Brillaloxazine liposome composition

Abstract

The present invention relates to a liposome composition comprising brillaroxazine incorporated into bilayer lipid vesicles. In one embodiment, the liposome composition comprises bilayer lipid vesicles encapsulating an aqueous solution, wherein the bilayer lipid vesicles contain one or more phospholipids, sterols, and brillaroxazine, and the aqueous solution contains maltodextrin. The present invention also relates to a pharmaceutical composition comprising a brillaroxazine liposome composition and a pharmaceutically acceptable carrier.

Description

Technical Field

[0001] The present invention relates to a liposomal composition comprising brexpiprazole or a pharmaceutically acceptable salt thereof. The liposomal composition comprises bilayer lipid vesicles encapsulating an aqueous solution, wherein the bilayer lipid vesicles contain brexpiprazole and the aqueous solution contains maltodextrin.

Background Art

[0002] Liposomes are particulate or colloidal carrier systems, usually with a diameter of 0.025 - 5.0 μm. Liposomes are composed of biodegradable and biocompatible components and provide a unique opportunity to deliver pharmaceuticals even within cells or individual cell compartments. Liposomes form spontaneously when lipids are hydrated in an aqueous medium at the transition temperature. Lipids are composed of natural and / or synthetic lipids (phospholipids and sphingolipids) and may contain other bilayer components such as cholesterol and hydrophilic polymer lipids. Figure 1 shows the general structure of a liposome.

[0003] The composition of liposomes determines their interaction with blood and tissues. The composition determines the net physicochemical properties of liposomes, namely membrane fluidity, charge density, and steric hindrance permeability. They have been found to be useful carriers for both hydrophilic and hydrophobic drugs. These drug delivery systems are used for the delivery of drugs with various lipid solubilities such that water-soluble drugs are encapsulated in the aqueous compartment and lipid-soluble drugs usually bind to the lipid bilayer or dissolve in the lipid phase.

[0004] Brillaloxazine (RP5063) is a polymorphic dopamine and 5-HT receptor modulator. Brillaloxazine exhibits high binding affinity to D2-4 and 5-HT1A receptors as a partial agonist, to 5-HT2A as a weak partial agonist or neutral agonist, and to 5-HT2B / 7 as an antagonist, while showing moderate affinity to the serotonin transporter (SERT). Brillaloxazine has established efficacy, safety, and pharmacokinetic profiles from phase 1 and 2 studies in healthy individuals and patients with schizophrenia. Preclinical studies also indicate that this drug inhibits the release of multiple inflammatory cytokines. [Brief explanation of the drawing]

[0005] [Figure 1] Figure 1 shows a diagram of the general structure of a liposome. [Figure 2] Figure 2 shows the particle size (Z-mean) and particle distribution index of brillaroxazine liposomes during liposome dispersion, as measured by DLS. [Figure 3] Figure 3 shows the HPLC chromatogram of brillaroxazine liposomes. [Figure 4] Figure 4 shows the HPLC chromatogram of the lipogel sample. The first peak is brillaroxazine, and the second peak is the excipient. [Figure 5] Figure 5 shows a graph of in vitro diffusion studies of lipogel through a membrane. [Modes for carrying out the invention]

[0006] Brillaloxazine liposomes The present invention relates to a liposome composition comprising brillaroxazine incorporated into bilayer lipid vesicles. In one embodiment, the liposome composition comprises bilayer lipid vesicles encapsulating an aqueous solution, wherein the bilayer lipid vesicles contain one or more phospholipids, sterols, and brillaroxazine, and the aqueous solution contains maltodextrin. Brillaloxazine (free base) is a basic and lipophilic molecule with a molecular weight of 450.36 g / mol. Its chemical structure is shown below.

[0007] [ka]

[0008] Brillaloxazine often forms an HCl salt with a molecular weight of 486.7 g / mol.

[0009] Maltodextrin consists of D-glucose units linked in a chain of variable length. These glucose units are primarily linked by α(1→4) glycosidic bonds. Maltodextrin typically comprises a mixture of chains ranging from 3 to 17 glucose units in length. Brillaloxazine liposomes encapsulating maltodextrin may offer a superior drug release profile compared to those without maltodextrin.

[0010] The lipids used in the formation of lipid vesicles typically consist of a lipid mixture mainly composed of phospholipids and sterols. A list of phospholipids commonly used in liposome preparation is provided on page 471 of Szoka et al. (Ann Rev Biophys Bioeng (1980) 9:467). The vesicles are formulated to contain negatively or positively charged lipids such as phosphatidic acid (PA) and phosphatidylglycerol (PG) to provide a desirable surface charge for the reagent vesicles. A small amount (0.1-1 mol%) of antioxidant, such as α-tocopherol, can be added to the lipid mixture to improve stability. A typical lipid mixture used in the formation of brillaroxazine liposomes in this invention includes phosphatidylcholine, cholesterol, and brillaroxazine.

[0011] Brillaloxazine liposomes are first prepared by dissolving vesicle-forming lipids (e.g., brillaloxazine, phosphatidylcholine, cholesterol) in an inert organic solvent or solvent system, such as chloroform and / or ethanol, to form an organic lipid phase solution. Generally, the inert organic solvent or solvent system is one in which the lipid components can be readily dissolved at a concentration range of approximately 0.5 to 50 mg lipid / mL. Next, the lipid solution is completely dried to remove the organic solvent and form a thin lipid film on the surface of the container. For example, the lipid solution can be dried at 45 to 50°C using a rotary evaporator under reduced pressure to remove the solvent. After all solvent removal, a thin film is formed in the round-bottom flask, and then the remaining components in the thin lipid film are completely removed under reduced pressure for 12 to 24 hours. After drying, the thin lipid film is then hydrated with an aqueous solution. In preferred embodiments, the aqueous solution contains maltodextrin.

[0012] In one embodiment, the brillaroxazine liposomes contain 10-40% or 20-30% brillaroxazine by weight.

[0013] In one embodiment, the brillaroxazine liposomes contain 20-60% or 30-45% maltodextrin by weight.

[0014] In one embodiment, the average particle size of brillaroxazine liposomes is 500-700 nm.

[0015] In one embodiment, the most prominent peak of the brillaroxazine liposome has a peak size of 900–1000 nM.

[0016] Pharmaceutical composition The present invention provides a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers and the brillaloxazine liposomes of the present invention. The amount of brillaloxazine or a pharmaceutically acceptable salt thereof in the pharmaceutical composition is generally about 0.01 to 20%, or 0.05 to 20%, or 0.1 to 20%, or 0.1 to 10%, or 0.1 to 5%, or 0.2 to 15%, or 0.2 to 10%, or 0.2 to 5%, or 0.2 to 2%, or 1 to 5% (w / w) in topical formulations, about 0.1 to 5% in injectable formulations, 0.1 to 5% in patch formulations, about 1 to 90% in tablet formulations, and 1 to 100% in capsule formulations.

[0017] In one embodiment, brillaroxazine liposomes are incorporated into any acceptable carrier, including creams, gels, lotions, or other types of suspensions, which can stabilize the active compound and deliver it to the affected area by topical administration. In another embodiment, the pharmaceutical composition may be in the form of tablets, capsules, granules, powders, syrups, suppositories, injections, patches, or similar dosage forms. The pharmaceutical composition can be prepared by conventional methods.

[0018] A pharmaceutically acceptable carrier, which is an inert component, can be selected by those skilled in the art using conventional criteria. A pharmaceutically acceptable carrier includes, but is not limited to, non-aqueous-based solutions, suspensions, emulsions, microemulsions, micelle solutions, gels, and ointments. Pharmaceutically acceptable carriers also include physiological saline and aqueous electrolyte solutions; ionic and nonionic osmotic agents such as sodium chloride, potassium chloride, glycerol, and dextrose; pH adjusters and buffers such as hydroxides, phosphoric acid, citric acid, acetic acid, boric acid, and salts of trolamine; antioxidants such as bisulfite, sulfite, metabisulfite, thiosulfite, ascorbic acid, acetylcysteine, cysteine, glutathione, butyrylhydroxyanisole, butyrylhydroxytoluene, tocopherol, and salts of ascorbyl palmitate, acids and / or bases; surfactants such as lecithin, phospholipids such as non-limited phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol; poloxamer and poloxamine, polysorbate 80, polysorbate The formula includes, but is not limited to, polysorbates such as T60 and polysorbate 20, polyethers such as polyethylene glycol and polypropylene glycol; polyvinyls such as polyvinyl alcohol and povidone; cellulose derivatives such as methylcellulose, hydroxypropylcellulose, hydroxyethylcellulose, carboxymethylcellulose and hydroxypropylmethylcellulose and their salts; petroleum derivatives such as mineral oil and white petrolatum; fats such as lanolin, peanut oil, palm oil and soybean oil; mono, di, and triglycerides; acrylic acid polymers such as carboxypolymethylene gel and hydrophobically modified crosslinked acrylic acid copolymers; polysaccharides such as dextran and glycosaminoglycans such as sodium hyaluronate. Other pharmaceutically acceptable carriers include xanthan gum, carrageenan, Avicel RC-591 (in combination with crystalline cellulose), and polyethylene glycol.Alternatively, the active compound may be dissolved or suspended in a pharmaceutically acceptable lipid preparation, such as vegetable oil, coconut oil, or castor oil, as shown by Kalepu et al. (Acta Pharmaceutica Sinica B, 3:361-372, 2013).

[0019] Such pharmaceutically acceptable carriers can be protected from bacterial contamination using well-known preservatives, which include, but are not limited to, benzalkonium chloride, ethylenediaminetetraacetic acid and its salts, benzethonium chloride, chlorhexidine, chlorobutanol, methylparaben, thimerosal, and phenylethyl alcohol, or can be formulated as non-preservative formulations for single or multiple use.

[0020] For example, tablet or capsule formulations of brillaroxazine liposomes may contain other excipients that are not bioactive and do not react with the active compound. Excipients for tablets or capsules may include fillers, binders, lubricants and flow enhancers, disintegrants, wetting agents, and release rate modifiers. Binders are important in tablet formulations as they promote particle adhesion in the formulation. Examples of excipients for tablets or capsules include, but are not limited to, carboxymethylcellulose, cellulose, ethylcellulose, hydroxypropylmethylcellulose, methylcellulose, karaya gum, starch, tragacanth gum, gelatin, magnesium stearate, titanium dioxide, polyacrylic acid, and polyvinylpyrrolidone. For example, tablet formulations may contain inert components such as colloidal silicon dioxide, crospovidone, hypromellose, magnesium stearate, crystalline cellulose, polyethylene glycol, sodium starch glycolate, and / or titanium dioxide. The capsule formulation may contain inert components such as gelatin, magnesium stearate, and / or titanium dioxide.

[0021] For example, a patch formulation of an active compound may contain several inactive ingredients such as 1,3 - butylene glycol, dihydroxyaluminum aminoacetate, disodium edetate, D - sorbitol, gelatin, kaolin, methylparaben, polysorbate 80, povidone, polypropylene glycol, propylparaben, sodium carboxymethylcellulose, sodium polyacrylate, tartaric acid, titanium dioxide, and purified water. The patch formulation may also contain a skin penetration enhancer such as a lactate ester or diethylene glycol monoethyl ether.

[0022] Topical formulations containing an active compound can take the form of gels, creams, lotions, liquids, emulsions, ointments, sprays, solutions, and suspensions. Inactive ingredients in the topical formulation include, for example, (skin softener / penetration enhancer), diethylene glycol monoethyl ether (skin softener / penetration enhancer), DMSO (solubility enhancer), silicone elastomer (rheology / texture modifier), caprylic / capric triglyceride, (skin softener), octisalate, (skin softener / UV filter), silicone fluid (skin softener / diluent), squalene (skin softener), sunflower oil (skin softener), and silicon dioxide (thickener), but are not limited thereto.

[0023] The present invention further provides a gel formulation comprising a blrilaxazine liposome composition, a gelling agent, and a water - retaining agent. In one embodiment, the gel formulation contains blrilaxazine in an amount of 0.005 - 10%, 0.01 - 5%, or 0.1 - 2% by weight. In one embodiment, the formulation has the appearance of a gel and the bilayer lipid vesicles are intact and stable in the gel. In one embodiment, the gelling agent is carbomer 940 and the water - retaining agent is glycerin.

[0024] Method of Use The present invention provides a method for treating psoriasis. The method comprises administering an effective dose of brillaroxazine liposomes to a subject in need. As used herein, “effective dose” means an amount effective in treating psoriasis by restoring the condition or reducing the symptoms of psoriasis. The method reduces one or more signs and symptoms selected from the group consisting of red spots of skin covered with thick, silvery scales; small, scaly spots; dry, cracked skin; itchy, burn, or painful skin; thickened, porosity or ridged nails; and enlarged, stiff joints.

[0025] The pharmaceutical compositions of the present invention can be administered by topical and systemic methods. Topical administration includes local administration. Commonly used conventional semi-fluid dosage forms have certain limitations in drug delivery due to the barrier properties of the skin. The skin is continuously involved in the efficient construction of a homeostatic barrier.

[0026] Brillaloxazine liposome gel formulations offer the following advantages: Liposomes are microscopic vesicles containing amphiphilic phospholipids arranged in one or more concentric bilayers that encapsulate an aqueous compartment. As a spherical shell, liposomes resemble biological membranes. Composed of biodegradable, biocompatible components, liposomes provide a drug delivery system that delivers pharmaceuticals into cells or individual cellular compartments. Therefore, brillaloxazine liposome gel formulations may be able to deliver brillaloxazine to deeper layers of the skin in severe psoriatic conditions.

[0027] Systemic administration methods include oral, parenteral (e.g., intravenous, intramuscular, subcutaneous, or rectal), and other systemic administration routes. In systemic administration methods, the active compound first reaches the plasma and then is distributed to the target tissue. Local administration and oral administration are preferred administration routes in this invention.

[0028] In one embodiment, the composition is applied topically to the affected area and rubbed in. The composition is administered topically at least once, twice, or three to four times a day, depending on whether the medical problem and the disease are chronic or acute. Generally, the topical composition contains about 0.01 to 10% (w / w) of the active compound. For example, the topical composition contains about 0.1 to 2% (w / w) of the active compound. Depending on the size of the affected area, 0.2 to 85 mL, typically 0.2 to 10 mL, of the topical composition is applied individually per dose. The active compound is delivered to the area of ​​discomfort by passing through the skin.

[0029] Those skilled in the art will recognize that a wide variety of delivery mechanisms are also suitable for the present invention.

[0030] This brillaroxazine liposome composition is useful for the treatment of mammals such as humans, horses, and dogs. The present invention is particularly useful for the treatment of humans.

[0031] The following embodiments further illustrate the present invention. These embodiments are intended merely to illustrate the present invention and should not be construed as limiting. [Examples]

[0032] Example 1: Preparation of brillaroxazine liposomes Table 1 shows the formulation composition of brillaroxazine liposomes.

[0033] [Table 1]

[0034] Brillaloxazine liposomes were prepared by lipid hydration. Briefly, phosphatidylcholine and cholesterol were dissolved in a suitable solvent (chloroform and / or ethanol), and brillaloxazine was dissolved in the same solvent. This drug-lipid solution was then dried under reduced pressure at 45-50°C in a rotary evaporator to completely remove the solvent. After all solvent was removed, a thin film formed in a round-bottom flask. The round-bottom flask containing this lipid film was kept under reduced pressure for 12-24 hours to completely remove any trace amounts of solvent present in the thin lipid film. After drying for 12-24 hours, the thin film was hydrated with 66 mL of maltodextrin solution at 60°C (concentration 39.57 mg / mL).

[0035] Example 2: Particle size and zeta potential analysis of brillaroxazine liposomes The liposomes prepared in Example 1 were observed under a light microscope at different magnifications to confirm the hydration process. To confirm the prepared spherical liposomal vesicles, light microscopy observations at different magnifications (10x, 20x, and 40x) were performed throughout the hydration process.

[0036] The particle size of the liposomes was analyzed using the DLS (dynamic light scattering) method for particle size analysis and drug content analysis. The Z-mean (particle size) was measured for the prepared liposomes. Figure 2 shows the particle size (Z-mean) and particle distribution index in the liposome dispersion, as measured by DLS of brillaroxazine liposomes.

[0037] The zeta potential was estimated from the experimentally determined electrophoretic mobility of the particles. The value of the Z potential indicates the stability of the colloidal dispersion.

[0038] Zeta potential (mV) value: • 0-5 - Rapid coagulation or aggregation • 10-30 - Initial unstable state 30-40 - Moderate stability 40-60 - Good stability ·>61—Excellent stability

[0039] Generally, colloidal dispersions with a zeta potential greater than 30mV positive or less than 30mV negative exhibit high stability. The higher the zeta potential (both positive and negative), the greater the stability.

[0040] Table 2 shows the particle size and zeta potential analysis of liposomes by DLS method.

[0041] [Table 2]

[0042] Example 3: Measurement of drug content by HPLC The liposome drug content was analyzed by HPLC.

[0043] HPLC system Shimadzu HPLC system (LC-2030C Plus, serial number: L21445711704 AE, Made in Japan), autosampler, UV detector, data acquisition system. An equivalent system can be used as a replacement.

[0044] HPLC column Shimadzu Shim-Pack GIST C18, 5μm, 250x4.6mm column or equivalent.

[0045] Reagent preparation Mobile phase A Dissolve 2.72g of KH2PO4 in 1000mL of ultrapure water (0.02M solution). Adjust the pH to 3.0 with phosphoric acid. Mix the above buffer solution with acetonitrile in a 90:10 ratio. • Filter through a membrane before use.

[0046] Mobile phase B Mix acetonitrile and ultrapure water in a 90:10 ratio. Adjust the pH to 3.0 with phosphoric acid. • Filter through a membrane before use.

[0047] Diluent • To optimize drug content and encapsulation efficiency, prepare a mixture of acetonitrile and phosphate buffer (Section 6.2.1.1) in a ratio of 85:15.

[0048] Sample preparation for drug content and incarnation efficiency Drug content Transfer the required amount of liposome sample (total dispersion) to a volumetric flask and add the required amount of diluent (6.2.3), mixing thoroughly. Maintain this mixture in a sonicator bath at 60°C for 30-45 minutes. Take the required amount of prepared sample and dilute to the required concentration. The assay / drug concentration is 20 μg / mL.

[0049] Import efficiency Place the required amount of liposome sample into a centrifuge tube and centrifuge the liposome dispersion at 10,000 rpm for 30 minutes at 20°C. Remove the supernatant and collect the pellet. Add the required amount of diluent to the liposome pellet and mix well. Incubate this mixture in a sonicator tank for 30-45 minutes at 60°C. Take the required amount of the prepared sample and dilute it to a drug concentration of 20 μg / mL.

[0050] Standard solution Prepare a 20 μg / mL standard solution using the above dilution.

[0051] analysis Configure the HPLC using the following parameters: Column: Shimadzu Shim-Pack GIST C18, 5μm, 250x4.6mm column or equivalent. Flow rate: 1mL / min Injection volume: 20μL Detection: UV@215nm Column temperature: 30℃ Execution time: 5-7 minutes HPLC conditions: Mobile phase A 25%, Mobile phase B 75%

[0052] Identification Compare the retention times of the standard / pure drug peak and the sample drug peak.

[0053] result The HPLC chromatogram is shown in Figure 3. The HPLC results indicate a drug content of 96% and a liposome uptake efficiency of 73%.

[0054] The final formulation composition was consistent with brillaroxazine (24.53%), lecithin (34.75%), cholesterol (2.97%), maltodextrin (37.74%), and purified water for lipid membrane hydration.

[0055] Example 4: Preparation of liposome gel formulation Liposome gels were prepared by incorporating a liposome dispersion into a gel formulation. First, a plain gel was prepared, then the liposome dispersion was added and thoroughly mixed to produce a liposome gel or lipogel. Lipogel formulations with various proportions (0.25–1.5% brillaroxazine) were prepared as needed. Table 3 shows the compositions of the lipogel formulations.

[0056] [Table 3]

[0057] The prepared liposome gel formulations were evaluated for their physical appearance and pH. All gel formulations were observed for intact liposomes under a light microscope. The liposome gel formulations had the appearance of a white cream gel, a pH of 5–6, and the presence of liposomes was confirmed by microscopic examination. Furthermore, the liposome particles were intact and stable in all gel formulations.

[0058] Example 5: Lipogel analysis by HPLC The lipogel sample was placed in a volumetric flask, and the diluent was added and mixed thoroughly. The brillaroxazine content was analyzed by HPLC according to the same protocol as in Example 3.

[0059] By comparing the retention times of the pure drug peak and the sample peak, the drug content was calculated to be 95.12%.

[0060] The HPLC chromatogram of the lipogel sample is shown in Figure 4. The first peak is brillaloxazine, and the second peak is the excipient. The brillaloxazine peak is distinct and separated from the excipient peak.

[0061] Example 6: In vitro diffusion / penetration study of lipogel The prepared lipogels were analyzed for drug diffusion / osmosis using a Franz diffusion cell. The Franz diffusion cell was filled with PBS buffer at pH 7.4. A regenerated cellulose dialysis membrane (MW cutoff: 12000-14000), surface-treated and neutralized with PBS at pH 7.4, was placed in the receptor compartment, and the weighed amount of lipogel was placed in the donor compartment. The receptor solution was agitated, and the skin temperature in the diffusion cell was maintained by temperature-controlled water circulation in an outer jacket. At different time intervals, the sample was removed through the sample port and replaced with an equal volume of plain PBS. The removed sample was mixed with an equal volume of HPLC diluent, and the drug content at different time intervals was analyzed. The drug diffusion rate, flux ratio, and osmosis coefficient were calculated for each lipogel formulation. The drug diffusion / release rate over time was plotted using GraphPad Prism version 6.01 software.

[0062] HPLC analysis of drug diffusion samples The HPLC analysis method, system, and column were the same as those described in Example 3, except that a mixture of acetonitrile and phosphate buffer (60:40) was used in the drug diffusion / permeation study.

[0063] Sample preparation for diffusion / permeation sample analysis Mix the diffusion / permeation sample solution recovered from the Franz diffusion apparatus with an equal volume of diluent. Mix thoroughly using a vortex mixer and filter the solution through a syringe filter.

[0064] result The HPLC chromatogram results of the lipogel diffusion sample show a clear and separated brillaroxazine peak.

[0065] Figure 5 shows the graph of the in vitro diffusion study of the lipogel through the membrane. The release profile showed steady and sustained brillaroxazine release from the formulation throughout the entire 8-hour study period.

[0066] Table 4 shows the results of in vitro diffusion studies of lipogel formulations.

[0067] [Table 4]

[0068] Liposome-containing liposomal formulations with maltodextrin exhibit superior drug release profiles and higher flux and osmotic values ​​in in vitro diffusion studies compared to liposome formulations without maltodextrin. This may be due to increased solubility of RP5063 in the liposome gel and an optimal particle size distribution within the formulation.

[0069] The foregoing describes preferred embodiments of the present invention, and it will be understood that modifications are possible without departing from the scope of the invention as described in the claims.

Claims

1. A liposome composition comprising bilayer lipid vesicles encapsulating an aqueous solution, wherein the bilayer lipid vesicles contain one or more phospholipids, sterols, and brillaroxazine, or pharmaceutically acceptable salts thereof, and the aqueous solution contains maltodextrin.

2. The liposome composition according to claim 1, wherein the phospholipid comprises phosphatidylcholine and sterols containing cholesterol.

3. The liposome composition according to claim 1, comprising 20 to 30% by mass of brillaroxazine.

4. The liposome composition according to claim 1, comprising 30 to 45% by mass of maltodextrin.

5. The liposome composition according to claim 1, wherein the average particle size of the lipid vesicles is 500 to 750 nm.

6. A pharmaceutical composition comprising the liposome composition of claim 1 and a pharmaceutically acceptable carrier.

7. A gel formulation comprising a liposome composition, a gelling reagent, and a water-retaining agent according to any one of claims 1 to 4, wherein the gel formulation contains 0.1 to 2% by mass of brillaroxazine, has a gel-like appearance, and the bilayer lipid vesicles are intact and stable in the gel.

8. The gel formulation according to claim 5, wherein the gelling reagent is carboner 940 and the water-retaining agent is glycerin.

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