Lipid-based topical ophthalmic formulation for enhancing the efficiency of the penetration of the eyeball barrier by the bioactives pilocarpine, the coumarin scopoletin and patuletin present in pilocarpus jaborandi extract and use thereof in preserving lens health

Abstract

The invention relates to a solution for enhancing the efficiency of the penetration and bioavailability of bioactives present in jaborandi extract in the scleral tissue of the eye ball, which is based on using advanced delivery systems and methods, such as lipid-based systems. Some of these lipid systems include lipid, micellar and liposome nanoparticles. The lipid nanoparticles are aqueous colloidal dispersions comprising solid lipids as matrix material, capable of encapsulating both hydrophilic and lipophilic agents, which ultimately yield thermodynamically stable compounds, with the encapsulated agent being protected. The compounds have controlled release and are biocompatible with the barriers of the eye ball, easy to produce and able to be sterilised in an autoclave. Accordingly, the lipid-based topical ophthalmic formulation comprising jaborandi extract represents an important advancement in enhancing the efficiency of the penetration and bioavailability of the bioactives thereof in scleral tissue (iris and lens) of the human eyeball.

Description

[0001] Lipid-based ophthalmic topical formulation to enhance the penetration of bioactive substances through the eyeball barriers.

[0002] PILOCARPINE, COUMARIN, SCOPOLETIN AND PATULETIN PRESENT IN THE EXTRACT OF PILOCARPUS JABORANDI AND THEIR USE IN PRESERVING THE HEALTH OF THE

[0003] CRYSTALLINE

[0004] FIELD OF INVENTION

[0005] The present invention relates to the fields of biotechnology and medicine. The invention consists of a lipid-based formulation for topical ophthalmic use to enhance the penetration of the anatomical, physiological, and dynamic barriers of the eyeball by the bioactives present in the Jaborandi extract (Pilocarpus Jaborandi), functioning as a delivery system for said bioactives to the white tissues within the eyeball (iris, ciliary body, and lens) in humans. The invention is also useful for preserving the health of the lens, its transparency, and its accommodative function.

[0006] BACKGROUND OF THE INVENTION

[0007] The lens is an essential ocular structure for vision, playing a key role in the refraction and focusing of light entering the eye. It is located in the eyeball, behind the iris and pupil, like a transparent biconvex lens. When we observe a cross-section of the eye, the lens appears as a kind of "lens" or "disc" lodged between the vitreous humor and the aqueous humor.

[0008] A distinctive feature of the lens is that it is enclosed in a thin, elastic capsule. This capsule is critical to the lens's function, allowing its shape to change during the accommodation process. The zonules, which are tiny ligament-like filaments, hold the lens in its proper position by connecting it to the ciliary body. This connection between the ciliary body and the lens is essential for the eye's ability to focus on objects at different distances.

[0009] Unlike most tissues in the human body, the lens has the unique characteristic of not containing blood vessels. This lack of vascularization is one of the reasons why the lens can maintain its transparency, a crucial aspect for its optical function. Furthermore, the lens is composed mostly of water and proteins, specifically a type of protein called crystallins. These proteins, which make up about 90% of all proteins in the lens, are responsible for its refractive index and clarity.

[0010] As we are born and grow, the lens also grows by adding new layers of cells. Interestingly, however, the oldest lens cells (those formed during embryonic development) don't disappear; instead, they compact toward the center of the lens. This process of growth and compaction gives rise to different regions or zones within the lens: the nucleus, the cortex, and the capsule.

[0011] The lens is a unique structure in the human eye, and its primary function is to refract and focus light onto the retina. Its physiology is complex and highly specialized to maintain its transparency and elasticity throughout life.

[0012] Proteins, especially crystalline proteins, account for about 90% of the dry matter of the lens. These proteins have an orderly structure that allows light to pass through the lens without interference. Over time, these proteins can change and aggregate, which can lead to clouding of the lens, known as a cataract. Cataracts are one of the leading causes of blindness and visual impairment worldwide. Although it is a condition that primarily affects older people, its impact goes beyond simple vision loss:

[0013] 1. Quality of life: Daily activities such as reading, driving, or simply recognizing faces become challenging, which can lead to frustration and reduce personal independence.

[0014] 2. Safety: Cataracts increase the risk of falls and home accidents due to blurred vision. For drivers, this can be especially dangerous, as reaction time and depth perception may be compromised.

[0015] 3. Psychological Impact: Blurry or cloudy vision can cause anxiety and depression in some people. Fear of blindness or dependence on others can lead to social isolation.

[0016] 4. Economics: Vision loss due to cataracts also has an economic impact. This includes the cost of medical care, but also potential job loss or reduced ability to work.

[0017] Currently, there are reports of preventive treatments for the development of cataracts, such as the use of vitamin C, although its use remains debated in the scientific community, to the point that the only treatment is cataract surgery and intraocular lens implantation. Therefore, there is a need for new treatments because, although safe, cataract surgery is not free of potential adverse events, is expensive, and requires a highly qualified structure and human capital for its execution. Furthermore, it is not a preventive treatment and is not compatible with maintaining the health of the lens.

[0018] An excellent alternative is the use of bioactives against oxidative stress.

[0019] Oxidative stress refers to the damage caused by free radicals, unstable molecules that can damage proteins, lipids, and DNA in cells. In the context of the eye, this oxidative damage can lead to alterations in lens proteins, leading to the formation of aggregates and, consequently, lens opacity. Given this fundamental role of oxidative stress in the development of cataracts, it has been theorized that antioxidants may offer a potential avenue for preventing or at least slowing this ocular disorder. Antioxidants are compounds that neutralize free radicals, thereby reducing the damage they can cause. Several studies have investigated the relationship between antioxidant intake and the risk of cataracts.Antioxidant-rich foods, such as fruits and vegetables containing vitamins C and E, as well as other antioxidant compounds, have shown some potential in protecting against cataract formation. Epidemiological studies have suggested that diets rich in these nutrients are associated with a reduced risk of cataracts.

[0020] Scopoletin, a bioactive contained in Jaborandi extract, has potent antioxidant activity that can neutralize free radicals and therefore protect lens proteins from oxidative damage. However, to date there is no efficient method for its administration, since the anatomical, physiological and functional barriers that the eyeball presents prevent its correct absorption to give a bio-presence in target tissues of the eyeball, specifically, the lens.

[0021] On the other hand, the lens's ability to change shape and adjust focus is known as accommodation. This process is made possible by the ciliary muscle and zonules that hold the lens in place. When the ciliary muscle contracts, the zonules relax, allowing the lens to increase its curvature and focus on near objects. As we age, the lens loses some of its elasticity, which can result in presbyopia, a condition that makes it difficult to focus on near objects. Presbyopia is estimated to affect more than one billion people over the age of 40 worldwide. Currently, the main treatment modalities for presbyopia are contact lenses, reading glasses, refractive surgery, and infraocular lens surgery.Each of these therapeutic options presents disadvantages and potential threats to ocular health, for example: the risk of corneal ulcers with contact lens use, discomfort and / or limitations in daily activities (such as sports) due to the use of over-the-air lenses, or the risks associated with ocular surgery, such as the development of corneal opacity due to refractive laser surgery or infraocular infections in lens surgery. Given the above, there is a growing need for effective, easy-to-use, and noninvasive approaches to maintain lens health throughout life and, thus, prevent the onset of pathological conditions such as loss of lens accommodation.

[0022] There are different treatment modalities for loss of accommodation (presbyopia). The most common is the use of glasses to compensate for the loss of accommodation. Other treatments include lens extraction and multifocal infraocular lens implantation, which is a surgically invasive procedure not without potential complications. Another option is pharmacological treatment, which represents an alternative to current therapies. Therefore, in recent years, novel pharmaceutical agents for topical ocular application have been investigated. The active ingredients currently used and / or investigated in the form of eye drops for the treatment of presbyopia are based on two main mechanisms of action: pupillary miosis by muscarinic stimulation and lens softening.

[0023] Parasympathomimetic pupillary miotics (carbachol, pilocarpine, and physostigmine) induce a pinhole effect in the patient and increase depth of field, while lens-targeted agents such as lipoic acid reduce protein oxidation, thereby preventing the lens rigidity characteristic of presbyopia. However, muscarinic stimulation by pupillary miotics can cause chronic inflammation and stimulation of the fixed pupil, posterior synechiae and spasmodic contractions of the iris, pigment dispersion, and myopic shift; lipoic acid, on the other hand, has little effect on near vision.

[0024] Pilocarpine, an alkaloid bioactive found in Jaborandi extract, is used topically to treat presbyopia, and the flavonoid patuletin inhibits aldose reductase in the lens. However, there is currently no efficient method for its administration, as the anatomical, physiological, and functional barriers present in the eyeball prevent its proper absorption, resulting in biopresence in the target tissues of the eyeball, specifically the lens.

[0025] In order to generate an effective, safe and natural therapeutic alternative for the preservation of lens health, we have developed a lipid-based formulation containing the bioactives pilocarpine, coumarin scopoletin and patuletin from the extract of jaborandi (Pilocarpus Jaborandi), a species of flowering plant belonging to the Rutaceae family, native to South America (Paraguay, Venezuela, Brazil to the Antilles). These bioactives have miotic, antioxidant and anti-inflammatory activity that can be used to preserve the transparency and accommodation of the lens.

[0026] While topical ophthalmic application is the gold standard for delivering drugs and bioactives to the ocular globe, this treatment modality faces the challenge of overcoming the various anatomical and physiological barriers of the eyeball that prevent bioactive agents from entering the eye. Many of these barriers are inherent and unique to ocular anatomy and physiology, making them a challenging task for innovations in drug delivery. Among the barriers found in the anterior segment of the eyeball are physical barriers such as the eyelid, cornea, conjunctiva, tenon, and blood-aqueous barrier, and physiological barriers such as conjunctival blood flow, lymphatic flow, and tearing. It is estimated that only 5% of the active ingredient in an ophthalmic formulation administered topically is internalized in the eye.

[0027] Previously, the use of lipid-based nanoparticles has been shown to be useful for the internalization of active agents in the posterior pole of the eyeball when applied topically via ophthalmic application. These nanosystems offer significant advantages over parenteral administration, such as increased bioavailability in the target tissue and greater comfort during administration, among others. Therefore, we decided to create a formulation of lipid-based nanoparticles for topical ophthalmic use that functions as a delivery system for the bioactive agents contained in Jaborandi extract to the target tissues within the eyeball (iris and lens) in humans, with the aim of improving the bioavailability of the bioactive agents within the eyeball. The invention is useful for preserving ocular health and reducing the development of lens opacification and rigidity, preserving its accommodative capacity, as well as preventing and treating presbyopia.

[0028] Justification for the development of the invention.

[0029] The loss of lens health has a significant impact on the quality of life of millions of people. The loss of lens transparency (cataract) represents one of the leading causes of blindness and visual impairment worldwide. Although it is a condition that primarily affects older adults, its impact goes beyond simple vision loss: it affects quality of life, safety, psychological impact, and economic impact. Likewise, presbyopia is a refractive condition that affects the global population over 40 years of age. Therefore, it represents a global eye health problem.

[0030] The current approach is not for the preservation of lens health and is far from preventative, as it is primarily corrective, consisting of lens extraction surgery and intraocular lens implantation (loss of lens transparency) and, in the case of loss of lens accommodation, it consists of the use of glasses, refractive surgery, and multiple intraocular lenses. However, these are still corrective methods, not preventive. They are even less associated with the preservation of lens health.

[0031] On the other hand, there are bioactive agents with miotic, antioxidant, and anti-inflammatory activity with the potential for positive effects on the lens. However, their administration faces the challenge of crossing the various anatomical and physiological barriers of the eyeball that prevent the entry of bioactive agents into the eye. Many of these barriers are inherent and unique to ocular anatomy and physiology, making them a challenging task for innovations in drug delivery. Among the barriers found in the anterior segment of the eyeball are physical barriers such as the eyelid, cornea, conjunctiva, tenon, and blood-aqueous barrier, and physiological barriers such as conjunctival blood flow, lymphatic flow, and tearing. It is estimated that only 5% of the active ingredient in an ophthalmic formulation administered topically is internalized in the eye.

[0032] Consequently, there is a need to create alternatives to make the penetration of the eyeball barriers of the bioactives pilocarpine, coumarin, scopoletin and patuletin present in the Pilocarpus Jaborandi extract more efficient and their use in preserving the health of the lens.

[0033] The presented invention (lipid-based formulation with bioactives from Jaborandi extract) which is for topical ophthalmic administration, has the ability to constitute itself as an alternative of natural origin, effective for the preservation of the health of the lens given that on the one hand, the bioactives of jaborandi such as; pilocarpine, scopoletin, patuletin, have parasympathomimetic and antioxidant biological activity that impact the physiology of the lens while, on the other hand, the lipid component of the formulation ensures greater penetration and bioavailability of the bioactives in the white ocular tissues (lens and iris).

[0034] Prior art.

[0035] Based on the analysis of prior art documents, the invention MX2017001880A was found with a priority date of February 10, 2017, which refers to a nutraceutical formulation for oral consumption, based on Jaborandi extract, fish oil (Omega-3 and Omega-6), recombinant human lactoferrin, Vitamin A and Vitamin E. The invention is within the field of ophthalmology and has been developed as an adjuvant to improve the signs of the ocular surface and symptoms associated with dry eye syndrome.This semi-liquid formulation, contained in soft gelatin capsules for easy consumption and to improve the absorption and bioavailability of its active compounds, contains an extract of natural origin (Pilocarpus Microphyllus) that stimulates the aqueous secretion of the tear film, also using eicosapentaenoic acid (EPA) omega 6 and docosahexaenoic acid (DHA) omega 3 obtained from fish oil, and which in conjunction with lactoferrin, vitamin A and vitamin E, an improvement in tear quality is achieved by understanding antimicrobial and anti-inflammatory effects. This formulation was designed to aid in the therapy of those subjects with symptoms associated with dry eye syndrome, which differs from our invention on the applications, administration method and bioactives in the field of ophthalmology.

[0036] On the other hand, the applications of Jaborandi extract according to the previous state-of-the-art analysis are in the field of skin solutions, hair loss, dandruff prevention, treatment of climacteric menopause, pilocarpine production, development of vaginal suppositories and treatment of xerostomia.

[0037] With respect to the above, an example is the invention WO9616667A1 with priority date of November 29, 1994, which refers to an improvement in a process for preparing cosmetic products intended for care, including cleaning, elimination of dandruff and excess oil accumulated in the pores of the scalp, preservation and recovery of hair and scalp care. This is done by applying cosmetic compositions that include a combination of natural extracts of Pilocarpus Jaborandi enriched with one of its constituent alkaloids isolated in the base form to reinforce the characteristics of the Jaborandi extract - pilocarpine, pilosine, isopilosine and / or epi - isopiloturin - the total extract and / or solutions made with the total extract of Aloe vera and water-soluble salts of quercetin.The specific compositions of the invention are presented as hair tonics, lotions, or shampoos in which the aforementioned natural products are mixed with other conventional materials used to prepare this type of cosmetic composition. This invention has a completely different application than the one developed and described herein.

[0038] Another example is invention CA2753288A1 with a priority date of February 25, 2009, which describes oral care compositions for promoting salivary flow; Methods for treating and preventing xerostomia are also disclosed. The oral care compositions may contain all-natural ingredients, including, for example, green tea polyphenols and Jaborandi extract.

[0039] PURPOSE OF THE INVENTION

[0040] The objective of this invention is to provide a lipid-based nanoparticle formulation for topical ophthalmic use that enhances the penetration of the bioactive agents present in Jaborandi extract to improve lens health. This invention functions as a delivery system for the bioactive agents contained in Jaborandi extract to the white tissues within the eyeball (iris and lens) in humans.

[0041] The formulation comprises a combination of Jaborandi, Labrasol® (PEG-8 caprylic / capric glycerides) as surfactant and lipid component of an oil-in-water nano emulsion, ethyl alcohol as organic solvent, Kolliphor® HS 15 (Polyethylene glycol (15) -hydroxystearate) as penetration enhancer, anhydrous citric acid and dehydrated sodium citrate as buffers and grade 2 purified water as inorganic solvent.

[0042] The technical details of the developed invention are described below.

[0043] BRIEF DESCRIPTION OF THE INVENTION.

[0044] The invention developed is a solution to make more efficient the penetration and bioavailability of bioactives present in the Jaborandi extract in the target tissues of the eyeball, it lies in 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 matrix material with the capacity to encapsulate both hydrophilic and lipophilic agents, which finally result in thermodynamically stable compounds, with protection to the encapsulated agent, of controlled release and biocompatible with the barriers of the eyeball, easy to produce and with the possibility of being sterilized in an autoclave.

[0045] Therefore, the topical ophthalmic formulation of lipid-based Jaborandi extract represents an innovation to make the penetration and bioavailability of its bioactives in target tissues (iris and lens) of the eyeball in humans more efficient.

[0046] DETAILED DESCRIPTION OF THE INVENTION

[0047] The characteristic details of the lipid-based formulation for topical ophthalmic use and its application in preserving lens health are clearly shown in the following description.

[0048] SYMBOLS:

[0049] • EX: Jaborandi Extract

[0050] • EL: Lipid emulsion

[0051] • EL-EJ: Lipid emulsion with Jaborandi Extract

[0052] • PIL: Pilocarpine concentration

[0053] • HPLC: High-performance liquid chromatography

[0054] Components. The compositions of the present invention contain an effective amount of Jaborandi extract (EJ). The concentration of Jaborandi extract in lipid-based formulations per 100 ml varies from 0.3 to 200 |d i.e. from 0.03 to 20% (volume / volume: v / v). Jaborandi extract has a large amount of bioactive agents such as pilocarpine (PIL), scopoletin and patuletin that have the potential to preserve ocular health and reduce the development of opacification and rigidity of the lens, preserve its accommodative capacity, as well as prevent and treat presbyopia. Labrasol® (PEG-8 caprylic / capric glycerides) is used as a surfactant and lipid base of a nanoemulsion at a concentration of 0.01 to 15% (weight / volume: w / v) and ethyl alcohol is used as an organic solvent for the generation of liposomes at a concentration of 0.7 to 2.1% (v / v).

[0055] 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).

[0056] On the other hand, the aqueous compositions of the present invention optionally comprise further 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, and benzalkonium chloride 0.1 mg is used as a preservative. Additionally, QS1.0 ml of grade 2 purified water is required.

[0057] Preparation method. The lipid-based compositions of the present invention can be prepared by conventional methods for preparing suspension pharmaceutical compositions.

[0058] According to the preferred method, the ethanolic extract of Jaborandi (0.3 to 20 |Lxl / ml) is first added to a lipid mixture containing PEG-8 caprylic / capric glycerides (0.01 to 15%), polyethylene glycol (15) -hydroxystearate (Kolliphor® HS15) (2.5 to 7.5%) and ethyl alcohol (0.7 to 2.1%) at 25 °C ± 2.5 °C, combining the components by stirring for 10 minutes ± 2.5 minutes.

[0059] On the other hand, a water-based mixture composed of grade 2 purified water (QS), anhydrous citric acid (0.04 to 0.16%), Sodium citrate dihydrate (0.23 to 0.69%) and Benzalkonium chloride (0.1 mg) is prepared at 25 °C ± 2.5 °C, combining the components by stirring for 10 minutes ±1 minute.

[0060] The aqueous mixture is added to the lipid mixture at 25 °C ± 2.5 °C and stirred for 5 minutes ± 1 minute to obtain the final formulation.

[0061] • Resulting formulation.

[0062] The resulting lipid-based formulation object of this invention is made up as follows per 100 ml:

[0063] Reagent Volume / Quantity

[0064] Jaborandi Extract From 0.3 to 20 ml i.e. from 0.03 to 2%

[0065] (volume / volume)

[0066] Polyethylene glycol (15)-hydroxystearate 2.5 to 7.5% (w / v)

[0067] PEG-8 caprylic / capric glycerides 0.01 to 15% (weight / volume)

[0068] Ethyl alcohol 0.7 to 2.1% (v / v)

[0069] Anhydrous citric acid 0.04 to 0.16% Sodium citrate dihydrate 0.23 to 0.69%

[0070] Benzalkonium chloride 0.9 to 0.1 1 mg

[0071] Purified water grade 2 QS I .O ml

[0072] Therefore, the resulting formulation and characteristics are as follows:

[0073] A lipid-based ophthalmic topical formulation to make more efficient the penetration of the eyeball barriers of the bioactives present in the Pilocarpus Jaborandi extract characterized in that it contains per 100 ml: a. Jaborandi extract from 0.3 to 20 ml, that is, from 0.03 to 2% (volume / volume), b. Polyethylene glycol (15) -hydroxystearate from 2.5 to 7.5% (weight / volume) which acts as a potent non-ionic solubilizer and emulsifying agent, with low toxicity to act as a permeability enhancer, c. PEG-8 caprylic / capric glycerides from 0.01 to 15% (weight / volume) which is used as a surfactant and lipid bases of a nano emulsion, d. Ethyl alcohol from 0.7 to 2.1% (vol / vol) 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, f. Sodium citrate dihydrate from 0.23 to 0.69% where this component is used as a buffer, g. Benzalkonium chloride with 0.09 to 0.1 1 mg which is used as a preservative, h. Purified water grade 2 with QS 1.0 ml.

[0074] The lipid-based ophthalmic topical formulation where the bioactives present in the Pilocarpus Jaborandi extract are pilocarpine, coumarin, scopoletin and patuletin.

[0075] In turn, the lipid-based ophthalmic topical formulation is characterized by:

[0076] • The formulation is used to preserve the health of the lens.

[0077] • The formulation is used to preserve accommodative function and lens transparency.

[0078] • The formulation is used to mitigate symptoms associated with presbyopia.

[0079] • The formulation is used to mitigate symptoms associated with lens opacity.

[0080] • The formulation is used in the prevention of the appearance and progression of lens opacity.

[0081] • The formulation is used in the prevention of the onset and progression of loss of accommodative function of the lens.

[0082] • Example.

[0083] The following example is intended to illustrate, but not to limit, the present invention [in terms of ingredient concentrations, these are presented in units of % weight / volume (% w / v) or % volume / volume (% v / v)]. The final concentration of Jaborandi Extract in the resulting nano suspension in this example is 0.3 mg / ml (0.03%). The composition of the lipid emulsion (LE) containing Jaborandi extract (LE), hereinafter LE-LE, is described below.

[0084] Reagent Volume / Quantity

[0085] Jaborandi Extract 0.3 LXI

[0086] Polyethylene glycol (15)-hydroxystearate 75 mg PEG-8 caprylic / capric glycerides 300 mg

[0087] Ethyl alcohol

[0088] Anhydrous citric acid 0.8 mg

[0089] Sodium citrate dihydrate 4,675 mg

[0090] Benzalkonium chloride 0.1 mg

[0091] Purified water grade 2 QSI .O ml

[0092] • Analysis of the resulting formulation.

[0093] The pH of the EL-EJ was analyzed using a potentiometer in triplicate at room temperature. Osmolarity was measured using a vapor pressure osmometer in triplicate at 33°C (the ocular surface temperature).

[0094] 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 s-1.

[0095] The particle size of lipid nanoparticles containing Jaborandi extract from EL-EJ was analyzed by dynamic light scattering and the zeta potential (ZS) was calculated by measuring the particle velocity using a laser Doppler velocimeter at 25°C (Zetasizer Nano ZS, Malvern Instruments, Malvern, UK). The particle size distribution (Z) (mean particle diameter) and the polydispersity index (PDI) were calculated from the particle size distribution.

[0096] The pH of EL-EJ was 6.9. The physiological pH range of tears is between 6.5 and 7.6. Furthermore, the viscosity of EL-EJ was 55.5 cP (high), which could favor its permanence on the ocular surface. The osmolarity was 408 mOsmol / l, within the isotonic range. The average particle diameter (z-average) was 79.85 nm, while its PDI was 12.8 and its zeta potential was 0.550. The physicochemical characteristics of EL-EJ are summarized in the following table.

[0097] Physicochemical characteristics of EL-EJ.

[0098] Formulation Physicochemical parameters pH Viscosity Osmolarity Z PDI

[0099] (cP) (mOsm / l) (nm) (nm)

[0100] EL-EJ 6.9 55.5 408 79.85 12.8 pH= Potential of hydrogen ions, cP= centipoise, PDI= polydispersity index

[0101] 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 of the cornea (6 mm in diameter) was placed between the upper and lower compartments of the diffusion chambers in order to act as a diffusion barrier for EL-EJ or EJ (Jaborandi extract). The upper compartment was filled with 180 pl of balanced salt solution (BSS) while the lower compartment was filled with either 200 pl of EL-EJ or 200 / 200 pl of EJ.

[0102] To prevent evaporation, the diffusion chambers were placed inside a humidified chamber at 37°C. To perform the EL-EJ diffusion analysis, the pilocarpine (PIL) concentration 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.

[0103] The determination of PIL concentration by HPLC was carried out using a Varian 920 LC (Aligent Technologies, Santa Clara, CA, USA) with a Zorbax Eclipse Plus C18, 4.6 x 100 mm, 3.5 pm column (Agilent, Santa Clara, CA, USA) at 30°C. Samples (20 l) 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 215 nm. The retention time and detection limit were 10 min and 0.008 pg / ml, respectively.

[0104] In vitro diffusion analysis showed that EL-EJ reached higher PIL concentrations compared to EJ. The results of the in vitro diffusion study are presented below.

[0105] Concentration of pilocarpine (PIL) in the upper compartment of the diffusion chamber (g / mL).

[0106] Time (hours) EL-EJ EJ

[0107] 2 0.42 0.21

[0108] 4 0.84 0.44

[0109] 6 0.96 0.53

[0110] 8 1.22 0.59

[0111] The values ​​represent the average of three measurements.

[0112] 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

CLAIMS Having sufficiently described my invention, I consider as a novelty and therefore claim as my exclusive property, the contents of the following clauses:

1. A lipid-based ophthalmic topical formulation to make the penetration of the eyeball barriers of the bioactives present in the Pilocarpus Jaborandi extract more efficient, characterized in that it contains per 100 ml: a. Jaborandi extract from 0.3 to 20 ml, that is, from 0.03 to 2% (volume / volume), b. Polyethylene glycol (15) -hydroxystearate from 2.5 to 7.5% (weight / volume) which acts as a potent non-ionic solubilizer and emulsifying agent, with low toxicity to act as a permeability enhancer, c. PEG-8 caprylic / capric glycerides from 0.01 to 15% (weight / volume) which is used as a surfactant and lipid bases of a nano emulsion, d. Ethyl alcohol from 0.7 to 2.1% (vol / vol) 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, f. Sodium citrate dihydrate from 0.23 to 0.69% where this component is used as a buffer, g. Benzalkonium chloride with 0.9 to 0.1 1 mg which is used as a preservative, h. Purified water grade 2 with QS 1.0 ml.

2. Lipid-based ophthalmic topical formulation to make the Penetration of the eyeball barriers by the bioactives present in the Pilocarpus Jaborandi extract according to claim 1, characterized in that the bioactives are pilocarpine, coumarin, scopoletin and patuletin.

3. The lipid-based ophthalmic topical formulation to make the penetration of the eyeball barriers of the bioactives present in the Pilocarpus Jaborandi extract more efficient according to claim 1, characterized in that the formulation is used in the preservation of the health of the lens.

4. The lipid-based ophthalmic topical formulation to make the penetration of the eyeball barriers of the bioactives present in the Pilocarpus Jaborandi extract more efficient according to claim 1, characterized in that the formulation is used to preserve the accommodative function and transparency of the lens.

5. The lipid-based ophthalmic topical formulation to make the penetration of the eyeball barriers of the bioactives present in the Pilocarpus Jaborandi extract more efficient according to claim 1, characterized in that the formulation is used in the mitigation of the symptoms associated with presbyopia.

6. The lipid-based ophthalmic topical formulation to make the penetration of the eyeball barriers of the bioactives present in the Pilocarpus Jaborandi extract more efficient according to claim 1, characterized in that the formulation is used in the mitigation of symptoms associated with lens opacity.

7. The lipid-based ophthalmic topical formulation to make the penetration of the eyeball barriers of the bioactives present in the Pilocarpus Jaborandi extract more efficient according to claim 1, characterized in that the formulation is used in the prevention of the appearance and progression of lens opacity.

8. The lipid-based ophthalmic topical formulation to make the penetration of the eyeball barriers of the bioactives present in the Pilocarpus Jaborandi extract more efficient according to claim 1, characterized in that the formulation is used in the prevention of the appearance and progression of the loss of the accommodative function of the lens.

9. The lipid-based ophthalmic topical formulation to make the penetration of the eyeball barriers of the bioactives present in the Pilocarpus Jaborandi extract more efficient according to claim 1, characterized in that it is prepared as follows for every 100 ml: a. According to the preferred method, the ethanolic extract of Jaborandi (0.3 to 20 |iL / mL) is first added to a lipid mixture containing PEG-8 caprylic / capric glycerides (0.01 to 15%), polyethylene glycol (15)-hydroxystearate (Kolliphor® HS15) (2.5 to 7.5%) and ethyl alcohol (0.7 to 2.1%) at 25 °C ± 2.5 °C, combining the components by stirring for 10 minutes ± 2.5 minutes, b. On the other hand, a water-based mixture composed of grade 2 purified water (QS), anhydrous citric acid (0.04 to 0.16%), Sodium citrate dihydrate (0.23 to 0.69%) and Benzalkonium chloride (0.1 mg) is prepared at 25 °C ± 2.5 °C, combining the components by stirring for 10 minutes ±1 minute, c. The aqueous mixture is added to the lipid mixture at 25 °C ± 2.5 °C and stirred for 5 minutes ± 1 minute to obtain the final formulation.

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