Lipid base formulation for topical ophthalmic use, containing piperite mint leaf extract to improve the penetration and bioavailability of its bioactive in white tissues of the posterior segment of the eyeball in human beings

Abstract

The present invention relates to a lipid-based formulation for topical ophthalmic use that functions as a liposomal delivery system for the bioactive agents contained in the Peppermint leaf extract in white tissues of the posterior segment of the eyeball in humans. Said formulation is helpful to preserve health and reduce the risk of developing chronic-degenerative processes in intraocular tissues, including those of the posterior segment of the eyeball, due to its anti-inflammatory, anti-angiogenic, anti-fibrotic, and antioxidant activity. Also, it has the potential to improve stability, make the penetration of the natural barriers of the eyeball more efficient, and increase the bioavailability of polyphenols in intraocular tissues.

Description

Lipid-based formulation for topical ophthalmic use, containing peppermint leaf extract to enhance penetration and bioavailability. OF ITS BIOACTIVES IN WHITE TISSUES OF THE POSTERIOR SEGMENT OF THE EYEBALL IN HUMANS FIELD OF INVENTION The present invention relates to the fields of biotechnology and medicine. It comprises a lipid-based formulation for topical ophthalmic use that functions as a liposomal delivery system for the bioactive agents contained in peppermint leaf extract to the white tissues of the posterior segment of the eyeball in humans. The invention is useful for maintaining eye health and reducing the risk of developing chronic degenerative processes in intraocular tissues, including those of the posterior segment of the eyeball, due to its anti-inflammatory, anti-angiogenic, anti-fibrotic, and antioxidant activity. BACKGROUND OF THE INVENTION The posterior segment of the eye is the part of the eyeball that includes the retina and its blood vessels, as well as the optic nerve head. It is in this tissue, and particularly in its central area called the macula, that the process of vision generation takes place. This structure is a complex tissue formed by multiple layers of neurons, which can be functionally divided into two parts: an outer and an inner part. The outer part comprises the retinal pigment epithelium. Ο77θηη / 77Π7 / β / YΙΛΙ The retina (RPE) and its basal lamina, known as Bruch's membrane, both of which maintain the integrity of the external blood-retinal barrier. The inner part, on the other hand, comprises the photosensitive layer of rods and cones and their nerve connections, which capture light and convert it into electrical nerve impulses, which are transmitted through the optic nerve. These tissues connect to the vascular structure that forms the internal blood-retinal barrier. These barriers are part of the anatomical and physiological barriers of the eyeball, limiting the penetration and bioavailability of bioactive agents in the retina when administered topically or systemically. The loss of structural integrity and / or function of the tissues of the posterior segment of the eyeball, secondary to proinflammatory and oxidative processes, leads to pathological states that negatively impact visual function. An extraordinary option for maintaining ocular health and preventing and / or controlling abnormal or pathological conditions is the use of bioactives with antioxidant, neuroprotective, antiangiogenic, and anti-inflammatory properties. Some of these elements are present in nature, particularly in extracts of some plants, which have become one of the main sources explored by experts in the search for new molecules, whose potential use may provide new prevention and / or treatment alternatives in conditions that affect the posterior segment of the eyeball and particularly the retina. In particular, the leaf extract of Peppermint has potent anti-inflammatory activity, inhibiting the production of nitric oxide (NO) and prostaglandins (PGE2). Likewise, it has been described as a mediator of the inflammatory response induced by macrophages. Several reports have shown 077θηη / 77Π7 / β / YILI that the use of bioactives contained in mint leaf extract such as rosmarinic acid (RA) and hesperidin (HP) have the potential to reduce the risk of developing chronic-degenerative processes in the retina through its neuroprotective and anti-angiogenic activity. Rosmarinic acid, a water-soluble phenolic compound, inhibits various important steps in the development of neovascularization, including proliferation, migration, and cell adhesion. In the case of the retina, this inhibition is through the cell cycle arrest pathway with an increase in the expression of p21 (WAF1) as well as the significant inhibition of endothelial cell proliferation.AR also decreases the levels of intracellular reactive oxygen species (ROS) dependent on H2O2, the expression of vascular endothelial growth factor (VEGF) and the release of interleukin-8 (IL-8) from endothelial cells, so its anti-angiogenic activity is also linked to its anti-oxidative activity. In addition, AR is associated with the inhibition of the activating protein kinase (AMPK: Adenine Monophosphate Activated Protein Kinase) of the consequent decrease in the production of metalloproteinases (MMP: Matrix MetalloProteinase), specifically MMP-2 (Matrix metalloproteinase-2 protein expression) and MMP-9 (Matrix metalloprotease 9) and the decrease in the adhesion molecules ICAM-1 and integrin B1. Last but not least, AR is an antagonist of the expression of the activation-dependent protein cyclooxygenase-2 (COX-2). The above is supported by the following scientific documents for reference: • Rosmarinic Acid Activates AMPK to Inhibit Metastasis of Colorectal Cancer. Frontiers in Pharmacology. 2018. Ο77θηη / 77Π7 / β / YILI • Rosmarinic Acid Antagonizes Activator Protein-1-Dependent Activation of Cycloxygenase-2 Expression in Human Cancer and Nonmalignant Cell Lines. The Journal of Nutrition. 2008. • Chemical Composition and Anti-inflammatory, Cytotoxic and Antioxidant Activities of Essential Oil from Leaves of Mentha piperita Grown in China. Píos One. 2014. • Antibacterial and antioxidant activities of Mentha piperita L. Arabian Journal of Chemistry. 2011. • The Hydroalcoholic Extract Obtained from Mentha piperita L. Leaves Attenuates Oxidative Stress and Improves Survival in Lipopolysaccharide-Treated Macrophages. Journal of Immunology Research. 2017. • Hollow fiber-based liquid phase microextraction combined with high-performance liquid chromatography for the analysis of flavonoids in Echinophora platyloba DC. and Mentha piperita. Food Chemistry. 2013:731-735. Ο77θηη / 77Π7 / β / ΥΙΛΙ • Total phenolic content and antioxidant activity of six wild Mentha species (lamiaceae) from northeast Algeria. Asian Pacific Journal of Topical Biomedicine. 2016;6(9):760-766. • Antioxidant activity of the flavonoid hesperidin in chemical and biological systems. J Agrie Food Chem. 2005. 15;53(12):4757-61. Furthermore, hesperidin, a flavonoid, provides potent cellular antioxidant protection by reducing free radicals (1,1-diphenyl-2-picrylhydrasyl) (DPPH). This is supported by the following scientific documents for reference: • Chemical Composition and Anti-inflammatory, Cytotoxic and Antioxidant Activities of Essential Oil from Leaves of Mentha piperita Grown in China. Píos One. 2014. • Antibacterial and antioxidant activities of Mentha piperita L. Arabian Journal of Chemistry. 2011. • The Hydroalcoholic Extract Obtained from Mentha piperita L. Leaves Attenuates Oxidative Stress and Improves Survival in Lypopolysaccharide-Treated Macrophages. Journal of Immunology Research. 2017. • Hollow fiber-based liquid phase microextraction combined with highperformance liquid chromatography for the analysis of flavonoids in Ο77θηη / 77Π7 / β / ΥΙΛΙ Echinophora platyloba DC. And Mentha piperita (Hollow-fiber liquid-phase microextraction combined with high-performance liquid chromatography for the analysis of flavonoids in Echinophora platyloba DC. and peppermint). Food Chemistry. 2013:731–735. • Total phenolic content and antioxidant activity of six wild Mentha species (lamiaceae) from northeast Algeria. Asian Pacific Journal of Topical Biomedicine. 2016;6(9):760-766. • Antioxidant activity of the flavonoid hesperidin in chemical and biological systems. J Agrie Food Chem. 2005. 15;53(12):4757-61. Likewise, hesperidin is an indirect mediator of the nuclear factor kappa light chain-enhancer of B cells (NF-kB), which is responsible for generating the inflammatory cascade (interleukins). HP (hesperidin) also enhances the expression of endogenous neuroprotective factors, resulting in improved neuronal growth factors and endogenous anti-oxidative defense mechanisms, inhibiting neuroinflammatory and apoptotic pathways. The above is scientifically supported by the following document for reference: • Hesperidin: a therapeutic agent for obesity. Drug Design, Development and Therapy. 2019. οζζοηη / ζζηζ / β / γΐΛΐ To administer these and other bioactive agents to the anterior segment of the eye, the ideal route of administration is topical, as it is easy for the patient to use, with few adverse effects and adequate control of the frequency used. However, the bioavailability of bioactive agents from herbal extracts following topical ophthalmic administration of conventional formulations is inefficient for target tissues of the posterior segment of the eyeball, as penetration is usually <5%, which constitutes a very important limitation and results in poor bioavailability. Therefore, conditions that affect the health of the white tissues in the posterior segment of the eyeball, such as the retina, are not candidates for this route of administration.Furthermore, the bioavailability of the bioactive agents present in Peppermint leaf extract in the retina, and in general in the different structures and tissues of the posterior segment of the eyeball, is seriously compromised by the poor penetration of molecules into the infraocular tissues. The characteristic resistance of the eyeball to the penetration of molecules and therapeutic agents is due to the presence of multiple natural barriers that prevent optimal concentrations of these agents. On the one hand, the tear film, cornea, conjunctiva, and sclera are highly efficient anatomical barriers that prevent the penetration of topically administered bioactive agents, and on the other hand, the blood-retinal barriers do the same with their systemic administration.Likewise, the tear film, tear drainage systems, the physiological aqueous cycle, and the formation of the vitreous gel are relevant physiological ocular barriers that prevent the correct distribution of molecules, including the bioactives present in the peppermint leaf extract. It is important to mention that the use of all these bioactives in an extract results in a synergistic effect by presenting different mechanisms of action that together result in a beneficial effect for the retina. Due to the high ozzonn / zznz / e / YiAi predisposition of several of these bioactives present in the peppermint leaf extract, they have a high predisposition to degradation, with a fairly low water solubility, so their bioavailability is consequently quite poor, therefore, a lipid-based formulation represents an advantage for the use of these bioactives topically. Our solution to increase the penetration and bioavailability of bioactives present in Peppermint leaf extract into the eyeball's target tissues is based on the use of advanced delivery methods and systems, such as lipid-based systems. Some of these lipid systems include lipid nanoparticles, micelles, and liposomes. Lipid nanoparticles are aqueous colloidal dispersions composed of solid lipids as a matrix material with the capacity to encapsulate both hydrophilic and lipophilic agents. This ultimately results in thermodynamically stable compounds that protect the encapsulated agent, offer controlled release, and are biocompatible with the eyeball's barriers. They are easy to produce and can be autoclaved. However, some of their limitations include a lower encapsulation and release capacity for both hydrophilic and lipophilic agents.Polymeric micelles (10–100 nm) comprise amphiphilic block copolymers above the critical micellar concentration, with a hydrophobic core and a hydrophilic shell. These micelles prevent or minimize the degradation of molecules, increase their penetration into infraocular tissues, and also increase their solubility. Finally, liposomes are biocompatible vesicles composed of a phospholipid bilayer with a structural similarity to the cell membrane, which form small spheroids capable of transporting both hydrophilic and lipophilic molecules. They are biocompatible, thermostable, and can present a high percentage of encapsulation efficiency for different compounds. In addition to peppermint, AR and HP are also found in many other plants classified in the Nepetoideae subfamily, Lamiaceae family, such as sage, mint, thyme, lemon balm, basil, oregano, and others. Other compounds have also been described for potential use in maintaining eye health and treating various ocular pathologies. Such is the case with vitamins A, C, D, and E, carotenoids such as zeaxanthin and lutein, and some minerals such as zinc and magnesium. Vitamin C, also known as ascorbic acid due to its chemical formulation, is a compound commonly found in fresh fruits and vegetables. This acidic compound undergoes oxidation, transforming into dehydroascorbic acid, thus participating as a cofactor in multiple hydroxylation reactions. It is thus involved in the biosynthesis of collagen, the main structural protein of connective tissue. This is the main reason, but not the only one, why it is associated with the prevention of capillary fragility; its daily requirement is 50 mg. Vitamin E is a fat-soluble vitamin with antioxidant effects and works uniquely by breaking the lipid peroxidation reaction chain. This action is crucial for stabilizing cell membranes and has been shown to prevent changes in the ocular surface and the retina. Vitamin E can regenerate other antioxidants such as ascorbic acid or glutathione. 077θηη / 77Π7 / β / YILI High concentrations of vitamin E similar to those found in the AREDS (AgeRelated Eye Disease Studies) formula may decrease the risk of progression of age-related macular degeneration. Vitamin E occurs naturally as eight distinct fat-soluble tocopherols and tocotrienols. Sunflower seeds and walnuts, as well as almonds, spinach, and dark leafy greens, are rich sources of vitamin E. Vitamin D has shown important antioxidant effects in retinal vasculopathies such as diabetic retinopathy. In the case of micronutrients such as magnesium and zinc, their use has expanded to include pathologies such as age-related macular degeneration. Magnesium is an essential cation, acting as a cofactor in enzymatic metabolic pathways that utilize phosphate. Hypomagnesemia, related to increased losses or insufficient intake, is closely associated with the metabolic syndrome observed in diabetes and its complications; in fact, it is considered the most frequent electrolyte imbalance in diabetic individuals. Since magnesium is predominantly an intracellular ion, plasma concentration may not reflect the body's overall magnesium status. Magnesium deficiencies can lead to alterations in tyrosine kinase enzyme activity and the number of insulin receptors, as well as increased intracellular calcium concentration, all of which are associated with insulin resistance. ozzonn / zznz / e / YiAi Other studies have linked low magnesium levels with elevated levels of tumor necrosis factor (TNF) and C-reactive protein (CRP), which could explain low-grade inflammatory processes. Zinc has the potential to have positive effects on retinal cells, so its absence is linked to dark adaptation defects. Equally important is the effect of carotenoids such as lutein and zeaxanthin, yellow pigments found in vegetables and other plants. Both have the potential to absorb excess light to prevent damage to the neurosensory retina, especially short-wavelength light rays like violet and blue. Their use is widely recognized in ophthalmology as preventative and / or conservative treatments for retinal health in patients with age-related macular degeneration. Regardless of their origin, these plant and mineral compounds could be administered for ophthalmological use, aiming for appropriate concentrations to preserve the health of the various structures of the eye and / or treat certain pathologies. Topical instillation is the preferred non-invasive route for administering drugs and molecules to treat diseases of the anterior segment of the eye, according to ophthalmologists worldwide. This approach is non-invasive, has few adverse effects, lower costs, and better patient adherence, as it does not require complex facilities or highly specialized personnel for its administration. However, the bioavailability of drugs administered via this route in the case of diseases affecting the posterior segment of the eye is almost nil. For drugs used to treat diseases affecting the retina, different routes of administration, such as intravitreal (intraocular) injections, periocular injections, and systemic administration, have been reported, without O77θηη / 77Π7 / β / YILI However, the presence of static barriers (cornea, sclera, Bruch's membrane and retinal pigment epithelium) prevent the passage of drugs and molecules into the vitreous cavity and the retina. Likewise, the dynamic barriers of the eye (blinking, tear film, tear duct drainage system, conjunctival, episcleral and choroidal dispersion, uveoscleral drainage, hydrostatic and osmotic pressure, as well as the different flow pumps and transporter proteins of the retinal pigment epithelium) facilitate the elimination of the poor concentrations that manage to reach the interior of the eyeball. Finally, metabolic barriers (cytochrome P450 and lysosomal enzymes) end up metabolizing the remainder of these concentrations. For all the above reasons, intravitreal injection is the most widely used delivery method by ophthalmology and retina specialists to deliver adequate and clinically effective concentrations for the treatment of diseases affecting the posterior segment of the eyeball. However, its use is associated with serious ocular adverse events such as endophthalmitis, retinal detachment, cataracts, and vitreous hemorrhage, among others. Furthermore, this treatment modality is expensive, painful, and inaccessible to the general population (as it requires hospital facilities and highly trained personnel for its administration), and generates little adherence and great discomfort for both the user (patients) and the medical team.In the case of systemic administration (oral route), which is the most commonly used for the use of herbal extracts (including Peppermint extract), high doses are required, which are often insufficient or even toxic with serious adverse events reported. ozzonn / zznz / e / YiAi On the other hand, blindness is currently one of the most significant disabilities in the world, as humans are primarily visual beings. Therefore, its development, in addition to being an individual catastrophe, represents a significant family, social, and economic burden. Conditions such as diabetic retinopathy have a high treatment cost. In the United States, the annual cost per eye for diabetic macular edema treatment can range from 14,000 to 25,000 USD, and in Mexico, it can range from 66,000 to 14,000 Mexican pesos. Age-related macular degeneration currently represents a 16 billion USD market.Additionally, the negative experience and discomfort generated by repeated drug injections, combined with the indispensable use of specialized facilities and human resources, result in poor adherence and access to treatment, as well as saturation of health services. It is essential to seek treatment alternatives and, above all, prevention. In summary, the eyeball presents significantly greater challenges than other organs for the delivery of drugs, molecules, and herbal extracts, given the large number of anatomical and physiological barriers it presents. Therefore, the greatest challenge for the release of drugs, molecules, and herbal extracts is to find a delivery method that is highly effective, with a high safety profile, that increases patient adherence, and that offers widespread access. Ο77θηη / 77Π7 / β / ΥΙΛΙ Previously, the research group that created this invention investigated a lipid-based formulation designed for the delivery of drugs, spheroids, proteins and peptides to infraocular tissues, which is referenced in the following publication: [Altamirano-Vallejo JC., Navarro-Partida., Santos A., et al. Characterization and Pharmacokinetics of Triamcinolone Acetonide-Loaded Liposomes Topical Formulations for Vitreoretinal Drug Delivery: Caracterización y farmacocinética de las formulaciones temacos de liposomes loaded with triamcinolone acetonide for the administration of vitreoretinal drugs. J of Ocul Pharmacol and Ther. 2018], However, said development did not include a nanosystem for the delivery of micronutrients present in foods useful for the preservation of ocular health, such as polyphenols present in herbal extracts, particularly in peppermint leaf extract. For this reason, having a topical delivery system for the polyphenols with bioactive properties contained in the peppermint leaf extract would be the ideal system. Justification for the development of the invention. Worldwide, it is estimated that approximately 1.3 billion people live with some form of visual impairment, according to the World Health Organization. Currently, the main causes of irreversible blindness are those affecting the white tissue of the posterior segment of the eyeball. Conditions such as Age-Related Macular Degeneration (AMD) and Diabetic Retinopathy (DR) comprise more than 80% of cases of irreversible blindness in the working-age group and in the elderly group in the Western world. Diabetic Retinopathy, in particular, is a O77θηη / 77Π7 / β / YILI is a problem in Latin American countries and in Mexico, given that it incapacitates economically active individuals, consequently impacting the economies of health systems and countries. This makes the field of ophthalmic therapeutics particularly interesting for researchers and entrepreneurs with a science and technology base. Unfortunately, administering therapies to the eye is extremely challenging due to the anatomical and physiological characteristics of the eyeball. Anatomical barriers include structures such as the corneal epithelium, conjunctiva, sclera, and both blood-retinal barriers; and physiological barriers encompass processes such as the tear film, its drainage systems, and blinking, as well as the aqueous humor production, flow, and absorption system and the composition of the vitreous gel. These barriers limit the systemic penetration of one or more bioactive agents present in an herbal extract. For administering bioactive agents to the anterior segment of the eye, the ideal route of administration is topical, as it is easy for the patient to use, with few adverse effects and adequate control of the frequency used. However, the bioavailability of bioactive agents from herbal extracts following topical ophthalmic administration of conventional formulations is inefficient for target tissues in the posterior segment of the eyeball, as penetration is usually less than 5%, a very significant limitation and results in poor bioavailability. Therefore, conditions that affect the health of target tissues in the posterior segment of the eyeball, such as the retina, are not candidates for this route of administration. Ο77θηη / 77Π7 / β / YΙΛΙ Because natural herbal extracts contain bioactive compounds with anti-inflammatory, anti-angiogenic, and antioxidant effects on certain target tissues, their use is becoming a viable option for conditions affecting retinal health. Extracts such as peppermint leaf extract, which have an additive effect of the bioactive compounds AR and HP, can be administered topically, achieving adequate penetration and bioavailability through the use of a lipid-based formulation. Currently, there is no evidence that commercially available natural products, such as herbal extracts, can help prevent and / or modulate the progression of these conditions, which are the main causes of irreversible blindness. Prior art. Based on the analysis of prior art documents, some inventions were found that involve peppermint leaf extract, such as invention US20080003310A1, dated September 12, 2007, which describes the preparation of perfume drops based on peppermint extract. However, said document does not involve the preparation of a lipid-based formulation for topical ophthalmic use, which contains peppermint leaf extract to enhance the penetration and bioavailability of its bioactives in target tissues of the posterior segment of the eyeball in humans. Therefore, this invention is different in content and application from the developed formulation. Ο77θηη / 77Π7 / β / ΥΙΛΙ US9278062B2, dated September 29, 2010, describes a method for preparing an eye drop and its composition in a transparent container that can mix a monoterpene and a polymer to give a viscosity of 500 to 100,000 mPa*s. The purpose of this document is to formulate a high-viscosity eye drop to prevent dry eye and to increase the performance of this drop. Based on the above, this invention is different from the developed formulation in both content and application. Invention W02003097027A1, with a priority date of May 22, 2002, describes a menthol emulsion in oil or water containing fatty acids and an emulsifier to ensure its stability for many hours after preparation. The formulation contains more than one fragrance, such as eucalyptus, lemon, and / or mint, to mask their chemical aroma. The invention is contraindicated in patients with cheiloma. Based on this, the invention differs from the previously developed formulation in both content and application, as it lacks a liposomal application to enhance the penetration and release of polyphenols and other elements into the target tissues of the posterior segment of the eye. The prior art review revealed no evidence of inventions related to the subject matter of the invention described herein. The technical details of the invention are described below. OBJECT OF THE INVENTION. O77Qnn / 77n7 / R / YIAI The object of this invention is to make available a lipid-based formulation for topical ophthalmic use that functions as a liposomal delivery system for the bioactive agents contained in peppermint (Mentha piperita) leaf extract to target tissues of the posterior segment of the eyeball in humans. The formulation comprises a combination of polyphenol-rich peppermint leaf extract as the active ingredient, polyethylene glycol (PEG-12) glyceryl dimyristate as a structural component of the liposomes, ethyl alcohol as an organic solvent for liposome generation, Kolliphor® HS 15 as a penetration enhancer, anhydrous citric acid and dehydrated sodium citrate as buffers, and grade 2 purified water as an inorganic solvent. Therefore, the formulations of the present invention are useful for preserving health and reducing the risk of developing chronic degenerative processes in the infraocular tissues, including those of the posterior segment of the eyeball, due to their anti-inflammatory, anti-angiogenic, and antioxidant activity. BRIEF DESCRIPTION OF THE INVENTION. In general, the invention comprises a topical ophthalmic formulation of lipid-based peppermint leaf extract where the primary value lies in allowing more efficient penetration and bioavailability of its bioactives in white tissues of the posterior segment of the eyeball in humans. It is important to mention that the use of natural extracts for beneficial purposes on eye health becomes relevant when considering that all bioactives The O77Qnn / 77n7 / R / VIAI contained therein will have a synergistic therapeutic effect, derived from the sum of the different mechanisms of action. Due to the high susceptibility of several of these bioactives present in the Peppermint leaf extract, they have a high predisposition to degradation, with a rather low water solubility, so their bioavailability is consequently quite poor. Therefore, this lipid-based formulation represents an advantage for the topical use of these bioactives; thus, the invention consists of a lipid-based formulation for topical ophthalmic use that functions as a liposomal delivery system for the bioactive agents contained in the Peppermint leaf extract to the target tissues of the posterior segment of the eyeball in humans. The invention is useful for preserving health and reducing the risk of developing chronic degenerative processes in intraocular tissues, including those of the posterior segment of the eyeball, due to its anti-inflammatory, anti-angiogenic and antioxidant activity. Therefore, and being an efficient option for use as a delivery system for polyphenol bioactives such as AR and HP, contained in the Peppermint leaf extract, a lipid-based ophthalmic topical formulation has the potential to be a release method that makes the penetration of the eyeball barriers more efficient, thus achieving an increase in the bioavailability of the bioactives in the white tissues of the posterior segment of the eyeball such as the retina, minimizing the risks associated with other forms of local and / or local administration. Systemic Ο77θηη / 77Π7 / β / ΥΙΛΙ. DETAILED DESCRIPTION OF THE INVENTION The characteristic details of the topical ophthalmic lipid-based formulation, containing peppermint leaf extract as the main component, to make the penetration and bioavailability of its bioactives in infra-ocular white tissues in humans more efficient, are clearly shown in the following description. • Components. The compositions of the present invention contain an effective amount of peppermint extract. The concentration of peppermint extract in the lipid-based formulations ranges from 0.3 to 2 mg, i.e., from 0.03 to 2% (w / v). Peppermint extract has a large amount of polyphenol bioactive agents such as AR and HP that have the potential to preserve health and reduce the risk of developing chronic degenerative processes in infraocular tissues, including those of the posterior segment of the eyeball, due to its anti-inflammatory, anti-angiogenic and antioxidant activity. Polyethylene glycol (PEG-12) glyceryl myristate is used as a structural component of liposomes at a concentration of 5-15% (w / v) and ethyl alcohol is used as an organic solvent for the generation of liposomes at a concentration of O77Qnn / 77n7 / R / YIAI 0.7 to 2.1% (v / v). In addition, the formulation contains polyethylene glycol (15) hydroxystearate or Kolliphor® HS15 at 2.5 to 7.5% (w / v), as a potent non-ionic solubilizer and emulsifying agent, with low toxicity proposed to act as a permeability enhancer. Kolliphor® HS15 also promotes the transport of molecules across cell membranes by increasing the rate of endocytosis and stimulates drug translocation through the paracellular pathway (affecting the organization of actin in the cellular cytoskeleton with subsequent opening of tight junctions between cells). Furthermore, the aqueous compositions of the present invention optionally comprise additional excipients selected from the group consisting of buffering agents, pH adjusting agents, and preservatives. Anhydrous citric acid (0.04 to 0.16%) and sodium citrate dihydrate (0.23 to 0.69%) are used as buffers. The lipid-based formulation may include micronutrients such as the following: a. Vitamin E which is a fat-soluble vitamin that has antioxidant effects and works uniquely by breaking the lipid peroxidation reaction chain. This action is crucial to stabilize cell membranes and has been shown to prevent changes in the ocular surface and retina, b. Vitamin D since it has shown important effects as an antioxidant in retinal vasculopathies such as diabetic retinopathy, O77Qnn / 77n7 / R / YIAI c. Magnesium and Zinc for pathologies such as age-related macular degeneration, d. Lutein and Zeaxanthin as preventive and / or preservative agents for retinal health in patients with age-related macular degeneration. The lipid-based compositions of the present invention can be prepared by conventional methods for preparing pharmaceutical suspension compositions. • Preparation method. According to the preferred method, peppermint extract (0.3 to 2 mg) is first added to a lipid mixture containing polyethylene glycol (PEG-12) (5-15%), polyethylene glycol (15)-hydroxystearate (trade name Kolliphor® HS15) (2.5 to 7.5%), and ethyl alcohol (0.7 to 2.1%) at 25 °C ±1 °C, combining the components by stirring for 10 minutes ±1 minute. Separately, an aqueous base mixture is prepared consisting of grade 2 purified water (QS), anhydrous citric acid (0.04 to 0.16%), and sodium citrate dihydrate (0.23 to 0.69%) at 25 °C ±1 °C, combining the components by stirring for 10 minutes ±1 minute. The aqueous mixture is added to the lipid mixture at room temperature and stirred for 5 min ±1 minute to obtain the final formulation. Resulting formulation. The resulting lipid-based formulation that is the subject of this invention is configured as follows: ozzonn / zznz / e / YiAi Peppermint extract 0.3 to 2 mg i.e. 0.03 to 2% (w / v) Polyethylene glycol (15) -hydroxystearate 2.5 to 7.5% (w / v) PEG-12 glyceryl myristate 5-15% (w / v) Ethyl alcohol 0.7 to 2.1% (v / v) Anhydrous citric acid 0.04 to 0.16% Sodium citrate dihydrate 0.23 to 0.69% Purified water grade 2 QS1.0 ml The formulation enhances the penetration and bioavailability of bioactive ingredients present in the peppermint extract into the target tissues of the eyeball due to the use of advanced delivery methods and systems, such as lipid-based systems. Some of these lipid systems include lipid nanoparticles, micelles, and liposomes. Lipid nanoparticles are aqueous colloidal dispersions composed of solid lipids as a matrix material with the capacity to encapsulate both hydrophilic and lipophilic agents, ultimately resulting in 15 thermodynamically stable compounds that protect the encapsulated agent, offer controlled release, are biocompatible with the barriers of the eyeball, are easy to produce, and can be autoclaved. However, some of their limitations include a lower encapsulation and release capacity for both hydrophilic and lipophilic agents. Polymeric micelles (10-100 nm) comprise amphiphilic block copolymers above the critical micellar concentration, with a hydrophobic core and a hydrophilic shell. Micelles prevent or minimize the degradation of molecules, increase their penetration into intraocular tissues, and also enhance their solubility. Finally, liposomes are biocompatible vesicles composed of a phospholipid bilayer structurally similar to the cell membrane, which form small spheroids capable of transporting both hydrophilic and lipophilic molecules. They are biocompatible, thermostable, and can exhibit high encapsulation efficiency for various compounds. The lipid-based formulation may include micronutrients such as the following: a. Vitamin E, which is a fat-soluble vitamin that has antioxidant effects and works uniquely by breaking the lipid peroxidation reaction chain. This action is crucial for stabilizing cell membranes and has been shown to prevent changes in the ocular surface and retina. b. Vitamin D, as it has shown important antioxidant effects in retinal vasculopathies such as diabetic retinopathy, c. Magnesium and Zinc for pathologies such as age-related macular degeneration, d. Lutein and Zeaxanthin as preventatives and / or preservers of retinal health in patients with age-related macular degeneration. Ο77θηη / 77Π7 / β / ΥΙΛΙ A lipid-based formulation has the potential to improve stability, make penetration of the eyeball's natural barriers more efficient, and increase the bioavailability of polyphenols in the infraocular tissues. Analysis of the resulting formulation. The pH of the MP-LF (Liposome Formulation Loaded with Peppermint Extract) was analyzed using a potentiometer in triplicate at room temperature. Osmolarity was measured using a vapor pressure osmometer in triplicate at 33°C (ocular surface temperature). Viscosity was also measured in triplicate at 33°C. Viscosity was measured using a thermostatically controlled rheometer when steady state was reached with shear rates increasing from 0 to 1000 s1. The particle size of the liposomes contained in the MP-LF was analyzed by dynamic light scattering and the zeta potential (ζ) was calculated by measuring the particle velocity using a laser Doppler velocity meter at 25°C (Zetasizer Nano ZS, Malvern Instruments, Malvern, UK). Ο77θηη / 77Π7 / β / ΥΙΛΙ The Z-diameter (mean particle diameter) and the polydispersity index (PDI) were calculated from the particle size distribution. The pH of the MP-LF was 6.9. The physiological pH range of tears is between 6.5 and 7.6. Furthermore, the viscosity value of TA-LF was 75 cP (high), which could favor its retention on the ocular surface. The osmolarity was 342 mOsmol / L, falling within the isotonic range. The average particle diameter (z-average) was 160 nm, while its PDI was 0.544. The physicochemical characteristics of MP-LF are summarized below. ___________Physicochemical characteristics of MP-LF·____________ Formulation Physicochemical parameters pH Viscosity Osmolarity Z PDI (cP) (mOsm / l) (nm) (nm) MP-LF 6.9 75 342 160 1 10 MP-LF = Liposome formulation loaded with Peppermint extract pH = Potential of hydrogen ions, cP = centipoise, PDI = polydispersity index O77Qnn / 77n7 / R / YIAI To evaluate the stability of MP-LF, a heat stress test was performed. The 10 MP-LF samples were maintained at 30°C, 40°C, and 60°C for three weeks. pH and encapsulation efficiency were assessed on days 0 (baseline), 7, 14, and 21 of thermal incubation. To evaluate encapsulation efficiency, we measured the concentration of 15-rosmarinic acid (15-AR) in the filtered solution obtained after MP-LF was extruded through a 0.22 µm polycarbonate membrane. The AR concentration in the MP-LF samples was subtracted from the concentration in the control samples (liposomal solution without peppermint extract). AR concentration was determined by high-performance liquid chromatography (HPLC) using a Varian 920 LC system (Agilent Technologies, Santa Clara, CA, USA) with a Zorbax Eclipse Plus C18 column (4.6 x 100 mm, 3.5 µm) at 30°C. Samples (20 μl) were eluted from the column in a mobile phase composed of water:methanol (30:70) at a flow rate of 1 ml / min. Detection was performed at 330 nm. The retention time and detection limit were 6.8 min and 0.004 mg / ml, respectively. The TA standard curve was linear from 0.004 to 0.100 mg / ml (correlation > 0.99). After 21 days of thermal stress, the MP-LF maintained its stable pH at 30 and 40°C and even improved encapsulation at higher temperatures (60°C). The results related to the stability tests are shown below. Results of the stability study MP-LF MP-LF MP-LF Parameter Time (days) 30°C 40°C 60°C 0 6.9 6.8 6.55 pH 7 6.85 6.85 6.45 14 6.85 6.85 6.45 21 6.9 6.9 6.5** Encapsulation 0 0.15 0.2 0.2 of extract of 7 0.19* 0.27** 0.31*' Peppermint 14 0.18 0.08 0.02 (%) 21 0.11 0.095 0.05 MP-LF; Liposome formulation loaded with peppermint extract. Asterisks indicate values ​​statistically different from day 0 values. *= P < 0.05, **= P < 0.01 Additionally, diffusion chambers and New Zealand white rabbit corneas were used to perform in vitro diffusion experiments (BW200S Chemotaxis Chambers, NeuroProbe, Gaithersburg, MD, USA). The central section 5 of the cornea (6 mm in diameter) was located between the upper and lower compartments of the diffusion chambers in order to act as a diffusion barrier for the MP-LF. The upper compartment was filled with 180 pl of balanced salt solution (BSS) while the lower compartment was filled with 200 pl of MP-LF. To prevent evaporation, the diffusion chambers were placed inside a humid chamber at 37 'C. To perform the diffusion analysis of In MP-LF, the concentration of rosmarinic acid (RA) contained in the peppermint extract was determined by high-performance liquid chromatography (HPLC) in the upper compartment at 2, 4, 6, and 8 hours after the start of the diffusion test. In relation to the in vitro diffusion analysis, it was observed that MP-LF 15 reached the highest PS concentrations after 8 hours of monitoring. The results of the in vitro diffusion study are presented below. Concentration of rosmarinic acid (RA) in the upper compartment of the diffusion chamber Time (hours) PS (pg / mL) 20.38 30.47 40.57 ozzonn / zznz / e / YiAi 50.79 60.88 71.15 81.32 The values ​​represent the average of three measurements. • Example of application of the formulation. The following example is intended to illustrate, but not limit, the present invention. The concentrations of the ingredients are presented in units of % weight / volume (% w / v) or % volume / volume (% v / v). The final concentration of peppermint extract in the resulting dispersion in the example is 0.3 mg / ml (0.03%). The composition of the liposome formulation loaded with peppermint extract (MP-LF), hereinafter referred to as MP-LF, is described below. ozzonn / zznz / e / Y MP-LF Formulation 0.03% Peppermint extract 0.3 mg Kolliphor® HS 15 50 mg Polyethylene glycol (PEG-12) glyceryl 100 mg myristate Ethyl alcohol 14 µL Anhydrous citric acid 0.8 mg Sodium citrate dihydrate 4.675 mg Purified water grade 2 QSIOml MP-LF; Liposome formulation loaded with peppermint extract The foregoing description of the disclosed definitions is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these definitions and / or implementations will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein, but should be accorded the broadest scope consistent with the following claims and the novel principles and features described herein.

Claims

1. A lipid-based formulation for topical ophthalmic use to make the penetration and bioavailability of its bioactives in infraocular white tissues in humans more efficient, characterized by comprising: a. Peppermint leaf extract of 0.3 to 2 mg, that is, 0.03 to 2% (w / v) where this component has a large amount of polyphenol bioactive agents such as anthocyanins, resveratrol and pterostilbene that have the potential to preserve health and reduce the risk of developing chronic-degenerative processes in infraocular tissues, b. Polyethylene glycol (15)-hydroxystearate of 2.5 to 7.5% (w / v) where this component is used as a powerful non-ionic solubilizer and emulsifying agent, with low toxicity proposed to act as a permeability enhancer, c. PEG-12 glyceryl dimistrate of 5-15% (w / v) where this component is used as a structural component of liposomes, d. Ethyl alcohol from 0.7 to 2.1% (v / v) where this component is used as an organic solvent for the generation of liposomes, e. Anhydrous citric acid from 0.04 to 0.16% where this component is used as a buffer, Ο77θηη / 77Π7 / β / YILI f. Sodium citrate dihydrate from 0.23 to 0.69% where this component is used as a buffer, g. Purified water grade 2 with QS 1.0 ml.

2. A method for preparing a lipid-based formulation for topical ophthalmic use to increase the penetration and bioavailability of its bioactive ingredients in the infraocular white tissues of humans according to claim 1, comprising the following steps: a. According to the preferred method, the extract of Peppermint (0.3 to 2 mg / mL) is first added to a lipid mixture containing PEG-12 glyceryl dimistrate (5 to 15%), polyethylene glycol (15)-hydroxystearate (2.5 to 7.5%) and ethyl alcohol (0.7 to 2.1%) at 25 °C ± 1 °C, the components being combined by stirring for 10 minutes ± 1 minute, b. On the other hand, a water-based mixture composed of grade 2 purified water (QS), anhydrous citric acid (0.04 to 0.16%) and sodium citrate dihydrate (0.23 to 0.69%) is prepared at 25 °C ±1 °C, combining the components by stirring for 10 minutes ±1 minute, c.The aqueous mixture is added to the lipid mixture at room temperature and stirred for 5 minutes ±1 minute to obtain the final formulation.

3. The lipid-based formulation for topical ophthalmic use to make the penetration and bioavailability of its bioactives in infraocular white tissues in humans more efficient according to claim 1, which has the property of: O77Qnn / 77n7 / R / VIAI a. Possessing the potential to improve stability and make the penetration of the natural barriers of the eyeball and in the infraocular tissues of the polyphenols contained in the Peppermint extract more efficient.

4. The lipid-based formulation for topical ophthalmic use to make the penetration and bioavailability of its bioactives in infraocular white tissues in humans more efficient according to claim 1, which may include micronutrients such as the following: a. Vitamin E which is a fat-soluble vitamin that has antioxidant effects and works in a unique way by breaking the reaction chain of lipid peroxidation. This action is crucial to stabilize cell membranes and has been shown to prevent changes in the ocular surface and in the retina, b. Vitamin D since it has shown important effects as an antioxidant in retinal vasculopathies such as diabetic retinopathy, c. Magnesium and Zinc for pathologies such as age-related macular degeneration, d. Lutein and Zeaxanthin as preventives and / or preservatives of retinal health in patients with age-related macular degeneration.