Solid-in-oil formulation

Abstract

To provide a Solid-in-Oil formulation that is suitable for ophthalmic instillation or intraocular administration.SOLUTION: A Solid-in-Oil formulation includes (i) a solid composite containing particles including a pharmaceutically acceptable target substance or target molecule and a pharmaceutically acceptable surfactant, and (ii) a pharmaceutically acceptable oily base. In the Solid-in-Oil formulation, the particles are typically coated with the surfactant. In the Solid-in-Oil formulation, the target substance or target molecule may be water-soluble.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present disclosure relates to solid-in-oil formulations. [Background technology]

[0002] Formulations for treating ophthalmic diseases have been developed. US2021 / 145736A, US2017 / 273901A, US2013 / 164285A, US2012 / 258163A, and US2016 / 0235674A disclose methods for intravitreal administration of drugs.

[0003] WO2021 / 049529A, EP3950071A, US2019 / 117777A, JP2018-70535A, EP2298282A, and US2010 / 298447A disclose dispersions of particles having a core-shell structure as formulations for transdermal administration. Summary of the Invention

[0004] The present disclosure relates to a solid-in-oil formulation. The solid-in-oil formulation comprises (i) a solid complex comprising particles containing a pharmaceutically acceptable target substance or target molecule and a pharmaceutically acceptable surfactant, and (ii) a pharmaceutically acceptable oil base. In a solid-in-oil formulation, the particles are typically coated with the surfactant. In a solid-in-oil formulation, the target substance or target molecule may be water-soluble. The present disclosure also provides a solid-in-oil formulation suitable for ophthalmic or intraocular administration. The present disclosure further provides a solid-in-oil formulation for ophthalmic or intraocular administration. In one aspect of the present disclosure, an ophthalmic formulation is provided, comprising (i) a solid complex comprising particles containing a pharmaceutically acceptable target substance or target molecule and a pharmaceutically acceptable surfactant, and (ii) a pharmaceutically acceptable oil base.

[0005] According to the present disclosure, there is provided an ophthalmic formulation comprising: (i) a solid complex comprising particles containing a pharmaceutically acceptable target substance or molecule and a pharmaceutically acceptable surfactant; and (ii) a pharmaceutically acceptable oily base.

[0006] According to the present disclosure, for example, the following inventions are provided. (1) An ophthalmic preparation, (i) a solid complex comprising particles containing a pharmaceutically acceptable target substance or target molecule and a pharmaceutically acceptable surfactant; (ii) a pharmaceutically acceptable oily base; Including, the particles are coated with the surfactant; The solid complex is dispersed in the oily base. Ophthalmic preparations. (2) The ophthalmic preparation according to (1) above, which is formulated as eye drops. (3) The ophthalmic preparation according to (1) above, which is formulated for intravitreal administration. (4) The ophthalmic preparation according to any one of (1) to (3) above, wherein the target substance or target molecule is water-soluble. (5) The ophthalmic preparation according to any one of (1) to (4) above, wherein the target substance or target molecule is sustained-released from the ophthalmic preparation after administration to a subject. (6) The ophthalmic preparation according to any one of (1) to (4) above, wherein the target substance or target molecule is sustainedly released from the ophthalmic preparation for at least 24 hours after administration to the vitreous of the subject. (7) The ophthalmic preparation according to any one of (1) to (3) above, wherein the target substance or molecule is sustainedly released from the ophthalmic preparation at a rate of 50% or less over 24 hours after administration to the vitreous body of the subject. (8) The ophthalmic preparation according to any one of (1) to (7) above, wherein the target substance or target molecule is a molecule selected from the group consisting of a protein, a nucleic acid, and an antibody. (9) The ophthalmic preparation according to any one of (1) to (8) above, wherein the average hydrodynamic particle size is 80 nm to 200 nm. (10) The ophthalmic preparation according to any one of the above (1) to (9), wherein the oily base is silicone oil. [Brief explanation of the drawings]

[0007] [Figure 1] FIG. 1 shows a schematic diagram of a test system that simulates the intravitreal environment. [Figure 2A] FIG. 2A shows the release behavior of antibody molecules from a formulation of the present disclosure in a test system that simulates the intravitreal environment. [Figure 2B] FIG. 2B shows particle formation and particle size distribution of the formed particles for a formulation of the present disclosure that includes an antibody as a target substance or molecule. [Figure 3A] FIG. 3A shows the release behavior of a protein molecule (lysozyme, for example) from a formulation of the present disclosure in a test system that simulates the intravitreal environment. [Figure 3B] FIG. 3B shows particle formation and the particle size distribution of the formed particles in a formulation of the present disclosure that includes a protein molecule (lysozyme) as a target substance or molecule. [Figure 4A] FIG. 4A shows the release profile of an anti-VEGF antibody from a formulation of the present disclosure in a test system that simulates the intravitreal environment. [Figure 4B] FIG. 4B shows the penetration of anti-VEGF antibodies from a formulation of the present disclosure into the retina and choroid after ocular administration to rabbits. [Figure 4C] FIG. 54 shows the vitreous accumulation of anti-VEGF antibodies from formulations of the present disclosure administered intravitreally to rabbits. [Figure 4D] FIG. 4D shows the penetration of anti-VEGF antibodies from formulations of the present disclosure into the retina and choroid after intravitreal administration to rabbits. [Figure 5] FIG. 5 shows the release profile of aflibercept from the formulation of the present disclosure in a test system simulating the intravitreal environment. [Figure 6] FIG. 6 shows a histological section (hematoxylin-eosin stained) of the eyeball of a mouse to which a formulation of the present disclosure was intravitreally administered. DETAILED DESCRIPTION OF THE INVENTION

[0008] Embodiments of the present invention will be described with reference to the drawings. The present invention is not limited to the following embodiments and drawings. In the following embodiments, the terms "have," "include," or "contain" also include the meaning of "consisting of" or "consisting of." The singular form means singular or plural.

[0009] As used herein, the term "subject" refers to a mammal, including humans, and preferably humans. Examples of mammals include, but are not limited to, humans, chimpanzees, and other primates; domestic animals such as dogs, cats, rabbits, horses, sheep, goats, cows, pigs, rats (including nude rats), mice (including nude mice and squirrel mice), and guinea pigs; pets, and laboratory animals.

[0010] As used herein, a "solid composite" refers to a composite consisting of solid components. A solid composite may contain water, but is a composite containing solid components. A solid composite may be water-free or have a low water content. For example, the water content may be 1% by weight or less, 0.9% by weight or less, 0.8% by weight or less, 0.7% by weight or less, 0.6% by weight or less, 0.5% by weight or less, 0.4% by weight or less, 0.3% by weight or less, 0.2% by weight or less, or 0.1% by weight or less, as measured by known methods (e.g., Karl Fischer method). A solid composite containing hydrophilic molecules and a surfactant may have a core-shell structure, in which the core contains hydrophilic molecules and the shell contains a surfactant, with the hydrophilic portion of the surfactant facing the core and the hydrophobic portion facing the outer surface of the solid composite. In this way, the surfactant preferably covers the surface of the hydrophilic drug, rendering the particle surface hydrophobic.

[0011] As used herein, a "solid particle-dispersed oil formulation" refers to a formulation comprising multiple solid particles and an oily base, with the solid particles dispersed in the oily base. The solid particles comprise a target substance or target molecule and a surfactant. The solid particles also contain no water or only a limited amount of water. The particle size is typically less than 1 μm. In a solid particle-dispersed oil formulation, the solid particles can be stably dispersed in the oily base. As used herein, a solid particle-dispersed oil formulation is also referred to as a "solid-in-oil formulation." As used herein, "solid particles" are also referred to as a "solid complex." The target substance or target molecule may be hydrophilic. In this case, the solid particle-dispersed oil formulation is prepared by dispersing a hydrophilic target substance or target molecule in an oily base using a surfactant. The target substance or target molecule may be, for example, a biomolecule. The target substance or target molecule may be, for example, an active pharmaceutical ingredient. The target substance or target molecule may be, for example, a therapeutic agent for an ophthalmic disease. The solid particle dispersed oil-based preparation preferably does not contain aggregates formed by aggregation of a plurality of particles (particularly 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more).

[0012] As used herein, a "surfactant" refers to a molecule having a relatively hydrophilic portion and a relatively hydrophobic portion. Surfactants are broadly classified into ionic surfactants and nonionic surfactants. Ionic surfactants include cationic surfactants and anionic surfactants. Typically, an ionic surfactant may have either a cation or an anion and a hydrophobic portion. A cationic surfactant may contain a cationic portion and a fatty chain (e.g., a long-chain fatty chain) such as an alkyl, alkenyl, or alkynyl. Anionic surfactants may contain an anionic portion and a fatty acid such as a long-chain fatty acid, an unsaturated fatty acid (e.g., monovalent or divalent), or a long-chain unsaturated fatty acid (e.g., monovalent or divalent). The term "long chain" refers to a compound having 14 to 22 carbon atoms. A fatty chain refers to a fatty chain having 8 to 22 carbon atoms, i.e., C8 to C9. 22 Alkyl, C8-C 22 Alkenyl, or C8-C 22 C8-C such as alkynyl22 The fatty chain may be a fatty chain. The fatty chain may also be a long fatty chain. Saturated means that the fatty chain has no double or triple bonds, and unsaturated means that the fatty chain has at least one double or triple bond. Fatty chains with double bonds include cis- and trans-types, with the cis-type being preferred from the viewpoint of biocompatibility. Nonionic surfactants are surfactants that do not have ionic groups in their molecules and have a nonionic hydrophilic portion and a hydrophobic portion. The hydrophobic portion may contain a fatty chain such as an alkyl, alkenyl, or alkynyl group (e.g., a long fatty chain).

[0013] Preferred nonionic surfactants are ester compounds derived from unsaturated fatty acids such as erucic acid and oleic acid. Examples of lipophilic nonionic surfactants include sucrose fatty acid esters (sucrose stearate, sucrose palmitate, sucrose myristate, sucrose oleate, sucrose laurate, sucrose erucate, and mixed sucrose fatty acid esters) with a high degree of esterification (i.e., a high ratio of di-, tri-, and polyester esters relative to monoesters), polyglycerol condensed ricinoleate, decaglycerol ester, glycerol fatty acid ester, polyglycerol fatty acid ester, polyoxyethylene glycerol fatty acid ester, sorbitan fatty acid ester, polyoxyethylene sorbit fatty acid ester, and polyoxyethylene castor oil / hydrogenated castor oil. Preferably, the nonionic surfactant is sucrose laurate. Surfactants may be used singly or in combination.

[0014] Examples of surfactants include polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 60, polyoxyethylene castor oil, polyoxyethylene (160) polyoxypropylene (30) glycol, polyoxyethylene (200) polyoxypropylene (70) glycol, polysorbate 80, macrogol 4000, macrogol 6000, aluminum monostearate, polyethylene glycol monostearate, glyceryl monostearate, nonoxynol-9, octoxynol-40, polyethylene glycol (PEG) / polypropylene glycol (PPG)-4 / 30 copolymer, poloxamer 188, poloxamer 407, polyoxyl 15 hydroxystearate, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, polyoxyl 40 stearate, and polysorbate 20.

[0015] A suitable example of a surfactant used for preparing the complex is a lipophilic (hydrophobic) nonionic surfactant having an HLB (Hydrophile-Lipophile Balance) value of 10 or less. The HLB of the nonionic surfactant is preferably 8 or less, more preferably 5 or less, and particularly preferably 3 or less.

[0016] As used herein, "antibody" refers to an immunoglobulin. The antibody may be of various isotypes, such as IgG. The antibody is preferably a monoclonal antibody. The antibody may be a human chimeric antibody, a humanized antibody, or a human antibody. A human chimeric antibody can be produced by replacing the constant region of a non-human antibody with the constant region of a human antibody. A humanized antibody can be produced by replacing the six CDRs of a human antibody with the six corresponding CDRs of a non-human antibody. A human antibody can be produced using an animal (e.g., a mouse) in which at least the heavy chain variable region of an immunoglobulin has been replaced with the corresponding region of a human locus. When the constant region is non-human, a human antibody can be obtained by replacing the constant region with the amino acid sequence of a human antibody. As used herein, the antibody is preferably a humanized antibody. As used herein, the antibody is preferably a human antibody. When produced intracellularly, an antibody has a signal peptide, but when secreted extracellularly, the signal peptide is cleaved. Therefore, when administered as a pharmaceutical, antibodies do not require a signal peptide.

[0017] As used herein, "CDR" refers to the complementarity determining region present in the heavy chain variable region and light chain variable region of an antibody. There are three CDRs in each of the heavy chain and light chain variable regions, and they are called CDR1, CDR2, and CDR3 from the N-terminus. CDRs can be determined, for example, based on the numbering system of Kabat et al. (Kabat, EA et al., Sequences of Proteins of Immunological Interest, 5th ed., 1991, Bethesda: US Dept. of Health and Human Services, PHS, NIH.).

[0018] As used herein, "antigen-binding fragment of an antibody" refers to a fragment of an antibody that maintains its ability to bind to an antigen. Examples of antigen-binding fragments include Fab, Fab', F(ab')2, Fv, scFv (single-chain Fv), diabody, and sc(Fv)2 (single-chain (Fv)2). For example, Fab can be obtained by digesting an antibody with papain. Alternatively, F(ab')2 can be obtained by digesting an antibody with pepsin, and Fab' can be obtained by further reducing this. Antigen-binding fragments of other antibodies can also be produced by methods well known to those skilled in the art. Such antigen-binding fragments of antibodies can be used in the present invention.

[0019] As used herein, "pharmaceutically acceptable" means that when administered to a subject as a pharmaceutical, it does not cause unacceptable toxicity.

[0020] As used herein, "intraocular administration" refers to invasive administration to the eyeball other than eye drops. Examples of intraocular administration include intravitreal administration, subconjunctival administration, suprachoroidal administration, and subretinal administration.

[0021] According to the present disclosure, there is provided a solid particle dispersed oil formulation (Solid-in-Oil formulation), (i) a solid complex comprising particles containing a pharmaceutically acceptable target substance or target molecule and a pharmaceutically acceptable surfactant; (ii) a pharmaceutically acceptable oily base; A formulation is provided comprising:

[0022] The solid particle dispersed oil-based formulation of the present disclosure may be free of water or have a low water content; for example, the water content may be 1% by mass or less, 0.9% by mass or less, 0.8% by mass or less, 0.7% by mass or less, 0.6% by mass or less, 0.5% by mass or less, 0.4% by mass or less, 0.3% by mass or less, 0.2% by mass or less, or 0.1% by mass or less, as measured by known methods.

[0023] In the solid particle-dispersed oil formulation of the present disclosure, the target substance or target molecule is hydrophilic. In the solid particle-dispersed oil formulation of the present disclosure, the target substance or target molecule can be a physiologically active substance, for example, a biomolecule. The biomolecule can be a molecule found in a living organism or its analog, for example, a protein, a peptide, an antibody or its antigen-binding fragment, a nucleic acid, or other biomolecule. The biomolecule can be chemically modified or not.

[0024] Ophthalmic solutions are the most widely used non-invasive drug administration form for treating anterior ocular diseases due to their non-invasiveness and convenience. Examples of anterior ocular diseases include glaucoma, allergic conjunctivitis, anterior uveitis, and cataract. The hydrophilic active ingredient in ophthalmic solutions is not particularly limited as long as it is a pharmaceutical ingredient that is administered to the eye.

[0025] The antibody may be, but is not limited to, an antibody that has benefits for absorption through the skin or intraocular (intravitreal) administration. Examples of antibodies include anti-VEGF antibodies (e.g., ranibizumab, bevacizumab, or aflibercept). The effectiveness of anti-VEGF antibodies has been confirmed for age-related macular degeneration, diabetic retinopathy (diabetic macular edema), retinal vein occlusion, pathological myopia (choroidal neovascularization), and the like. The antibody or its antigen-binding fragment retains its antigen-binding ability in a solid complex or a solid particle-dispersed oil formulation and exhibits antigen-specific binding after administration. The protein or peptide retains its antigen-binding ability in a solid complex or a solid particle-dispersed oil formulation and exerts at least a portion of its function after administration. Examples of proteins or peptides include lysozyme, insulin, albumin, ovalbumin, and neutrophin 4. The nucleic acid includes one or more selected from the group consisting of DNA, RNA, and modified nucleic acids. The nucleic acid may be a single-stranded nucleic acid or a double-stranded nucleic acid. Nucleic acid can be antisense oligo.RNA includes artificial RNA for gene silencing such as siRNA and shRNA, non-coding RNA such as microRNA (miRNA), aptamer, and natural RNA such as mRNA.Nucleic acid or RNA includes aptamer (for example, anti-VEGF aptamer).These RNAs can be modified to be stabilized in vivo.

[0026] Modified nucleic acids include, for example, fluorescent dye-modified nucleic acids, biotinylated nucleic acids, and nucleic acids into which a cholesteryl group has been introduced. To enhance the stability of RNA, bases may be modified with 2'-O-methyl, 2'-fluoro, or 2'-methoxyethyl (MOE), and the phosphodiester bond in the nucleic acid backbone may be replaced with a phosphorothioate bond. Artificial nucleic acids include nucleic acids in which the oxygen atom at the 2' position and the carbon atom at the 4' position are cross-linked. Such artificial nucleic acids include, for example, locked nucleic acids, which are cross-linked DNA in which the oxygen atom at the 2' position and the carbon atom at the 4' position are cross-linked via a methylene. bridged nucleic acids (BNA) such as LNA (Likely a nucleotide), ENA (where the oxygen atom at the 2' position and the carbon atom at the 4' position are bridged via ethylene), BNACOC (where the oxygen atom at the 2' position and the carbon atom at the 4' position are bridged via -CH2OCH2-), and BNANC (where the oxygen atom at the 2' position and the carbon atom at the 4' position are bridged via -NR-CH2- (where R is a methyl or hydrogen atom)); and cMO (where the oxygen atom at the 2' position and the carbon atom at the 4' position are bridged via -CH2(OCH3)-). Examples include E, cEt in which the oxygen atom at the 2' position and the carbon atom at the 4' position are bridged via -CH2(CH3)-, AmNA in which the carbon atoms at the 2' and 4' positions are bridged via an amide, scpBNA in which the oxygen atom at the 2' position and the carbon atom at the 4' position are bridged via a methylene and a cyclopropane is formed at the 6' position, and peptide nucleic acid (PNA) in which the polymer backbone is made of N-(2-aminoethyl)glycine amide-linked instead of deoxyribose or ribose.

[0027] Any pharmaceutically acceptable surfactant may be used without particular limitation, including nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, and bile salts.

[0028] Examples of nonionic surfactants include polyglycerol condensed ricinoleate esters, decaglycerol esters, glycerol fatty acid esters, polyglycerol fatty acid esters, polyoxyethylene glycerol fatty acid esters, sorbitan fatty acid esters, polyoxyethylene sorbit fatty acid esters, polyoxyethylene castor oil / hydrogenated castor oil, sucrose fatty acid esters (sucrose stearate, sucrose palmitate, sucrose myristate, sucrose oleate, sucrose laurate, sucrose erucate, sucrose mixed fatty acid esters), etc. One of these surfactants may be selected for use, or two or more may be used in combination.

[0029] These nonionic surfactants are preferably ester compounds made from unsaturated fatty acids such as erucic acid and oleic acid, more preferably sucrose erucate, sucrose oleate, and sucrose mixed fatty acid esters. Alternatively, one or more surfactants selected from the group consisting of glycerin fatty acid esters, polyglycerin fatty acid esters, polyoxyethylene glycerin fatty acid esters, sorbitan fatty acid esters, sucrose fatty acid esters, polyoxyethylene sorbit fatty acid esters, polyoxyethylene castor oil, and hydrogenated castor oil can be used.

[0030] The HLB value indicates the degree of hydrophilicity and hydrophobicity of a surfactant. A smaller HLB value indicates a higher hydrophobicity. In the present disclosure, the surfactant is not particularly limited, but it is preferable to use a surfactant with a high hydrophobicity, with an HLB value of 10 or less. This is because it can facilitate dissolution or dispersion of the antigen-containing complex in the oil phase. In the present disclosure, surfactants with an HLB value of 10 or less, 8 or less, 6 or less, 5 or less, 4 or less, or 3 or less can be preferably used. In the present disclosure, a smaller HLB value is preferable.

[0031] When a water-soluble target substance or molecule is mixed with a surfactant in an oil phase (preferably a volatile oil phase), particles with a core-shell structure are formed, with the water-soluble target substance or molecule in the core and the surfactant in the shell. The particles are dispersed in the oil phase. If the oil phase used here is a volatile oil phase (e.g., cyclohexane), the solvent can be easily evaporated, and by removing the solvent by drying (e.g., lyophilization), particles with a core-shell structure containing the water-soluble target substance or molecule in the core and the surfactant in the shell can be obtained as solid particles (also referred to as "solid composites"). By dispersing the solid particles in an oily base, a solid particle-dispersed oil formulation or a solid-in-oil formulation can be obtained.

[0032] In the present disclosure, the solid particles may have an average hydrodynamic particle size of 50 nm to 200 nm as determined by dynamic light scattering. In some embodiments, the average hydrodynamic particle size may be 100 to 160 nm. In some embodiments, the average hydrodynamic particle size may be 80 to 120 nm.

[0033] In the present disclosure, the oily base can be any oil that is acceptable for use in pharmaceutical preparations administered to the eye. In the present disclosure, the oily base can preferably be an oil that is liquid at room temperature (25°C). In the present disclosure, the oily base can be either a natural or synthetic product. In the present disclosure, the oily base can be, for example, vegetable oils such as soybean oil, cottonseed oil, rapeseed oil, sesame oil, corn oil, peanut oil, safflower oil, sunflower oil, olive oil, castor oil, rapeseed oil, perilla oil, fennel oil, lanolin, lanolin oil, lanolin alcohol, refined lanolin, mineral oil, cocoa oil, cinnamon oil, peppermint oil, eucalyptus oil, and bergamot oil, or animal oils such as beef tallow, lard, and fish oil. The oily base may be a neutral lipid such as glyceride, triolein, trilinolein, tripalmitin, tristearin, trimyristin, or triarachidonin, or a synthetic lipid. The oily base may also be a sterol derivative such as cholesteryl oleate, cholesteryl linoleate, cholesteryl myristate, cholesteryl palmidate, or cholesteryl arachidate, or a long-chain fatty acid ester such as isopropyl myristate, octyldodecyl myristate, cetyl myristate, ethyl oleate, ethyl linoleate, isopropyl linoleate, isopropyl palmitate, or butyl stearate. The oil phase may also be a carboxylic acid ester such as ethyl lactate, cetyl lactate, triethyl citrate, diisopropyl adipate, diethyl sebacate, diisopropyl sebacate, or cetyl 2-ethylhexanoate, or a hydrocarbon such as petrolatum, white petrolatum, liquid paraffin, squalane, or vegetable squalane, or a silicone oil. The oily base may be used alone or in combination of two or more. Preferred oily bases include squalane oil, castor oil, sesame oil, white petrolatum, and liquid paraffin, and more preferred oily bases include castor oil, isopropyl myristate, and silicone oil.

[0034] In one embodiment, the oily base is castor oil, and the surfactant can be any one or more surfactants selected from the group consisting of glyceryl dioleate, polyoxyethylene hydrogenated castor oil 10, sorbitan sesquioleate, and sucrose laurate.

[0035] According to the present disclosure, the solid particle dispersed oil formulation (Solid-in-Oil formulation) is formulated as an ophthalmic formulation (e.g., eye drops or intraocular formulation). Thus, according to the present disclosure, an ophthalmic formulation (e.g., eye drops or intraocular formulation) comprising: (i) a solid complex comprising particles containing a pharmaceutically acceptable target substance or target molecule and a pharmaceutically acceptable surfactant; (ii) a pharmaceutically acceptable oily base; In a preferred embodiment, the ophthalmic formulation is an eye drop. In a preferred embodiment, the ophthalmic formulation is an intravitreal formulation.

[0036] According to the present disclosure, a solid-in-oil formulation can sustainably release the target substance or molecule. In one embodiment of the present disclosure, the solid-in-oil formulation can sustainably release the target substance or molecule in the eye (e.g., the vitreous body) for at least 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 9 hours, 12 hours, 18 hours, 24 hours (2 days), 3 days, 4 days, 5 days, 6 days, 7 days (1 week), or 2 weeks. The solid-in-oil formulation of the present disclosure can sustainably release the target substance or molecule into the ocular irrigation solution for at least 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 9 hours, 12 hours, 18 hours, 24 hours (2 days), 3 days, 4 days, 5 days, 6 days, 7 days (1 week), or 2 weeks. According to the present disclosure, a solid-in-oil formulation sustains release of the target substance or molecule at a sustained-release reference value or less. The sustained-release reference value is defined as a sustained-release of 50% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, or 5% or less of the target substance or molecule contained in the formulation within a sustained-release reference time. The sustained-release reference time may be, for example, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 9 hours, 12 hours, 18 hours, 24 hours (2 days), 3 days, 4 days, 5 days, 6 days, 7 days (1 week), or 2 weeks. The sustained release preferably occurs after administration into the vitreous of a subject.

[0037] The solid-in-oil formulation of the present disclosure can deliver the target substance or molecule to one or more anterior ocular tissues selected from the group consisting of the cornea, conjunctiva, aqueous humor, iris, ciliary body, and lens, and to one or more posterior ocular tissues selected from the group consisting of the sclera, choroid, retinal pigment epithelium, neural retina, optic nerve, vitreous membrane, and vitreous humor. The solid-in-oil formulation of the present disclosure can deliver the target substance or molecule to one or more of the above tissues over an extended period of time.

[0038] The eye drops of the present disclosure are dispersed in an oily base and are less likely to be excreted by tears than hydrophilic substances. The eye drops of the present disclosure can also pass through the corneal epithelium, which exhibits hydrophobic properties, and be suitable for the penetration of drugs into ocular tissues and posterior ocular tissues. Furthermore, the eye drops of the present disclosure can have tissue permeability. The formulation for intravitreal administration of the present disclosure is dispersed in an oily base and is less likely to diffuse or be excreted by vitreous humor than hydrophilic substances. It is believed that the form of the solid particle-dispersed oily formulation can prevent the active ingredient from diffusing prematurely into body fluids, allowing for sustained release of the active ingredient.

[0039] According to the present disclosure, there is provided a method for treating an ophthalmic disease in a subject. The method comprises administering to the subject a therapeutic agent for ophthalmic disease of the present disclosure. The therapeutic agent for ophthalmic disease may be a solid-in-oil formulation containing the therapeutic agent for ophthalmic disease as a drug. Administration may be by eye drop or intraocular administration, preferably intravitreal administration.

[0040] According to the present disclosure, there is provided a method for administering a therapeutic agent for an ophthalmic disease to a subject, wherein the therapeutic agent for an ophthalmic disease is a solid particle dispersed oil formulation (solid-in-oil formulation) containing the therapeutic agent for an ophthalmic disease as a drug.

[0041] According to the present disclosure, there is provided a solid-in-oil formulation for use in the methods of the present disclosure. [Example]

[0042] Example 1: Preparation of eye drop preparation and intravitreal preparation In this example, a formulation in which solid particles of a drug coated with a surfactant are dispersed in oil (solid-in-oil type formulation or solid particle dispersed oil formulation) was applied to the eye.

[0043] The drugs used were from various pharmaceutical modalities such as proteins, antibodies, and nucleic acids.

[0044] An experimental system was constructed to investigate the application of solid-in-oil formulations as ophthalmic formulations. Specifically, a jacketed, static Franz diffusion cell (Cosmedy) for membrane permeation tests was divided into upper and lower sections by a polycarbonate membrane (pore size: 0.1 μm, Merck). The solid-in-oil formulation was added to the upper section, and the lower section was filled with ocular irrigation solution (Opeguard MA ocular irrigation solution, Senju Pharmaceutical Co., Ltd.). The temperature was maintained at 36°C, the temperature of the ocular surface. When the resulting solid-in-oil formulation was added to the upper section, drug transfer to the ocular irrigation solution in the lower section was confirmed.

[0045] (1) Antibody A human whole IgG antibody was used as the antibody. An aqueous solution containing 1 mg / mL IgG and 0.3 mg mannitol was mixed with a cyclohexane solution containing 25 mg / mL surfactant and stirred at 26,000 rpm for 2 minutes using a homogenizer. The surfactant was selected from glyceryl dioleate (DGMO), sorbitan sesquioleate (SO), polyoxyethylene hydrogenated castor oil 10 (HCO), and sucrose laurate (L195). The resulting emulsion was lyophilized for 24 hours to remove the solvent and yield solid particles containing the drug coated with the surfactant. The resulting solid particles were dispersed in 1 mL of castor oil (Fujifilm Wako Pure Chemical Industries, Ltd.) to obtain a solid-in-oil formulation containing the IgG antibody. The particle size distribution of the solid particles in the resulting solid-in-oil formulation was measured using a Zetasizer (Zetasizer Nano ZS, Marvern). The particle size distribution results are shown in Figure 2B. The average hydrodynamic particle size was approximately 110 to 170 nm. Specifically, the average hydrodynamic particle size of particles with HCO as the surfactant was 120 ± 9 nm, with a PDI of 0.564 to 0.787. The average hydrodynamic particle size of particles with DGMO as the surfactant was 114 ± 7 nm, with a PDI of 0.307 to 0.399. The average hydrodynamic particle size of particles with SO as the surfactant was 122 ± 12 nm, with a PDI of 0.345 to 0.693. The average hydrodynamic particle size of particles with L195 as the surfactant was 163 ± 8 nm, with a PDI of 0.604 to 0.816.

[0046] Using the above experimental system, we confirmed the transfer of IgG to the ocular irrigation fluid in the lower part of the sample when the solid-in-oil formulation obtained in the upper part was added. The results are shown in Figure 2A. As shown in Figure 2A, IgG transferred slowly over time to the ocular irrigation fluid in the lower part of the sample for both surfactant-containing particles.

[0047] (2) Protein Cy5-labeled lysozyme hydrochloride was used as the protein. 1 mg of Cy5-lysozyme hydrochloride was dissolved in 1 mL of distilled water to obtain a 1 mg / mL lysozyme aqueous solution. The 1 mg / mL lysozyme aqueous solution was mixed with a cyclohexane solution containing 25 mg / mL of surfactant (sucrose erucate (ER-290) or sucrose laurate (L195)) and stirred at 26,000 rpm for 2 minutes using a homogenizer. The resulting emulsion was lyophilized for 24 hours to remove the solvent and obtain solid particles composed of the drug coated with the surfactant. The resulting solid particles were dispersed in 1 mL of castor oil (Fujifilm Wako Pure Chemical Industries, Ltd.) to obtain a solid-in-oil formulation containing lysozyme as the active ingredient. The particle size distribution of the solid particles in the resulting solid-in-oil formulation was measured using a Zetasizer (Zetasizer Nano ZS, Marvern). The particle size distribution results are shown in Figure 3B.

[0048] Using the above experimental system, we confirmed that lysozyme migrated to the ocular irrigation fluid below when the solid-in-oil formulation obtained above was added. The results are shown in Figure 3A. As shown in Figure 3A, lysozyme (LYZ) migrated slowly over time into the ocular irrigation fluid below for particles prepared using either surfactant.

[0049] (3) Anti-VEGF antibody The antibody used was a commercially available mouse-derived anti-VEGF antibody (Recombinant Mouse VEGF 164) was used. 100 μg of a commercially available antibody lyophilized with trehalose and PBS was dissolved in 500 μL of Milli-Q water. 50 μL of the antibody solution was mixed with 2 mL of a cyclohexane solution containing 4 mg / mL of surfactant and stirred at 26,000 rpm for 2 minutes using a homogenizer. The resulting emulsion was lyophilized for 24 hours to remove the solvent and yield solid particles of the drug coated with surfactant. The resulting solid particles were dispersed in 50 μL of castor oil to obtain a solid-in-oil formulation containing an anti-VEGF antibody as the active ingredient. The particle size distribution of the solid particles in the resulting solid-in-oil formulation was measured using a Zetasizer (Zetasizer Nano ZS, Marvern). The particle size distribution results are shown in Figure 4A.

[0050] The drug's post-administration kinetics were confirmed using rabbits. The resulting solid-in-oil formulation was administered intraocularly or intravitreally to rabbits at a dose of 0.4 μg per eye. The aqueous humor, vitreous, and retina / choroid were collected 24, 72, and 120 hours after administration, and the antibody levels in each were measured. As shown in Figure 4B, after intraocular administration of the resulting solid-in-oil formulation, a portion of the anti-VEGF antibody was detected in the retina / choroid up to 120 hours after administration. Furthermore, as shown in Figures 4C and 4D, after intravitreal administration of the resulting solid-in-oil formulation, most of the antibody migrated to the retina / choroid and maintained its concentration for a long period of time.

[0051] The antibody used was the anti-VEGF antibody aflibercept. Aflibercept was purified. Specifically, product additives from Eylea® intravitreal injection solution were removed using a PD-10 (Cytiva) according to the manufacturer's instructions. Subsequently, mannitol was added to the resulting antibody to a final concentration of 0.1%, and Milli-Q water was added to the resulting antibody to achieve an antibody concentration of 1 mg / mL, yielding an aqueous antibody solution. The aqueous antibody solution was mixed with a cyclohexane solution containing 25 mg / mL of surfactant and stirred at 26,000 rpm for 2 minutes using a homogenizer. The resulting emulsion was lyophilized for 24 hours to remove the solvent, yielding solid particles of the drug coated with surfactant. The resulting solid particles were dispersed in 1 mL of isopropyl myristate (Tokyo Chemical Industry Co., Ltd.) to yield a solid-in-oil formulation containing the anti-VEGF antibody as the active ingredient.

[0052] In the above experimental system, when 0.2 mL of the resulting solid-in-oil formulation was added to 5 mL of Opeguard MA, the drug migration into the lower ocular irrigation fluid was confirmed. In this experiment, the solid particles were dispersed in castor oil. The results are shown in Figure 5. As shown in Figure 5, the anti-VEGF antibody migrated slowly into the lower ocular irrigation fluid from both solid-in-oil formulations using surfactants. In contrast, aflibercept disappeared quickly. ELISA confirmed that the antibody released from the solid-in-oil formulation had the ability to bind to VEGF.

[0053] Example 2: Toxicity Testing Mice were used in the study. 100 μL of solid-in-oil formulation was intravitreally injected into mice (Balb / cJJcl, 10-week-old, female) (see STAR Protocols 1, 100094, September 18, 2020). Two weeks later, the eyes were harvested and histological sections were examined microscopically. The histological sections were 20 μm thick and stained with hematoxylin and eosin. The toxicity of the oil-based formulations, castor oil, isopropyl myristate, or silicone oil (KF96), was evaluated.

[0054] The results are shown in Figure 6 and Table 1. No significant toxicity was observed in the eye for any of the oil phases.

[0055] [Table 1]

[0056] As a way to allow drugs (especially water-soluble drugs) to penetrate the skin, preparations have been developed in which solid particles of the drug coated with a surfactant are dispersed in oil (solid-in-oil preparations) (WO2006 / 025583A). Because the skin is highly hydrophobic, hydrophilic drugs cannot penetrate the skin as they are. In contrast, by coating an aqueous preparation with a surfactant to form solid particles and dispersing them in an oil phase, the aqueous preparation can penetrate the skin and reach the bloodstream.

[0057] In this disclosure, solid-in-oil formulations were administered intravitreally and intraocularly. The solid-in-oil formulations demonstrated excellent long-term sustained drug release performance when administered intravitreally and intraocularly. The solid-in-oil formulations also had advantages such as high drug delivery and / or accumulation in the retina / choroid. Solid-in-oil formulations, in which solid particles are dispersed in oil, have high tissue penetration, suggesting their effectiveness in various intraocular administrations, not just intravitreal administration.

Claims

1. 1. An ophthalmic formulation comprising: (i) a solid complex comprising particles containing a pharmaceutically acceptable target substance or molecule and a pharmaceutically acceptable surfactant; (ii) a pharmaceutically acceptable oily base; and Including, the particles are coated with the surfactant; The solid complex is dispersed in the oily base. Ophthalmic preparations.

2. 10. The ophthalmic preparation of claim 1 formulated as eye drops.

3. 10. The ophthalmic preparation of claim 1, formulated for intravitreal administration.

4. The ophthalmic preparation according to any one of claims 1 to 3, wherein the target substance or target molecule is water-soluble.

5. The ophthalmic formulation according to any one of claims 1 to 4, wherein the target substance or molecule is sustained-released from the ophthalmic formulation after administration to a subject.

6. The ophthalmic formulation of any one of claims 1 to 4, wherein the target substance or molecule is sustainedly released from the ophthalmic formulation for at least 24 hours after administration to the vitreous of a subject.

7. The ophthalmic formulation according to any one of claims 1 to 3, wherein the target substance or molecule is sustainedly released by 50% or less over 24 hours from the ophthalmic formulation after administration to the vitreous of a subject.

8. The ophthalmic preparation according to any one of claims 1 to 7, wherein the target substance or target molecule is a molecule selected from the group consisting of a protein, a nucleic acid, and an antibody.

9. 9. The ophthalmic formulation of any one of claims 1 to 8, having a mean hydrodynamic particle size of 80 nm to 200 nm.

10. The ophthalmic preparation according to any one of claims 1 to 9, wherein the oily base is silicone oil.

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