Nanosuspension containing progerinin and method for producing the same

Abstract

The present invention relates to a nanosuspension formulation suitable for poorly soluble progerinin drugs and a method for preparing the same. Specifically, the progerinin nanosuspension according to the present invention is a vehicle composition for improving the dispersion stability of poorly soluble progerinin drugs (a), and is prepared by using hydroxypropylmethylcellulose (HPMC) (b) or hydroxypropylcellulose (HPC) (b'), TPGS (c) or dioctyl sodium succinate (DOSS) (c') and potassium sorbate (d), and subjecting the mixture to wet ball milling in a dyno mill chamber to prepare a white nanosuspension containing a progerinin drug with an average particle size of 100 to 300 nm. Such a nanosuspension has uniform dispersion of progerinin drug particles, minimized size change of drug particles even at low temperatures or in harsh environments, and excellent bioabsorption efficiency of the drug. Therefore, the nanosuspension containing progerinin according to the present invention is widely used as an oral nanosuspension formulation for confirming the therapeutic effect of premature aging.

Description

[Technical Field]

[0001] The present invention relates to a nanosuspension containing the poorly soluble drug progerinin and a method for producing the same. [Background technology]

[0002] Many useful drugs are hydrophobic, with low solubility in aqueous media, making them difficult to formulate into suspensions in aqueous vehicles. Due to these characteristics, wetting agents are often required to facilitate the suspension of hydrophobic drug particles in aqueous media. Surface-active wetting agents (i.e., surfactants), such as sodium lauryl sulfate, are known to reduce the interfacial tension between drug particles and the suspension vehicle, allowing the suspension vehicle to penetrate drug aggregates and / or drug particle pores, at least in part, thereby increasing the suspendability of hydrophobic drugs in aqueous media. However, in addition to beneficial drug-suspension effects, the use of surfactants in suspensions can also result in the undesirable result of solubilized and / or dissolved free drug.

[0003] Because solubilized and / or dissolved drugs are vulnerable to chemical degradation and / or interactions with other components, suspensions containing free drug are chemically unstable. Another undesirable consequence of using relatively high amounts of surfactant to promote the suspension of low-solubility drugs is that air entrained during homogenization or shaking of such suspensions tends to remain trapped because the surfactant stabilizes air bubbles. Such trapped air changes the volume of the suspension, which varies depending on the stirring force, stirring duration, and stirring time, making it difficult or impossible to administer a uniform dose over time.

[0004] If a low-water-solubility drug is administered as a suspension, it is desirable for the suspension to exhibit slow settling to provide adequate dose uniformity. Conversely, if rapid settling occurs, as in the case of a vehicle, the suspension must be shaken before each administration to achieve dose uniformity. Other factors being equal (e.g., drug particle size, uniformity, and density), as the viscosity of a particular suspension vehicle increases, the rate of settling of drug particles decreases. Therefore, it is desirable for a suspension to be appropriately viscous to prevent or slow the settling of drug particles. However, while such increased viscosity promotes physical stability, it also makes the suspension difficult to pour or administer.

[0005] Meanwhile, progerinin having the structure of the following chemical formula 1 exhibits excellent effects of inhibiting progerin expression and inhibiting the binding of progerin to lamin A, and is a drug that has the effect of extending the survival time of an animal model in which progeria is induced, and is used as a pharmaceutical composition for preventing or treating progeria.

[0006] [ka]

[0007] The compound name of the progerinin is (7S)-(+)-8,8-Dimethyl-7-(3-phenylallyloxy)-7,8-dihydro-6H-pyrano[3,2-g]chromen-2-one (which will be named "SLC-D011").

[0008] Progerinin is a drug that has been shown to be effective against premature aging, and is a BCS class II molecule that exhibits high apparent permeability and low solubility in aqueous media according to the BCS (Biopharmaceutical Classification System) criteria.

[0009] The low aqueous solubility of progerinin drugs severely limits their therapeutic applications. To overcome these limitations, numerous approaches have been explored, including the use of polymeric nanoparticles, solid lipid nanoparticles, self-emulsifying drug delivery systems, nanoemulsions, liposomes, nanosuspensions, and nanofibers. Nanosuspensions are colloidal dispersions of drug particles stabilized by polymers, surfactants, or both. Nanosuspensions are used to deliver drug substances with low aqueous and lipid solubility. The small particles of nanosuspensions provide a significantly larger drug surface area, increasing the dissolution rate of insoluble drugs. As a result, BCS Class II and IV compounds exhibit improved bioavailability, rapid activity, and other desirable biopharmaceutical effects.

[0010] Under these circumstances, the present inventors have been able to conduct animal experiments using a mixture of monoolein and tricaprin, an oil-based solution, but have found that the use of a dosage form dissolved in oil is not possible due to the characteristics of the disease, which requires high doses and long-term administration, and have recognized the need to develop a new dosage form that allows long-term administration. In particular, progerinin is so poorly soluble in water that its solubility is close to zero, and it also has very low solubility in existing ingestible solvents, so the introduction of new technology is needed to develop a dosage form that allows long-term administration and absorption by the human body.

[0011] Therefore, the present inventors attempted to develop a nanosuspension dosage form suitable for a progerinin drug having the effect of preventing or treating premature aging, and a manufacturing method thereof. Specifically, the inventors optimized the particle size of the progerinin drug, which has excellent bioabsorption efficiency, and selected water-soluble polymers, excipients, etc. for the preparation of the nanosuspension, thereby developing an oral progerinin nanosuspension formulation containing a progerinin drug with excellent dispersibility or uniformity, improved bioavailability, and improved stability. Summary of the Invention [Problem to be solved by the invention]

[0012] The present invention provides a method for preparing a nanosuspension containing a progerinin drug with excellent bioavailability. Another object of the present invention is to provide a dosage form that is easy to take orally and has improved stability as well as bioavailability of the drug. [Means for solving the problem]

[0013] To achieve the above object, the present invention provides a progerinin-containing nanosuspension comprising: (a) 1 to 10 wt% of a progerinin compound represented by the following chemical formula 1; (b) 0.5 to 5 wt% of a polymeric suspending agent, hydroxypropyl methylcellulose (HPMC); (c) 0.5 to 5 wt% of a solubility enhancer, D-α-tocopherol polyethylene glycol succinate (TPGS); and (d) 0.1 to 0.5 wt% of a preservative, potassium sorbate; wherein the progerinin compound is in the form of drug particles formed by wet-type ball milling to have an average particle size (D50) of 100 to 300 nm.

[0014] To achieve the other object, the present invention provides a progerinin-containing nanosuspension comprising: (a) 1 to 10 wt % of a progerinin compound represented by the following chemical formula 1; (b') 0.5 to 5 wt % of a polymeric suspending agent, hydroxypropyl cellulose (HPC); (c') 0.1 to 0.5 wt % of a surfactant, dioctyl sodium succinate (DOSS); and (d) 0.1 to 0.5 wt % of a preservative, potassium sorbate; wherein the progerinin compound is drug particles formed by wet ball milling to have an average particle size (D50) of 100 to 300 nm.

[0015] To achieve yet another object, the present invention provides a method for producing a nanosuspension containing progerinin, comprising the steps of: (i) mixing hydroxypropyl methylcellulose (HPMC) (b) and D-α-tocopherol polyethylene glycol succinate (TPGS) (c) with purified water to prepare a vehicle solution, and then adding progerinin (a) represented by the following Chemical Formula 1 and mixing to prepare a suspension; (ii) wet ball milling the suspension to prepare a nanosuspension; and (iii) mixing potassium sorbate (d) with the nanosuspension to finally prepare a nanosuspension with improved stability; wherein the progerinin compound in the nanosuspension of step (iii) is in the form of drug particles having an average particle size (D50) of 100 to 300 nm.

[0016] To achieve yet another object, the present invention provides a method for producing a nanosuspension containing progerinin, comprising the steps of: (i) mixing hydroxypropyl cellulose (HPC) (b') and dioctyl sodium succinate (DOSS) (c') with purified water to prepare a vehicle solution, and then adding and mixing progerinin (a) represented by the following Chemical Formula 1 to prepare a suspension; (ii) wet ball milling the suspension to prepare a nanosuspension; and (iii) mixing potassium sorbate (d) with the nanosuspension to finally prepare a nanosuspension with improved stability; wherein the progerinin compound in the nanosuspension of step (iii) is in the form of drug particles having an average particle size (D50) of 100 to 300 nm.

[0017] [ka] [Effects of the Invention]

[0018] The present invention relates to a nanosuspension dosage form suitable for the poorly soluble progerinin drug and a method for preparing the same. The nanosuspension according to the present invention is prepared by mixing a micronized progerinin drug with a water-soluble polymer, hydroxypropyl methylcellulose (HPMC) or hydroxypropyl cellulose (HPC); a dissolution enhancer, d-α-tocopherol polyethylene glycol succinate (TPGS) or a surfactant, dioctyl sodium succinate (DOSS); and a preservative, potassium sorbate, and ball milling the mixture in a Dyno-Mill chamber for a predetermined period of time. The nanosuspension contains drug particles with an average particle size of 200 nm or less, and exhibits excellent bioavailability (Example 1-1 and FIG. 1).

[0019] As described above, the nanosuspension containing progerinin as an active ingredient according to the present invention has uniform dispersibility and stability. Experimental results confirmed that the nanosuspension maintains particle uniformity of 200 nm or less for 20 days or more (up to 70 days) at 4°C, demonstrating excellent dispersion stability and bioabsorption efficiency. [Brief explanation of the drawings]

[0020] [Figure 1] 1 shows the pharmacokinetic analysis results of progerinin suspension. [Figure 2] 1 shows the results of measuring the solubility of solid dispersions produced using hot melt extrusion (HMT). [Figure 3] 1 shows the results of measuring the solubility of solid dispersions prepared using a hot melt extrusion process using polymers at various ratios (1:1). [Figure 4]Pharmacokinetic analysis results for solid dispersions prepared using a hot melt extrusion process. [Figure 5] The results show the solubility of amorphous solid dispersions measured using a spray drying process (A: pH 1.2, B: pH 6.8). [Figure 6] Pharmacokinetic analysis results for solid dispersions prepared using a spray drying process. [Figure 7] The shape of the nanosuspension prepared by wet ball milling using beads (top) and the results of measuring the average particle size of the progerinin drug (bottom). [Figure 8] 1 is a diagram illustrating a process for preparing a nanosuspension according to the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0021] The present invention will now be described in more detail.

[0022] The inventors have completed a method for producing a progerinin nanosuspension by mixing micronized progerinin drug with a vehicle solution containing hydroxypropyl methylcellulose (HPMC), d-α-tocopherol polyethylene glycol succinate (TPGS), and potassium sorbate, and then wet ball milling the mixture in a Dyno-Mill chamber for a predetermined period of time. The resulting nanosuspension contains particles with an average particle size (D50) of 200 nm or less, has uniform particle dispersion, and maintains its dispersion for more than 20 days at 4°C, resulting in stability and improved bioavailability.

[0023] Thus, the present invention provides nanosuspensions comprising progerinin with improved bioavailability.

[0024] Specifically, as one example, the nanosuspension according to the present invention contains: (a) 1 to 10 wt % of a progerinin compound represented by the following chemical formula 1; (b) 0.5 to 5 wt % of hydroxypropylmethylcellulose (HPMC), a polymeric suspending agent; (c) 0.5 to 5 wt % of D-α-tocopherol polyethylene glycol succinate (TPGS), a solubility enhancer; and (d) 0.1 to 0.5 wt % of potassium sorbate, a preservative.

[0025] In another embodiment, the nanosuspension according to the present invention comprises: (a) 1 to 10 wt % of a progerinin compound represented by the following chemical formula 1; (b') 0.5 to 5 wt % of a polymeric suspending agent, hydroxypropyl cellulose (HPC); (c') 0.1 to 0.5 wt % of a surfactant, dioctyl sodium succinate (DOSS); and (d) 0.1 to 0.5 wt % of a preservative, potassium sorbate.

[0026] [ka]

[0027] The progerinin (a) is a decursin derivative, (7S)-(+)-8,8-dimethyl-7-(3-phenyl-allyloxy)-7,8-dihydro-6H-pyrano[3,2-g]chromen-2-one. In the present invention, progerinin can also be named "SLC-D011."

[0028] The progerinin drug exhibits excellent inhibitory effects on progerin expression and on the binding of progerin and lamin A, and is effective in extending the survival time of an animal model in which progeria is induced, and is used as a pharmaceutical composition for preventing or treating progeria, an age-related disease. Progerinin is poorly soluble in water, with a solubility close to zero. However, the present invention provides a dosage form that improves solubility and allows for long-term administration and absorption by the human body.

[0029] Specifically, in the present invention, hydroxypropylmethylcellulose (HPMC) (b) or hydroxypropylcellulose (HPC) (b'), which corresponds to a water-soluble polymer as a polymeric suspending agent, is a component that helps disperse the progerinin drug and other ingredients in the solution. The polymeric suspending agent is preferably contained in an amount of 0.5 to 5 wt % of the total nanosuspension, more preferably 1 to 3 wt %.

[0030] If the polymer suspending agent is contained in an amount less than the above range, the settling rate of the suspension will be poor, and if the polymer suspending agent is contained in an amount exceeding the above range, stirring will be difficult, making manufacturing difficult. In particular, if the polymer suspending agent is contained in an amount exceeding the above range, stirring will not be uniform and the dispersibility of the suspension will be low, which will result in an increase in the amount of defective products due to under-achieving the intended content during production.

[0031] In the present invention, the solubility enhancer D-α-tocopherol polyethylene glycol succinate (TPGS) (c) is a component that improves the solubility of poorly soluble drugs. The TPGS is preferably contained in an amount of 0.5 to 5 wt % of the total nanosuspension, more preferably 1 wt %. If the amount of the solubility enhancer is less than this range, the solubility will decrease, and if the amount of the solubility enhancer is more than this range, the acceptable toxicity limit may be exceeded. Therefore, it is preferable to use the solubility enhancer within this range.

[0032] In addition, in the present invention, a surfactant, dioctyl sodium succinate (DOSS) (c'), can be used instead of the solubility enhancer. In this case, the surfactant is preferably used in an amount of 0.1 to 0.5 wt %, more preferably 0.25 wt %. If the surfactant is contained in an amount less than this range, the solubility will decrease, and if the surfactant is contained in an amount exceeding this range, the acceptable toxicity limit may be exceeded, so it is preferable to use the surfactant within this range.

[0033] The poorly soluble drug progerinin in the suspension preferably has an average particle size (D50) of 100 to 300 nm. More preferably, the average particle size is 150 to 250 nm, and even more preferably, the average particle size is 200 nm or less. When the average particle size is within this range, the solubility is excellent, and the nanoparticles have a larger surface area and are more easily dissolved, thereby maximizing bioavailability or bioabsorption efficiency. This was confirmed in Example <1-1> below. Furthermore, a nanosuspension having the above average particle size can be prepared by wet ball milling using a Dyno-Mill.

[0034] In the present invention, "stability" may include any of the homogeneity, dispersibility, precipitation stability, and storage stability of drug particles. In particular, the nanosuspension according to the present invention has uniform dispersibility for 20 days or more at 4°C, and has an advanced effect in that it has improved the ease of manufacture and storage stability of the suspension.

[0035] Meanwhile, "sinking rate" refers to the speed and degree to which particles settle in a fluid, and in the case of a suspension, a certain level of sinking rate must be met (0.9 or higher, where 1 is the maximum) in order to be approved for sale as a pharmaceutical. The nanosuspension of the present invention has the advantage of an excellent sinking rate due to the use of a vehicle solution containing hydroxypropyl methylcellulose (HPMC), TPGS or dioctyl sodium succinate (DOSS), and potassium sorbate, which are suitable for progerinin drugs.

[0036] The pH of the suspension of the present invention is 5.5 to 8.5, and preferably 6.0 to 7.0.

[0037] The nanosuspension has a viscosity of 2 to 8.5 mPa·s, preferably 7 to 8.5 mPa·s.

[0038] The nanosuspension of the present invention may further comprise one or more ingredients selected from the group consisting of sweeteners, preservatives, flavoring agents, pH adjusters, and coloring agents.

[0039] The term "sweetener" as used herein refers to a food additive that provides (or reproduces) a sweet taste but generally has a lower calorie content, and includes both natural and synthetic sweeteners. The sweetener may be any sweetener commonly used in the food and pharmaceutical fields and is applicable to the present invention without limitation.

[0040] The term "preservative" as used herein refers to a pharmaceutically acceptable substance that prevents decomposition due to microbial growth or undesired chemical changes. The preservative has bactericidal and / or fungicidal or antioxidant properties, and in the present invention, potassium sorbate (d) is preferably used at 0.1 to 0.5 wt%, more preferably 0.2 wt%. Within this range, the shelf life can be improved without impairing the efficacy of the drug. If added before milling, the content decreases, so it is recommended to add it after milling to complete the nanosuspension.

[0041] The term "flavor," "flavor," or "aroma" as used herein includes flavoring ingredients or compositions commonly used in the food industry, whether of natural or synthetic origin, including single compounds or mixtures.

[0042] The term "pH adjusting agent" as used in the present invention refers to an excipient used to adjust the pH of the suspension to a desired value, such as, but not limited to, citric acid, sodium citrate, or ascorbic acid, and any excipient commonly used in the technical field of the present invention may be used in the present invention without limitation.

[0043] The term "colorant" as used in the present invention refers to a substance that is included to change the color of the suspension, and can be used to prepare a suspension with a desired color by being included in a dosage form. Any substance commonly used in the technical field of the present invention is included in the present invention without limitation.

[0044] The present invention also provides a method for preparing a nanosuspension containing a poorly soluble decurcin derivative drug with improved bioavailability and stability.

[0045] Specifically, as one embodiment, the method for producing a nanosuspension according to the present invention includes the steps of: (i) mixing hydroxypropyl methylcellulose (HPMC) (b) and D-α-tocopherol polyethylene glycol succinate (TPGS) (c) with purified water to produce a vehicle solution, and then adding and mixing progerinin (a) represented by the following chemical formula 1 to produce a suspension; (ii) wet ball milling the suspension to produce a nanosuspension; and (iii) mixing potassium sorbate (d) with the nanosuspension to finally produce a nanosuspension with improved stability.

[0046] In another embodiment, a method for preparing a nanosuspension according to the present invention includes the steps of: (i) mixing hydroxypropyl cellulose (HPC) (b') and dioctyl sodium succinate (DOSS) (c') with purified water to prepare a vehicle solution, and then adding progerinin (a) represented by the following chemical formula 1 and mixing to prepare a suspension; (ii) wet ball milling the suspension to prepare a nanosuspension; and (iii) mixing potassium sorbate (d) with the nanosuspension to finally prepare a nanosuspension with improved stability.

[0047] [ka]

[0048] The role of each component and the amount used are the same as those described above, so additional explanations will be omitted. Figure 8 is a flowchart showing a method for preparing a nanosuspension containing a poorly soluble decurcin derivative drug with improved bioavailability and stability according to the present invention.

[0049] First, in the present invention, step (i) is a process of mixing a micronized progerinin drug with a vehicle solution, specifically, a step of mixing purified water with a polymeric suspending agent, hydroxypropyl methylcellulose (HPMC) or hydroxypropyl cellulose (HPC) (b'), and D-α-tocopherol polyethylene glycol succinate (TPGS) (c) or dioctyl sodium succinate (DOSS) (c'). In this case, purified water having a temperature of 50 to 70°C can be used, preferably 60°C. The resulting coarse suspension is cooled to room temperature and then subjected to the next step to prepare a nanosuspension.

[0050] Next, in step (ii), the suspension of step (i) is subjected to wet ball milling to prepare a nanosuspension. Here, a Dyno-Mill can be used for wet ball milling. The ball milling is preferably performed until the average particle size of the drug particles reaches 100 to 300 nm. More preferably, the average particle size is 150 to 250 nm, and even more preferably, the average particle size is 200 nm. When the average particle size is within this range, the drug has the best solubility and can maximize the bioavailability or bioabsorption efficiency.

[0051] In the present invention, ball milling is a wet ball milling method for preparing a nanosuspension, and is an important step in the preparation of a progerinin nanosuspension. Selection of appropriate bead size, grinding media, and API ratio in the chamber, temperature control of the mill chamber, etc. are considered to be important.

[0052] Specifically, when preparing the nanosuspension, wet ball milling can be performed using the suspension from step (i) and zirconia beads with an average particle size of 0.1 to 1 mm, preferably 0.02 to 0.04 mm. The suspension from step (i) and zirconia beads can be used in a volume ratio of 1:1 to 1:5, but this can be adjusted as needed to suit the conditions as long as the average particle size (D50) of the drug can be adjusted to 200 nm or less.

[0053] Next, in step (iii), potassium sorbate is added to the nanosuspension prepared in step (ii) to produce a nanosuspension with improved stability. The addition of potassium sorbate completely disperses the drug particles, further improving the stability of the nanosuspension.

[0054] Meanwhile, all of the above processes include a process of mixing the poorly soluble drug with a solvent, a water-soluble polymer or a surfactant, and the nanosuspension can be prepared at a mixing speed of 500 to 1,000 rpm.

[0055] The present invention will be described in detail below with reference to examples to aid in understanding the present invention. However, the following examples are provided to more completely explain the present invention to those skilled in the art and to illustrate the content of the present invention, but the scope of the present invention is not limited to the following examples.

[0056] Example 1: Preparation and Limitations of Lipid-Based Progerinin Solution To overcome the poor bioavailability of progerinin drugs, the present inventors prepared progerinin solutions using a lipid-based formulation (monoolein:tricaprylin = 2:1) in preclinical studies. However, lipid-based formulations were deemed unsuitable for use in clinical settings due to inadequate drug loading and the high lipid intake expected at the proposed clinical dose. Limited attempts to produce amorphous solid dispersions (ASDs) have shown that ASDs suffer from rapid crystallization when dispersed in aqueous media. Furthermore, such lipid-based formulations are deemed unsuitable for use in clinical settings due to inadequate drug loading and the high lipid intake expected at the proposed clinical dose.

[0057] Therefore, the present inventors intend to prepare a nanosuspension of progerinin, a poorly soluble drug, and select a vehicle composition that improves the stability of the drug to realize a nanosuspension optimized for clinical use.

[0058] The nanosuspension manufacturing process is thermodynamically unstable. Therefore, stabilizers such as polymers and surfactants are essential to maintain a physically stable state. Among the polymers, HPMC and HPC occupy adsorption sites on the surface of drug particles, preventing drug molecules from forming a crystalline lattice in solution and providing a mechanical barrier to crystallization. However, if the polymer concentration is insufficient, crystals grow rapidly and aggregate. As the polymer concentration continues to increase, particle size increases due to the thick layer on the particle surface, and diffusion between the solvent and semisolvent is suppressed during the aggregation process. Furthermore, as the polymer concentration increases, osmotic pressure increases, resulting in increased attraction between colloidal particles, leading to particle growth. On the other hand, surfactants adsorb at the solid-liquid interface, reducing interfacial surface tension and increasing the rate of nucleation, resulting in an initial decrease in particle size. Moreover, surfactant adsorption reduces hydrophobic interactions and coagulation, making resveratrol particles less hydrophobic and reducing particle growth. In particular, when the ionic surfactant SLS is adsorbed onto the particle surface, the particle surface becomes negatively charged. This increases the repulsive force between particles, increasing the energy barrier and thereby preventing particle growth and aggregation. As a result, the addition of an appropriate concentration of a stabilizing agent can reduce the excessively high surface energy of the nanoparticles produced. Furthermore, the size of drug particles in a nanosuspension is an important factor in determining bioabsorption efficiency.

[0059] Therefore, the inventors determined that the selection of stabilizers such as polymers and surfactants, and the particle size within the suspension are important factors in preparing nanosuspensions of poorly soluble drugs. First, because the size of drug particles within a drug is an important factor in determining bioabsorption efficiency, the average particle size of progerinin (SLC-D011), a drug with excellent bioabsorption efficiency, was selected, and the polymers, surfactants, preservatives, etc. for preparing the nanosuspension were optimized.

[0060] <1-1> Determination of the mean particle size (D50) of progerinin drug As mentioned above, the size of drug particles in a drug is an important factor in determining bioabsorption efficiency. Therefore, the inventors conducted pharmacokinetic (PK) analysis of the compound using mice to design the diameter size that facilitates bioabsorption of the drug progerinin. Specifically, a microsuspension containing micro-sized progerinin (D50 = 1.5 mm), nanosuspension 1 containing nano-sized progerinin (Nanosuspension 1 = 200 nm (D = 50)), and nanosuspension 2 containing nano-sized progerinin (Nanosuspension 2 = 350 nm (D = 50)) were prepared. The microsuspension was prepared using Vivapur Power, and the two nanosuspensions were prepared using zirconia beads. The resulting suspensions were orally administered to mice at 10 mg / kg, 30 mg / kg, and 100 mg / kg, and the pharmacokinetic (PK) analysis of the suspensions in vivo was performed (Figure 1).

[0061] As shown in Figure 1, it was determined that a drug can be smoothly absorbed in the body when the average particle size is 200 nm or less. Based on this, the inventors decided to develop a suspension formulation containing drug nanoparticles with an average particle size of 200 nm or less.

[0062] In order to devise a manufacturing method that can meet this dosage form, the inventors tried a hot melt extrusion process, a spray drying process, a process using an ultra-high pressure homogenizer, and a process using wet ball milling with beads, and tried to select the most suitable manufacturing method for progerinin nanosuspension from among these.

[0063] <1-2> Manufacturing of amorphous solid dispersions using the hot melt extrusion (HME) process First, solid dispersions were prepared using various polymers (HPMCAS, HPC, HPMC, PVP VA64, Eudragit EPO, and AEA) using hot-melt extrusion. The drug / polymer ratio was 1:3. The solubility of the solid dispersions corresponding to 100 mg of progerinin was tested using 300 mL of pH 1.2 buffer (Eudragit EPO, AEA) or 300 mL of pH 6.8 buffer (HPMCAS, HPC, HPMC, and PVP VA64) as the dissolution medium (Figures 2 to 4). The dissolution conditions were as follows: paddle method, temperature: 37°C, rotation speed: 150 rpm.

[0064] As shown in Figure 2, it was confirmed that the solubility was most improved when HPMCAS was used as the polymer.

[0065] Next, the HPMCAS with the highest solubility was reselected and solid dispersions were prepared at various ratios (1:2, 1:2.5, 1:3, and 1:4). A solubility test was conducted on the solid dispersions equivalent to 100 mg of progerinin using 300 mL of pH 6.8 buffer as the eluent (Figure 3). The elution conditions were as follows: paddle method, temperature: 37°C, rotation speed: 150 rpm.

[0066] As shown in Figure 3, the highest solubility was observed when the drug to HPMCAS ratio was 1:3. However, the HME method actually produced a light yellow powder, confirming that the solid remained amorphous. Furthermore, as shown in Figure 4, the powder obtained using the HME method immediately precipitated when mixed with water, and when bioabsorption was confirmed using mice, there was almost no absorption in the body after oral administration.

[0067] Therefore, it was concluded that it is currently impossible to develop a dosage form of progerinin, the main component of the drug, using the hot melt extrusion process.

[0068] <1-3> Manufacturing amorphous solid dispersions using the spray drying process Amorphous solid dispersions were prepared using the polymers described in Examples 1-2 above, using a spray-drying process. Solid dispersions were prepared for various polymers (HPMCAS, HPC, HPMC, PVP VA64, Eudragit EPO, and AEA) using a spray-drying process. The drug / polymer ratio was 1:3. The solubility of the solid dispersions corresponding to 100 mg of progerinin was tested using 300 mL of pH 1.2 buffer (Eudragit EPO, AEA) and 300 mL of pH 6.8 buffer (HPMCAS, HPC, HPMC, and PVP VA64) as dissolution media (Figure 5). The dissolution conditions were as follows: paddle method, temperature: 37°C, rotation speed: 150 rpm.

[0069] As shown in FIG. 5, when HPMCAS was used as the polymer, as in the solid dispersion prepared by hot melt extrusion, the solubility of the polymer was most improved at pH 1.2 and pH 6.8.

[0070] However, when bioabsorption (PK analysis) was confirmed using mice, it was found that, similar to HME, there was almost no absorption in the body after oral administration (Figure 6). Furthermore, when the stability of other polymers was confirmed in a high temperature and humid environment for two weeks, recrystallization of progerinin particles was observed, resulting in unsatisfactory results (Table 1).

[0071] [Table 1]

[0072] Therefore, it was determined that it is currently impossible to develop a dosage form of progerinin, the main component of the drug, using the spray drying method.

[0073] <1-4> Manufacturing nanosuspensions using an ultra-high pressure homogenizer As another method for producing nanosuspensions, the inventors prepared nanosuspensions using an ultra-high-pressure homogenizer. Specifically, a polymer (HPMC 3cp) and a surfactant (TPGS) were dissolved in a 100 mL beaker containing distilled water, and then the drug (SLC-D011) was weighed and added. The mixture was stirred for 2 hours to produce a suspension. The suspension was then placed in an ultra-high-pressure homogenizer (pressure: 40,000 psi, 1 hour) to produce a nanosuspension.

[0074] [Table 2]

[0075] Table 2 shows the measurement of drug particle size, and it was confirmed that the particle size increased after 3 days. Therefore, the preparation of nanosuspension using an ultra-high pressure homogenizer was limited by the increase in particle size over time. This also does not appear to be a suitable method for preparing progerinin nanosuspension.

[0076] <1-5> Preparation of nanosuspension using wet ball milling with beads Alternatively, the inventors prepared a nanosuspension using wet ball milling with beads. Specifically, a polymer (HPMC 3cp) and a surfactant (TPGS) were dissolved in a 100 mL beaker containing distilled water, and then progerinin was weighed and added. The mixture was stirred for two hours to prepare a suspension. The tube containing the beads and suspension was then placed in a DeltaVita® Netzsch (Zentrimix 380R) device to prepare a nanosuspension (speed: 1200 rpm, temperature: -10°C, 6 hours). Figure 7 shows the shape (top) and average particle size (bottom) of the nanosuspension prepared using this method.

[0077] As shown in Figure 7, a nanosuspension containing drug nanoparticles with an average particle size of 170 nm was prepared. It was confirmed that when the average particle size of drug particles is 200 nm or less, the stability of the nanosuspension is most excellent, and the nanoparticles have a larger surface area and are more easily dissolved, maximizing bioavailability.

[0078] Therefore, the inventors prepared nanosuspensions using wet ball milling with beads, and determined that it was necessary to select a vehicle composition suitable for preparing nanosuspensions in which the average particle size of the drug is 200 nm or less. They attempted to select optimized polymers, surface stabilizers such as surfactants, and preservatives.

[0079] Example 2: Selection of an optimized vehicle solution <2-1> Preparation of progerinin nanosuspension a) Suspension Preparation (Step (i)) TPGS solution is prepared by dissolving TGPS in 60°C hot water and allowing it to cool to room temperature. HPMC E3 solution is prepared by dissolving HPMC E3 in 60°C hot water and allowing it to cool to room temperature. The two solutions are then thoroughly mixed by stirring. To prepare a suspension, progerinin (API) is gradually added to the mixed solution while continuing to stir.

[0080] b) Nano Milling (Step (ii)) The parameters of the Agitator Bead Mill, a mixing and wet grinding equipment, are set appropriately to begin wet ball milling, and the constant pressure pump is adjusted to circulate the suspension. The suspension exiting the Agitator Bead Mill is recycled back to the suspension container while being maintained at a preset temperature of 35°C or less. The drug particles continue to undergo wet ball milling in the Dyno Mill until the PSD (D50≦200 nm) is reached. Once the target PSD (D50≦200 nm) is reached, the pump is turned off and the milling suspension line is separated from the suspension container.

[0081] c) Preparation of nanosuspension with improved stability (step (iii)) The nanomilled suspension is mixed thoroughly with potassium sorbate as a preservative and filled into sterilized glass bottles. The bottle filling process is 100% visually inspected and checked by fill weight. After filling, the bottles are sealed with a rubber finish and an aluminum flip-off seal.

[0082] <2-2> Selection of optimized vehicle solutions Oral progerinin nanosuspensions were prepared using the same method as in Example <2-1>. Specifically, Prototype 1 consisted of 10.0 wt% progerinin, 3.0 wt% HPMC E3, 1.0 wt% surfactant TPGS, and 0.2 wt% potassium sorbate. TPGS and HPMC solutions were mixed and progerinin was added to prepare a suspension. The resulting mixture was mixed with 0.3 mm VHD ZrO beads in a volume ratio of 1:1 to 5 and ball milled. The container temperature was closely monitored during the manufacturing process. Furthermore, the particle size distribution was monitored during milling until the target average particle size (D50) reached 200 nm or less. Prototypes 2 to 5 were also prepared using the same method. The particle diameter, zeta potential, drug content, flowability, and viscosity of the nanosuspensions were measured using the following methods.

[0083] (1) Particle diameter (D50) measurement: Approximately 2 μL of nanosuspension was dispersed in 1 mL of water, placed in a sample tube, and measured using a Zeta Potential & Particle Sizer (ZPPS) (Nicomp 380 / ZLS, Nicomp) used for aqueous suspensions. (2) Zeta potential measurement: This is a measurement of the electrical charge on the particle surface, which indicates the physical stability of the colloidal system. The zeta potential was measured using the Laser-Doppler method. (3) Drug content measurement: The content of progerinin relative to the total weight of the nanosuspension was measured. (4) Measurement of dispersibility: Dispersibility was evaluated by visual observation based on the presence or absence of precipitation in the suspension. (5) Viscosity measurement: Using a Brookfield viscometer (TC-550MX-230), the torsional moment was 32.9%, the temperature was 25°C, the probe rotation speed was 200 rpm, and the viscosity was 13.03 dyne / cm 2 Shear force, 264.0 S -1 The shear rate was measured for 1 minute.

[0084] [Table 3]

[0085] As shown in Table 3, the mean particle size (D50) of drug particles for Prototype 1 and Prototype 3 was 165.8 nm and 146.3 nm, respectively. Both had mean particle sizes of 200 nm or less, and showed drug content greater than 95% without detectable impurities. Therefore, Prototype 1 and Prototype 3 were confirmed to be the most suitable vehicle compositions in terms of dispersibility, stability, and drug efficacy. On the other hand, Prototype 4 had limitations in terms of drug efficacy and could not be selected as a suitable vehicle composition. Prototype 2 tended to increase in particle size over time, making it unsuitable as a candidate. In the case of Prototype 5, it was impossible to form a nanosuspension.

[0086] <Example 3> Preparation of suspension using roller mixer and Dyno-Mill and comparison of physical properties <3-1> Manufacturing suspension using a roller mixer A nanosuspension of prototype 1 with a target concentration of 100 mg / g was prepared using a roller mixer. Specifically, the nanosuspension of prototype 1 was prepared and mixed with zirconia beads shown in Table 2 below in a 1:1 weight ratio. The PSD was monitored daily to observe the size reduction trend, as shown in Table 4.

[0087] [Table 4]

[0088] As shown in Table 4, after 8 days of milling, the particle size remained at D90 = 500 nm and did not decrease further. Therefore, it was concluded that it would be difficult to produce a nanosuspension containing drug particles of 200 nm or less using a roller mixer.

[0089] <3-2> Preparation of suspension using Dyno-Mill and evaluation of its physical properties Based on the results of <3-2> above, we attempted to produce a nanosuspension of Prototype 1 with a target concentration of 100 mg / g using a Dyno-Mill (5 g API scale). Specifically, 45 mL of 0.5 mm zirconia beads were loaded into the Dyno-Mill milling chamber. The prepared nanosuspension of Prototype 1 (5 g, 10% w / w HPMC E3, 3% / TPGS 1%) was topped into the milling chamber. The machine was operated at 2,200 rpm for 5 hours with a circulating cooling liquid at -19°C. A white nanosuspension (FR00497-3-190911-100) was collected as the final nanosuspension.

[0090] Additionally, similar methods were used to prepare 6 mg / g Prototype 1 nanosuspension (FR00497-3-190910-6) and 20 mg / g Prototype 1 nanosuspension (FR00497-3-190910-20). The three nanosuspensions collected in this manner were checked for appearance, PSD, HPLC, and syringeability, and the results are shown in Table 5 below.

[0091] [Table 5]

[0092] The results in Table 5 confirm that the nanosuspension of Prototype 1 satisfied all of the physical properties, including particle size and syringeability. Additionally, the low-temperature stability of the three nanosuspensions was observed at 4°C for two weeks. Specifically, stability was evaluated by measuring particle size to determine whether the average particle size (D50) was 200 nm or less. Syringeability was evaluated by determining whether the nanosuspension could easily pass through a 20 GA syringe. The results are shown in Table 6 below.

[0093] [Table 6]

[0094] The results in Table 6 confirm that all three nanosuspensions are stable at a low temperature of 4°C without significant changes in particle size and have excellent syringeability. In the present invention, a mean particle size (D50) of 200 nm or less is the most ideal requirement in terms of particle dispersion stability and bioavailability of the progerinin drug.

[0095] <3-3> Scale-up production of nanosuspension To scale up the process, the inventors attempted to produce a nanosuspension with a target concentration of 100 mg / g using a DynoMill at a 30 g API scale. Specifically, 45 mL of 0.3 mm zirconia beads was loaded into the DynoMill chamber. 30 g of the prepared progerinin was first suspended in a vehicle solution (30 g, 10% w / w in the vehicle HPMC E3 3% / TPGS 1% in water) to prepare a suspension. The suspension was then poured into the milling chamber from above. The equipment was operated at 2,200-2,500 rpm for 20 hours at a low temperature of -19°C with a circulating cooling liquid. The PSD was monitored over time as shown in Table 7, and a white nanosuspension was finally collected. The obtained white nanosuspension was then refrigerated for 20-70 days to evaluate its stability (Table 8).

[0096] [Table 7]

[0097] From the results of Table 7, it was confirmed that progerinin drug particles with an average particle size (D50) of 200 nm or less can be successfully produced even when a nanosuspension is produced by scaling up using a Dynomill.

[0098] [Table 8]

[0099] From the results of Table 8, it was confirmed that the nanosuspension according to the present invention maintains stability for 20 days or more, specifically 20 to 70 days, under refrigerated conditions.

[0100] Example 4: Selection of optimized excipients To evaluate compatibility with excipients, progerinin was mixed with HPMC E3, TPGS, and potassium sorbate in a ratio of 1:10, and the effectiveness of the API component was measured for 2 weeks at 40°C / 75% RH. The results are shown in Tables 9 and 10.

[0101] [Table 9]

[0102] [Table 10]

[0103] API:Active Pharmaceutical Ingredient(SLC-D001) The results of Tables 9 and 10 confirm that adding HPMC E3, TPGS, and potassium sorbate to the progerinin drug does not affect the drug. In particular, in the case of preservatives, if added before milling, the content decreases, so it is recommended to add them after milling. Such compatibility studies of pre-excipients suggest that the drug is compatible with the excipients.

[0104] <Example 5> Confirmation of stability Further stabilization studies of progerinin nanosuspensions were conducted for Prototype 1 and Prototype 3, which showed excellent particle stability during the vehicle solution selection process. Specifically, evaluations were conducted for three weeks at 2-8°C, 40°C / RH 75%, or under 1.2M lux exposure. The stabilization study of progerinin nanosuspensions was conducted for three weeks at room temperature (25±5°C, 60% RH). The progerinin nanosuspensions were characterized in relation to particle size and drug content. The results are shown in Tables 11 and 12.

[0105] [Table 11]

[0106] [Table 12]

[0107] From the results of Tables 11 and 12, it was confirmed that the average particle size was 200 nm or less even at low temperatures or under harsh conditions, and the nano drug particles were stable for more than 3 weeks.

[0108] Example 6: Stability of optimized progerinin nanosuspension in simulated intestinal fluid (SIF) and simulated gastric fluid (SGF) A nanosuspension of prototype 1 was prepared using the same method as in Example <2-1>, but with the ingredients and contents shown in Table 13 below. The dispersion stability of the nanosuspension thus prepared was confirmed in simulated intestinal fluids (SIF) and simulated gastric fluids (SGF). The specific experimental method is as follows.

[0109] [Table 13]

[0110] a) Simulated intestinal fluid (SIF) Monobasic potassium phosphate (6.8 gm) and sodium hydroxide (0.616 gm) were added to a 1000 mL volumetric flask with 250 mL of distilled water, swirled until the ingredients were dissolved, and then an additional 700 mL of distilled water was added and the pH was measured. The pH was adjusted to pH 6.8 + / - 0.1 by adding 0.2 N sodium hydroxide or 0.2 N hydrochloric acid.

[0111] b) Simulated gastric fluid (SGF) To a 1000 mL volumetric flask was added sodium chloride (2 gm), 750 mL distilled water, and 7.0 mL concentrated hydrochloric acid, followed by 1000 mL distilled water, and the components were mixed by swirling until the pH of the solution was 1.2.

[0112] c) Preparation of the dispersion An appropriate amount of prototype 1 nanosuspension was added to a translucent HDPE vial (25 mL) with an HDPE lid. It was diluted with 15 mL of simulated gastric or intestinal fluid to a final progenin concentration of 0.5 mg / mL. A dispersant was then added, and the resulting dosage form was shaken until completely dispersed. The vial was placed in a 37°C oil bath until flocculation was observed. The size of the precipitated particles was measured using a Horiba-LA-910 particle analyzer. The material was then incubated in fasted human gastric fluid for approximately 3 hours.

[0113] d) Particle size measurement When measuring beads coated with bead cores containing insoluble particles, the weight of the bead cores was calculated in SIF or SGF experiments, and the bead cores were dispersed in the appropriate volume of SIF or SGF. 120 g of distilled water was poured into the Horiba LA-910 chamber, and the instrument was drained to empty the chamber. Then, 120 g of distilled water was poured into the Horiba LA-910 chamber, and the entire volume of the cultured formulation (in 15 mL of SGF or SIF) was poured into the chamber, and the average particle size was measured.

[0114] [Table 14]

[0115] As shown in Table 14, the progenin drug (Prototype 1) with an average particle size (D50) of 200 nm or less maintained an average particle size (D50) of 190-220 nm even in simulated intestinal fluid or simulated gastric fluid, confirming that the dispersion stability and bioavailability of the drug were maintained.

[0116] Although the specific details of the present invention have been described above, it is obvious to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the present invention, and the substantial scope of the present invention is defined by the claims and their equivalents.

Claims

1. (i) preparing a vehicle solution by mixing hydroxypropyl methylcellulose (HPMC) (b) and D-α-tocopherol polyethylene glycol succinate (TPGS) (c) with purified water, and then adding and mixing progerinin (a) represented by the following Chemical Formula 1 to prepare a suspension; (ii) wet ball milling the suspension in a Dyno-Mill at 2,200 to 2,500 rpm for 8 to 20 hours to prepare a nanosuspension; (iii) mixing potassium sorbate (d) with the nanosuspension to finally prepare a nanosuspension having improved stability; The method for producing a nanosuspension containing progerinin, wherein the progerinin compound in the nanosuspension in step (iii) is in the form of drug particles having an average particle size (D50) of 100 to 200 nm. 【Chemistry 1】

2. A method for producing a nanosuspension containing progerinin as described in claim 1, characterized in that the nanosuspension in step (iii) contains (a) 1 to 10 wt % of the progerinin compound represented by chemical formula 1, (b) 3 wt % of hydroxypropyl methylcellulose (HPMC) as a polymeric suspending agent, (c) 1 wt % of D-α-tocopherol polyethylene glycol succinate (TPGS) as a solubility enhancer, (d) 0.1 to 0.5 wt % of potassium sorbate as a preservative, and (e) the remaining amount of purified water.

3. (i) preparing a vehicle solution by mixing hydroxypropyl cellulose (HPC) (b'); and dioctyl sodium succinate (DOSS) (c'); with purified water, and then adding progerinin (a) represented by the following formula 1 and mixing to prepare a suspension; (ii) wet ball milling the suspension in a Dyno-Mill at 2,200 to 2,500 rpm for 8 to 20 hours to prepare a nanosuspension; (iii) mixing potassium sorbate (d) with the nanosuspension to finally prepare a nanosuspension having improved stability; The method for producing a nanosuspension containing progerinin, wherein the progerinin compound in the nanosuspension in step (iii) is in the form of drug particles having an average particle size (D50) of 100 to 200 nm. 【Chemistry 2】

4. A method for producing a nanosuspension containing progerinin as described in claim 3, characterized in that the nanosuspension in step (iii) contains (a) 1 to 10 wt % of the progerinin compound represented by chemical formula 1, (b') 1 wt % of hydroxypropyl cellulose (HPC) which is a polymeric suspending agent, (c') 0.25 wt % of dioctyl sodium succinate (DOSS) which is a surfactant, (d) 0.1 to 0.5 wt % of potassium sorbate which is a preservative, and (e) the remaining amount of purified water.

5. The method for preparing a nanosuspension containing progerinin according to claim 2 or 4, wherein the purified water in step (i) is at a temperature of 50 to 70°C.

6. 5. The method for producing a nanosuspension containing progerinin according to claim 2 or 4, wherein the nanosuspension is produced by mixing the suspension of step (i) with zirconia beads in a volume ratio of 1:1 to 1:5 during wet ball milling in step (ii).

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