Nanoemulsion formulation with improved tacrolimus stability and skin penetration.

Description

1 / 56 Nanoemulsion formulation with improved tacrolimus stability and skin penetration. DESCRIPTION

[001] The present invention relates to a composition comprising an oil-in-water nanoemulsion and a highly lipophilic macrolide lactone as an active agent, such as tacrolimus, dissolved in the nanoemulsion. In this formulation, tacrolimus can be fully dissolved, rather than suspended, and shows enhanced stability in terms of active ingredient content, pH, particle size and particle size homogeneity. Background of the invention

[002] Dispersions are colloidal systems, which include micelles, liposomes, virosomes, emulsions and micro-, nanoemulsions, suspensions and polymeric solutions. Emulsions or microemulsions can be oil-in-water, water-in-oil or intermediate-phase dispersions, generally containing surfactants as emulsifiers. Nanoemulsions are a subgroup of emulsions that contain very fine oil-in-water dispersions. Nanoemulsions are highly homogeneous, transparent and slightly opalescent. The dispersed droplets (liquid) or vesicles in such emulsions are composed of a lipid core surrounded by at least one monolayer of surfactant or emulsifier. Nanoemulsions are characterized by an average particle or vesicle size smaller than 200 nm, often smaller than 100 nm, and a narrow monodisperse particle or vesicle size distribution.

[003] Although nanoemulsions are generally more thermodynamically stable than conventional emulsions, they are often not stable under stress conditions, such as high temperatures or freezing conditions. Nanoemulsions can be in a metastable state and the Petition 870250090034, dated 02 / 10 / 2025, pp. 388 / 465 2 / 56 The structure often depends on the manufacturing process, which makes them complicated to formulate into a pharmaceutical composition with a long shelf life under different storage conditions. If destabilized, they can become heterogeneous, milky, and / or exhibit phase separation. On the other hand, nanoemulsions can provide useful applications in skin care, as they can exhibit good textural and sensual properties due to the very fine size of droplets or globules.

[004] Nanoemulsions are often manufactured by the mechanical fragmentation of an oil phase in an aqueous phase in the presence of a surfactant. The very small size of the oil globules is often achieved by at least one pass through a high-pressure homogenizer or a sonicator.

[005] Tacrolimus (also referred to in the present invention as TC) is a macrolide lactone molecule cultivated from the soil bacterium Streptomyces tsukubaensis. In pharmaceutical medicine, it is described as a calcineurin inhibitor with immunosuppressive capacity. It is applied topically to treat immune-mediated skin conditions such as atopic dermatitis or psoriasis. TC is a molecule with a molecular weight of 804.03 g / mol and highly lipophilic properties (logP > 3), i.e., six orders of magnitude more lipophilic than ALA. Due to its highly lipophilic nature, TC has been formulated in mixtures of mineral oil, paraffin, propylene carbonate, white petrolatum, and white wax. TC in aqueous compositions has been formulated as suspensions and was previously considered to be weakly stable in aqueous formulations (approximately 3 months at room temperature or up to 9 months at 5 °C).

[006] Liquid TC formulations in predominantly water-based systems (such as nanoemulsions) have not been marketed as Petition 870250090034, dated 02 / 10 / 2025, pp. 389 / 465 3 / 56 finished pharmaceutical products produced by pharmaceutical companies to date are likely hampered by the challenges of solubilization and stabilization in such formulations. With the pharmaceutical use of a topical TC formulation, there are two additional challenges. The first is its high lipophilicity, which can hinder its release from a fat-based formulation into the skin. The other is its poor distribution capacity in the aqueous compartments of the skin (such as living cells). Summary of the invention

[007] A first aspect of the invention relates to a formulation comprising (a) a nanoemulsion comprising: (i) at least one aqueous component; (ii) a carrier component comprising: (1) at least one lipophilic component, (2) at least one surfactant and (3) at least one alcohol; and (b) an active agent, wherein the active agent is a highly lipophilic macrolide lactone.

[008] Another aspect of the invention relates to the formulation of the first aspect for use in medicine.

[009] Another aspect of the invention relates to the formulation of the first aspect for use as a method of treating or preventing a dermatological, ophthalmic or autoimmune disease or condition, or for preventing organ rejection after transplantation. Detailed description

[010] Before the present invention is described in detail below, it should be understood that this invention is not limited to the particular methodology, protocols, and reagents described in the present invention, as these may vary. It should also be understood that the terminology used in the present invention is for the purpose of describing particular embodiments and is not intended to limit the scope of the present invention, which will be limited only by the claims. Petition 870250090034, dated 02 / 10 / 2025, pp. 390 / 465 4 / 56 attached. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meanings as commonly understood by a person skilled in the art.

[011] Preferably, the terms used in the present invention are defined as described in “A multilingual glossary of biotechnological terms: (IUPAC Recommendations)”, Leuenberger, HGW, Nagel, B. and Kolbl, H. eds. (1995), Helvetica Chimica Acta, CH-4010 Basel, Switzerland).

[012] Several documents are cited throughout the text of this descriptive report. Each of the documents cited in the present invention (including all patents, patent applications, scientific publications, manufacturer specifications, instructions, etc.), whether above or below, is incorporated into the present invention by reference in its entirety.

[013] The elements of the present invention will be described below. These elements are listed with specific embodiments; however, it should be understood that they can be combined in any way and in any number to create additional embodiments. The various examples described and the preferred embodiments should not be interpreted as limiting the present invention only to the embodiments explicitly described. This description should be understood to support and encompass embodiments that combine the explicitly described embodiments with any number of the disclosed and / or preferred elements. Furthermore, any permutations and combinations of all the elements described in this application should be considered disclosed by the description of the present application, unless the context indicates otherwise.

[014] Throughout this descriptive report and in the claims that follow, unless the context requires otherwise, the word “understand” and variations such as “understands” or “understanding” should be understood Petition 870250090034, dated 02 / 10 / 2025, pp. 391 / 465 5 / 56 as implying the inclusion of a whole number or step or group of whole numbers or steps, but not the exclusion of any other whole number or step or group of whole numbers or steps. As used in this descriptive report and the accompanying claims, the singular forms of “a”, “an” and “the” include the plural references, unless the wording clearly dictates otherwise.

[015] The formulation of a pharmaceutical composition is a highly complex process that needs to take into account different aspects, such as pH, solubility, polymorphism, applicability, and overall stability of the pharmaceutical composition. Additionally, it is necessary to consider the benefits and limitations of the active pharmaceutical ingredient (API), the excipients, the interaction of all components, and the manufacturing process. In oil-in-water nanoemulsion formulations, there are two different phases that may need to be stabilized and patient-compatible: one is the hydrophobic carrier component, which is usually the carrier and needs to stabilize and release the API, and the other is the aqueous component. All these aspects lead to complex formulations with a large number of ingredients. A very attractive property of oil-in-water nanoemulsion formulations is their ability to increase the penetration of active ingredients.

[016] The state of the art describes aqueous pharmaceutical formulations of TC as suspensions with short stability (approximately 3 months at room temperature or up to 9 months at 5 °C).

[017] The state of the art fails to provide an aqueous pharmaceutical composition capable of solubilizing tacrolimus, a highly lipophilic active agent. Furthermore, the state of the art fails to provide aqueous formulations (in solution or suspension) for tacrolimus with extended stability to Petition 870250090034, dated 02 / 10 / 2025, pp. 392 / 465 6 / 56 long of 24 months.

[018] As described above, nanoemulsions tend to coalesce under certain circumstances, such as exposure to extreme temperature differences, leading to larger droplet sizes and impairing nanoemulsion quality.

[019] These aspects make it clear that the design and formulation of a composition with high penetration and an improved impurity profile are highly desirable.

[020] The terms “active agent” and “active ingredient” are used interchangeably in the present invention. As used in the present invention, “active agent” includes an active pharmaceutical agent (also referred to in the present invention as “active pharmaceutical agent” or “active pharmaceutical ingredient”, “API”) and an active cosmetic agent (also referred to in the present invention as “active cosmetic agent”). As used in the present invention, an active pharmaceutical agent is defined as the chemical, biological, mineral or any other entity or component responsible for the therapeutic effects (pharmacological, physiological, physical, etc.) in a product. As used in the present invention, an active cosmetic agent is defined as the chemical, biological, mineral or any other entity or component responsible for the cosmetic effects of a product. The active agent may be a plant extract. The active agent may be present as a pharmaceutically acceptable salt.The active ingredient may be present as a cosmetically acceptable salt.

[021] The formulations of the invention comprise two phases: (I) an aqueous phase or aqueous component, (II) a lipid phase or carrier component.

[022] A first aspect of the invention relates to a Petition 870250090034, dated 02 / 10 / 2025, pp. 393 / 465 7 / 56 formulation comprising (a) a nanoemulsion comprising: (i) at least one aqueous component; (ii) a carrier component comprising: (1) at least one lipophilic component, (2) at least one surfactant and (3) at least one alcohol; and (b) an active agent, wherein the active agent is a highly lipophilic macrolide lactone.

[023] In the context of this descriptive report, the term “highly lipophilic” refers to a compound having a logP value of at least 3, where P is the octanol-water partition coefficient.

[024] Preferably, the active agent has a logP value of 3 or higher, such as 3.0 to 7.0 or 3.0 to 5.0.

[025] In the context of this descriptive report, the term "macrolide lactone" refers to a compound comprising or consisting of a macrocyclic lactone ring. One or more deoxy sugars may be attached to the lactone ring. Preferably, the macrocyclic lactone ring comprises at least one half-cyclic acetal, a 1,2-dicarbonyl substructure, as well as a piperidine substructure. Preferably, the macrocyclic lactone ring comprises two methoxy ligands at positions 14 and 16. Preferably, the macrolide lactone is not halogenated.

[026] Preferably, the active agent has immunosuppressive capacity.

[027] It is particularly preferred that the active agent be Tacrolimus, Pimecrolimus, Everolimus or Sirolimus, preferably Tacrolimus, a derivative, an isomeric form, a tautomeric form, a precursor, a Petition 870250090034, dated 02 / 10 / 2025, pp. 394 / 465 8 / 56 metabolite, hydrate and / or a pharmaceutically acceptable salt thereof.

[028] Tacrolimus, also referred to as TC in the present invention, is identified by CAS number 104987-11-3 and has the following chemical formula: cr HO

[029] Tacrolimus is a macrolide lactone molecule cultivated from the soil bacterium Streptomyces tsukubaensis. In pharmaceutical medicine, it is described as a calcineurin inhibitor with immunosuppressive capacity. It is applied topically to treat immune-mediated skin conditions such as atopic dermatitis or psoriasis. TC is a molecule with a molecular weight of 804.03 g / mol and highly lipophilic properties (logP > 3), i.e., six orders of magnitude more lipophilic than 5-aminolevulinic acid (ALA). Due to its highly lipophilic nature, TC has been formulated in mixtures of mineral oil, paraffin, propylene carbonate, white petrolatum, and white wax. Previously, TC was found to be poorly stable in aqueous formulations (approximately 90 days at room temperature).

[030] Formulations of TC in predominantly water-based semi-solid systems (such as emulsions) have not yet been put into pharmaceutical use, probably hampered by the challenges of solubilizing and stabilizing it in such formulations. With the pharmaceutical use of TC, there are two challenges. The first is its high lipophilicity, which can hinder its release from a fat-based formulation onto the skin. The other is its poor ability to Petition 870250090034, dated 02 / 10 / 2025, pp. 395 / 465 9 / 56 distribution in the aqueous compartments of the skin (such as living cells).

[031] A precursor of TC is, for example, Pre-Tacrolimus, which has the following chemical formula:

[032] Sirolimus is identified by CAS number 53123-88-9 and has the

[033] A precursor of Sirolimus is, for example, Pre-Sirolimus, which has the following chemical formula:

[034] Pimecrolimus is identified by CAS number 137071-32-0. Petition 870250090034, dated 02 / 10 / 2025, pp. 396 / 465 10 / 56

[035] Everolimus is identified by CAS number 159351-69-6.

[036] In preferred embodiments, the formulation is a pharmaceutical formulation.

[037] In some embodiments, the formulation is a lotion, a spray, a foam, an emulsion, a nanoemulsion, a gel, or a cream. In some embodiments, the formulation is a lotion. In the context of this descriptive report, a lotion is a low-viscosity topical preparation intended for application to the skin. A lotion has a lower viscosity than a cream or a gel due to its higher water content. In some embodiments, the lotion has a viscosity of < 8 Pa s (pascal-second), < 6 Pa s, < 5 Pa s, < 4 Pa ​​s, < 3 Pa s, < 1.0 Pa s, or < 0.5 Pa s.

[038] A person skilled in the art knows appropriate methods for determining viscosity. Preferably, viscosity is determined as described in the examples section.

[039] The formulation may be for topical, ophthalmic, or systemic use. In preferred modalities, the formulation is for topical use.

[040] In preferred embodiments of the formulations described in the present invention, the aqueous component comprises an aqueous phase or forms an aqueous phase.

[041] In preferred embodiments of the formulations described in the present invention, the carrier component comprises or consists of nanovesicles. The carrier component may also be referred to as the lipid phase of the nanoemulsion. Preferably, the active agent is dissolved in the lipid phase of the nanovesicles. In other words, the active agent is dissolved in the lipid phase of the nanoemulsion.

[042] The active agent may be present as a salt, hydrate or derivative. Petition 870250090034, dated 02 / 10 / 2025, pp. 397 / 465 11 / 56

[043] The small size of the nanovesicles and their high homogeneity give them advantageous properties that distinguish them from conventional emulsions: The nanoemulsions and formulations comprising nanoemulsions of the present invention are transparent. Additionally, the nanoemulsion and formulations comprising nanoemulsions of the present invention can carry active agents such as tacrolimus more efficiently and thus become increasingly important in the field of medicine and pharmacy.

[044] “Aging”, as used in the present invention, refers to the alteration, disintegration and / or degradation of the formulation, affecting chemical and physical stability during storage, particularly under stressful conditions. Such physical or chemical changes due to storage may include, but are not limited to, Ostwald ripening, flocculation, coalescence and / or breakage, which may lead to a change in vesicle size or polydispersity index.

[045] The inventors discovered that tacrolimus can surprisingly be dissolved, rather than suspended, in the aqueous formulations of the present invention containing nanoemulsion.

[046] The inventors have further discovered that the formulations of the present invention are surprisingly stable and resistant to aging. In particular, the formulations of the present invention are stable in terms of TC content and particle size and particle size distribution, even after storage, for example, for 24 months at 2 to 8 °C.

[047] In the context of this descriptive report, whenever a duration is described as “one month, two months, three months”, etc., this should include modalities in which the duration is “at least one month, at least two months, at least three months”, etc. Petition 870250090034, dated 02 / 10 / 2025, pp. 398 / 465 12 / 56

[048] As used in the present invention, a “nanovesicle emulsion” or a “nanoemulsion” is an oil-in-water dispersion (oil-in-water dispersion, oil-in-water emulsion, O / W emulsion). The nanoemulsion may be monophasic, transparent and / or slightly opalescent. The nanoemulsions of the present invention may be colloidal systems, which include dispersed nanovesicles comprising a lipid core surrounded by at least one surfactant or emulsifier monolayer. The nanoemulsions and formulations comprising the nanoemulsions of the present invention are characterized by an average particle or nanovesicle size of less than 500 nm, less than 200 nm or less than 100 nm. The nanoemulsions and formulations comprising the nanoemulsions of the present invention have a narrow (homogeneous) nanovesicle size distribution, for example, a nanovesicle size distribution characterized by a polydispersity index less than or equal to 0.4.

[049] As used in the present invention, “nanovesicle”, “nanovesicle”, “lipid vesicles”, “oil droplets”, “droplets” and “oil globules” are interchangeable and refer to small oil droplets in an oil-in-water emulsion. A lipid vesicle of an average size (see above, for example, below 500 nm, 200 nm, 100 nm) that is compiled from a monolayer of a surfactant and a lipid core. In the present invention, the nanovesicles may have a size less than or equal to 500 nm, or less than or equal to 300 nm, preferably in the range of 5 nm to 200 nm, more preferably in the range of 5 nm to 100 nm.

[050] As used in the present invention, the term “nanoparticle” or “nanoparticle” is distinguished from “nanovesicles” and refers to solid particles, which are not described in this invention. The formulation of the present invention may be a formulation that is essentially free of Petition 870250090034, dated 02 / 10 / 2025, pp. 399 / 465 13 / 56 nanoparticles. Essentially free of nanoparticles means that the formulation comprises less than or equal to 2% by weight, or less than or equal to 1% by weight of, or does not comprise nanoparticles. Nanoparticles are primarily solid inorganic lipids or solid polymeric particles that may have a size below 100 nm, below 200 nm, or below 500 nm. The size can be determined by the methods described in the present invention. For example, the formulation may be essentially free of nanoparticles with a diameter smaller than 100 nm, as determined by dynamic light scattering.

[051] As used in the present invention, topical use or topical treatment of the formulation of the invention describes an application to a specific location on the body, in particular the human body. This includes, but is not limited to, administration of the formulation to body surfaces such as the skin or mucous membranes. Topical use may be epicutaneous, meaning that the formulation is administered directly to the skin. In particular, topical use is a pharmaceutical use.

[052] As used in the present invention, systemic use or systemic treatment of the formulation of the invention describes an application in which the active agent is distributed throughout the body via the blood or lymphatic system, for example, after an injection or oral ingestion.

[053] As used in the present invention, the stability of a formulation comprising nanovesicles, as described in the present invention, includes, but is not limited to, physical and chemical stability. In particular, in the present invention, a formulation is stable if the integrity of the nanovesicles is considered stable. A measure known to those skilled in the art for describing the integrity of nanovesicles is size, as for example determined by dynamic light scattering, as described in Petition 870250090034, dated 02 / 10 / 2025, pages 400 / 465 14 / 56 present invention. The nanovesicles produced according to the invention may have a size below 100 nm, preferably below 50 nm, more preferably in the range of 20 nm to 30 nm, immediately after manufacture. For example, the formulation, as described in the present invention, is stable if the nanovesicles in the formulation of the present invention have a size (or diameter) less than or equal to 500 nm or less than or equal to 300 nm, preferably in the range of 5 nm to 200 nm, more preferably in the range of 5 nm to 100 nm.

[054] Stability may also refer to the absence of processes described above such as aging, leading to loss of functionality or pharmaceutical quality. The composition described in this invention is functional or pharmaceutically functional, provided that the vesicle size is less than or equal to 500 nm or less than or equal to 300 nm, preferably in the range of 5 nm to 200 nm, more preferably in the range of 5 nm to 100 nm.

[055] Furthermore, stability may refer to the stable content of the active agent, in particular tacrolimus. During storage, the content of the active agent is, for example, considered stable if at least 70%, at least 80% or at least 90% of the content of the active agent is still present when stored, for example, under stressful conditions, as described in the present invention.

[056] In one formulation of the invention, the content of the active agent may be (i) at least 90% after storage for 1 month at 40 °C, and / or (ii) at least 90% after storage for 3 or 6 months at 25 °C, and / or (iii) at least 90% after storage for 12, 18 or 24 months at 2 to 8 °C.

[057] In the present invention, the active agent of the invention can be stable for at least one month, at least 3 months, at least 6 months, at least Petition 870250090034, dated 02 / 10 / 2025, pp. 401 / 465 15 / 56 months, at least 12 months or at least 24 months, at 2 to 8 °C, or at about 5 °C.

[058] In the present invention, the formulation of the invention can be stable for at least one month, at least 3 months, at least 6 months, at least 9 months, at least 12 months or at least 24 months, at 2 to 25 °C, at 2 to 8 °C, at 15 to 25 °C, at 25 °C or at about 5 °C.

[059] The formulation of the present invention may have a nanovesicle size less than or equal to 500 nm, or less than or equal to 300 nm, or less than or equal to 200 nm, preferably in the range of 5 nm to 200 nm, more preferably in the range of 5 nm to 100 nm, when stored for 3 months at 40 °C.

[060] The formulation of the present invention may have a nanovesicle size less than or equal to 500 nm, or less than or equal to 300 nm, or less than or equal to 200 nm, preferably in the range of 5 nm to 100 nm, when stored for 24 months at 25 °C.

[061] Preferably, the active agent has a content greater than or equal to 80%, preferably greater than or equal to 85%, more preferably greater than or equal to 90% when (a) stored for one month, two months, three months, six months at 2 to 25 °C; or (b) stored for one month, two months, three months, six months, twelve months, eighteen months or twenty-four months at 2 to 8 °C.

[062] Preferably, the nanoemulsion comprises nanovesicles, wherein the nanovesicles have a size less than or equal to 500 nm, preferably less than or equal to 200 nm, more preferably in the range of 5 nm to 100 nm when (a) stored for one month, two months, three months, six months, twelve Petition 870250090034, dated 02 / 10 / 2025, pp. 402 / 465 (a) stored for 16 / 56 months, eighteen months or twenty-four months at 2 to 25 °C; or (b) stored for one month, two months, three months, six months, twelve months, eighteen months, twenty-four months, thirty months or thirty-six months at 2 to 8 °C.

[063] Preferably, the polydispersity index of the formulation is less than or equal to 0.4 when (a) stored for one month, two months, three months, six months, twelve months, eighteen months or twenty-four months at 25 °C. (b) stored for one month, two months, three months, six months, twelve months, eighteen months, twenty-four months, thirty months, thirty-six months at 2 to 8 °C.

[064] The size or diameter of the nanovesicles, as described in the present invention, can be expressed as the Z-average (also referred to as “z-average”). The size distribution of the nanovesicles can be characterized by the polydispersity index. These parameters are well known to those skilled in the art and are widely used in the art to characterize particles or vesicles in polymeric emulsions, suspensions and / or solutions.

[065] In the present invention, the size of nanovesicles (e.g., average z in nm) and / or the heterogeneity of nanovesicle formulations (characterized by the polydispersity index) can be determined by dynamic light scattering (also referred to as Photon Correlation Spectroscopy (PCS) or Quasi-Elastic Light Scattering (QELS)). Dynamic light scattering is well known in the art and well established for determining the size of nano or microparticles or vesicles in polymer emulsions, suspensions and / or solutions with a laser.

[066] In the formulation, as described in the present invention, the total aqueous component may be present in an amount of 50% to Petition 870250090034, dated 02 / 10 / 2025, pp. 403 / 465 17 / 56 99% w / w, based on the total weight of the nanoemulsion (a), preferably 70% to 95% (w / w), and more preferably 75% to 95% (w / w), or 80% to 95%.

[067] As used in the present invention, “weight for weight”, “weight / weight” or “w / w” means the weight concentration or mass concentration of a component in a formulation described in the present invention. The weight or mass of a component is expressed as a percentage of a reference formulation. For example, the weight or mass of a component may be expressed as a percentage of the total weight or mass of the formulation of the invention, or as a percentage of the total weight or mass of the nanoemulsion (a).

[068] The aqueous component may comprise at least one pH buffering agent. Any suitable buffering agent may be used. Suitable buffering agents are known to those skilled in the art. For example, at least one pH buffering agent may be selected from the group consisting of citrate, phosphate, acetate, and carbonate.

[069] The pH of the aqueous component may be in the range of 2 to 9. The pH of the aqueous component may also preferably be in the range of 2 to 6, such as 2, 3, 4, 5 or 6, more preferably in the range of 3 to 6, such as 3, 4, 5 or 6, or 3 to 5, such as 3, 4 or 5. In other embodiments, the pH of the aqueous component may also be in the range of 4 to 10, or 5 to 7, preferably around 7.4.

[070] In cases where the formulation is topical or oral, the pH of the formulation is 2 to 7, preferably 2 to 6, more preferably 3 to 5. In cases where the formulation is a parenteral or ophthalmic formulation, the pH of the formulation is 4 to 10, or 5 to 7, preferably around 7.4.

[071] In the formulation, as described in the present invention, at least one lipophilic component may be selected from triglycerides. Petition 870250090034, dated 02 / 10 / 2025, pp. 404 / 465 18 / 56 and mixtures thereof.

[072] Preferably, the at least one lipophilic component is a lipid, a synthetic oil, a vegetable oil and / or an animal oil. Suitable lipids according to the present invention are physiologically acceptable lipids, such as ceramide, mono-, di- and triacylglycerol (triglycerides). In particular, the at least one lipophilic component is a triglyceride, preferably a triglyceride comprising a C8-10 fatty acid, or a mixture thereof. More particularly, the at least one lipophilic component is a caprylic and / or capric triglyceride and / or a mixture thereof, particularly preferably Miglyol (such as Miglyol 812, available, for example, from IOI Oleochemical) or Myritol (such as Myritol 318, available, for example, from BASF). Suitable vegetable and animal oils include, for example, sunflower oil, soybean oil, peanut oil, rapeseed oil, fish oil, and / or cetacean oil.

[073] In the formulation, as described in the present invention, at least one lipophilic component may be present in an amount of 0.1% to 30% (w / w) based on the total weight of the nanoemulsion (a), preferably 0.25% to 15% (w / w), preferably 0.25% to 10% (w / w) and more preferably 0.5% to 8% (w / w) or 3% to 8% (w / w). It is also preferred that at least one lipophilic component be present in an amount of 10% to 30% (w / w) based on the total weight of the nanoemulsion (a), more preferably 15% to 30% or 10% to 20%.

[074] In the formulation, as described in this invention, at least one surfactant may be any suitable surfactant known to those skilled in the art.

[075] Surfactants, also referred to as surface-active agents or emulsifiers, are well known in the state of the art and include any Petition 870250090034, dated 02 / 10 / 2025, pp. 405 / 465 19 / 56 agents that bind oil and water in the composition to form an emulsion. They decrease the surface tension of two liquids and are amphiphilic. In emulsions, they are referred to as emulsifiers and coat the droplets, preventing coalescence. Emulsifiers can be described by their hydrophilic / lipophilic balance (HLB), which expresses their affinity for water or oil. Low HLB (e.g., HLB=1) refers to lipophilic emulsifiers and high (e.g., HLB=20) to hydrophilic emulsifiers. In general, lipophilic emulsifiers are used for water-in-oil emulsions and hydrophilic emulsifiers for oil-in-water emulsions. Skilled individuals will identify which emulsifiers or mixtures thereof are suitable for the preferred vehicles and the purpose of the composition. In certain emulsions, combinations of emulsifiers may be advantageous.

[076] A suitable membrane-forming surfactant is a phospholipid, a lysophospholipid, a ceramide and / or a mixture thereof. Preferably, the phospholipid is lecithin or cephalin from soy or chicken eggs. Preferably, at least one surfactant is a phospholipid, more preferably lecithin, most preferably soy lecithin.

[077] In the formulation, as described in the present invention, the phospholipid, in particular phosphatidylcholine, lysophospholipid, ceramide and / or a mixture thereof may be present in an amount of 0.1% to 10% (w / w), based on the total weight of the nanoemulsion(a), preferably 0.15% to 5% (w / w), and more preferably 0.2% to 3% (w / w) or 0.2% to 4% (w / w), most preferably 2.5% to 4% (w / w).

[078] Preferably, the lecithin has a phosphatidylcholine content of at least 80% by weight, more preferably at least 90% by weight and most preferably at least 94% by weight. The quality of the lecithin, i.e. its phosphatidylcholine content, plays a crucial role in the size of the Petition 870250090034, dated 02 / 10 / 2025, pages 406 / 465 20 / 56 vesicles of the nanoemulsion. The higher the phosphatidylcholine content of the lecithin, the smaller the size of the nanoemulsion vesicles.

[079] As O / W emulsion-forming surfactants, anionic, nonionic, cationic and / or amphoteric surfactants are suitable, as are block copolymers. Suitable anionic surfactants are soaps, alkylbenzene sulfonates, alkanes sulfonates, alkyl sulfates and / or alkyl ether sulfates. Suitable cationic surfactants are quaternary ammonium compounds, preferably having one or two hydrophobic groups (e.g., cetyltrimethylammonium bromide and cetyltrimethylammonium chloride) and / or salts of long-chain primary amines. A suitable amphoteric surfactant is N-(acylamidoalkyl)betaine, N-alkyl-e-aminopropionate, alkylammonium phosphate compounds and / or amine-N-oxide. A suitable copolymer building block, for example, is propylene oxide. In the present invention, a nonionic surfactant is particularly preferred as an O / W emulsion-forming surfactant.

[080] In preferred embodiments, the surfactant is a polyoxyethylene type surfactant. In the formulation as described in the present invention, at least one surfactant may be any polyoxyethylene type surfactant. A suitable nonionic surfactant may be selected from the group consisting of fatty alcohol polyglycol ether, alkylphenol polyglycol ether, alkylpolyglucoside, fatty acid glucamide, fatty acid polyglycol ether, ethylene oxide-propylene oxide block polymer, polyglycerol fatty acid ester, fatty acid alkanolamide and sorbitan fatty acid ester (ethoxylated) (sorbitan). A particularly preferred ethoxylated sorbitan fatty acid ester is polyoxyethylene sorbitan monooleate, more preferably Polysorbate 80.

[081] At least one surfactant, such as a type of surfactant Petition 870250090034, dated 02 / 10 / 2025, pp. 407 / 465 21 / 56 polyoxyethylene, may be present in an amount of 0.1% to 10% (w / w), based on the total weight of the nanoemulsion (a), more preferably 0.2% to 5% (w / w) and most preferably 1% to 5% (w / w) or 0.5% to 5% (w / w).

[082] The formulation of the invention may comprise at least one hydrophilic surfactant with an HLB of 9 to 17, more preferably 12 to 16, particularly polysorbate 80 to form a nanoemulsion.

[083] At least one surfactant may be a sugar-based surfactant. Sugar-based surfactants are a group of nonionic surfactants that use hydrophilic sugars to which hydrophobic tails are attached. A common substance in this class is n-dodecyl-D-maltoside, a member of the maltoside surfactants, so named because the sugar unit used is maltose. An example of a pyranoside surfactant is n-octyl-D-thioglucopyranoside. This class uses pyranose as the sugar unit. Examples of glycoside surfactants are octylglucoside, decylglucoside, and laurylglucoside. An example of a polysugar surfactant is digitonin.

[084] Another very important group of sugar-based surfactants are the Tween surfactants, the most notable being Tween 20 (also referred to in the present invention as Polysorbate 20) and Tween 80 (also referred to in the present invention as Polysorbate 80). These surfactants are based on sorbitan sugar, which is why they are commonly referred to as polysorbate surfactants. Three oligo(ethylene glycol) side groups of varying lengths are attached to the sugar, increasing the hydrophilicity of the main group. This structure forms the core of all Tween surfactants. They deviate at the hydrophobic tail, which is a fatty acid coupled via an ester to four oligo(ethylene glycol) tails. In Tween 20 this fatty acid is lauric acid; in Tween 80 it is oleic acid.

[085] In some embodiments, at least one surfactant is Petition 870250090034, dated 02 / 10 / 2025, pp. 408 / 465 22 / 56 selected from the group consisting of a phospholipid, in particular phosphatidylcholine, a lysophospholipid, a ceramide and / or a mixture thereof. In some embodiments, at least one surfactant is a polyoxyethylene type surfactant. In some embodiments, at least one surfactant is phosphatidylcholine. In some embodiments, the formulation comprises a phospholipid as a surfactant and a polyoxyethylene type surfactant. In some embodiments, the formulation comprises phosphatidylcholine as a surfactant and a polyoxyethylene type surfactant. In some embodiments, the formulation comprises phosphatidylcholine and polysorbate 80 as surfactants.

[086] In some embodiments, at least one surfactant is phosphatidylcholine.

[087] In some embodiments, the formulation comprises 0.5% to 5% (w / w), preferably 1% to 4% (w / w), more preferably 1.2% to 3.5% (w / w) of phosphatidylcholine.

[088] In some embodiments, the formulation comprises 0.1% to 10% (w / w), preferably 0.15% to 5% (w / w), more preferably 0.25% to 4.5% (w / w) of phosphatidylcholine.

[089] In preferred embodiments, at least one alcohol comprises at least three carbon atoms.

[090] In the formulation as described in the present invention, the at least one alcohol preferably has, independently, 3 to 5 (i.e., not more than 5) or 3 or 4 (i.e., not more than 4) carbon atoms. The at least one alcohol may be at least one monohydric alcohol. Particularly suitable alcohols having 5 carbon atoms are 1-pentanol and / or 4-methyl-2-pentanol. Suitable alcohols having 4 carbon atoms are 1-butyl alcohol, isobutyl alcohol (2-methyl-1-propanol), tert-butyl alcohol (2-methyl-2-propanol). Petition 870250090034, dated 02 / 10 / 2025, p. 409 / 465 23 / 56 propanol) and / or sec-butyl alcohol (2-butanol). The alcohol is not propylene glycol.

[091] Preferably, at least one alcohol has 3 carbon atoms, that is, it is selected from the group consisting of 1-propanol or 2-propanol (isopropyl alcohol) and mixtures thereof. A preferred alcohol is 2-propanol.

[092] In the formulation, as described in the present invention, the alcohol may be present in an amount of 0.1% to 10% w / w based on the total weight of the nanoemulsion (a), preferably 0.5% to 5% (w / w) and more preferably 1% to 2% (w / w).

[093] The formulation, as described in the present invention, may comprise a gelling agent. Any suitable gelling agent may be used. Suitable gelling agents and mixtures thereof are known to those skilled in the art. In the formulation, as described in the present invention, the gelling agent may be selected from the group consisting of poloxamer, xanthan gum, bentonite, sodium carboxymethylcellulose, hydroxymethylcellulose, carbomer, hydroxypropylcellulose, gellan gum, guar gum, pectin, poly(ethylene) oxide, polycarbophil, alginate, tragacanth, povidone, gelatin and mixtures thereof.

[094] It is preferable that the gelling agent be selected from poloxamer, xanthan gum and / or mixtures thereof.

[095] It is also preferable that the gelling agent be xanthan (xanthan gum).

[096] It is also preferable that the gelling agent be a poloxamer.

[097] Poloxamers are nonionic triblock copolymers composed of a central hydrophobic polyoxypropylene (poly(propylene oxide)) chain flanked by two hydrophilic polyoxyethylene (poly(oxide)) chains. Petition 870250090034, dated 02 / 10 / 2025, pp. 410 / 465 24 / 56 ethylene). Poloxamer 407 and Poloxamer 188 are commercially available. Poloxamer 407 may have an average molecular weight of about 12,600 Daltons. Poloxamer 188 may have an average molecular weight of about 8,400 Daltons. In the formulation as described in the present invention, a preferred poloxamer is Poloxamer 407.

[098] In the formulation, as described in the present invention, the gelling agent may be present in an amount of 0.1% to 10% (w / w), based on the total weight of the formulation, preferably 0.25% to 5% (w / w), and more preferably 0.5% to 4% (w / w) or 1% to 4% (w / w).

[099] The formulation, as described in the present invention, may comprise one or more preservatives. Any suitable preservative or a mixture thereof may be used. Suitable preservatives are known to those skilled in the art. The preservative may be selected from benzoate, tocopherol or derivatives and any mixtures thereof, citric acid, EDTA, potassium sorbate, vitamin C and / or derivatives and any mixtures thereof, wherein the preservative is preferably sodium benzoate. Suitable aqueous mixtures of sodium benzoate and potassium sorbate are commercially available, for example, preservative Euxyl™ K 712 (Ashland). These components may form part of the aqueous component and / or the nanovesicles.

[100] The preservative may be present in the formulation, as described in the present invention, in an amount of 0.01% to 10% (w / w), 0.01% to 7% (w / w), 0.01% to 5% (w / w), 0.01% to 3% (w / w) based on the total weight of the formulation, preferably 0.2% to 2% (w / w) or 0.1% to 2% (w / w) and more preferably 0.2% to 1.5% (w / w).

[101] The formulation of the present invention may be a gel formulation. As used in the present invention, a “gel” is a colloidal material. Petition 870250090034, dated 02 / 10 / 2025, pp. 411 / 465 25 / 56 a two-phase elastic material, consisting of a dispersed liquid incorporated into the solid phase, frequently consisting of a gelling agent. Suitable gelling agents, such as xanthan gum, are described in the present invention.

[102] The formulation of the present invention may be supplied in a container or dispenser. Suitable dispensers and containers are known to those skilled in the art. For example, the dispenser may be a compression tube comprising the formulation as described in the present invention. The compression tube may contain the gel formulation as described in the present invention. The dispenser may also be a metered dose dispenser or a foam dispenser or a spray dispenser.

[103] The container or dispenser may comprise a propellant, wherein the propellant is supplied to pressurize the container or dispenser. Any suitable propellant may be used. Suitable propellants and mixtures thereof are known to those skilled in the art. Preferably, the propellant is selected from propane, isobutane, n-butane and mixtures thereof.

[104] In some embodiments, the formulation comprises a propellant and is contained in a pressurized container.

[105] Surprisingly, the formulation of the present invention comprised in a pressurized container has a high foaming capacity and forms a stable foam (long collapse time) when released from the pressurized container, even in the absence of a fatty alcohol or other foaming aid.

[106] Surprisingly, the formulation of the present invention contained in a pressurized container is highly stable with respect to nanovesicle size, even in the absence of a gelling agent or Petition 870250090034, dated 02 / 10 / 2025, pp. 412 / 465 26 / 56 in the presence of only low concentrations of a gelling agent.

[107] Surprisingly, the formulation of the present invention contained in a pressurized container is highly stable with respect to the concentration of the active agent, even in the absence of a gelling agent or in the presence of only low concentrations of a gelling agent.

[108] The formulation of the present invention comprised in a pressurized container is surprisingly resistant to aging under stress conditions with respect to API content and nanoemulsion vesicle size, compared to a formulation. In the context of this descriptive report, the term petrolatum relates to a semi-solid mixture of hydrocarbons derived from petroleum distillation. The hydrocarbons that make up petrolatum mainly comprise at least 25 carbon atoms. The CAS number of petrolatum is 8009-03-8. Preferably, the formulation of the first aspect of the invention essentially does not comprise any petrolatum.

[109] The formulation of the present invention can be prepared as a foamable formulation.

[110] The formulation of the present invention can be prepared as a pressurized formulation, wherein a propellant is provided to pressurize the formulation. Any propellant as described in the present invention can be used.

[111] The formulation of the present invention can be prepared as a pressurized and foamable formulation, wherein a propellant is provided to pressurize the formulation. Any propellant as described in the present invention can be used.

[112] The formulation of the invention can be supplied in a foam dispenser, as described in the present invention. The foam dispenser Petition 870250090034, dated 02 / 10 / 2025, pp. 413 / 465 27 / 56 comprises a container, wherein said container comprises the formulation as described in the present invention, and a propellant. The propellant is provided to pressurize the foam dispenser. Any suitable propellant may be used as described in the present invention. A foam generating device is mounted in the container. In particular, the formulation is prepared as a foamable formulation. Said foam generating device may comprise a valve for releasing and dispensing the formulation and a push button to actuate the valve. By actuating the push button, the formulation may be released and form a foam. Suitable dispensers are known to those skilled in the art.

[113] The formulation of the invention may be provided in a spray dispenser, as described in the present invention. The spray dispenser comprises a container, wherein said container comprises the formulation, as described in the present invention, and a propellant. The propellant is provided to pressurize the spray dispenser. Any suitable propellant may be used, as described in the present invention. A spray generating device is mounted on the container. In particular, the formulation is prepared as a sprayable formulation.

[114] The invention also provides a foam, comprising the formulation of the present invention, as described in the present invention.

[115] In preferred embodiments, the active agent in the formulation of the invention is a highly lipophilic active agent, such as tacrolimus.

[116] The active agent may be present in an amount of 0.001% to 25%, 20%, 15%, 10% or 5% w / w, based on the total weight of the formulation. In particular, the active agent may be present in an amount of 0.001% to 10% w / w, based on the total weight of the formulation, 0.005% to 5%, or 0.01% to 0.5% w / w, based on the total weight of the formulation. Petition 870250090034, dated 02 / 10 / 2025, pp. 414 / 465 28 / 56

[117] In some embodiments, the formulation is a topical formulation and the active agent is present in an amount of 0.001% to 0.5% w / w, preferably 0.005% to 0.2% w / w, more preferably 0.01% to 0.1% w / w, based on the total weight of the formulation.

[118] In some embodiments, the formulation is a systemic formulation and the active agent is present in an amount of 0.005% to 25% w / w, preferably 0.01% to 10% w / w, more preferably 0.01% to 5% w / w, even more preferably 0.01% to 2% w / w, based on the total weight of the formulation. The systemic formulation may be an injection formulation or an oral formulation. In some embodiments, the systemic formulation is diluted with a pharmaceutically acceptable buffer before injection, resulting in a final concentration of 0.01% to 1% w / w based on the total weight of the formulation.

[119] In some embodiments, the formulation is an ophthalmic formulation, in particular eye drops, and the active agent is present in an amount of 0.001% to 0.5% w / w, preferably 0.005% to 0.2% w / w, more preferably 0.01% to 0.1% w / w, based on the total weight of the formulation.

[120] In preferred embodiments, the formulation of the invention essentially does not comprise any fatty alcohol.

[121] In the context of this descriptive report, the expression “does not essentially comprise any” “is essentially free of” specifies that the formulation is free of a compound or comprises less than 0.5% (w / w), less than 0.4% (w / w), less than 0.3% (w / w), less than 0.2% (w / w), less than 0.1% (w / w), less than 0.08% (w / w), less than 0.07% (w / w), less than 0.06% (w / w), less than 0.05% (w / w), less than 0.04% (w / w), less than 0.03% (w / w), less than 0.02% (w / w) or less than 0.01% (w / w) of a compound based on the total weight of the formulation. Petition 870250090034, dated 02 / 10 / 2025, pp. 415 / 465 29 / 56

[122] Fatty alcohols have been described as acting as foaming agents. They are used in prior art formulations, in particular in foamable formulations.

[123] Preferably, the formulation comprises essentially no fatty alcohols and essentially no fatty acids.

[124] In the context of this descriptive report, the term “fatty alcohol” relates to alcohols having at least 6 carbon atoms, typically 6 to 28 carbon atoms. Fatty alcohols may be saturated or unsaturated and branched or unbranched. Fatty alcohols are typically linear chain primary alcohols. The term fatty alcohol, as used in the present invention, relates to fatty alcohols in their standalone form and does not include esters comprising fatty alcohols.

[125] In the context of this descriptive report, the term fatty acids relates to carboxylic acids with an aliphatic chain of at least 6 carbon atoms, typically 6 to 28 carbon atoms. Fatty acids can be saturated or unsaturated and branched or unbranched. Most naturally occurring fatty acids have an unbranched chain of carbon atoms. The term fatty acid, as used in the present invention, relates to fatty acids in their standalone form and does not include esters comprising fatty acids.

[126] In preferred embodiments, the formulation is free of a fatty alcohol (in its standalone form, or in other words, as an isolated molecule) or comprises less than 0.5% (w / w), less than 0.4% (w / w), less than 0.3% (w / w), less than 0.2% (w / w), less than 0.1% (w / w), less than 0.08% (w / w), less than 0.07% (w / w), less than 0.06% (w / w), less than 0.05% (w / w), less than 0.04% (w / w), less than 0.03% (w / w), less than 0.02% (w / w) or less than 0.01% (w / w) of a fatty alcohol (in its standalone form, or in other words, as an isolated molecule) Petition 870250090034, dated 02 / 10 / 2025, pp. 416 / 465 30 / 56 isolated molecule) based on the total weight of the formulation.

[127] In particular, in preferred embodiments, the formulation is free of a fatty acid and a fatty alcohol (in their standalone form, or in other words, as isolated molecules) or comprises less than 0.5% (w / w), less than 0.4% (w / w), less than 0.3% (w / w), less than 0.2% (w / w), less than 0.1% (w / w), less than 0.08% (w / w), less than 0.07% (w / w), less than 0.06% (w / w), less than 0.05% (w / w), less than 0.04% (w / w), less than 0.03% (w / w), less than 0.02% (w / w) or less than 0.01% (w / w) of a fatty acid and a fatty alcohol (in their standalone form, or in other words, as isolated molecules) based on the total weight of the formulation.

[128] More preferably, the formulation essentially does not comprise any foaming aid. In particular, the formulation may be foaming aid-free or comprise less than 0.5% (w / w), less than 0.4% (w / w), less than 0.3% (w / w), less than 0.2% (w / w), less than 0.1% (w / w), less than 0.08% (w / w), less than 0.07% (w / w), less than 0.06% (w / w), less than 0.05% (w / w), less than 0.04% (w / w), less than 0.03% (w / w), less than 0.02% (w / w) or less than 0.01% (w / w) of a foaming aid based on the total weight of the formulation.

[129] In the context of this descriptive report, the term “foaming agent” relates to compounds capable of increasing the foaming capacity of a formulation and / or stabilizing a foam. In particular, the term “foaming agent” relates to fatty acids and fatty alcohols having at least 6 carbon atoms. In particular, the formulation of the present invention may comprise or consist of: (a) a nanoemulsion comprising (i) an aqueous component, present in an amount of 70% to 95% w / w, based on the total weight of the nanoemulsion (a). Petition 870250090034, dated 02 / 10 / 2025, pp. 417 / 465 31 / 56 (ii) nanovesicles, comprising (1) 1 to 5% of at least one phospholipid, based on the total weight of the nanoemulsion (a); (2) 2 to 10% of at least one polyoxyethylene type surfactant, based on the total weight of the nanoemulsion (a); (3) 1 to 5% C3 to C5 alcohol, based on the total weight of the nanoemulsion (a); and (4) 2 to 10% triglycerides, based on the total weight of the nanoemulsion (a); (b) 0.01 to 1% of a highly lipophilic macrolide lactone, based on the total weight of the formulation; and (c) 0.1 to 10%, preferably 0.1 to 5%, more preferably 0.1 to 2% of at least one preservative, based on the total weight of the formulation; (d) optionally, 0.1 to 30%, preferably 0.1 to 20%, more preferably 0.1 to 10% of at least one gelling agent, based on the total weight of the formulation; wherein the formulation preferably has a pH of 2 to 7. Preferably, the formulation may comprise two surfactants, most preferably soy lecithin and Polysorbate 80.

[130] In this formulation, at least one phospholipid, at least one polyoxyethylene type surfactant, the C3 to C5 alcohol, the triglycerides and at least one preservative can be selected independently according to the disclosure described in the present invention.

[131] In particular, the formulation of the present invention may comprise, consist essentially of, or consist of: (a) a nanovesicle emulsion comprising (i) an aqueous component, present in an amount of 70% to Petition 870250090034, dated 02 / 10 / 2025, pp. 418 / 465 32 / 56 95% w / w, based on the total weight of the nanoemulsion (a). (ii) nanovesicles, comprising (1) 1 to 5% of at least one phospholipid, based on the total weight of the nanoemulsion (a); (2) 2 to 10% of at least one polyoxyethylene type surfactant, based on the total weight of the nanoemulsion (a); (3) 1 to 5% C3 to C5 alcohol, based on the total weight of the nanoemulsion (a); and (4) 2 to 10% triglycerides, based on the total weight of the nanoemulsion (a); (b) 0.01 to 1% of a highly lipophilic macrolide lactone, preferably tacrolimus, based on the total weight of the formulation; (c) 0.1 to 10%, preferably 0.1 to 5%, more preferably 0.1 to 2% of at least one preservative, based on the total weight of the formulation; (d) optionally, 0.1 to 30%, preferably 0.1 to 20%, more preferably 0.1 to 10% of at least one gelling agent, based on the total weight of the formulation; where the formulation preferably has a pH of 2 to 7.

[132] Preferably, the formulation may comprise two surfactants, most preferably soy lecithin and Polysorbate 80.

[133] In this formulation, at least one phospholipid, at least one polyoxyethylene type surfactant, C3 to C5 alcohol, triglycerides and at least one preservative may be independently selected according to the disclosure described in the present invention. This formulation may further comprise components selected from EDTA, α-tocopheryl acetate and citric acid. These components may form part of the aqueous component and / or nanovesicles. Petition 870250090034, dated 02 / 10 / 2025, pp. 419 / 465 33 / 56

[134] In preferred embodiments, the formulation of the present invention may comprise or consist of: (a) a nanoemulsion comprising (i) an aqueous component, present in an amount of 70 to 95% w / w, based on the total weight of the nanoemulsion (a). (ii) nanovesicles, comprising (1) 1 to 5% of at least one phospholipid, based on the total weight of the nanoemulsion (a); (2) 2 to 10% of at least one polyoxyethylene type surfactant, based on the total weight of the nanoemulsion (a); (3) 1 to 5% C3 to C5 alcohol, based on the total weight of the nanoemulsion (a); and (4) 2 to 10% triglycerides, based on the total weight of the nanoemulsion (a); (b) 0.01 to 1% of a highly lipophilic, pharmaceutically acceptable macrolide lactone, preferably a calcineurin inhibitor, such as tacrolimus, based on the total weight of the formulation; (c) optionally 0.5 to 6% of at least one gelling agent, based on the total weight of the formulation; (d) optionally 0.1 to 2% of at least one preservative, based on the total weight of the formulation; and (e) a propellant, wherein the formulation is comprised in a pressurized container and wherein the propellant is supplied to pressurize the container to provide a pressurized formulation.

[135] The present invention also relates to a nanovesicle, comprising, consisting essentially of, or consisting of Petition 870250090034, dated 02 / 10 / 2025, pp. 420 / 465 34 / 56 (i) 16 to 19% w / w soy lecithin, (ii) 32 to 36% w / w Polysorbate 80, (iii) 32 to 36% w / w caprylic / capric triglycerides and (iv) 12 to 16% w / w isopropyl alcohol.

[136] A preferred nanovesicle of the invention comprises, consists essentially of, or consists of (i) 17% w / w soy lecithin (ii) 34% w / w Polysorbate 80 (iii) 35% w / w caprylic / capric triglycerides (iv) 14% w / w isopropyl alcohol.

[137] The present invention also relates to a nanoemulsion comprising, consisting essentially of or comprising (a) 1.6 to 3.6% w / w of soy lecithin (b) 3.3 to 6.9% w / w of Polysorbate 80 (c) 3.3 to 7.0% w / w of caprylic / capric triglycerides (d) 1.3 to 2.9% w / w of isopropyl alcohol (e) aqueous phosphate buffer, for example aqueous phosphate buffer of 5 to 100 mM, pH 2 to 8, preferably pH 2 to 7, more preferably pH 2 to 5, up to 100%.

[138] A preferred nanoemulsion of the present invention comprises, consists essentially of, or consists of: (a) 1.7% w / w soy lecithin (b) 3.4% w / w Polysorbate 80 (c) 3.5% w / w caprylic / capric triglycerides (d) 1.4% w / w isopropyl alcohol (e) aqueous phosphate buffer (5 mM to 100 mM, preferably 10 mM to 50 mM), pH 6, ad 100%. Petition 870250090034, dated 02 / 10 / 2025, pp. 421 / 465 35 / 56

[139] This nanoemulsion is designated “BF200” in the present invention. The BF200 nanoemulsion can be obtained by contacting a mixture of ingredients (a)-(d) in a total amount of 10% w / w 90% w / w of an aqueous phosphate buffer at 10 mM, pH 6, under conditions that allow the formation of a nanoemulsion, thus forming the nanoemulsion. An exemplary method for manufacturing the BF200 formulation is described in Example 1.

[140] Another preferred nanoemulsion of the present invention comprises, consists essentially of, or consists of: (a) 2 to 3% w / w soy lecithin (b) 4.5 to 5.5% w / w Polysorbate 80 (c) 4.5 to 5.5% w / w caprylic / capric triglycerides (d) 2 to 3% w / w isopropyl alcohol (e) 10 mM aqueous phosphate buffer, pH 6 or less, ad 100%.

[141] This nanoemulsion is designated “BF215” in the present invention. The BF215 nanoemulsion can be obtained by contacting a mixture of ingredients (a)-(d) in a total amount of 15% w / w 85% w / w of an aqueous phosphate buffer at 10 mM, pH 6, under conditions that allow the formation of a nanoemulsion, thus forming the nanoemulsion. An exemplary method for manufacturing the BF215 formulation is described in Example 1.

[142] Yet another preferred nanoemulsion of the present invention comprises, consists essentially of, or consists of: (a) 3 to 4% w / w soy lecithin (b) 6 to 7% w / w Polysorbate 80 (c) 6 to 8% w / w caprylic / capric triglycerides (d) 2 to 4% w / w isopropyl alcohol (e) 10 mM aqueous phosphate buffer, pH 6, ad 100%.

[143] This nanoemulsion is referred to as “BF220” in the present invention. A Petition 870250090034, dated 02 / 10 / 2025, pp. 422 / 465 36 / 56 Nanoemulsion BF220 can be obtained by contacting a mixture of ingredients (a)-(d) in a total amount of 20% w / w 80% w / w of an aqueous phosphate buffer at 10 mM, pH 6, under conditions that allow the formation of a nanoemulsion, thus forming the nanoemulsion. An exemplary method for manufacturing the BF220 formulation is described in Example 1.

[144] All definitions and embodiments described for the first aspect are also provided for all other aspects described in the present invention, where applicable.

[145] Yet another aspect of the present invention relates to the formulations, as described in the present invention, for use in medicine.

[146] Yet another aspect of the present invention relates to the formulations as described in the present invention for use in medicine.

[147] Yet another aspect of the present invention relates to the formulations, as described in the present invention, for use in a method of treating and / or preventing a dermatological, ophthalmic or autoimmune disease or condition, or for preventing organ rejection after transplantation.

[148] The dermatological disease or condition to be treated with the formulation, as described in the present invention, may include, but is not limited to, diseases or conditions of the skin, skin appendages or mucous membranes.

[149] The dermatological disease or condition to be treated with the formulation, as described in the present invention, may be selected from the group consisting of inflammatory, neoplastic, proliferative, infectious and / or autoimmune diseases or conditions, and / or the cutaneous manifestation thereof, and / or diseases associated with single lesions or fields of lesions, neoplastic, proliferative and / or inflammatory changes.

[150] The inflammatory dermatological disease or condition to be treated with Petition 870250090034, dated 02 / 10 / 2025, pp. 423 / 465 37 / 56 The formulation, as described in the present invention, may be selected from the group consisting of dermatitis, contact dermatitis, acne, atopic dermatitis, eczema, pustular dermatitis, seborrheic dermatitis, perioral dermatitis, chronic wound, urticaria, skin ulcer, rosacea, rash, drug eruptions, toxic epidermal necrolysis; erythema multiforme, erythema nodosum, granuloma annulare and other cutaneous manifestations of inflammation.

[151] The dermatological disease or condition may be an autoimmune dermatological disease or condition. The autoimmune dermatological disease or condition, or the cutaneous manifestation of the autoimmune condition to be treated with the formulation, as described in the present invention, may be selected from the group consisting of psoriasis, pemphigus, systemic lupus erythematosus, lichen planus, morphea, scleroderma, epidermolysis bullosa, dermatomyositis, graft-versus-host syndrome.

[152] Additional diseases or conditions to be treated with the formulation, as described in the present invention, may be selected from the group consisting of organ rejections after organ transplants (such as heart, kidney, liver and lung transplants).

[153] The ophthalmic disease or condition to be treated with the formulation, as described in the present invention, may be selected from the group consisting of keratoconjunctivitis (AKC), vernal keratoconjunctivitis (VKC), dry eye, endothelial corneal rejection after corneal transplantation.

[154] Yet another aspect of the present invention is a method for preparing the formulation, as described in the present invention, comprising the following steps: (a) mixing at least one lipophilic component, at least one surfactant and at least one alcohol having at least three carbon atoms, Petition 870250090034, dated 02 / 10 / 2025, pp. 424 / 465 38 / 56 (b) mix an active agent into the lipid phase and (c) place the mixture obtained in step (a) in contact with an aqueous component, under conditions that allow the formation of a nanoemulsion.

[155] In step (b), the conditions that allow the formation of the nanoemulsion may include mixing both phases at an appropriate temperature and agitation so as to form nanovesicles. The person skilled in the art knows the appropriate temperature and agitation conditions. A vesicle size less than or equal to 500 nm or less than or equal to 300 nm can be obtained, preferably in the range of 5 nm to 200 nm, more preferably in the range of 5 nm to 100 nm. In particular, the nanoemulsion of the present invention according to step (b) can be prepared without the use of high-energy methods, which are well known in the state of the art. The high-energy method includes high-pressure homogenization, microfluidization and ultrasonication (Prev Nutr Food Sci. September 2019; 24 (3): 225-234).

[156] The method comprises a step of adding a lipophilic active agent to the lipid phase until complete solution. Preferably, the active agent is tacrolimus.

[157] The method for preparing the pharmaceutical formulation of the present invention may further comprise: (i) add a gelling agent and / or (ii) add a preservative.

[158] Yet another aspect of the present invention is a dispensing product or container product, comprising the formulation as described in the present invention. In the dispensing product or container product, the formulation of the present invention is provided in a container or dispenser. Suitable dispensers and containers are known to those skilled in the art. For example, the dispenser may be a tube of Petition 870250090034, dated 02 / 10 / 2025, pp. 425 / 465 39 / 56 compression, comprising the formulation as described in the present invention. The compression tube may contain the gel formulation as described in the present invention. The dispenser may also be a metered dose dispenser, a foam dispenser, or a spray dispenser.

[159] The foam container or dispenser or spray dispenser may comprise a propellant, wherein the propellant is provided for pressurizing the foam container or dispenser or spray dispenser. Any suitable propellant may be used. Suitable propellants and mixtures thereof are known to those skilled in the art. Preferably, the propellant is selected from propane, isobutane, n-butane and mixtures thereof.

[160] The dispensing product may be a foam dispenser or a spray dispenser, comprising a foam dispenser or a spray dispenser, as described in the present invention, the foam dispenser or spray dispenser comprising a container, wherein said container comprises the formulation, as described in the present invention, and a propellant. The propellant is provided to pressurize the foam dispenser or the spray dispenser. Any suitable propellant may be used, as described in the present invention. A foam generating device or spray generating device is mounted on the container. In particular, the formulation is prepared as a foamable formulation.

[161] Yet another aspect of the present invention is the use of the formulation of the present invention, as described in the present invention, for the manufacture of a medicament for topical or systemic treatment and / or prevention of a dermatological, ophthalmic or autoimmune disease or condition in an individual or for the prevention of organ rejection after transplantation. Petition 870250090034, dated 02 / 10 / 2025, pp. 426 / 465 40 / 56

[162] Yet another aspect of the present invention is a method of treating and / or preventing a dermatological disease or condition in an individual, said method comprising administering to the individual a pharmaceutically effective amount of the formulation as described in the present invention. In particular, the dermatological disease is a dermatological disease or condition as described in the present invention.

[163] The invention also belongs to the following items: 1. A formulation comprising (a) a nanoemulsion, said emulsion comprising: (i) at least one aqueous component; (ii) a carrier component comprising: (1) at least one lipophilic component, (2) at least one surfactant and (3) at least one alcohol; and (b) an active agent, wherein the active agent is a highly lipophilic macrolide lactone, preferably tacrolimus. 5. The formulation of any of the above items, wherein at least one alcohol comprises at least three carbon atoms, preferably 3, 4 or 5 carbon atoms. 7. The formulation of any of the preceding items, in which the active agent is present in an amount of 0.001% to 5% (w / w), preferably 0.005% to 1% (w / w), more preferably 0.01% to 1.0% (w / w), based on the total weight of the formulation. 12. The formulation of any of the above items, where the formulation is a pharmaceutical formulation. 13. The drafting of any of the above items, where the drafting is for topical or oral use. Petition 870250090034, dated 02 / 10 / 2025, pp. 427 / 465 41 / 56 14. The formulation of any of the above items, in which the active agent is dissolved in the carrier component. 15. The wording of any of the above items, where the active agent has a logP value of 3 or higher. 16. The formulation of any of the preceding items, wherein the nanoemulsion comprises nanovesicles, wherein the nanovesicles have a size less than or equal to 500 nm, preferably in the range of 5 nm to 200 nm, preferably from 5 nm to 100 nm when stored for one month, two months or three months at 40 °C. 17. The formulation of any of the preceding items, wherein the nanoemulsion comprises nanovesicles, wherein the nanovesicles have a size less than or equal to 500 nm, preferably in the range of 5 nm to 200 nm, more preferably from 5 nm to 100 nm when stored for six months, twelve months, eighteen months or 24 months at 25 °C. 18. The formulation of any of the preceding items, in which the aqueous component is present in an amount of 50% to 99% w / w, based on the total weight of the nanoemulsion (a), preferably 70% to 95% (w / w), and more preferably 80% to 95% (w / w). 19. The formulation of any of the above items, in which the aqueous component comprises at least one pH buffering agent. 20. The formulation of item 19, in which the pH buffering agent is selected from the group consisting of citrate, phosphate, acetate, and carbonate. 21. The topical or oral formulation of any of the above items, having a pH of 2 to 7. 22. The parenteral or ophthalmic formulation of any of the above items, having a pH of 5 to 9. Petition 870250090034, dated 02 / 10 / 2025, pp. 428 / 465 42 / 56 23. The formulation of any of the preceding items, in which at least one lipophilic component is selected from triglycerides and mixtures thereof. 24. The formulation of any of the above items, in which at least one lipophilic component comprises a caprylic and / or capric triglyceride or a mixture thereof. 25. The formulation of any of the preceding items, in which at least one lipophilic component is present in an amount of 0.1% to 30% (w / w), based on the total weight of the nanoemulsion (a), preferably 0.25% to 10% (w / w), and more preferably 0.5% to 8% (w / w) or 3% to 8% (w / w). 26. The formulation of any of the above items, wherein at least one surfactant comprises (a) a phospholipid, a lysophospholipid, a ceramide and / or a mixture thereof, and / or (b) a polyoxyethylene type surfactant. 27. The formulation of any of the above items, in which at least one surfactant comprises lecithin, preferably soy lecithin. 28. The nanoemulsion of item 27, in which the lecithin has a phosphatidylcholine content of at least 80% by weight. 29. The formulation of any of items 26 to 28, in which the phospholipid, lysophospholipid, ceramide and / or mixture thereof is present in an amount of 0.1% to 10% (w / w), based on the total weight of the nanoemulsion (a), preferably 0.15% to 5% (w / w) and more preferably 0.2% to 3% (w / w). 30. The formulation of any of items 26 to 29, in which the polyoxyethylene type surfactant comprises Polysorbate 80. 31. The wording of any of items 26 to 30, in which the surfactant of Petition 870250090034, dated 02 / 10 / 2025, pp. 429 / 465 43 / 56 type polyoxyethylene is present in an amount of 0.1% to 10% (w / w), based on the total weight of the nanoemulsion (a), more preferably from 0.2% to 5% (w / w) and most preferably from 0.5% to 5% (w / w). 32. The formulation of any of the above items, in which at least one alcohol is selected from the group consisting of 1-propanol or 2-propanol and a mixture thereof. 33. The formulation of any of the preceding items, in which at least one alcohol is present in an amount of 0.1% to 10% (w / w), based on the total weight of the nanoemulsion (a), preferably 0.5% to 5% (w / w) and more preferably 1% to 2% (w / w). 34. The formulation of any of the above items, comprising at least one gelling agent. 35. The formulation of any of the preceding items, wherein the gelling agent is selected from the group consisting of poloxamer, xanthan gum, bentonite, sodium carboxymethylcellulose, hydroxymethylcellulose, carbomer, hydroxypropylcellulose, gellan gum, guar gum, pectin, poly(ethylene) oxide, polycarbophil, alginate, tragacanth, povidone, gelatin, and mixtures thereof. 36. The formulation of item 34 or 35, in which the gelling agent is selected from poloxamer, xanthan gum and / or mixtures thereof. 37. The formulation of any of the preceding items, in which the gelling agent is present in an amount of 0.1% to 10% (w / w), based on the total weight of the formulation, preferably 0.25% to 5% (w / w), and more preferably 1% to 4% (w / w). 38. The formulation of any of the preceding items, additionally comprising at least one preservative. 39. The formulation in item 38, where the preservative is benzoate, Petition 870250090034, dated 02 / 10 / 2025, pp. 430 / 465 44 / 56 preferably sodium benzoate. 40. The formulation of item 38 or 39, in which the preservative is present in an amount of 0.01% to 3% w / w, based on the total weight of the formulation, preferably 0.2% to 2% (w / w), and more preferably 0.2% to 1.5% (w / w). 41. The formulation of any of the above items, which is essentially paraben-free. 42. The formulation of any of the preceding items, characterized by a polydispersity index less than or equal to 0.8, where the polydispersity index is determined by dynamic light scattering. 43. The formulation of any of items 1 to 42, comprising: (a) a nanovesicle emulsion comprising (i) an aqueous component, present in an amount of 70% to 95% w / w, based on the total weight of the nanoemulsion (a).

[164] (ii) nanovesicles, comprising (1) 1 to 5% of at least one phospholipid, based on the total weight of the nanoemulsion (a); (2) 2 to 10% of at least one polyoxyethylene type surfactant, based on the total weight of the nanoemulsion (a); (3) 1 to 5% C3 to C5 alcohol, based on the total weight of the nanoemulsion (a); and (4) 2 to 10% triglycerides, based on the total weight of the nanoemulsion (a); (b) 0.01 to 1% of a highly lipophilic macrolide lactone or a combination thereof, based on the total weight of the formulation; (c) 0.1 to 10% of at least one preservative, based on the total weight of the formulation and where the formulation preferably has a pH of 2 to 7. Petition 870250090034, dated 02 / 10 / 2025, pp. 431 / 465 45 / 56 44. The formulation of any of the preceding items, supplied in a container, wherein the container further comprises a propellant, and wherein the propellant is supplied to pressurize the container and / or to pressurize the formulation in the container. 45. The formulation of any of the above items, for use in medicine. 46. ​​The formulation of any of the above items, for use in a topical or systemic treatment and / or prevention of a dermatological, ophthalmic, or autoimmune disease or condition in an individual, or for the prevention of organ rejection after transplantation. 47. The formulation for use of item 46, in which the ophthalmic disease is selected from the group consisting of keratoconjunctivitis (AKC), vernal keratoconjunctivitis (VKC), dry eye, endothelial rejection of the cornea after corneal transplantation. 48. The formulation for use in item 46, where the dermatological disease or condition includes diseases or conditions of the skin, skin appendages, or mucous membranes. 49. The formulation for use of any of items 46 or 48, wherein the dermatological disease or condition is selected from the group consisting of inflammatory, neoplastic, proliferative, infectious and / or autoimmune diseases or conditions, and / or the cutaneous manifestation thereof, and / or diseases associated with single lesions or fields of lesions, neoplastic, proliferative and / or inflammatory changes. 50. The formulation for use in item 49, where the inflammatory dermatological disease or condition is selected from the group consisting of dermatitis, contact dermatitis, acne, atopic dermatitis, eczema, pustular dermatitis, seborrheic dermatitis, perioral dermatitis, chronic wound, urticaria, skin ulcer, rosacea, rash, drug eruptions, necrolysis. Petition 870250090034, dated 02 / 10 / 2025, pp. 432 / 465 46 / 56 toxic epidermal; erythema multiforme, erythema nodosum, granuloma annulare and other cutaneous manifestations of inflammation. 51. The formulation for use of item 49, in which the autoimmune dermatological disease or condition, or the cutaneous manifestation of the autoimmune condition, is selected from the group consisting of psoriasis, pemphigus, systemic lupus erythematosus, lichen planus, morphea, scleroderma, epidermolysis bullosa, dermatomyositis, graft-versus-host syndrome. 52. A dispensing product comprising the formulation of any one of items 1 to 51. 53. A container comprising the formulation of any one of items 1 to 51. 54. Use of the formulation according to any of items 1 to 51, for the manufacture of a medicament for the treatment and / or prevention of a dermatological disease or condition in an individual. 55. A method of treating and / or preventing a dermatological disease or condition in an individual, said method comprising administering to the individual a pharmaceutically effective amount of the formulation of any of items 1 to 57.

[165] The present invention is further illustrated by the following figures and examples. Figure captions

[166] Figure 1: Tacrolimus solubility. Top corner: Tacrolimus solubility tests (time point 0). From left to right: TC2 (1 mg TC / mL aqueous phosphate buffer), TC3 (20 mg TC / g lipid phase or carrier component), TC4 (2 mg TC / mL BF200 nanoemulsion), TC5 (1 mg TC / mL BF200 nanoemulsion); Bottom corner: Tacrolimus solubility tests after 6 months of storage at 5 °C and 25 °C. From left Petition 870250090034, dated 02 / 10 / 2025, pp. 433 / 465 47 / 56 to the right: TC4 stored at 5 °C, TC4 stored at 25 °C, TC5 stored at 5 °C, TC5 stored at 25 °C.

[167] Figure 2: Tacrolimus assay in nanoemulsion formulations with different nominal TC contents (0.1%, 0.01%) after storage at 2 to 8 °C, 25 °C or 40 °C.

[168] Figure 3: Particle size in nanoemulsion formulations with different nominal TC content (0.1%, 0.01%) after storage at 2 to 8 °C, 25 °C or 40 °C.

[169] Figure 4: Polydispersity index (PDI) of formulations with different nominal TC content (0.1%, 0.01%) after storage at 2 to 8 °C, 25 °C or 40 °C.

[170] Figure 5: In Vitro Release (SUPAC-SS) after 2.6 hours of tacrolimus nanoemulsion formulations compared with commercially available tacrolimus ointments.

[171] Figure 6: Epidermal penetration of the tacrolimus nanoemulsion formulation compared to commercially available tacrolimus ointment.

[172] Some data shown in the figures result from grouped experiments. Examples Example 1: Preparation of Nanoemulsions BF200, BF215 and BF220 Table 1: Lipophilic content of the nanoemulsions used in the Examples BF200 10% lipophilic content BF215 15% lipophilic content BF220 20% lipophilic content

[173] The qualitative and quantitative compositions of nanoemulsions BF200, BF215 and BF220 are given in Table 2. Table 2: Composition of nanoemulsions BF200, BF215 and BF220 Petition 870250090034, dated 02 / 10 / 2025, pp. 434 / 465 48 / 56 Ingredient Content (% w / w) BF200 Content (% w / w) BF215 Content (% w / w) BF220 Function Quality 1 Soy lecithin 1.7 2.0 to 3.0 3.0 to 4.0 surfactant >90% phosphatidylcholine, for pharmaceutical use, USP 2 Polysorbate 80 (polyoxyethylene sorbitol monooleate) 3.4 4.5 to 5.5 6.0 to 7.0 surfactant Ph. Eur. 3 Caprylic / capric triglycerides 3.5 4.5 to 5.5 6.0 to 8.0 lipid core Ph. Eur. 4 Isopropyl alcohol 1.4 2.0 to 3.0 2.0 to 4.0 solvent Ph. Eur. Content (% w / w) ingredients 1 to 4 10.0 15.0 20.0 Phosphate buffer ad 100.00 ad 100.00 ad 100.00 solvent Water: Ph. Eur.; Disodium phosphate and sodium hydrogen phosphate: Ph. Eur.

[174] The manufacturing process of nanoemulsions in a typical batch size consists of the following steps 1 to 4: Step 1: Preparation of phosphate buffer (aqueous component)

[175] Phosphate buffer (1000 g) was prepared and optionally sterilized. Step 2: Preparation of the carrier component (lipid phase) containing the lipophilic component, surfactants, and alcohol. Table 3: Carrier component Ingredient Weight (g) Soy lecithin 17 Polysorbate 80 (polyoxyethylene sorbitol monooleate) 34 Caprylic / capric triglycerides 35 Isopropyl alcohol 14

[176] Soy lecithin (17 g) was weighed into a suitable receptacle, isopropyl alcohol (14 g) was added, and the receptacle was covered to prevent the Petition 870250090034, dated 02 / 10 / 2025, pp. 435 / 465 49 / 56 alcohol evaporation. Soy lecithin was dissolved under continuous stirring with a suitable stirrer at room temperature. Caprylic / capric triglycerides (35 bg) and Polysorbate 80 (34 g) were weighed and added to the soy lecithin solution. The mixture was stirred with a suitable stirrer at room temperature until a homogeneous and transparent solution was obtained. This solution is the carrier phase to be included in the nanoemulsion containing all the emulsifiers and lipid components of nanoemulsion BF200. According to this procedure, BF215 and BF220 were prepared by adjusting the quantity of the components (see Table 2).

[177] In all examples shown in the present invention, the BF200 nanoemulsion was used. Step 3: Manufacturing the nanoemulsion by mixing the aqueous component from Step 1 and the carrier from Step 2 to a lipid content of 10% (BF200).

[178] Fabrication of an emulsion by mixing 900 g of phosphate buffer (from Step 1) and 100 g of carrier (from Step 2). First, the aqueous component comprising the phosphate buffer was heated to approximately 45 to 60 °C in a suitable receptacle. Then, the carrier (concentrate) from Step 2 was heated to approximately 45 to 60 °C. Subsequently, the carrier was poured into the phosphate buffer under continuous stirring with a propeller mixer, resulting in the formation of a stable trombe (or nozzle) having the maximum possible diameter, without causing foaming or splashing. The resulting nanoemulsion was stirred for 15 min. Finally, the nanoemulsion was cooled to room temperature.

[179] In nanoemulsion BF215, 850 g of phosphate buffer (from Step 1) and 150 g of carrier were mixed. In nanoemulsion BF220, 800 g of phosphate buffer (from Step 1) and 200 g of carrier were mixed. Step 4: Preparation of the final formulation and primary packaging Petition 870250090034, dated 02 / 10 / 2025, pp. 436 / 465 50 / 56

[180] Optionally, the nanoemulsion can be sterilized.

[181] Depending on the purpose of the nanoemulsion, adjuvants and / or excipients and / or active ingredients may be added (at the appropriate step according to the description) and / or the nanoemulsion may be diluted in order to obtain a suitable pharmaceutical formulation, for example, by adding water, a suitable buffer or an additional aqueous gel base with, for example, poloxamer 407 or xanthan gum. Example 2: Preparation of a nanoemulsion formulation (BF220) containing 0.1% or 0.01% tacrolimus (TC) and 4% Poloxamer 407

[182] The formulations were prepared according to Example 1 with the addition of the appropriate amount of an aqueous gel base with poloxamer 407 in step 4. TC in an amount of 0.1% or 0.01% was added to the lipophilic component of step 2 of example 1. Example 3: Determination of the solubility properties of tacrolimus in nanoemulsion formulations.

[183] ​​Tacrolimus was dissolved by mechanical techniques (stirring). The determination of a complete solution of a substance was based on visual observation. A complete dissolution was defined as a clear solution, without signs of turbidity or precipitation. Pure solvents were used as a reference. Example 4: Determination of vesicle size and polydispersity index by dynamic light scattering

[184] The size of the nanovesicles, expressed as average z-size (e.g., in nm), and the homogeneity of the nanovesicle formulations, expressed as polydispersity index, were determined by dynamic light scattering (sometimes referred to as Photon Correlation Spectroscopy (PCS) or Quasi-Elastic Light Scattering (QELS)). The technique is well known. Petition 870250090034, dated 02 / 10 / 2025, pp. 437 / 465 51 / 56 in the state of the art and well established for determining the size of nano or microparticles or vesicles in emulsions, suspensions or polymer solutions with a laser. The measurements were conducted with a Zetasizer Nano ZS (Malvern Instruments Ltd, Malvern, Worcestershire, United Kingdom). The measurement was performed according to the manufacturer's instructions.

[185] The Zetasizer Nano ZS is instrumented with a 633 nm green laser and 173° scattering detection angle optics for size measurement. The device can be operated under vacuum for measurements, but in these cases, vacuum was not applied to the samples for size and homogeneity measurements. Example 5: In Vitro Release (SUPAC-SS) of tacrolimus formulations with nanoemulsion and tacrolimus ointments.

[186] The in vitro release method is based on an open-chamber diffusion cell system, such as a Franz cell system. The Franz cylindrical glass cell is a diffusion chamber comprising an upper and a lower part between which the synthetic membrane (e.g., EMD Millipore MF membrane; 0.025 µm) is fixed. The lower part (approx. 7 mL) is filled with an acceptor medium (EtOH / H2O 75:25% v / v) maintained at a temperature of about 32 °C at which the API has sufficient solubility. The acceptor medium is stirred (at approximately 400 rpm) to ensure partitioning and dissolution of the API. The diffusion area of ​​the membrane is approximately 1.8 cm2 (25 mm in diameter). The diffusion of tacrolimus from a topical product to and along the membrane was monitored (up to 2.6 hours) by assay using high-performance liquid chromatography (HPLC-UV). Example 6: Epidermal penetration of tacrolimus nanoemulsion formulation compared to commercially available tacrolimus ointment. Petition 870250090034, dated 02 / 10 / 2025, pp. 438 / 465 52 / 56

[187] The nanoemulsion formulation with 0.1% tacrolimus was compared to conventional 0.1% ointment in a skin penetration study, evaluating tacrolimus deposition in a layered assay using ex vivo facial skin. An established ex vivo model was used to explore drug penetration into human skin from routine facial cosmetic surgeries. The penetration test is divided into the following steps: I. Skin sampling: Samples were collected from blepharoplasty (upper and lower eyelids) and rhytidoplasty (cheek / periauricular). II. Sample preparation: a. A non-occlusive dermal application to ex vivo skin samples 6 mm in diameter. b. Approximately 28.3 mg (100 mg / cm2) of test items c. After incubation at approximately 37 °C and 5% CO2 in a humidified atmosphere, the test formulation was washed off the samples with 70% ethanol. The samples were frozen in freezing medium at -80 °C. d. Cryosection at -28 °C with a thickness of 10 μm e. Extraction of tacrolimus with ACN at approximately 20 °C for 24 hours at 450 rpm followed by centrifugation at 20 °C. III. Tacrolimus content determined by assay using high-performance liquid chromatography (HPLC-UV). Example 7: Determination of viscosity

[188] Viscosity was measured by rotation (measurement geometry: cone / plate) with a constant shear rate of 90.0 s-1 at 20 °C. Example A Tacrolimus (TC) content over time in BF220 with PX=4% as a gelling agent at different temperatures.

[189] Nanoemulsion formulations BF220 TC=0.1% and BF220 TC=0.01% Petition 870250090034, dated 02 / 10 / 2025, pp. 439 / 465 53 / 56 were prepared as described in Example 2. The nanoemulsion formulations were stored at 2 to 8 °C, 25 °C, and 40 °C. The TC content was determined at 0 (baseline) and at various time points during the 24-month storage period. The results are shown in Figure 2.

[190] Conclusion: At 2 to 8 °C: The formulation (Example 2) preserves the API (TC) content better than conventional aqueous formulations. The TC in the Example 2 formulation proved to be stable after 24 months of storage at 2 to 8 °C. At 40 °C, the TC in the Example 2 formulation is stable for at least 1 month. Example B Particle size and particle size distribution over time in BF220 with PX = 4% as gelling agent at different temperatures.

[191] Nanoemulsion formulations BF220 TC=0.1% and BF220 TC=0.01% were prepared as described in Example 2. The nanoemulsion formulations were stored at temperatures of 2 to 8 °C, 25 °C, and 40 °C. Particle size, particle size distribution, and pH were determined at 0 (baseline) and at various times during the 24-month storage period. The results are shown in Figure 3 and Figure 4.

[192] Conclusion: At 2 to 25 °C: The formulations (Example 2) containing API (TC) proved to be stable for at least 24 months of storage at 2 to 25 °C. At 40 °C, the formulations are stable for at least 3 months. Example C In Vitro Release (SUPAC-SS) of tacrolimus formulations with nanoemulsion and tacrolimus ointments Petition 870250090034, dated 02 / 10 / 2025, pp. 440 / 465 54 / 56

[193] From both test formulations, a significantly greater amount of tacrolimus is released and can penetrate the skin and through it if the formulations are applied. The properties of the nanoemulsion formulation contribute to a more effective release of tacrolimus than that of an ointment, where the diffusion of tacrolimus is additionally inhibited by the ointment.

[194] Using the tacrolimus nanoemulsion release test method, no tacrolimus was released from the reference ointments with tacrolimus contents of 0.1% and 0.03%. From the test formulation with BF220 nanoemulsion and 0.03% tacrolimus, 28.39% of the applied amount of tacrolimus was released. From the test formulation with BF220 nanoemulsion and 0.1% tacrolimus, 42.81% of the applied amount of tacrolimus was released. The data are shown in Figure 5. Example D Solubility of tacrolimus in nanoemulsion formulations

[195] The samples were prepared as explained in Example D. The compound was dissolved in 10 mM phosphate buffer (TC2) at the defined concentration (see table below). For samples TC3, TC4, and TC5, the lipid (intermediate) phase of the BF200 nanoemulsion was used as a solvent. In a first step, Tacrolimus was dissolved in the lipid phase, followed by the preparation of the nanoemulsion by combining the aqueous phase with the lipid phase (containing Tacrolimus). Samples TC4 and TC5 were stored at 5 °C and 25 °C for a period of 6 months. The results of the solubility tests for the 0 and 6 month time points are shown in Figure 1 and Table 4. Table 4: Solubility test results Petition 870250090034, dated 02 / 10 / 2025, pp. 441 / 465 55 / 56 Tacrolimus Concentration Solvent Test Solution Name Solution Appearance (T0) Solution Appearance (T6) Solubility 1 mg / mL Phosphate Buffer TC2 Turbid, white precipitate Turbid, white precipitate Insoluble Lipid phase 20 mg / g of nanoemulsion TC3 Clear, yellow color Clear, yellow color Soluble BF200 100% of Clear, Clear, 2 mg / mL nanoemulsion TC4 Slightly slightly Soluble BF200 opalescent opalescent 50% of Clear, Clear, 1 mg / mL nanoemulsion TC5 Slightly slightly Soluble BF200 opalescent opalescent

[196] Conclusion: Tacrolimus is highly soluble in the nanoemulsion aqueous (at least 90% water content) at a concentration of at least 2 mg / mL. Example E Epidermal penetration of the tacrolimus nanoemulsion formulation compared to commercially available tacrolimus ointment.

[197] The results indicate that the tacrolimus from the conventional lipophilic ointment containing 0.1% tacrolimus remains mainly on the skin surface, presumably associated with its formulation, and does not penetrate through the epidermis, whereas a greater deposition of tacrolimus in the deeper layers of the skin can be achieved by the formulation with nanoemulsion (BF220) containing 0.1% tacrolimus. This result is in agreement with the results of Example C.

[198] After 24 h of incubation, differences in the deposition profile of samples treated with nanoemulsion (BF220) containing 0.1% tacrolimus and conventional lipophilic ointment containing 0.1% were observed. It was found that the amount of TC after treatment with nanoemulsion (BF220) containing 0.1% tacrolimus was distributed very uniformly. Petition 870250090034, dated 02 / 10 / 2025, pp. 442 / 465 56 / 56 throughout all three layers of the skin, with a tendency to accumulate towards the deeper dermis. During treatment with conventional lipophilic ointment containing 0.1%, the highest amount of tacrolimus is found in the upper layer of the skin, with the concentration decreasing sharply with depth of the layer. The data are shown in Figure 6. Petition 870250090034, dated 02 / 10 / 2025, pp. 443 / 465

Claims

1 / 5 CLAIMS 1. Formulation, characterized in that it comprises (a) a nanoemulsion comprising: (i) at least one aqueous component; (ii) a carrier component comprising: (1) at least one lipophilic component, (2) at least one surfactant and (3) at least one alcohol; and (b) an active agent, wherein the active agent is a highly lipophilic macrolide lactone.

2. Formulation according to claim 1, characterized in that the macrolide lactone has a logP value of 3 or higher.

3. Formulation according to claim 1 or 2, characterized in that the active agent is dissolved in the carrier component of the nanoemulsion.

4. Formulation, according to any one of claims 1 to 3, characterized in that the active agent is Tacrolimus, Pimecrolimus, Everolimus or Sirolimus, preferably Tacrolimus, a derivative, an isomeric form, a tautomeric form, a precursor, a metabolite, hydrate and / or a pharmaceutically acceptable salt thereof.

5. Formulation, according to any one of claims 1 to 4, characterized in that the active agent is present in an amount of 0.001% to 25% (w / w), preferably 0.005% to 1% (w / w), most preferably 0.01% to 1% (w / w) based on the total weight of the formulation.

6. Formulation, according to any one of claims 1 to 5, characterized in that the aqueous component is present in an amount of 70% to 95% (w / w), and more preferably 80% to 95% (w / w) based on the total weight of the nanoemulsion.

7. Formulation, according to any one of claims 1 to 6, characterized in that it comprises a total aqueous component in an amount of at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75% or at least 80% (w / w), based on the total weight of the formulation, preferably from 50% to 99% (w / w), from 70% to 95% (w / w), more preferably from 75% to 95% (w / w), even more preferably from 80% to 90% (w / w).

8. Formulation according to any one of claims 1 to 7, characterized in that (a) the at least one alcohol comprises at least three carbon atoms, preferably 3, 4 or 5 carbon atoms, preferably wherein the at least one alcohol is selected from the group consisting of 1-propanol or 2-propanol and mixtures thereof; (b) the at least one lipophilic component is selected from triglycerides and mixtures thereof, preferably wherein the at least one lipophilic component is a caprylic and / or capric triglyceride or a mixture thereof; and / or (c) the at least one surfactant is selected from the group consisting of a phospholipid, a lysophospholipid, a ceramide and / or a mixture thereof, and / or the at least one surfactant is a polyoxyethylene type surfactant.

9. Formulation according to any one of claims 1 to 8, characterized in that at least one surfactant is phosphatidylcholine.

10. Formulation, according to any one of claims 1 to 9, characterized in that it comprises from 0.5% to 5% (w / w), preferably from 1% to 4% (w / w), more preferably from 1.2% to 3.5% (w / w) of phosphatidylcholine.

11. Formulation, according to any one of claims 1 to 10, characterized in that it comprises a propellant and is contained in a pressurized container.

12. Formulation, according to any one of claims 1 to 11, characterized in that it essentially comprises no fatty alcohols.

13. Formulation, according to any one of claims 1 to 12, characterized in that it does not essentially comprise any emollient selected from a monoester or diester comprising an alcohol and a fatty acid.

14. Formulation according to any one of claims 1 to 13, characterized in that it comprises at least one gelling agent.

15. Formulation, according to any one of claims 1 to 14, characterized in that the active agent has a content greater than or equal to 80%, preferably greater than or equal to 85%, more preferably greater than or equal to 90% when (a) stored for one month, two months, three months, six months at 2 to 25 °C; or (b) stored for one month, two months, three months, six months, twelve months, eighteen months or twenty-four months at 2 to 8 °C.

16. Formulation, according to any one of claims 1 to 15, characterized in that the nanoemulsion comprises nanovesicles, wherein the nanovesicles have a size less than or equal to 500 nm, preferably less than or equal to 200 nm, more preferably in the range of 5 nm to 100 nm when (a) stored for one month, two months, three months, six months, twelve months, eighteen months or twenty-four months at 2 to 25 °C; or (b) stored for one month, two months, three months, six months, twelve months, eighteen months, twenty-four months, thirty months or thirty-six months at 2 to 8 °C.

17. Formulation according to any one of claims 1 to 16, characterized in that it has a polydispersity index less than or equal to 0.4 when (a) stored for one month, two months, three months, six months, twelve months, eighteen months or twenty-four months at 2 to 25 °C; (b) stored for one month, two months, three months, six months, twelve months, eighteen months, twenty-four months, thirty months, thirty-six months at 2 to 8 °C.

18. Formulation, according to any one of claims 1 to 17, characterized in that it is a topical formulation, an ophthalmic formulation, a parenteral formulation or an oral formulation.

19. Formulation, according to any one of claims 1 to 18, characterized in that it is a lotion, a spray, a foam, an emulsion, a nanoemulsion, a gel or a cream.

20. Formulation according to any one of claims 1 to 19, characterized in that it is for use in medicine.

21. A formulation, according to any one of claims 1 to 20, characterized in that it is for use in a method of treating or preventing a dermatological, ophthalmic, or autoimmune disease or condition, or for preventing organ rejection after transplantation.

22. Foam dispensing container or product or spray dispenser, characterized in that it comprises a container comprising a formulation, wherein said formulation comprises a nanoemulsion comprising: (i) at least one aqueous component; (ii) a carrier component comprising: (1) at least one lipophilic component, (2) at least one surfactant and (3) at least one alcohol; and (iii) an active agent, wherein the active agent is a highly lipophilic macrolide lactone; wherein the container further comprises a propellant, wherein the propellant is provided to pressurize the container.

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