ocular implant, or as a preservative for extracted corneas.

12. 11. The ophthalmic composition of claim 10, further characterized in that the excipient is selected from water, a buffer or sodium chloride solution, a surfactant or cyclodextrin, an emulsion, a liposome, an ophthalmically acceptable gel or suspension, or a combination thereof.

13. 11. The ophthalmic composition of claim 10, further characterized in that the ophthalmically acceptable vehicle is an ophthalmic nanoemulsion.

14. 14. The ophthalmic composition of claim 13, further characterized by comprising an oil phase having an average particle size of 5 to 999 nm nanoemulsified in an aqueous phase.

15. 15. The ophthalmic composition of claim 14, further characterized by comprising an oil phase having an average particle size of 20 to 300 nm.

16. 15. The ophthalmic composition of claim 14, further characterized in that the oil phase comprises an oil, a surfactant, and an ophthalmically acceptable organic solvent.

17. 17. The ophthalmic composition of claim 16, further characterized in that the oil of the oil phase is a triacylglyceride oil.

18. 18. The ophthalmic composition of claim 17, further characterized in that the oils in the oil phase are a glycerol derivative and three fatty acids.

19. 18. The ophthalmic composition of claim 17, further characterized in that the oil in the oil phase is a mixture of medium chain triacylglycerides.

20. 17. The ophthalmic composition of claim 16, further characterized in that the surfactant is amphiphilic and of synthetic or natural origin.

21. 21. The ophthalmic composition of claim 20, further characterized in that the surfactant is selected from phosphatidylcholines and mono- and diacylglycerides.

22. 17. The ophthalmic composition of claim 16, further characterized in that the ophthalmically acceptable organic solvent is an alcohol.

23. 23. The ophthalmic composition of claim 22, further characterized in that the alcohol has 1 to 3 carbon atoms.

24. 24. The ophthalmic composition of claim 23, further characterized in that the alcohol is ethanol.

25. 15. The ophthalmic composition of claim 14, further characterized in that the aqueous phase comprises deionized water and glycerol as a stabilizer.

26. 26. An ophthalmic composition according to any one of claims 1 to 25 for use in the treatment and / or prevention of eye diseases caused by alterations in the normal lens structure, primarily conditions that modify or alter the proteins that form the lens.

27. 27. The ophthalmic composition of claim 26, further characterized in that the ocular disease is selected from cataract, presbyopia, lens sclerosis, retinal degeneration, Refsum's disease, Smith-Lemli-Opitz syndrome (SLOS), drusen, Schneider's corneal dystrophy (SCD), abetalipoproteinemia (ABL), familial hypobetalipoproteinemia (FHBL), aging, macular degeneration, and diabetic retinopathy.

28. 27. The ophthalmic composition of claim 26, further characterized in that the eye disease is treated or prevented by at least one application of the composition.

29. 27. The ophthalmic composition of claim 26, further characterized in that when the disease is cataract, the composition of the present invention is capable of reversing to a minimum the opacity caused by the disease.

30. 27. The ophthalmic composition of claim 26, further characterized in that the eye disease is treated or prevented by up to two intravitreal applications of the composition.

31. Use of a polyphenolic compound in combination with a terpenoid or a salt or a pharmaceutically acceptable crystalline form thereof for the manufacture of a medicament for the treatment and / or prevention of eye diseases caused by alterations in the normal lens structure, including conditions that modify or alter the proteins that form the lens.

32. 32. The use according to claim 31 , wherein the polyphenolic compound is selected from natural curcuminoids or derivatives thereof.

33. 33. The use according to claim 32, wherein the polyphenolic compound is selected from curcumin, bisdemethoxycurcumin, demethoxycurcumin and synthetically derived bis-o-demethylcurcumin and / or other demethylated curcuminoids, or stable isomers thereof.

34. 33. The use according to claim 32, wherein the polyphenolic compound is natural curcumin.

35. 32. The use according to claim 31, wherein the terpenoid is selected from lanosterol and its steroid derivatives.

36. The terpenoids are selected from the group consisting of lanosterol, dihydrolanosterol, 4,4-dimethylcholesta-8(9),14,24-trien-3β-ol, 4,4-dimethylcholesta-8,24-dien-3β-ol, 4,4-dimethylcholesta-8-en-3β-ol, 4,4-dimethylcholesta-8(9),14-dien-3β-ol, 14-desmethyllanosterol, lathosterol, Δ7,24-cholestadienol, cholesterol, cholesta-7-enol, cholesterol esters, 7-dehydrocholesterol, desmosterol, 7-dehydrodesmosterol, di 36. The use according to claim 35, wherein the compound is selected from mosterol; 27-hydroxycholesterol; cholesta-7,24-dien-3-β-ol; cholesta-8(9)-en-3-β-ol; 5α-cholestan-3β-6-one; 5-cholesten-3β,25-diol; 5-cholesten-3β,25-OSO3H (5-cholesten-3β,25-sulfate); 5-cholesten-3β-OSO3H,25-ol (5-cholesten-3β-sulfate,25-ol); 5-cholesten-3β,25-diol; disulfates and / or esters thereof; or stable isomers thereof.

37. 36. The use of claim 35, wherein the terpenoid is lanosterol.

38. 35. The use of claim 34, wherein the curcumin is derived from the rhizome of the perennial herb Curcuma longa.

39. The use according to claim 37, wherein the terpenoid is lanosterol, an amphiphilic tetracyclic terpenoid synthesized from a wax such as squalene or lanolin.

40. 32. The use of claim 31, wherein the ocular disease is selected from cataract, presbyopia, lens sclerosis, retinal degeneration, Refsum's disease, Smith-Lemli-Opitz syndrome (SLOS), drusen, Schneider's corneal dystrophy (SCD), abetalipoproteinemia (ABL), familial hypobetalipoproteinemia (FHBL), aging, macular degeneration, and diabetic retinopathy.

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