WO2024085100A1

WO2024085100A1 - Solid particle dispersed oily preparation

Info

Publication number: WO2024085100A1
Application number: PCT/JP2023/037321
Authority: WO
Inventors: 桃子北岡; 航平石濱
Original assignee: ＮＯＶＩＧＯＰｈａｒｍａ株式会社
Priority date: 2022-10-17
Filing date: 2023-10-16
Publication date: 2024-04-25

Abstract

The present disclosure relates to a solid particle dispersed oily preparation. The present disclosure also provides a solid particle dispersed oily preparation suited to eyedrops or intraocular administration. The solid particle dispersed oil preparation contains (i) solid complexes that contain a pharmaceutically acceptable surfactant and particles containing a pharmaceutically acceptable target substance or target molecule and (ii) a pharmaceutically acceptable oily base. The particles in the solid particle dispersed oily preparation are typically covered with the surfactant. Also, the target substance or target molecule in the solid particle dispersed oily preparation can be water-soluble.

Description

Solid particle dispersion oil-based preparation

The present disclosure relates to solid particle dispersion oil-based formulations.

Formulations for treating ophthalmic diseases are being developed. US2021/145736A, US2017/273901A, US2013/164285A, US2012/258163A, and US2016/0235674A disclose methods for intravitreal administration of drugs.

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.

The present disclosure relates to a solid particle dispersion oil formulation. The solid particle dispersion oil formulation includes (i) a solid complex including particles containing a pharma- ceutically acceptable target substance or target molecule (e.g., a nucleic acid or a biopharmaceutical) and a pharma- ceutically acceptable surfactant, and (ii) a pharma- ceutically acceptable oil base. In the solid particle dispersion oil formulation, the particles are typically coated with the surfactant. In the solid particle dispersion oil formulation, the target substance or target molecule may be water-soluble. The present disclosure also provides a solid particle dispersion oil formulation suitable for ophthalmic or intraocular administration. The present disclosure further provides a solid particle dispersion oil formulation for ophthalmic or intraocular administration. In one aspect of the present disclosure, an ophthalmic formulation is provided, the formulation including (i) a solid complex including particles containing a pharma- ceutically acceptable target substance or target molecule and a pharma- ceutically acceptable surfactant, and (ii) a pharma- ceutically acceptable oil base.

In accordance with the present disclosure, there is provided an ophthalmic formulation comprising: (i) a solid complex comprising particles containing a pharma- ceutically acceptable target substance or molecule and a pharma-ceutically acceptable surfactant; and (ii) a pharma-ceutically acceptable oil base.

According to the present disclosure, for example, the following inventions are provided.
(1) An ophthalmic preparation comprising:
(i) a solid complex comprising particles containing a pharma- ceutically acceptable target substance or molecule and a pharma-ceutically acceptable surfactant;
(ii) a pharma- ceutically acceptable oil base;
the particles are coated with the surfactant,
The solid complex is dispersed in the oil base.
Ophthalmic preparations.
(2) The ophthalmic preparation according to (1) above, which is formulated as an eye drop.
(3) The ophthalmic preparation according to (1) above, which is for intravitreal administration or 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 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 molecule is sustained 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 sustained-released from the ophthalmic preparation at 50% or less over 24 hours after administration to the vitreous of a 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 (1) to (9) above, wherein the oily base is silicone oil.

FIG. 1 shows a schematic diagram of a test system simulating the intravitreal environment. FIG. 2A shows the release profile of antibody molecules from a formulation of the present disclosure in a test system simulating the intravitreal environment. FIG. 2B shows particle formation and particle size distribution of the particles formed for a formulation of the present disclosure that includes an antibody as a substance or molecule of interest. FIG. 3A shows the release behavior of a protein molecule (eg, lysozyme) from a formulation of the present disclosure in a test system simulating the intravitreal environment. FIG. 3B shows particle formation and the 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. FIG. 4A shows the release profile of an anti-VEGF antibody from a formulation of the present disclosure in a test system simulating the intravitreal environment. FIG. 4B shows the transfer of anti-VEGF antibody from a formulation of the present disclosure to the retina and choroid following ocular administration to rabbits. FIG. 54 shows the vitreous accumulation of anti-VEGF antibodies from a formulation of the present disclosure administered intravitreally to rabbits. FIG. 4D shows the translocation of anti-VEGF antibody from a formulation of the present disclosure to the retina and choroid following intravitreal administration in rabbits. FIG. 5 shows the release profile of aflibercept from a formulation of the present disclosure in a test system simulating the intravitreal environment. FIG. 6 is a histological section (hematoxylin and eosin stained) of the eye of a mouse to which a formulation of the present disclosure was administered intravitreally.

The following describes an embodiment of the present invention with reference to the drawings. Note that the present invention is not limited to the following embodiment and drawings. Note that in the following embodiment, the expressions "have", "include" and "contain" also include the meaning of "consisting of" or "consisting of". The singular form means singular or plural.

In this specification, a "subject" refers to a mammal, including a human, and preferably a human. Examples of mammals include, but are not limited to, humans, chimpanzees, and other primates; dogs, cats, rabbits, horses, sheep, goats, cows, pigs, rats (including nude rats), mice (including nude mice and skid mice), guinea pigs, and other livestock animals, pets, and laboratory animals.

As used herein, a "solid composite" is a composite made of solids. 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, a water content of 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 a known method (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, and the surfactant may have a hydrophilic portion facing the core and a hydrophobic portion facing the outer surface of the solid composite. In this way, the surfactant preferably covers the surface of the hydrophilic drug and turns the particle surface hydrophobic.

In this specification, a "solid particle-dispersed oil preparation" is a preparation that includes a plurality of solid particles and an oil base, and the solid particles are dispersed in the oil base. The solid particles include a target substance or target molecule (e.g., a nucleic acid or a biopharmaceutical) 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 the solid particle-dispersed oil preparation, the solid particles can be stably dispersed in the oil base. In this specification, the "solid particle" is also referred to as a "solid complex." The target substance or target molecule may be hydrophilic, and in this case, the solid particle-dispersed oil preparation is a preparation in which a hydrophilic target substance or target molecule is dispersed in an oil 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, a pharmaceutical active ingredient. The target substance or target molecule may be, for example, a therapeutic drug for an ophthalmic disease. The solid particle dispersion oil preparation preferably does not contain aggregates formed by aggregation of multiple 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). A biopharmaceutical is a pharmaceutical product that contains an active pharmaceutical ingredient produced by cells.

As used herein, a "surfactant" is a molecule that has a relatively hydrophilic portion and a relatively hydrophobic portion in the molecule. 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 fatty chain) such as an alkyl, alkenyl, or alkynyl. An anionic surfactant 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). "Long chain" is a term given to compounds that have 14 to 22 carbon atoms. The fatty chain may be a fatty chain with 8 to 22 carbon atoms, i.e., a C ₈ to C ₂₂ fatty chain such as a C ₈ _to C 22 alkyl, C ₈ to C ₂₂ alkenyl, or C ₈ to C ₂₂ alkynyl. The fatty chain may 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. The fatty chain having a double bond may be a cis type or a trans type, and the cis type is preferred from the viewpoint of biocompatibility. A nonionic surfactant is a surfactant that does not have an ionic group in the molecule, and has a nonionic hydrophilic portion and a hydrophobic portion. The hydrophobic portion may include a fatty chain such as an alkyl, alkenyl, or alkynyl (e.g., a long fatty chain).

As the nonionic surfactant, an ester compound made from unsaturated fatty acids such as erucic acid and oleic acid is preferred. Examples of lipophilic nonionic surfactants include sucrose fatty acid esters (sucrose stearate, sucrose palmitate, sucrose myristic acid, sucrose oleate, sucrose laurate, sucrose erucate, sucrose mixed fatty acid esters) with a high degree of esterification (i.e., a high ratio of di-, tri-, and polyester to monoester), polyglycerol condensed ricinoleate, decaglycerol ester, glycerol fatty acid ester, polyglycerol fatty acid ester, polyoxyethylene glycerol fatty acid ester, sorbitan fatty acid ester, polyoxyethylene sorbitol fatty acid ester, and polyoxyethylene castor oil/hydrogenated castor oil. The nonionic surfactant is preferably sucrose laurate. The surfactant may be used alone or in a mixture of two or more.

Surfactants include, for example, 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.

A suitable example of a surfactant used to prepare the complex is a lipophilic (hydrophobic) nonionic surfactant with 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.

As used herein, "antibody" refers to an immunoglobulin. The antibody may be of various isotypes, e.g., IgG. The antibody may preferably be a monoclonal antibody. The antibody may be a human chimeric antibody, a humanized antibody, or a human antibody. A human chimeric antibody may be produced by replacing the constant region of a non-human antibody with the constant region of a human antibody. A humanized antibody may be produced by replacing the six CDRs of a human antibody with the six corresponding CDRs of a non-human antibody. A human antibody may 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 may preferably be a humanized antibody. As used herein, the antibody may preferably be a human antibody. An antibody has a signal peptide when produced intracellularly, but the signal peptide is removed when secreted extracellularly. Therefore, when administered as a pharmaceutical, the antibody does not require a signal peptide.

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

In the present specification, "antigen-binding fragment of antibody" means a fragment of an antibody that maintains binding 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 prepared by methods well known to those skilled in the art. Such antigen-binding fragments of antibodies can be used in the present invention.

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

In this specification, "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.

According to the present disclosure, there is provided a solid particle dispersion oil formulation, comprising: (i) a solid complex comprising particles containing a pharma- ceutically acceptable target substance or molecule and a pharma-ceutically acceptable surfactant;
(ii) a pharma- ceutically acceptable oily base.

The solid particle dispersion oil 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.

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

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 cataracts. There are no particular limitations on the hydrophilic active ingredient in eye drops, so long as it is a medicinal ingredient that is administered to the eye.

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 the antibody include, for example, anti-VEGF antibodies (e.g., ranibizumab, bevacizumab, aflibercept, or brolucizumab). Anti-VEGF antibodies have been confirmed to be effective against age-related macular degeneration, diabetic retinopathy (diabetic macular edema), retinal vein occlusion, pathological myopia (choroidal neovascularization), and the like. The antibody or antigen-binding fragment thereof does not lose its antigen-binding ability in a solid complex or in a solid particle-dispersed oil formulation, and exhibits antigen-specific binding after administration. The protein or peptide does not lose its antigen-binding ability in a solid complex or in a solid particle-dispersed oil formulation, and exerts at least a part of its function after administration. Examples of the protein or peptide include lysozyme, insulin, albumin, ovalbumin, neutrophin 4, and the like. The nucleic acid includes one or more selected from the group consisting of DNA, RNA, and modified nucleic acid. The nucleic acid may be a single-stranded or double-stranded nucleic acid. The nucleic acid may be an antisense oligo. The RNA may include artificial RNA for gene silencing, such as siRNA and shRNA, non-coding RNA, such as microRNA (miRNA), aptamers, and natural RNA, such as mRNA. The nucleic acid or RNA may include aptamers (e.g., anti-VEGF aptamers). These RNAs may be modified to be stabilized in vivo.

Modified nucleic acids include, for example, nucleic acids modified with fluorescent dyes, biotinylated nucleic acids, and nucleic acids into which cholesteryl groups have been introduced. In order to enhance the stability of RNA, bases may be modified with 2'-O-methyl, 2'-fluoro, or 2'-methoxyethyl (MOE), and the phosphodiester bonds in the nucleic acid backbone may be replaced with phosphorothioate bonds. Artificial nucleic acids include nucleic acids in which the oxygen atom at the 2' position is crosslinked with the carbon atom at the 4' position. Examples of such artificial nucleic acids include locked nucleic acid (LNA), which is a bridged DNA in which the oxygen atom at the 2' position and the carbon atom at the 4' position are bridged via methylene; ENA, in which the oxygen atom at the 2' position and the carbon atom at the 4' position are bridged via ethylene; BNACOC, in which the oxygen atom at the 2' position and the carbon atom at the 4' position are bridged via -CH ₂ OCH ₂ -; bridged nucleic acid (BNA), such as BNANC, in which the oxygen atom at the 2' position and the carbon atom at the 4' position are bridged via -NR-CH ₂ - (where R is a methyl or hydrogen atom); cMOE, in which the oxygen atom at the 2' position and the carbon atom at the 4' position are bridged via -CH ₂ (OCH ₃ ₎ - _; )-bridged cEt; AmNA in which the 2'- and 4'-carbon atoms are bridged via an amide; scpBNA in which the 2'-position oxygen atom and the 4'-position carbon atom are bridged via a methylene and a cyclopropane is formed at the 6'-position; and peptide nucleic acid (PNA) in which the main chain of the polymer is formed by amide-bonding N-(2-aminoethyl)glycine instead of deoxyribose or ribose.

　Any surfactant can be used without particular restrictions as long as it is medicamentally acceptable. Examples of surfactants include nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, and bile salts.

Nonionic surfactants include polyglycerol condensed ricinoleate, decaglycerol ester, glycerol fatty acid ester, polyglycerol fatty acid ester, polyoxyethylene glycerol fatty acid ester, sorbitan fatty acid ester, polyoxyethylene sorbitol fatty acid ester, polyoxyethylene castor oil, hydrogenated castor oil, sucrose fatty acid ester (sucrose stearate, sucrose palmitate, sucrose myristate, sucrose oleate, sucrose laurate, sucrose erucate, sucrose mixed fatty acid ester), etc. One of these may be selected for use, or a mixture of two or more may be used.

As these nonionic surfactants, ester compounds made from unsaturated fatty acids such as erucic acid and oleic acid are preferred, and more preferred are sucrose erucic acid ester, sucrose oleic acid ester, and sucrose mixed fatty acid ester. Alternatively, one or more surfactants selected from the group consisting of glycerin fatty acid ester, polyglycerin fatty acid ester, polyoxyethylene glycerin fatty acid ester, sorbitan fatty acid ester, sucrose fatty acid ester, polyoxyethylene sorbit fatty acid ester, polyoxyethylene castor oil, and hydrogenated castor oil can be used.

The HLB value indicates the degree of hydrophilicity and hydrophobicity of a surfactant. A smaller HLB value means a higher hydrophobicity. In the present disclosure, the surfactant is not particularly limited, but it is preferable to use one with high hydrophobicity, with an HLB value of 10 or less. This is because it can easily dissolve or disperse the antigen-containing complex in the oil phase. In the present disclosure, surfactants having an HLB value of 10 or less, 8 or less, 6 or less, 5 or less, 4 or less, or 3 or less may be preferably used. In the present disclosure, the smaller the HLB value, the more preferable.

When a water-soluble target substance or molecule is mixed with a surfactant in an oil phase (preferably a volatile oil phase), particles having a core-shell structure are formed, the core containing the water-soluble target substance or molecule and the shell containing the surfactant. The particles are dispersed in the oil phase. If the oil phase used here is a volatile oil phase (e.g., cyclohexane), it is easy to subsequently evaporate the solvent, and by removing the solvent by drying (e.g., freeze-drying), the core-shell particles containing the water-soluble target substance or molecule and the shell containing the surfactant can be obtained as solid particles (also called "solid composites").
By dispersing the solid particles in an oily base, a solid particle-dispersed oil-based preparation can be obtained.

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.

In the present disclosure, the oil base may be any oil that is acceptable for use in pharmaceutical preparations administered to the eye (i.e., ophthalmic preparations such as eye drops or preparations for intravitreal administration). In the present disclosure, the oil base may be preferably an oil that is liquid at room temperature (25°C). In the present disclosure, the oil base may be either natural or synthetic. In the present disclosure, the oil base may 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, cacao oil, cinnamon oil, peppermint oil, eucalyptus oil, and bergamot oil, or animal oils such as beef tallow, lard, and fish oil. The oil base may be a neutral lipid such as glyceride, triolein, trilinolein, tripalmitin, tristearin, trimyristin, triarachidonin, or a synthetic lipid. The oil base may 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 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 oil-based base material may be used alone or in a mixture of two or more. Preferred oil-based base materials include squalane oil, castor oil, sesame oil, white petrolatum, and liquid paraffin, and more preferred oil-based base materials include castor oil, isopropyl myristate, and silicone oil.

In one embodiment, the oil 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.

According to the present disclosure, the solid particle-dispersed oil-based 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) is provided,
(i) a solid complex comprising particles containing a pharma- ceutically acceptable target substance or molecule and a pharma-ceutically acceptable surfactant;
(ii) A pharma- ceutically acceptable oil base is provided. In a preferred embodiment, the ophthalmic formulation is an eye drop. In a preferred embodiment, the ophthalmic formulation is an intravitreal formulation.

According to the present disclosure, the solid particle dispersion oil formulation can release the target substance or target molecule in a sustained manner. In one embodiment of the present disclosure, the solid particle dispersion oil formulation can release the target substance or target molecule in the eye of a living body (e.g., in 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 particle dispersion oil formulation of the present disclosure can release the target substance or target molecule in 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, the solid particle dispersion oil formulation releases the target substance or target molecule only at or below the sustained release standard value. The sustained release standard value is a standard time in which the target substance or target molecule contained in the formulation is released at 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. The sustained release standard time can 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 to the vitreous of the subject.

The solid particle dispersion oil formulation of the present disclosure can deliver the target substance or molecule to one or more of the anterior ocular tissues selected from the group consisting of the cornea, conjunctiva, aqueous humor, iris, ciliary body, and lens, and the 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 particle dispersion 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.

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

According to the present disclosure, there is provided a method of treating an ophthalmic disease in a subject. The method includes administering to the subject a therapeutic agent for ophthalmic disease of the present disclosure. The therapeutic agent for ophthalmic disease may be a solid particle dispersion oily formulation containing the therapeutic agent for ophthalmic disease as a drug. The administration may be by eye drops or intraocular administration, preferably intravitreal administration. The amount administered may be a therapeutically effective amount.

The present disclosure provides a method for administering a therapeutic drug for ophthalmic disease to a subject, the therapeutic drug for ophthalmic disease being a solid particle dispersion oil formulation containing the therapeutic drug for ophthalmic disease as a drug.

According to the present disclosure, there is provided a solid particle dispersion oil formulation for use in the method of the present disclosure. According to the present disclosure, there is provided the use of a solid particle dispersion oil formulation or a target substance or target molecule in the manufacture of a medicament for use in the method of the present disclosure.

Example 1: Preparation of eye drop preparation and intravitreal preparation In this example, a preparation in which solid particles formed by coating a drug with a surfactant are dispersed in oil (solid particle-dispersed oil-based preparation) was applied to the eye.

Drugs used included those from a variety of pharmaceutical modalities, including proteins, antibodies, and nucleic acids.

An experimental system was constructed to examine the application of solid particle dispersion oil preparations as ophthalmic preparations. Specifically, a stationary jacketed Franz diffusion cell for membrane permeation tests (manufactured by Cosmedy) was divided into the top and bottom by a polycarbonate membrane (pore size: 0.1 μm, manufactured by Merck), the solid particle dispersion oil preparation was added to the top, and the bottom was filled with ocular irrigation fluid (Opeguard) MA ocular irrigation fluid, manufactured by Senju Pharmaceutical Co., Ltd.). The temperature was maintained at 36°C, which is the eyeball surface temperature. When the obtained solid particle dispersion oil preparation was added to the top, the transfer of the drug to the ocular irrigation fluid in the bottom was confirmed.

(1) Antibody A human total 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 the mixture was stirred for 2 minutes at 26,000 rpm using a homogenizer. As the surfactant, one selected from glyceryl dioleate (DGMO), sorbitan sesquioleate (SO), polyoxyethylene hydrogenated castor oil 10 (HCO), and sucrose laurate (L195) was used. The obtained emulsion was freeze-dried for 24 hours to remove the solvent, and solid particles in which the drug was coated with the surfactant were obtained. The obtained solid particles were dispersed in 1 mL castor oil (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) to obtain a solid particle dispersion oil formulation containing an IgG antibody. The particle size distribution of the solid particles in the obtained solid particle dispersion oil formulation was measured using a Zetasizer (Zetasizer Nano ZS, manufactured by Marvern). The particle size distribution results were as shown in FIG. 2B. The average hydrodynamic particle size was about 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, and 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.

In the above experimental system, the transfer of the drug, IgG, to the ocular irrigation fluid at the bottom was confirmed when the solid particle dispersion oil preparation obtained was added to the top. The results are shown in Figure 2A. As shown in Figure 2A, IgG transferred slowly over time to the ocular irrigation fluid at the bottom for particles using either surfactant.

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

Using the above experimental system, we confirmed that lysozyme migrated to the ocular irrigation fluid at the bottom when the solid particle dispersion oil preparation obtained at the top was added. The results were as shown in Figure 3A. As shown in Figure 3A, lysozyme (LYZ) migrated slowly over time into the ocular irrigation fluid at the bottom for particles using either surfactant.

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

The kinetics of the drug after administration was confirmed using rabbits. The obtained solid particle dispersion oil formulation was administered by eye drop or intravitreally to rabbits so that the antibody amount was 0.4 μg per single eye. The aqueous humor, vitreous body, and retina/choroid of the rabbits were collected 24 hours, 72 hours, and 120 hours after administration, and the amount of antibody contained in each was measured. As shown in Figure 4B, after the obtained solid particle dispersion oil formulation was administered by eye drop, some of the anti-VEGF antibodies were detected in the retina/choroid up to 120 hours after administration. In addition, as shown in Figures 4C and 4D, after the obtained solid particle dispersion oil formulation was administered intravitreally, most of the antibodies migrated to the retina/choroid and maintained their concentration for a long period of time.

The antibody used was aflibercept, an anti-VEGF antibody. Aflibercept was purified. Specifically, the product additives of Eylea (registered trademark) intravitreal injection solution were removed using PD-10 (Cytiva) according to the manufacturer's manual, and then mannitol was added to the obtained antibody to a final concentration of 0.1%, and Milli-Q water was added to the obtained antibody to an antibody concentration of 1 mg/mL to obtain an antibody aqueous solution. The antibody aqueous solution was mixed with a cyclohexane solution containing 25 mg/mL surfactant, and stirred at 26,000 rpm for 2 minutes using a homogenizer. The obtained emulsion was freeze-dried for 24 hours to remove the solvent, and solid particles in which the drug was coated with the surfactant were obtained. The obtained solid particles were dispersed in 1 mL of isopropyl myristate (manufactured by Tokyo Chemical Industry Co., Ltd.) to obtain a solid particle dispersion oil formulation containing an anti-VEGF antibody as an active ingredient.

In the above experimental system, when 0.2 mL of the obtained solid particle dispersion oil preparation was added to the top of 5 mL of Opeguard MA, the migration of the drug into the lower ocular irrigation fluid of IgG was confirmed. In this experiment, the solid particles were dispersed in castor oil. The results were as shown in Figure 5. As shown in Figure 5, the anti-VEGF antibody slowly migrated to the lower ocular irrigation fluid from the solid particle dispersion oil preparations using both surfactants. In contrast, aflibercept disappeared quickly. ELISA confirmed that the antibody released from the solid particle dispersion oil preparation had the ability to bind to VEGF.

Example 2: Toxicity Test Mice were used in the test. Mice (Balb/cJJcl, 10 weeks old, female) were intravitreally injected with 100 μL of solid particle dispersion oil formulation (see STAR Protocols 1, 100094, September 18, 2020), and the eyeballs were collected after 2 weeks and tissue sections were observed under a microscope. The tissue sections were 20 μm thick and were subjected to hematoxylin-eosin staining. The toxicity of any of the oil bases, castor oil, isopropyl myristate, or silicone oil (KF96), was evaluated.

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

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 particle-dispersed oil preparations) (WO2006/025583A). The skin is highly hydrophobic, and hydrophilic drugs cannot penetrate the skin as is. In contrast, by coating an aqueous preparation with a surfactant, forming it into solid particles, and dispersing them in the oil phase, the aqueous preparation can penetrate the skin and reach the blood.

In the present disclosure, the solid particle-dispersed oil formulation was administered by eye drop and intravitreal administration. The solid particle-dispersed oil formulation was excellent in long-term sustained release performance of the drug when administered by eye drop and intravitreal administration. The solid particle-dispersed oil formulation also had advantages such as high delivery and/or accumulation of the drug to the retina/choroid. It is suggested that the preparation in which solid particles are dispersed in oil (solid particle-dispersed oil formulation) has high tissue permeability and is effective not only in intravitreal administration but also in various intraocular administrations.

Claims

1. An ophthalmic formulation comprising:
(i) a solid complex comprising particles containing a pharma- ceutically acceptable target substance or target molecule and a pharma-ceutically acceptable surfactant;
(ii) a pharma- ceutically acceptable oil base;
the particles are coated with the surfactant,
The solid complex is dispersed in the oil base.
Ophthalmic preparations.
The ophthalmic preparation according to claim 1, which is formulated as an eye drop.
The ophthalmic preparation of claim 1, which is for intravitreal administration or is formulated for intravitreal administration.
The ophthalmic preparation according to any one of claims 1 to 3, wherein the target substance or target molecule is water-soluble.
The ophthalmic preparation according to any one of claims 1 to 4, wherein the target substance or target molecule is sustained-released from the ophthalmic preparation after administration to a subject.
The ophthalmic formulation according to any one of claims 1 to 4, wherein the target substance or molecule is released from the ophthalmic formulation for at least 24 hours after administration to the vitreous of the subject.
The ophthalmic preparation according to any one of claims 1 to 3, wherein the target substance or molecule is released from the ophthalmic preparation at a sustained rate of 50% or less over a 24-hour period following administration to the vitreous of a subject.
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 proteins, nucleic acids, and antibodies.
The ophthalmic preparation according to any one of claims 1 to 8, wherein the average hydrodynamic particle size is 80 nm to 200 nm.
The ophthalmic preparation according to any one of claims 1 to 9, wherein the oily base is silicone oil.