Viscous composition for manufacturing growth hormone microneedles, growth hormone microneedle structure manufactured using same, and microneedle patch and microarray comprising microneedle structure

Abstract

The present invention relates to a viscous composition for manufacturing microneedles for loading growth hormone, a microneedle structure, a microneedle patch, and a microneedle array. The growth hormone microneedle structure manufactured using the viscous composition for manufacturing microneedles, the microneedle patch, and the microneedle array, according to the present invention, exhibit improved content storage stability and / or purity storage stability compared to a liquid growth hormone raw material.

Description

Mucus composition for manufacturing growth hormone microneedles, growth hormone microneedle structures manufactured using the same, microneedle patches and microarrays including the microneedle structures

[0001] The present invention relates to a mucus composition for manufacturing microneedles for loading growth hormones, a microneedle structure, a microneedle patch, and a microneedle array.

[0002] Although numerous drugs and therapeutic agents have been developed to treat diseases, there are still issues that need to be improved in terms of drug delivery into the body, such as the passage of biological barriers (e.g., skin, oral mucosa, and blood-brain barrier) and the efficiency of drug delivery.

[0003] Drugs are typically administered orally in tablet or capsule form. However, many drugs cannot be effectively delivered using this method alone due to factors such as digestion or absorption in the gastrointestinal tract or clearance through hepatic mechanisms. Furthermore, some drugs cannot effectively diffuse through the intestinal mucosa. Furthermore, patient compliance is also a concern, particularly for critically ill patients who require medication at specific intervals or are unable to take medication.

[0004] Another common drug delivery technique is the use of conventional needles. While more effective than oral administration, this method carries the risk of pain at the injection site, local skin damage, bleeding, and infection at the injection site.

[0005] Growth hormone injections are expected to be effective in treating growth failure in children due to pituitary growth hormone deficiency, idiopathic hypogrowth in children, and growth failure in children diagnosed with Turner syndrome. However, these growth hormone injections require daily administration, which can be difficult to administer, and the device itself poses a significant cost burden.

[0006] Accordingly, the present invention aimed to develop a novel growth hormone-loaded microneedle formulation for the treatment of hypogonadism, utilizing microneedles that are less painful and easier to apply. It is crucial to develop such a novel formulation in an optimized form, taking into account not only the feasibility of microneedle manufacturing but also the stability and bioavailability of growth hormone.

[0007] As a prior art, Patent Document 1 discloses a microneedle band for promoting fingernail and toenail growth, in which microneedles containing human growth hormone are formed on one side of a band that is adhesive so that it can be detachably attached to the skin. However, there is no mention of a stabilizing formulation for manufacturing microneedles with improved storage stability and purity storage stability of growth hormone content.

[0008] [Prior Art Literature]

[0009] [Patent Document]

[0010] (Patent Document 1) Republic of Korea Publication No. 10-2020-0110256

[0011] Accordingly, the inventors of the present invention aimed to develop a microneedle patch through research into the formulation of microneedles loaded with recombinant human growth hormone. Accordingly, the present invention aimed to develop a novel formulation capable of maintaining drug stability and to deliver the drug stably into the body via microneedles.

[0012] Accordingly, an object of the present invention is to provide a mucus composition for manufacturing microneedles loaded with growth hormone.

[0013] Another object of the present invention is to provide a method for manufacturing a microneedle structure using the above mucus composition and a growth hormone microneedle structure manufactured therefrom.

[0014] Another object of the present invention is to provide a microneedle patch comprising the growth hormone microneedle structure.

[0015] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

[0016] To solve the above-described problem, the present invention provides a mucus composition for manufacturing growth hormone microneedles, comprising growth hormone, a water-soluble polymer, and an additive.

[0017] In the present invention, the growth hormone can be concentrated by centrifugal concentration or freeze-drying.

[0018] In the present invention, the centrifugal concentration method can be performed by filtering the liquid raw material of growth hormone through a membrane filter having a molecular weight cutoff of 3 to 20 kDa.

[0019] In the present invention, the freeze-drying method can be performed in the following steps a) to c):

[0020] a) A step of filtering a liquid raw material of growth hormone through a membrane filter having a molecular weight cutoff of 3 to 20 kDa;

[0021] b) adding sucrose to the filtered concentrate; and

[0022] c) A step of performing freeze-drying.

[0023] In the present invention, the growth hormone may be human growth hormone and may be included in an amount of 10 to 45 wt% with respect to the entire mucus composition.

[0024] In the present invention, the water-soluble polymer may be hyaluronic acid and may be included in an amount of 10 to 45 wt% with respect to the entire mucus composition.

[0025] In the present invention, the additive is sucrose, trehalose, oligosaccharide, maltose, lactose, cellobiose, glycine, mannitol, calcium D-gluconate, arginine, alginic acid, pectin, carrageenan, chondroitin sulfate, dextran sulfate, chitosan, polylysine, collagen, gelatin, carboxymethyl chitin, fibrin, agarose, polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), polyethylene glycol (PEG), From the group consisting of polymethacrylate, hydroxypropylmethylcellulose (HPMC), ethylcellulose (EC), hydroxypropylcellulose (HPC), carboxymethylcellulose, cyclodextrin, gentiobiose, alkyltrimethylammonium bromide, hexadecyltrimethylammonium bromide, gentian violet, benzethonium chloride, docusate sodium salt, a SPAN-type surfactant, polysorbate, sodium dodecyl sulfate, benzalkonium chloride and glyceryl oleate. There may be one or more of the selected ones,It may be included in an amount of 10 to 45 wt% with respect to the total mucus composition.

[0026] In the present invention, the additives may be sucrose and polysorbate, and the sucrose may be included in an amount of 9.95 to 40 wt% based on the total mucus composition, and the polysorbate may be included in an amount of 0.05 to 5 wt% based on the total mucus composition.

[0027] In the present invention, the mucus composition can constitute the core layer in a growth hormone microneedle structure including a base layer formed on a support; a core layer formed on the base layer; and a shell layer formed on the base layer to cover the core layer.

[0028] The present invention also provides a method for manufacturing a growth hormone microneedle structure using the mucus composition for manufacturing various types of growth hormone microneedles described above, and a growth factor microneedle structure manufactured therefrom.

[0029] In the present invention, the mucus composition may constitute part or all of the microneedle structure.

[0030] In the present invention, the microneedle structure:

[0031] It may include a base layer formed on a support; a core layer formed on the base layer and composed of a mucus composition for manufacturing growth hormone microneedles of various shapes as described above; and a shell layer formed on the base layer to cover the core layer.

[0032] In the present invention, the base layer and the shell layer do not contain growth hormones and may be composed of biodegradable or biocompatible materials.

[0033] In the present invention, the growth hormone microneedle structure can be manufactured in a cone-shaped, quadrangular pyramid-shaped, sphere-shaped, short-headed, wedge-shaped, blade-shaped or candle-shaped shape.

[0034] In the present invention, when the growth hormone microneedle structure is manufactured in a candle-shaped shape, the maximum diameter of the vertical cross-section with respect to the protrusion direction of the microneedle structure may be 200 to 400 μm.

[0035] In the present invention, the growth hormone microneedle structure can be inserted into the skin in a patch or implant form.

[0036] In the present invention, the growth hormone microneedle structure can exhibit improved content storage stability and / or purity storage stability compared to the liquid growth hormone raw material.

[0037] In the present invention, the method for manufacturing the growth hormone microneedle structure may include a step of forming a mucus composition for manufacturing the growth hormone microneedle; a step of forming the mucus composition into a microneedle structure; and a step of solidifying the mucus composition into the shape of the microneedle structure.

[0038] Furthermore, the present invention provides a microneedle patch or microneedle array comprising various types of growth hormone microneedle structures described above.

[0039] The microneedle structure manufactured using the mucus composition for manufacturing growth hormone microneedles according to the present invention exhibits excellent insertion rates and storage stability. Furthermore, the disposable patch formulation is expected to result in cost savings compared to conventional refrigerated cartridges and needle replacement-required injections. Furthermore, it can be conveniently applied to the skin without the pain associated with injections, making it effective for treating hypogonadism.

[0040] FIG. 1 is a photograph showing the shape of a microneedle structure according to one embodiment of the present invention.

[0041] FIG. 2 is a perspective view of a microneedle structure according to one embodiment of the present invention.

[0042] Fig. 3 is a cross-sectional view showing a cross-section of a microneedle structure along line AA of Fig. 2.

[0043] Figure 4 is a photograph of the shape of microneedles manufactured using composition 4 of [Table 2] observed under a microscope.

[0044] Figure 5 is a photograph (A) showing changes in the shape of microneedles observed under a microscope according to differences in the composition of hGH / polymer used in the core formulation when manufacturing microneedles, and a photograph (B) showing the results of application to pig skin.

[0045] Figure 6 shows the HPLC profile results for the liquid raw material of hGH performed to verify the self-content test method.

[0046] Figures 7a to 7d show the purity test peaks (A) for the liquid raw material of hGH and the purity test peaks (B) for the polymer matrix performed to verify the in-house purity test method.

[0047] Figure 8 shows the results of evaluating the storage stability of a microneedle patch containing composition 2 of [Table 4] as a core layer after manufacturing the same. A represents the results of the content storage stability evaluation, and B represents the results of the purity storage stability evaluation.

[0048] Figures 9a and 9b show microscopic photographs of the shape of microneedles manufactured using a centrifugal concentration method formulation (top of Figure 9a), heat map analysis results for each application time of the manufactured microneedle patch (bottom of Figure 9a), and analysis results for dissolution rate for each application time (Figure 9b).

[0049] Figure 10 shows the results of shape improvement of microneedles manufactured using a freeze-dried formulation, and is a photograph of the shape before improvement (A) and after improvement (B) observed under a microscope.

[0050] Figure 11 shows microscopic photographs of microneedles (A) before shape improvement and (B) after shape improvement, manufactured using a freeze-dried formulation, after 5 minutes of application to pig skin.

[0051] Figure 12 shows the results of the skin insertion evaluation of microneedles with improved shape, and is a photograph of the microneedles observed under a microscope before application (A), after 5 minutes of application (B), and after 10 minutes of application (C), respectively.

[0052] Figure 13 shows microscopic photographs of the shape of microneedles before (A) and after (B) increasing hGH content, and the specifications of each microneedle.

[0053] Figure 14 shows the results of an in vivo rat skin insertion evaluation after increasing hGH content, and shows the results of an application and delivery rate experiment, showing microneedle photographs and skin delivery rates before application (A), after 5 minutes of application (B), and after 10 minutes of application (C).

[0054] Figure 15 is a graph comparing the PK profile and bioavailability of a solution containing dissolved hGH microneedles and a liquid raw material of hGH administered to rats through the same route.

[0055] Figure 16 is a graph comparing the PK profiles that appear after subcutaneously administering an hGH microneedle patch and an hGH liquid raw material to rats.

[0056] Hereinafter, the present invention will be described in more detail.

[0057] The terminology used herein is for the purpose of description only and should not be construed as limiting. The singular expression includes the plural expression unless the context clearly dictates otherwise. In this specification, terms such as "comprise" or "have" should be understood to indicate the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but do not preclude the possibility of the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof. Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the embodiments pertain. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning they have in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined in this application.

[0058] As described above, there is no known optimized formulation for manufacturing microneedles loaded with growth hormone, considering not only the possibility of manufacturing into microneedles and skin solubility, but also the stability and bioavailability of growth hormone. Accordingly, the inventors of the present invention screened a mucus composition that enables the manufacture of microneedles with a uniform shape and exhibits suitable solubility by dissolving even the core layer containing growth hormone when applied to the skin, and confirmed that when microneedles are manufactured with the screened mucus composition, they exhibit significantly improved storage stability compared to liquid raw materials for growth hormone. In addition, it was confirmed that the delivery rate and bioavailability of growth hormone are significantly improved by controlling the head diameter of the microneedle, the number of microneedles, and the concentration multiple of the growth hormone.

[0059] Accordingly, the first aspect of the present invention relates to a mucus composition for manufacturing growth hormone microneedles, comprising growth hormone, a water-soluble polymer, and an additive.

[0060] In this regard, the present invention provides the use of a mucus composition comprising a growth hormone, a water-soluble polymer and an additive for use in the manufacture of a growth hormone microneedle structure.

[0061] Furthermore, the present invention provides the use of a mucus composition comprising a growth hormone, a water-soluble polymer and an additive for use in the manufacture of a microneedle patch or microneedle array comprising the microneedle structure.

[0062] In this specification, growth hormone may be used with the same meaning as “Recombinant Human Growth Hormone”, “hGH”.

[0063] In the present invention, the mucus composition for manufacturing the growth hormone microneedle contains a high-concentration hGH raw material, and the high-concentration hGH raw material can be manufactured by concentrating the growth hormone.

[0064] In the present invention, the method for concentrating the growth hormone may be a centrifugal concentration method or a freeze-drying method, but is not limited thereto, and the growth hormone may be concentrated to a specific concentration or higher using a known concentration method.

[0065] In the present invention, the centrifugal concentration method can be performed by filtering the liquid raw material of growth hormone through a membrane filter having a molecular weight cutoff of 3 to 20 kDa.

[0066] In the present invention, the freeze-drying method can be performed by the steps of a) filtering a liquid raw material of growth hormone through a membrane filter having a molecular weight cutoff of 3 to 20 kDa; b) adding sucrose to the filtered concentrate; and c) performing freeze-drying.

[0067] In the present invention, the growth hormone may be a human growth hormone protein, and the human growth hormone protein may include an amino acid sequence disclosed in an open database such as NCBI, for example, may include an amino acid sequence disclosed in GenBank: QKG82153.1, GenBank: AAA72260.1, GenBank: AML27053.1, GenBank: AIA66930.1, etc. or a part thereof, but may include without limitation any amino acid sequence that exhibits substantially the same or corresponding effect as the protein. The human growth hormone protein may also include an isoform or a precursor thereof.

[0068] In the present invention, the human growth hormone may be included in an amount of 10 to 45 wt%, preferably 15 to 45 wt%, and most preferably 15 to 35 wt%, relative to the total mucus composition.

[0069] In the present invention, the water-soluble polymer may be a biocompatible water-soluble polymer, and may be at least one selected from the group consisting of hyaluronic acid, hydroxyethyl starch, hydroxypropyl methyl starch, gelatin, pullulan, dextran, sodium chondroitin sulfate, sodium hyaluronate, carboxymethylcellulose, polyvinylpyrrolidone, polyoxyethylene polyoxypropylene glycol, polyethylene glycol, and polyvinyl alcohol. Preferably, the water-soluble polymer may be hyaluronic acid.

[0070] In the present invention, the water-soluble polymer may be included in an amount of 10 to 45 wt%, preferably 15 to 25 wt%, and more preferably 15 to 20 wt%, relative to the total mucus composition.

[0071] In the present invention, if the water-soluble polymer is included in less than 10 wt% of the total mucus composition, it may be difficult to form the composition into a solid microneedle structure, and the physical strength of the microneedle structure may be insufficient to penetrate the skin. If the water-soluble polymer is included in more than 45 wt% of the total mucus composition, it may be difficult to homogenize the water-soluble polymer and growth hormone, and the release rate of the growth hormone loaded in the microneedle may be significantly reduced.

[0072] In the present invention, the additive is sucrose, trehalose, oligosaccharide, maltose, lactose, cellobiose, glycine, mannitol, calcium D-gluconate, arginine, alginic acid, pectin, carrageenan, chondroitin sulfate, dextran sulfate, chitosan, polylysine, collagen, gelatin, carboxymethyl chitin, fibrin, agarose, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), polymethacrylate, At least one selected from the group consisting of hydroxypropyl methylcellulose (HPMC), ethyl cellulose (EC), hydroxypropyl cellulose (HPC), carboxymethyl cellulose, cyclodextrin, gentiobiose, alkyltrimethylammonium bromide, hexadecyltrimethylammonium bromide, gentian violet, benzethonium chloride, docusate sodium salt, a SPAN-type surfactant, polysorbate, sodium dodecyl sulfate, benzalkonium chloride, and glyceryl oleate. Can be.Preferably, the additive may be sucrose and / or polysorbate.

[0073] In the present invention, the additives may be two or more types, and may be included in an amount of 10 to 45 wt% with respect to the entire mucus composition.

[0074] According to one embodiment of the present invention, when there are two types of additives, the first additive may be included in an amount of 9.95 to 40 wt% based on the entire mucus composition, and the second additive may be included in an amount of 0.05 to 5 wt% based on the entire mucus composition. For example, the first additive may be sucrose, and the second additive may be polysorbate. In this case, the polysorbate may include polysorbate 20 (Tween 20), polysorbate 60 (Tween 60), polysorbate 65 (Tween 65), or polysorbate 80 (Tween 80).

[0075] The mucus composition for manufacturing growth hormone microneedles of the present invention can be manufactured by mixing each component in a solid state, and the weight % included based on the solid content can be changed as follows: Based on the solid content for manufacturing the mucus composition for manufacturing growth hormone microneedles, the growth hormone is 10 to 30 wt%, the water-soluble polymer is 40 to 85 wt%, and the additive is 10 to 30 wt%.

[0076] In the present invention, when the growth hormone, water-soluble polymer, and additive are used as solids, each solid may be dissolved in a solvent and mixed. Preferred solvents include, but are not limited to, water, ionized water, saline solution, distilled water, purified water, sterilized purified water, phosphate buffered saline, C1-4 alcohol, or mixtures of any one or more thereof.

[0077] The mucus composition for manufacturing growth hormone microneedles of the present invention may include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is one commonly used in formulations, and includes, but is not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above components, the growth hormone microneedle composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like.

[0078] The appropriate dosage of the mucus composition for manufacturing growth hormone microneedles of the present invention can be determined in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity.

[0079] In the present invention, when the combination and ratio of water-soluble polymers and additives additionally included in addition to the growth hormone are optimized, the stability of the growth hormone increases and the solid form can be maintained, thereby stably maintaining the content and purity.

[0080] In one embodiment of the present invention, the mucus composition for manufacturing growth hormone microneedles may include additional excipients. For example, it may include additional additives for maintaining the stability of the growth hormone and additional polymers for maintaining the solid form.

[0081] Exemplary excipients may include, for example, buffers, carbohydrates, polymers, amino acids, peptides, surfactants, proteins, nonvolatile non-aqueous solvents, acids, bases, antioxidants, and saccharin.

[0082] One or more buffers may be used as part of the one or more excipients. The buffers typically function to stabilize the pH during the microneedle manufacturing process. The specific buffer used can be appropriately selected by those skilled in the art based on the content of the growth hormone of the present invention.

[0083] One or more carbohydrates, including mixtures of carbohydrates, may be used as part of the one or more excipients. The carbohydrates may be sugars, including monosaccharides, disaccharides, and polysaccharides, and may include, for example, non-reducing sugars such as raffinose, stachyose, sucrose, and trehalose; and reducing sugars such as monomers and disaccharides. Exemplary monomers may include apiose, arabinose, digitoxose, fucose, fructose, galactose, glucose, gulose, hamamellose, idose, lyxose, mannose, ribose, tagatose, sorbitol, xylitol, and xylose. Exemplary disaccharides may include, for example, sucrose, trehalose, cellobiose, gentiobiose, lactose, lactulose, maltose, melibiose, primerose, rutinose, sylabiose, sophorose, turanose, and bicyanose. In another embodiment, sucrose, trehalose, fructose, maltose, or a combination thereof may be used. All optical isomers (D, L, and racemic mixtures) of the exemplified sugars are also encompassed by the present invention.

[0084] Polysaccharides include, for example, starches, such as hydroxyethyl starch, pregelatinized corn starch, pentastarch, dextrins, dextran or dextran sulfate, gamma-cyclodextrin, alpha-cyclodextrin, beta-cyclodextrin, glucosyl-alpha-cyclodextrin, maltosyl-alpha-cyclodextrin, glucosyl-beta-cyclodextrin, maltosyl-beta-cyclodextrin, 2-hydroxy-beta-cyclodextrin, 2-hydroxypropyl-beta-cyclodextrin, 2-hydroxypropyl-gamma-cyclodextrin, hydroxyethyl-beta-cyclodextrin, methyl-beta-cyclodextrin, sulfobutylether-alpha-cyclodextrin, sulfobutylether-beta-cyclodextrin, and Sulfobutyl ether-gamma-cyclodextrin may be included. In embodiments, hydroxyethyl starch, dextrin, dextran, gamma-cyclodextrin, beta-cyclodextrin, or a combination thereof may be used. In embodiments, dextran having an average molecular mass of 35,000 to 76,000 may be used.

[0085] The one or more carbohydrates may be cellulose. Suitable celluloses may include, for example, hydroxyethyl cellulose (HEC), methyl cellulose (MC), microcrystalline cellulose, hydroxypropyl methyl cellulose (HPMC), hydroxyethylmethyl cellulose (HEMC), hydroxypropyl cellulose (HPC), and mixtures thereof.

[0086] One or more amino acids may be used for at least a portion of the one or more excipients. Suitable amino acids may include, for example, lysine, histidine, cysteine, glutamate, lysine acetate, sarcosine, proline, threonine, asparagine, aspartic acid, glutamic acid, glutamine, isoleucine, leucine, methionine, phenylalanine, serum tryptophan, tyrosine, valine, alanine, arginine, and glycine. In many cases, a salt form of the amino acid may be used to increase the aqueous solubility of the amino acid in an aqueous medium or formulation.

[0087] One or more peptides may be used for at least a portion of the one or more excipients. The amino acids constituting the peptides may be identical, or at least some may differ from each other. Suitable polyamino acids (identical amino acids) may include, for example, polyhistidine, polyaspartic acid, and polylysine.

[0088] One or more proteins may be used for at least a portion of the one or more excipients. Suitable proteins may include, for example, human serum albumin and bioengineered human albumin.

[0089] One or more saccharins may be used for at least a portion of said one or more excipients. For example, the saccharin is saccharin sodium dihydrate.

[0090] One or more lipids may be used for at least a portion of the one or more excipients. For example, the lipid may be dipalmitoylphosphatidylcholine (DPPC).

[0091] One or more acids and / or bases may be used for at least a portion of the one or more excipients. For example, one or more weak acids, weak bases, strong acids, strong bases, or some combination thereof may be used. The acids and bases may serve the purpose of solubilizing or stabilizing the growth hormone and / or dose-extending component. Such acids and bases may be referred to as counterions. Such acids and bases may be organic or inorganic. Exemplary weak acids include, for example, acetic acid, propionic acid, pentanoic acid, citric acid, succinic acid, glycolic acid, gluconic acid, glucuronic acid, lactic acid, malic acid, pyruvic acid, tartaric acid, tartronic acid, fumaric acid, glutamic acid, aspartic acid, malonic acid, butyric acid, crotonic acid, diglycolic acid, and glutaric acid. Exemplary strong acids include, for example, hydrochloric acid, hydrobromic acid, nitric acid, sulfonic acid, sulfuric acid, maleic acid, phosphoric acid, benzene sulfonic acid, and methane sulfonic acid. Exemplary weak bases include, for example, ammonia, morpholine, histidine, lysine, arginine, monoethanolamine, diethanolamine, triethanolamine, tromethamine, methylglucamine, and glucosamine. Exemplary strong bases include, for example, sodium hydroxide, potassium hydroxide, calcium hydroxide, and magnesium hydroxide.

[0092] One or more surfactants may be used for at least a portion of the one or more excipients. The one or more surfactants may be amphoteric, cationic, anionic, or nonionic. Suitable surfactants may include, for example, lecithin, polysorbates (such as, for example, polysorbate 20, polysorbate 40, and polysorbate 80), glycerol, sodium lauroamphoacetate, sodium dodecyl sulfate, cetylpyridinium chloride (CPC), dodecyltrimethyl ammonium chloride (DoTAC), sodium desoxycholate, benzalkonium chloride, sorbitan laurate, and alkoxylated alcohols (such as laureth-4).

[0093] One or more inorganic salts may be used for at least a portion of the one or more excipients. Suitable inorganic salts may include, for example, sodium chloride and potassium chloride.

[0094] Non-volatile, non-aqueous solvents may also be used for at least a portion of one or more of the excipients. Examples include propylene glycol, dimethyl sulfoxide, glycerin, 1-methyl-2-pyrrolidinone, N,N-dimethylformamide, and the like.

[0095] One or more antioxidants may be used for at least a portion of the one or more excipients. Suitable antioxidants may include, for example, sodium citrate, citric acid, ascorbic acid, methionine, sodium ascorbate, and combinations thereof.

[0096] In one embodiment of the present invention, if the combination and ratio conditions of the additives are not satisfied, the solution may not dissolve properly in the skin, may not be properly dispensed due to high viscosity, or may not be homogeneous.

[0097] According to one embodiment of the present invention, the mucus composition for manufacturing growth hormone microneedles may include a viscosity modifying agent for the purpose of controlling viscosity.

[0098] For example, the thickener is a thickener commonly used in the art, such as hyaluronic acid and its salts, polyvinylpyrrolidone, cellulose polymers (e.g., hydroxypropyl methylcellulose, hydroxyalkyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, ethyl hydroxyethyl cellulose, alkyl cellulose, and carboxymethyl cellulose), dextran, gelatin, glycerin, polyethylene glycol, polysorbate, propylene glycol, povidone, carbomer, gum ghatti, guar gum, glucomannan, glucosamine, dammer resin, rennet casein, locust bean gum, microfibrillated cellulose, psyllium seed gum, It may include, but is not limited to, xanthan gum, arabino galactan, gum arabic, alginic acid, gelatin, gellan gum, carrageenan, karaya gum, curdlan, chitosan, chitin, tara gum, tamarind gum, tragacanth gum, furcelleran, pectin or pullulan.

[0099] In the present invention, the mucus composition for manufacturing growth hormone microneedles may additionally contain a biocompatible and / or biodegradable material.

[0100] As used herein, the term "biocompatible material" refers to a material that is substantially non-toxic to the human body, chemically inert, and non-immunogenic. As used herein, the term "biodegradable material" refers to a material that can be decomposed in a living body by body fluids or microorganisms.

[0101] Biocompatible and / or biodegradable materials that can be used in the present invention include, for example, polyesters, polyhydroxyalkanoates (PHAs), poly(α-hydroxyacid), poly(β-hydroxyacid), poly(3-hydroxybutyrate-co-valerate; PHBV), poly(3-hydroxyproprionate; PHP), poly(3-hydroxyhexanoate; PHH), poly(4-hydroxyacid), poly(4-hydroxybutyrate), poly(4-hydroxyvalerate), poly(4-hydroxyhexanoate), poly(esteramide), polycaprolactone, polylactide, polyglycolide, poly(lactide-co-glycolide;PLGA), polydioxanone, polyorthoester, polyetherester, polyanhydride, poly(glycolic acid-co-trimethylene carbonate), polyphosphoester, polyphosphoester urethane, poly(amino acid), polycyanoacrylate, poly(trimethylene carbonate), poly(iminocarbonate), poly(tyrosine carbonate), polycarbonate, poly(tyrosine arylate), polyalkylene oxalate, polyphosphazenes, PHA-PEG, ethylene vinyl alcohol copolymer (EVOH), polyurethane, silicone, polyester, polyolefin, polyisobutylene and ethylene-alphaolefin copolymer, styrene-isobutylene-styrene triblock copolymer, acrylic polymers and copolymers, vinyl halide polymers and copolymers, polyvinyl chloride, polyvinyl ether, polyvinyl methyl ether, polyvinylidene halides, Polyvinylidene fluoride, polyvinylidene chloride, polyfluoroalkene, polyperfluoroalkene, polyacrylonitrile, polyvinyl ketone, polyvinyl aromatics, polystyrene, polyvinyl ester, polyvinyl acetate, ethylene-methyl methacrylate copolymer, acrylonitrile-styrene copolymer, ABS resin and ethylene-vinyl acetate copolymer, polyamide, alkyd resin, polyoxymethylene, polyimide, polyether, polyacrylate, polymethacrylate, polyacrylic acid-co-maleic acid, chitosan, dextran, cellulose, heparin, hyaluronic acid, alginate, inulin, starch or glycogen, and preferably polyester, polyhydroxyalkanoate (PHAs), poly(α-hydroxyacid), poly(β-hydroxyacid), Poly(3-hydroxybutyrate-co-valerate; PHBV), poly(3-hydroxyproprionate; PHP), poly(3-hydroxyhexanoate; PHH), poly(4-hydroxyacid), poly(4-hydroxybutyrate), poly(4-hydroxyvalerate), poly(4-hydroxyhexanoate), poly(esteramide), polycaprolactone, polylactide, polyglycolide, poly(lactide-co-glycolide;PLGA), polydioxanone, polyorthoester, polyetherester, polyanhydride, poly(glycolic acid-co-trimethylene carbonate), polyphosphoester, polyphosphoester urethane, poly(amino acid), polycyanoacrylate, poly(trimethylene carbonate), poly(iminocarbonate), poly(tyrosine carbonate), polycarbonate, poly(tyrosine arylate), polyalkylene oxalate, polyphosphazenes, PHA-PEG, chitosan, dextran, cellulose, heparin, hyaluronic acid, alginate, inulin, starch or glycogen.;

[0102] In the present invention, the mucus composition for manufacturing growth hormone microneedles can be used to manufacture a microneedle structure, and preferably can constitute a core layer of the microneedle structure.

[0103] According to a specific embodiment of the present invention, as shown in FIGS. 2 and 3, the microneedle structure (1) having the core layer (30) may include, for example, a base layer (20) formed on a support (10); a core layer (30) formed on the base layer (20); and a shell layer (40) formed on the base layer (20) to cover the core layer (30).

[0104] The second aspect of the present invention relates to a microneedle structure (1) manufactured using the aforementioned mucus composition for manufacturing growth hormone microneedles.

[0105] In the present invention, the mucus composition for manufacturing the growth hormone microneedle can constitute part or all of the microneedle structure (1), and since the description of the composition characteristics of the mucus composition for manufacturing the growth hormone microneedle is the same as described above, the description thereof is omitted.

[0106] Specifically, as illustrated in FIGS. 2 and 3, the microneedle structure (1) may include a base layer (20) formed on a support (10); a core layer (30) formed on the base layer (20) and composed of a mucus composition for manufacturing growth hormone microneedles of various forms as described above; and a shell layer (40) formed on the base layer (20) to cover the core layer (30). The shell layer (40) may include a cover layer (41) and a tip portion (42).

[0107] The above cover layer (41) is intended to protect the growth hormone of the core layer (30) and can completely cover the core layer (30) so that it is not exposed to the outside.

[0108] The tip portion (42) may be formed with a sharp tip to facilitate insertion into the skin. At this time, the shell layer (40) may be made of a material having high physical strength or may be manufactured using a manufacturing process for this purpose.

[0109] In this way, the microneedle structure (1) according to one embodiment of the present invention may only include the base layer (10) and the shell layer (40) in appearance because the cores (30) are not exposed to the outside.

[0110] In the present invention, the base layer (20) and the shell layer (40) do not contain growth hormones and may be composed of biodegradable and / or biocompatible materials. The biodegradable and / or biocompatible materials that can be used in the base layer (20) and the shell layer (40) are the same as those described above, and therefore, their description is omitted.

[0111] In addition, the base layer (20) and shell layer (40) can use all the same components used in the core layer (30) except for growth hormone.

[0112] In the present invention, the support (10) may be a polymer film. For example, (i) a plastic sheet or film selected from the group consisting of polyethylene, polyurethane, polyvinyl alcohol, polypropylene, polyethylene terephthalate, polystyrene (GPPS), polylactide, polyglycolide, polycarboxylic acid, polyethylene glycol, polyanhydride, polyamide, polyesteramide, polyorthoester, polydioxanone, polyacetal, polyketal, polycarbonate, polyorthocarbonate, polyphosphazene, polyhydroxybutyrate, polyhydroxyvalerate, polyalkylene oxalate, polyalkylene succinate, poly(malic acid), poly(amino acid), poly(methyl vinyl ether), chitin, chitosan and copolymers thereof, terpolymers, carboxymethylcellulose, hyaluronic acid, polyvinylpyrrolidone and ethylene acetate vinyl copolymer (EVA); (ii) a paper sheet selected from the group consisting of sterilized paper, cellophane, non-woven fabric and woven fabric; (iii) a silicone resin film by spraying or applying; or (iv) a fluorine oil film by spraying or applying, but is not limited thereto.

[0113] In the present invention, the growth hormone microneedle structure (1) can be manufactured in various shapes at regular intervals on the support. For example, it can be a cone shape, a square pyramid shape, a sphere shape, a short-headed shape, a wedge shape, a blade shape, or a candle shape, but is not limited thereto.

[0114] In the present invention, when the microneedle structure (1) is manufactured in a candle shape, the maximum diameter of the vertical cross-section with respect to the protrusion direction of the microneedle structure (1) may be 200 to 400 μm. If the diameter is outside the above range, the core layer containing the growth hormone may not be completely dissolved when applied to the skin.

[0115] In the present invention, the growth hormone microneedle structure (1) can be inserted into the skin in a patch or implant form.

[0116] As used herein, the term “patch” refers to a formulation that is attached to the skin to deliver a drug into the body.

[0117] In one embodiment of the present invention, when the microneedle structure (1) is inserted into the skin in the form of a patch, the support (10) may have adhesiveness to the skin to enhance adhesion to the skin.

[0118] As one form for the adhesiveness of the above support (10), a support (10) coated with an adhesive material, i.e., a support (10) coated with an adhesive, can be used.

[0119] As the above adhesive material, an adhesive commonly used in patch preparations can be used, and for example, a grade having wet surface adhesiveness of an acrylic, silicone, or rubber adhesive is preferable.

[0120] As another form for the adhesiveness of the above support (10), the support (10) may be water-soluble. By using a low molecular weight water-soluble film such as polyvinylpyrrolidone (PVP), carboxymethylcellulose (CMC), or polyvinyl alcohol (PVA), adhesiveness can be exhibited by the moisture of the skin. In this case, it is preferable to further laminate a water-insoluble polymer film on the surface facing the opposite side of the water-soluble support (10) so as not to adhere to the opposite side of the skin to which it is to be applied.

[0121] In one embodiment of the present invention, when the microneedle structure (1) is inserted into the skin in an implantable manner, the implantable microneedle structure (1) can be formed on a support (10) on which a coating layer is formed, and can be easily implanted into the body by separating the coating layer by an external stimulus.

[0122] In the present invention, the growth hormone microneedle structure can exhibit improved content storage stability and / or purity storage stability compared to the liquid growth hormone raw material.

[0123] Intravenous administration of growth hormone via microneedles, as described in the present invention, offers numerous advantages, including solving cold chain issues, reducing dose, and facilitating self-administration. Furthermore, because microneedles are solid, they can maintain the activity of growth hormone more stably than liquid injections.

[0124] A third aspect of the present invention relates to a microneedle patch or microneedle array comprising the various types of growth hormone microneedle structures described above.

[0125] In the present invention, the microneedle patch refers to a form in which a microneedle structure is inserted into the skin in the form of a patch, and since the description thereof is the same as described above, the description thereof is omitted.

[0126] In the present invention, the microneedle array can be manufactured in a form in which a plurality of microneedle structures are formed at a certain interval on a substrate layer.

[0127] When the above microneedle array is applied to the skin, the microneedle structure can reach the mucosa or skin, dissolve, and release growth hormone. The substrate of the microneedle array can adhere to the contours of the mucosa or skin under a high-humidity environment, thereby inducing a high delivery effect of growth hormone.

[0128] When applying a microneedle array to the skin, in order to make it difficult to break with an appropriate hardness and to facilitate penetration of growth hormone, the microneedle array can be formed using a mixture of a high molecular weight polymer material having a weight average molecular weight of 100,000 or more and a low molecular weight polymer material having a weight average molecular weight of 50,000 or less. The weight average molecular weight of the high molecular weight polymer material is preferably 50,000 or more and 2,000,000 or less. In addition, the weight average molecular weight of the low molecular weight polymer material is preferably 1,000 or more and 50,000 or less. In one embodiment, the weight average molecular weight can be measured by gel permeation chromatography (GPC).

[0129] In one embodiment of the present invention, the microneedle array can be attached to a support layer.

[0130] That is, the support layer can be integrated with the back surface of the microneedle array using an adhesive or pressure-sensitive adhesive. The size of the microneedle array and the support layer can be the same, but it is more preferable that the support layer be manufactured larger than the microneedle array to enhance the adhesion of the microneedle array to the mucosa, skin, etc.

[0131] The above support layer can be manufactured in a size and shape that is easy to handle, depending on the application area. For example, it is preferable to make it approximately 3 to 20 mm larger than the outer edge of the microneedle array. The thickness of the support layer can be the same as, thinner than, or thicker than the thickness of the microneedle array substrate. The support layer can be manufactured to be flexible and thin enough to support the microneedle array, and also to have a thickness that is easy to handle during use.

[0132] The fourth aspect of the present invention relates to a method for manufacturing a growth hormone microneedle structure using the aforementioned mucus composition for manufacturing growth hormone microneedles.

[0133] Specifically, the manufacturing method may include: forming a mucus composition for manufacturing the growth hormone microneedle; forming the mucus composition into a microneedle structure; and solidifying the mucus composition into the shape of the microneedle structure.

[0134] In the manufacturing method of the present invention, the step of forming the mucus composition is a step of mixing growth hormone, a water-soluble polymer, and an additive to prepare a mucus composition for manufacturing growth hormone microneedles according to the first aspect.

[0135] At this time, the description of the types and contents of the growth hormone, water-soluble polymer, and additives is the same as described above, so the description thereof is omitted.

[0136] In the manufacturing method of the present invention, the forming step may be characterized by at least one selected from the group consisting of fluidization, molding, centrifugal lithography, and blowing tension methods.

[0137] In the manufacturing method of the present invention, the microneedle structure can be formed into various shapes such as a cone shape, a square pyramid shape, a sphere shape, a short-headed shape, a wedge shape, a blade shape, or a candle shape, but is not limited thereto.

[0138] In a specific embodiment of the present invention, a microneedle structure can be manufactured including a first dispensing step of dispensing a first composition onto a support; a drying step of drying the first composition to form a base layer; a second dispensing step of dispensing a second composition containing a growth hormone onto the base layer; a third dispensing step of dispensing a third composition onto the base layer to cover the second composition; and a molding step of forming a core layer using the second composition and a shell layer using the third composition using a fluidization and centrifugal lithography technique. The first composition can include 65% (w / v) of Sodium Hyaluronate 30 kDa, which is a water-soluble polymer, using PBS as a solvent, and the second composition containing a growth hormone can be applied onto the base layer. Sodium hyaluronate 30 kDa 65% (w / v) can be applied as a third composition using PBS as a solvent to cover the second composition, and then a core layer using the second composition and a shell layer using the third composition can be formed through fluidization and centrifugal lithography steps.

[0139] Hereinafter, the composition and effects of the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention, and the scope of the present invention is not limited by these examples.

[0140] [Example 1]

[0141] Confirmation of the feasibility of manufacturing microneedles with hGH loaded in the core layer

[0142] 1-1. Formability evaluation and skin application test according to polymer type

[0143] Four types of viscous solutions in [Table 1] below were prepared, and the uniformity and dischargeability of the solutions were evaluated to confirm their suitability for molding.

[0144] Composition hGH concentration polymer polymer concentration Additive 1 Additive 1 concentration Additive 2 Additive 2 concentration 130% HA 30kDa 15% Sucrose 30% TWEEN 200.2% 230% HA HMW 2% 330% FG (Fish Gelatin) 30% 430% CMC 5%

[0145] Composition mixing, degassing solution uniformity discharge possibility evaluation result 1○X△Solution uniformity is poor 2○○XHigh viscosity, so dischargeability is low 3○○XHigh gel property, so dischargeability is low 4○△△Uniformity is poor, but discharge is possible

[0146] The evaluation criteria in [Table 2] above are as follows.

[0147] [Stirring, defoaming]

[0148] ○: Uniform appearance when observed with the naked eye.

[0149] △: Some irregularities in appearance are observed with the naked eye.

[0150] X: The appearance is uneven when observed with the naked eye.

[0151] [Solution uniformity]

[0152] ○: When discharging is repeated 5 times under the same discharging conditions, the relative standard deviation of the weight of the discharged material is 20% or less.

[0153] △: When discharging 5 times under the same discharging conditions, the relative standard deviation of the weight of the discharged material is 20% or more and 40% or less.

[0154] X: When discharging is repeated 5 times under the same discharging conditions, the relative standard deviation of the weight of the discharged material is 40% or more.

[0155] [Possibility of discharge]

[0156] ○: When dispensing viscous solution on the support, more than 55 out of 61 are good products.

[0157] △: When discharging a viscous solution on the support, 30 or more and 55 or fewer out of 61 are good products.

[0158] X: Less than 30 good products when dispensing viscous solution on the support

[0159] According to the results in [Table 2], a microneedle patch containing hGH in the core layer was manufactured using Composition 4 (CMC 5%) using a centrifugal molding method. For specific microneedle patch manufacturing methods, refer to Korean Patent Publication No. 10-2014-0131879 and Korean Patent Publication No. 10-2022-0003800.

[0160] The formability of the manufactured microneedles was determined through microscopic observation.

[0161] As shown in Fig. 4, microscopic observation results confirmed that microneedles with a constant diameter and length were produced, and that uniformly shaped microneedles could be formed.

[0162] Next, the microneedle patch manufactured with composition 4 was applied to a pig skin section to evaluate the skin insertion rate and the solubility of the hGH core layer. However, it was confirmed that the hGH core layer was not completely dissolved even after 40 hours of application.

[0163] 1-2. Formability evaluation and skin application test by core layer composition

[0164] After confirming that the skin solubility of composition 4 was not suitable, microneedle patches were manufactured by changing the composition of the viscous solution as shown in Table 3 below, and then the formability evaluation and skin application test were conducted in the same manner.

[0165] hGH / HA 30kDa GH / CMC 90kDa Content 272ug / patch 265ug / patch Core layer main composition hGH 15%, HA 20%, trehalose 15% hGH 15%, CMC 10%, trehalose 15% Microneedle composition HA 50%, PVP 5% HA 50%, PVP 5%

[0166] As a result of microscopic observation, as shown in A of Fig. 5, it was confirmed that the microneedles manufactured with the two compositions of Table 3 were manufactured with a constant diameter and length, making it possible to form microneedles with a uniform shape.

[0167] Next, the two types of fabricated microneedles were applied to pig skin sections to evaluate the insertion rate and the solubility of the hGH core layer. As confirmed in Fig. 5B, when applied to the pig skin section for 30 minutes, the hGH / HA core layer dissolved, but the hGH / CMC core layer remained.

[0168] Based on the above results, it was confirmed that the dissolution rate varies depending on the type of polymer used in the hGH core formulation, and that the dissolution rate is faster when HA is used than when CMC is used.

[0169] [Example 2]

[0170] Formulation screening for development of stabilized formulations

[0171] Based on the results of the possibility of manufacturing microneedles in Example 1, nine types of formulations shown in [Table 4] below were manufactured to develop a stabilized formulation of the core layer.

[0172] Composition hGH Concentration Polymer 1 Polymer 1 Concentration Polymer 2 Polymer 2 Concentration Additive 1 / Concentration Additive 2 / Concentration 1 30% HA 30kDa 10%--Sucrose / 30% TWEEN 20 / 0.20% 2 HA 30kDa 20%--3 HA 30kDa 30%--4 HA 30kDa 10% PVP K 175% 5 HA 30kDa 20% PVP K 175% 6 HA 30kDa 20% PVP K 1710% 7 HA 1000kDa 2%--8 Fish Gelatin 30%--9 CMC 250 kDa 5%--

[0173] To determine the suitability of the nine formulations described above for formability, the solution uniformity and dispensability were evaluated. Furthermore, microneedle patches containing hHG in the core layer were fabricated using each of the nine formulations described above, and then applied to pig skin sections to evaluate the solubility of the hGH core layer. The results of the formability and skin application tests are shown in [Table 5].

[0174] Composition Stirring, Degassing Solution UniformityDispensing UniformityDissolution Evaluation Results After Application1○XX-Solution uniformity is poor due to low viscosity2○○○○Dispensing uniformity is high, and core dissolution is confirmed when applied to the skin3○XX-Solution uniformity is poor due to high viscosity4○XX-Solution uniformity is poor due to low viscosity5○○X-Dispensing uniformity is poor6○○X-Dispensing uniformity is poor7○○X-Dispensing uniformity is poor8○○X-Dispensing uniformity is poor9○○○XDispensing uniformity is high, but core dissolution is not possible when applied to the skin

[0175] The evaluation criteria in [Table 5] are the same as in [Table 2] above. Based on the results in [Table 5], Composition 2 was determined as the core layer composition.

[0176] [Example 3]

[0177] Establishment of content test method and purity test method

[0178] 3-1. Establishment of content testing method

[0179] We established our own content test method by referring to the hGH content test method of the United States Pharmacopeia (USP), and measured the content of the hGH liquid raw material (15 mg / mL of recombinant human growth hormone) by performing HPLC under the conditions of [Table 6] below.

[0180] HPLC Conditions Wavelength 214 nm Column Packing L33 (XBridge Protein BEH SEC Column, 200Å, 3.5 μm, 7.8 mm X 300 mm) Column Temperature Room Temperature Flow Rate 0.6 mL / min Run Time 25 min Retention Time 15 min Pressure 800-900 psi Injection Volume 20 μL Results Resolution Solution Resolution < 0.4 (Dimer and Monomer Peaks) Tailing Factor < 1.7 (Monomer Peak) Concentration Cs (ru / rs); Cs = STD Concentration, ru = Sample Peak Area, rs = STD Peak Area

[0181] Based on the HPLC peaks in Fig. 6 and the content test results in [Table 7], the in-house content test method was verified by confirming that the theoretical concentration and the actual concentration were consistent.

[0182] Theoretical concentration (mg / ml)Csr u r s Concentration (mg / ml) 1130881835310604710.990.10.1289237427541660.11

[0183] 3-2. Establishment of purity test method

[0184] In order to qualitatively and quantitatively evaluate the denaturation of drugs during the microneedle manufacturing process, a purity test method was established by using hGH:USP29-NF24 chromatographic purity to confirm whether microneedle analysis was possible under the conditions of [Table 8].

[0185] For polymer matrix samples containing 40% HA, 15% trehalose (Tre), and 10% PVP, chromatographic purity tests were performed to measure purity. As shown in Fig. 7a, some peaks occurred within the hGH analysis time, and thus the corresponding areas were calculated and removed to measure purity.

[0186] Chromatographic purity test conditions Wavelength 220 nm Column Packing L26 (XBridge Protein BEH C4 Column, 300Å, 3.5 μm, 4.6 mm X 250 mm) Column temperature 45℃ Sample temperature 5℃ Flow rate 0.5 mL / min Run time 60 min Retention time 43 min Pressure 1800-1900 psi Injection volume 20 μL Result (Resolution solution) Resolution < 1.0 Tailing factor: 0.9-1.8 Impurity < 6.0%

[0187] As a result of the purity test of the polymer matrix used in the formulation of hGH microneedles, as shown in Figures 7b to 7d, it was confirmed that there was no interference with the hGH peak. Accordingly, it was decided to proceed with a storage stability test using the purity test method established in this example. [Example 4]

[0188] Storage stability assessment for development of stabilized formulations

[0189] 4-1. Content storage stability evaluation

[0190] To evaluate the content stability under different storage conditions through quantitative evaluation within microneedles, microneedle patches containing Composition 2 of [Table 4] as a core layer were manufactured, and hGH content analysis was performed via HPLC according to the storage stability test plan of [Table 9] below. The quantitative goal was to maintain 80% or more of the initial hGH content for 12 months under refrigerated storage, 6 months under room temperature storage, and 3 months under accelerated storage. Liquid hGH raw material (recombinant human growth hormone 15 mg / mL) was used as a control.

[0191] Long-term preservation and accelerated test_Content and flexible substances_Each MN and drug were diluted to 1 mL per vial of drug, and the content and flexible substances were evaluated respectively. MN Packaging conditions: Sealed packaging (protected from light / moisture) Storage conditions Refrigerated storage Room temperature storage (25℃ / 60%RH) Accelerated storage (40℃ / 75%RH) MN Total drug Total 4℃ MN hGH MN hGH Starting MN 3 / Drug 1, (In case of drug, repeat analysis 3 times) 312 weeks 31311 months 33131922 months 3131623 months 33131926 months 331 61 Stability Single lot required sample number 369

[0192] As confirmed in A of Fig. 8, in the 6-month content stability results, the microneedles maintained the hGH content both in room temperature storage (25°C) and accelerated storage (40°C), but the liquid raw material was measured to be 96% and 76% of the initial hGH content in room temperature storage and accelerated storage, respectively, confirming that the hGH content storage stability of the microneedles was superior to that of the liquid raw material.

[0193] The specific storage stability results of microneedle hGH content are shown in [Table 10].

[0194] Storage Conditions Start 0.5 months 1 month 6 months AVGSDRSD(%) Refrigerated storage (4℃) 256.93 37.43 10.0 34.5 11.13 04.23 55.7 276.73 29.0 AVG 279.33 40.7 SD 23.7 13.7 % of start content 100% 122% Room temperature storage (25℃ / 60%RH) 256.93 53.23 32.33 16.33 9.6 12.5 304.23 58.23 47.6 276.73 48.7 346.7 AVG 279.33 53.43 42.2 SD 23.7 4.7 8.6 % of start content %100%127%123%Accelerated storage (40℃ / 75%RH)256.9281.6341.8327.1303.536.211.9304.2257.2352.2329.2276.7308.5352.8327.4AVG279.3282.4348.9327.9 SD23.725.66.21.1% compared to initial content100%101%125%117%

[0195] 4-2. Purity storage stability evaluation

[0196] In order to evaluate the purity stability under different storage conditions through quantitative evaluation within the microneedle, analysis of flexible substances was performed using the purity test method established in Example 4 according to the storage stability test plan of Table 9 above, and the results are shown in B of Fig. 8. Similarly, the same hGH liquid raw material as above was used as a control group.

[0197] As confirmed in B of Fig. 8, the impurities of the microneedles were all measured below the standard when stored in the refrigerator, and when stored at room temperature, only 1 out of 3 samples measured above the standard (8.0%). In contrast, the impurities of the liquid raw material were measured at 100% in both room temperature storage and accelerated storage, confirming that the hGH purity stability of the microneedles according to the present invention is significantly higher than that of the liquid raw material.

[0198] Specific microneedle hGH purity storage stability results are shown in [Table 11].

[0199] Storage conditions 2 months 6 months AVGSDRSD(%) Refrigerated storage (4℃) 0.21.21.61.167.51.80.42.71.3 AVG(%) 1.60.9 SD 1.33.1 Room temperature storage (25℃ / 60%RH) 0.36.81.31.296.90.53.83.03.9 AVG(%) 1.34.8 SD 1.52.1 Accelerated storage (40℃ / 75%RH) 9.28.01.113.58.3 6.6 AVG(%) 8.0 SD 1.3

[0200] [Example 5]

[0201] Evaluation of microneedle insertion rate and dissolution rate according to concentration method

[0202] Since high-concentration hGH raw material is required for microneedle formulation, Composition 2 of [Table 4] was concentrated using a centrifugal concentration method or a freeze-drying method, and a microneedle patch containing the concentrated hGH raw material as a core layer was manufactured. The skin insertion rate and dissolution rate were then evaluated.

[0203] 5-1. Centrifugal concentration method

[0204] 10 mL of hGH liquid raw material was concentrated using a centrifugal filter (Amicon Ultra-15 centrifugal filter units, MWCO, 10 kDa; Merck Millipore, Billerica, MA) (4500 rpm, 2 h, 4°C).

[0205] The microneedle patch was manufactured using the concentrated formulation as a core layer. The upper part of Fig. 9a shows the microneedle shape of the manufactured microneedle patch. The manufactured microneedle patch was applied to the skin of a hairless SD rat (Coatech Co., Ltd.), and the skin penetration rate was evaluated through heat map analysis according to attachment time and shape analysis after patch removal. The lower part of Fig. 9a shows the results of heat map analysis according to attachment time. As confirmed by the results, the microneedle penetration rate was 100% with only 10 minutes of application, indicating that all microneedles pierced the skin.

[0206] Next, the hGH dissolution rate [=(content-residual amount) / content] was measured by checking the hGH remaining amount in the patch according to the attachment time. As a result of the dissolution rate analysis, as shown in Fig. 9b, the dissolution rates for attachment times of 30 minutes, 1 hour, and 3 hours were confirmed to be 13%, 34%, and 79%, respectively, indicating that the dissolution rate also increases as the attachment time increases.

[0207] 5-2. Freeze-drying method

[0208] To enhance the skin insertion and delivery rate of microneedles, the concentration method was changed to freeze-drying, and the shape of the microneedles was improved. The specific freeze-drying method is as follows.

[0209] 1) Concentrate 20 mL of hGH liquid raw material using a centrifugal filter (10 kDa Cutoff) (4℃, 4500 rpm, 2 h)

[0210] 2) Add 150 mg of sucrose to the centrifugally concentrated hGH concentrate (approximately 2 mL) and freeze at -80°C.

[0211] 3) Freeze-drying (-80℃, <50 mTorr, overnight)

[0212] Microneedles were manufactured in the form of patches using the concentrated formulation as a core layer, and, to improve the shape, the diameter of the thickest head portion of the microneedle was changed from 450 μm to 270 μm, as confirmed in Fig. 10. The specifications of the microneedle before and after improvement are shown in [Table 12].

[0213] Before improvement Microneedle After improvement Microneedle head diameter 450 ㎛ 270 ㎛ Length 850 ㎛ 850 ㎛ Number of microneedles 61 MN / patch 61 MN / patch Capacity 208 μg / patch 120 μg / patch

[0214] The fabricated microneedle patch was applied to the skin of a mini pig to evaluate its penetration rate, and compared with a microneedle patch of identical specifications before shape improvement. As shown in Figure 11, after 5 minutes of skin application, the microneedle before improvement appeared to dissolve outside the skin due to skin fluids. However, after improvement, the core layer containing the hGH of the microneedle was confirmed to dissolve inside the skin.

[0215] Additionally, the skin insertion of microneedles after shape improvement was evaluated in vivo in rats. As shown in Fig. 12, unlike the results of mini-pig skin application, most of the core layer containing hGH remained after 5 minutes of application (Fig. 12 B: residual amount 83 μg, delivery rate 29%), and it was confirmed that it was completely dissolved after 10 minutes of application (Fig. 12 C: 60 μg, delivery rate 49%).

[0216] [Example 6]

[0217] Evaluation of skin insertion with increasing concentration and number of microneedles

[0218] The decrease in hGH content due to improved microneedle geometry was overcome by increasing the concentration multiple and the number of microneedles. To this end, the concentration multiple of the original solution was increased from 10 to 20 times, and the number of microneedles was also increased from 61 to 91, producing microneedles. As shown in Figure 13, the diameter of the microneedle head was changed to 350 μm after the content was increased. The microneedle specifications before and after the content increase are shown in [Table 13].

[0219] Before content increase, microneedle After content increase, microneedle head diameter 270 ㎛ 350 ㎛ Length 850 ㎛ 850 ㎛ Number of microneedles 61 MN / patch 91 MN / patch Capacity 120 μg / patch 628 μg / patch

[0220] The skin insertion evaluation of the microneedle patch fabricated as described above was conducted in vivo in rats. As shown in Figure 14, the core layer containing hGH remained after 5 minutes of application, and was completely dissolved after 10 minutes of application. Analysis of the remaining hGH content within the patch after application revealed a 41% skin delivery rate after 10 minutes, demonstrating a significantly improved skin delivery rate of hGH.

[0221] [Example 7]

[0222] Pharmacokinetic evaluation

[0223] To determine the effects of polymers and additives contained in microneedles on pharmacokinetics, solutions containing dissolved microneedles and liquid raw hGH were administered to animals and the PK profiles of each formulation were analyzed.

[0224] 7-1. Animal Experiment Design

[0225] Eight-week-old SD rats (Orient Bio) were used as experimental animals. For growth hormone administration, the liquid hGH raw material was diluted in PBS and administered at a concentration of 365.4 μg / 300 μL, and the number of microneedles per animal was adjusted to be as close to 365.4 μg. Animals to which the microneedle patches were to be applied were depilated 1 hour before administration. According to the experimental group information, sample administration and blood collection were performed using EDTA-coated tubes, and the collected blood samples were centrifuged to separate the plasma, and the plasma was analyzed using an ELISA Kit (ALPCO, 25-HGHHU-E01). The plasma samples from the hGH microneedle patch experimental group and the hGH solution experimental group were diluted 100-fold in PBS containing 0.1% BSA and analyzed, and the dilution factor of 100 was reflected in the analysis.

[0226] 7-2. PK profile and bioavailability calculation results

[0227] As confirmed in the results of Fig. 15 and [Table 14] below, the bioavailability (BA) of the solution containing dissolved microneedles compared to the liquid raw material was confirmed to be 81.36% upon subcutaneous injection, confirming that the hGH structure within the MN was maintained to some extent.

[0228] Sample hGH solution subcutaneous injection Reconstituted hGH microneedle solution subcutaneous injection Dosage (μg / head) 365.4 365.4 AUC~6hr (min*ng / ml) 1587.5 129 1.6 Cmax (ng / ml) 732.24 32.3 Tmax (hr) 0.5 0.5 BA (%) compared to dose conversion 100.00 8 1.36

[0229] [Example 8]

[0230] Final formulation PK test results

[0231] A comparison of the pharmacokinetic properties of a shape-improved human growth hormone (hGH) microneedle (MN) patch and a liquid raw material subcutaneous injection was performed. Table 15 below shows the conditions of each experimental group used for the comparison.

[0232] Experimental group, route of administration, number of experimental animals, hGH dose (μg), hair removal (electric razor), blood collection time, hGH MN patch ID (patch), SD rat 4441O0 / 0.5 / 1 / 2 / 3 / 6h (6 points), hGH solution subcutaneous injection, SD rat 3441X0 / 0.5 / 1 / 2 / 3 / 6h (6 points)

[0233] The hGH content was confirmed using the content test method established in Example 3-1, and plasma analysis was performed using an ELISA Kit (ALPCO, 25-HGHHU-E01). As a result of the analysis, hGH was detected at 0 and 6 hours in the experimental group to which the microneedle patch was applied, and this was corrected in the PK calculation process. As shown in Fig. 16 and [Table 16] below, the BA of the experimental group to which the microneedle patch was applied was calculated to be 62% compared to the experimental group to which the liquid raw material was subcutaneously injected. The BA value of 62%, which is significantly higher than that confirmed in the previous experiment, was confirmed.

[0234] Sample hGH solution subcutaneous injection hGH microneedle patch Dosage (μg / head) 441 441 AUC~6hr (min*ng / ml) 776.8 485.7 Cmax (ng / ml) 270.1 266.7 Tmax (hr) 10.5 BA (%) compared to dose conversion 100 61.7

Claims

1. A mucus composition for manufacturing growth hormone microneedles, comprising growth hormone, a water-soluble polymer, and an additive.

2. A mucus composition for manufacturing growth hormone microneedles, characterized in that the growth hormone in the first paragraph is concentrated by a centrifugal concentration method or a freeze-drying method.

3. A mucus composition for manufacturing growth hormone microneedles, characterized in that in the second paragraph, the centrifugal concentration method is performed by filtering the growth hormone liquid raw material through a membrane filter having a molecular weight cutoff of 3 to 20 kDa.

4. In the second paragraph, a mucus composition for manufacturing growth hormone microneedles, characterized in that the freeze-drying method is performed in the following steps a) to c): a) A step of filtering a liquid raw material of growth hormone through a membrane filter having a molecular weight cutoff of 3 to 20 kDa; b) adding sucrose to the filtered concentrate; and c) A step of performing freeze-drying.

5. A mucus composition for manufacturing growth hormone microneedles, characterized in that in the first paragraph, the growth hormone is human growth hormone and is included in an amount of 10 to 45 wt% with respect to the entire mucus composition.

6. A mucus composition for manufacturing growth hormone microneedles, characterized in that in paragraph 1, the water-soluble polymer is hyaluronic acid and is included in an amount of 10 to 45 wt% based on the total mucus composition.

7. In the first paragraph, the additive is sucrose, trehalose, oligosaccharide, maltose, lactose, cellobiose, glycine, mannitol, calcium D-gluconate, arginine, alginic acid, pectin, carrageenan, chondroitin sulfate, dextran sulfate, chitosan, polylysine, collagen, gelatin, carboxymethyl chitin, fibrin, agarose, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), polymethacrylate, At least one selected from the group consisting of hydroxypropyl methylcellulose (HPMC), ethyl cellulose (EC), hydroxypropyl cellulose (HPC), carboxymethyl cellulose, cyclodextrin, gentiobiose, alkyltrimethylammonium bromide, hexadecyltrimethylammonium bromide, gentian violet, benzethonium chloride, docusate sodium salt, a SPAN-type surfactant, polysorbate, sodium dodecyl sulfate, benzalkonium chloride, and glyceryl oleate,A mucus composition for manufacturing growth hormone microneedles, characterized in that it comprises 10 to 45 wt% of the total mucus composition.

8. A mucus composition for manufacturing growth hormone microneedles, characterized in that in paragraph 7, the additives are sucrose and polysorbate, the sucrose is included in an amount of 9.95 to 40 wt% based on the total mucus composition, and the polysorbate is included in an amount of 0.05 to 5 wt% based on the total mucus composition.

9. In the first paragraph, the mucus composition A mucus composition for manufacturing a growth hormone microneedle, characterized in that the core layer is formed in a growth hormone microneedle structure including a base layer formed on a support; a core layer formed on the base layer; and a shell layer formed on the base layer to cover the core layer.

10. Base layer formed on the support; A core layer comprising a mucus composition for manufacturing growth hormone microneedles according to any one of claims 1 to 9; and A growth hormone microneedle structure comprising a shell layer formed to cover the core layer on the base layer.

11. A growth hormone microneedle structure according to claim 10, characterized in that the base layer and the shell layer do not contain growth hormone and are composed of a biodegradable or biocompatible material.

12. In the 10th paragraph, the growth hormone microneedle structure is characterized in that it is manufactured in a shape of a cone, a square pyramid, a sphere, a short head, a wedge, a blade, or a candle.

13. In the 12th paragraph, when the growth hormone microneedle structure is manufactured in a candle-shaped shape, the maximum diameter of the vertical cross-section with respect to the protrusion direction of the microneedle structure is characterized in that it is 200 to 400 ㎛.

14. A growth hormone microneedle structure according to claim 10, characterized in that the growth hormone microneedle structure is inserted into the skin in a patch or implant form.

15. A growth hormone microneedle structure according to claim 10, characterized in that the growth hormone microneedle structure exhibits improved content storage stability and / or purity storage stability compared to a liquid growth hormone raw material.

16. A microneedle patch or microneedle array comprising the growth hormone microneedle structure of Article 10.

17. A method for manufacturing a growth hormone microneedle structure using a mucus composition for manufacturing growth hormone microneedles according to any one of claims 1 to 9.

18. In paragraph 17, A step of forming a mucus composition for manufacturing the growth hormone microneedle; A step of forming the above mucus composition into a microneedle structure; and A method for manufacturing a growth hormone microneedle structure, comprising the step of solidifying the above mucus composition into a microneedle structure shape.

19. Use of a mucus composition according to any one of claims 1 to 9 for use in the manufacture of a growth hormone microneedle structure, or a microneedle patch or microneedle array comprising the microneedle structure.

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