Composition for alleviating or treating hair loss having hair loss drug loaded in lipid nanoparticles

Abstract

The present invention relates to a composition for alleviating or treating hair loss in which a hair loss drug is loaded in lipid nanoparticles. The lipid nanoparticles contain 0.30 mol% or more of a hydrophobic drug such as finasteride or dutasteride relative to the lipid nanoparticles and are loaded with the hydrophobic drug at an encapsulation rate of 90.0% or more, and thus have the advantage of being able to maintain water dispersibility for a long period of time while increasing transdermal delivery. Therefore, the present invention can more effectively alleviate symptoms of male pattern hair loss on the scalp where localized hair loss or thinning of hair occurs.

Description

Composition for improving or treating hair loss containing hair loss drug loaded in lipid nanoparticles

[0001] The present invention relates to a composition for improving or treating hair loss, wherein a hair loss drug is loaded into lipid nanoparticles, and is characterized in that the encapsulation rate of the hydrophobic hair loss drug in the lipid nanoparticles is increased to 90% or more.

[0002]

[0003] Minoxidil, dutasteride, and finasteride are the most commonly used hair loss treatments. While minoxidil can be taken orally, dutasteride and finasteride are primarily used as topical treatments. Based on their mechanism of action, minoxidil is primarily used to treat stress-related hair loss and female pattern baldness, while dutasteride and finasteride are used to treat male pattern baldness.

[0004] Among these, dutasteride and finasteride are hydrophobic drugs, and various lipid nanoparticle technologies have been developed to increase their skin delivery ability, but there has been a drawback in that the drug content in lipid nanoparticles is very low, less than 10%.

[0005] Therefore, there is an urgent need to develop a carrier composition that can maintain dutasteride and finasteride, hydrophobic hair loss drugs with low solubility in aqueous solutions and low encapsulation rate in lipid nanoparticles, in a stable, long-term dispersed state while increasing water dispersibility without using an organic solvent such as ethanol, so that they can be used as topical skin agents.

[0006]

[0007] [Prior Art Literature]

[0008] [Patent Document]

[0009] Japanese Patent No. 6944061 (Nanoliposome-microbubble complex encapsulating a complex of Cas9 protein, guide RNA that suppresses the expression of the SRD5A2 gene, and cationic polymer, and a composition containing the same for improving or treating hair loss)

[0010] Korean Patent No. 10-1918250 (Nanoliposome-microbubble complex encapsulating a drug for hair loss treatment and a composition containing the same for improving or treating hair loss)

[0011]

[0012] The purpose of the present invention is to provide a composition for improving or treating hair loss in which a hair loss drug is loaded into lipid nanoparticles, and is characterized in that the encapsulation rate of the hydrophobic hair loss drug in the lipid nanoparticles is increased to 90% or more.

[0013]

[0014] The present invention relates to a composition for improving or treating hair loss, wherein a hair loss drug is loaded into lipid nanoparticles.

[0015] The above hair loss medication can be selected from Dutasteride and Finasteride.

[0016] The lipid nanoparticles are characterized by comprising anionic lipids, neutral lipids, and fatty acids having 10 to 22 carbon atoms.

[0017] It is preferable that the above lipid nanoparticles contain 15 to 45 mol% of anionic lipid, 30 to 50.5 mol% of neutral lipid, and 15 to 35 mol% of fatty acid having 10 to 22 carbon atoms.

[0018] The drug is loaded at 0.3 mol% or more relative to the lipid nanoparticles, and the final encapsulation rate of the drug is 90.0% or more, and the encapsulation rate is according to the following equation 1.

[0019] More preferably, the drug is loaded at 0.3 to 0.75 mol% relative to the lipid nanoparticle, and the final encapsulation rate of the drug may be about 90.0 to 99.0%.

[0020] [Formula 1]

[0021] Encapsulation rate (%) = (molar ratio of drug loaded into lipid nanoparticles / molar ratio of drug injected into lipid nanoparticles) x 100

[0022] The particle size of the above lipid nanoparticles may be 10 to 600 nm, more preferably 10 to 400 nm, and most preferably 200 to 400 nm.

[0023] The above anionic lipids are,

[0024] DSPG (1,2-distearoyl-sn-glycero-3-phosphatidylglycerol),

[0025] DPPG (dipalmitoylphosphatidylglycerol),

[0026] DOPG (dioleoylphosphatidylglycerol),

[0027] SOPG (1-stearoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol),

[0028] Soy PG (L-α-phosphatidylglycerol (Soy)),

[0029] Egg PG (L-α-phosphatidylglycerol (Egg, Chicken)),

[0030] DLPG (1,2-dilauroyl-sn-glycero-3-phosphatidylglycerol),

[0031] Cardiolipin (1',3'-bis[1,2-dioleoyl-sn-glycero-3-phospho]-glycerol),

[0032] DMPG (dimyristoylphosphatidylglycerol),

[0033] POPG (palmitoyloleoylphosphatidylglycerol),

[0034] DOPS (dioleoylphosphatidylserine),

[0035] DTPA (diethylenetriamine pentaacetic acid),

[0036] DPTGA (1,4-dipalmitoyl-tartarate-2,3-diglutaric acid),

[0037] DSTSA (1,4-disteroyl-tartarate-2,3-disuccinic acid),

[0038] CHHDA (2-carboxyheptadecanoyl heptadecylamide),

[0039] DMPS (dimyristoylphosphatidylserine),

[0040] DPPS (dipalmitoylphosphatidylserine),

[0041] POPS (palmitoyloleoylphosphatidylserine),

[0042] DMPA (dimyristoylphosphatidic acid),

[0043] DPPA (dipalmitoylphosphatidic acid),

[0044] DOPA (dioleoylphosphatidic acid),

[0045] POPA (palmitoyl-oleoylphosphatidic acid),

[0046] CetylP(Cetyl phosphate) and

[0047] It is characterized by being a lipid comprising at least one selected from the group consisting of CHEMS (cholesterol hemisuccinate) or a salt thereof.

[0048] 상기 중성지질은 HSPC(hydrogenated soy phosphatidylcholine),

[0049] DSPC(1,2-distearoyl-sn-glycero-3-phosphocholine),

[0050] DOPC(1,2-dioleoyl-sn-glycero-3-phosphocholine),

[0051] POPE(1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine),

[0052] DSPE(1,2-distearoyl-sn-glycero-3-phosphoethanolamine),

[0053] DPPC(1,2-dipalmitoyl-sn-glycero-3-phosphocholine),

[0054] POPC(1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine),

[0055] DOPE(1,2-dioleoyl-sn-glycero-3-phosphoethanolamine),

[0056] SM(N-palmitoyl-D-erythro-sphingosylphosphorylcholine),

[0057] DLPE(1,2-dilauroyl-sn-glycero-3-phosphoethanolamine),

[0058] DPPE(1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine),

[0059] 포스파티딜 콜린(phosphatidyl choline),

[0060] 디미리스톨글리세롤(dimyristoylglycerol),

[0061] succinoyl-diacylglycerol,

[0062] phosphatidyl ethanolamine,

[0063] tetraether lipid,

[0064] sphingolipid,

[0065] diacryl glycerol and

[0066] It may include one or more selected from the group consisting of glycerides or a salt thereof.

[0067] The fatty acid having 10 to 22 carbon atoms may include both saturated fatty acids and unsaturated fatty acids, and preferably may include at least one fatty acid or a salt thereof selected from the group consisting of capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, γ-linolenic acid, α-linolenic acid, docosahexaenoic acid, stearidonic acid, eicosapentaenoic acid, docosapentaenoic acid, and eicosatetraenoic acid.

[0068] The lipid nanoparticles are characterized by having stable water dispersibility for more than 8 weeks in an aqueous solution state having a solid content of 0.1 to 2.0 wt%, and maintaining a polydispersity index (PDI) of 0.30 or less. More preferably, the lipid nanoparticles maintain a polydispersity index of 0.15 to 0.30 in an aqueous solution state having a solid content of 0.1 to 2.0 wt% for 8 to 12 weeks.

[0069] The above lipid nanoparticles have a 5α-dihydrotestosterone (5α-DHT) reducing effect, and are preferably characterized by a reduction of about 30 to 40% compared to the control group.

[0070] The above lipid nanoparticles may be nanoliposomes.

[0071] The fatty acid provides morphological stability to the lipid filling within the lipid nanoparticles, and is dispersed in the core and surface of the nanoparticles to enhance the stability of the nanoparticles.

[0072] The neutral lipid refers to a nonionic surfactant that is uncharged or has a neutral zwitterionic form within a pH range of 4.0 to 8.0. In addition, the neutral lipid may include any neutral lipid known to those skilled in the art.

[0073] The above anionic lipid refers to any amphipathic lipid having at least one negative charge within the range of pH 4.0 to pH 8.0 as an anionic surfactant. In addition, the above anionic lipid may include any anionic lipid known to those skilled in the art.

[0074] The above lipid nanoparticles may have a particle size of 10 to 600 nm. If the lipid nanoparticles have a size of less than 10 nm or greater than 600 nm, particle stability may be reduced, which is not desirable.

[0075] In the lipid nanoparticles, if the anionic lipid is less than 15 mol% or more than 45 mol%, the neutral lipid is less than 30 mol% or more than 50.5 mol%, or the fatty acid is less than 15 mol% or more than 35 mol%, particle stability or water dispersibility may not be maintained. In addition, although they may appear to be stable for a short period of time, they may not be able to maintain their state for a long period of time, more than 2 months after manufacture, which is not desirable. In particular, the fatty acid must be at least 15 mol% or more for their stability.

[0076] In the composition of the present invention, the drug is preferably loaded in an amount of 0.30 to 0.75 mol% relative to the lipid nanoparticles, and the final encapsulation rate of the drug is 90.0 to 99.0%, most preferably, 0.35 to 0.75 mol% is loaded, and the final encapsulation rate of the drug may be 95.0 to 99.0%.

[0077] It is preferable that the composition containing the above lipid nanoparticles be a topical skin preparation formulation.

[0078] The present invention also relates to a method for producing the lipid nanoparticles loaded with a hair loss drug.

[0079] Preferably, (step 1) a step of preparing a mixed composition by mixing at least one lipid selected from anionic lipids, neutral lipids, and fatty acids having 10 to 22 carbon atoms; and a hair loss drug; in an organic solvent phase; and,

[0080] (Step 2) A step of preparing the above mixed composition in a dispersion state;

[0081] may include.

[0082] At this time, in the first step, when the total of each raw material is 100 parts by weight, 100 to 5000 parts by weight of organic solvent can be mixed.

[0083] After the above 2 steps, if necessary, the dispersion may be subjected to high pressure homogenization.

[0084] In addition, the above dispersion can be purified by column purification to obtain a purified product and then subjected to sterilization filtration. The filter used for the above sterilization filtration is characterized by having a pore size of 0.1 to 0.4 μm.

[0085] The above high-pressure homogenization can be performed at 20,000 to 30,000 psi at 50 to 60°C and can be repeated 1 to 3 times. The dispersion liquid before homogenization can be heated to 50 to 60°C and used.

[0086] The above organic solvent may be selected from methanol, ethanol, benzene, butanol, butyl acetate, t-butyl alcohol, carbon tetrachloride, chloroform, cyclohexane, dichloroethane, dichloromethane, diethyl ether, diisopropyl ether, ethyl acetate, heptane, hexane, isooctane, methyl ethyl ketone (MEK), methyl t-butyl ether (MTBE), pentane, toluene, trichloroethylene, xylene, and mixed solvents thereof.

[0087] In the second step, a dispersion can be prepared by sonication or freeze-thawing. At this time, the dispersion can be prepared by a method of hydrating a freeze-dried composition, film, cake, etc. during freeze-thawing.

[0088] In particular, when the process of freezing and thawing the mixed composition is performed, it can preferably be repeated 1 to 12 times. By repeating the steps of freezing and thawing the mixed composition, lipid nanoparticles of more uniform size can be formed, and the formation efficiency of lipid nanoparticles can be increased. Since the formation of lipid nanoparticles of uniform size may be reduced if the number of times exceeds 12, it is preferable to repeat the process 12 or less.

[0089] At this time, the dispersion in the second step can be prepared in a separate reactor. By repeating the freezing and thawing process of the mixed composition within the reactor, lipid nanoparticles can be formed.

[0090] When freezing the lipid nanoparticles, sucrose, glycerol, and polyethylene glycol can be used as cryopreservatives, and preferably polyethylene glycol can be used. Among these, one selected from PEG-200, PEG-400, PEG-600, PEG-800, PEG-1000, PEG-1500, PEG-2000, PEG-4000, PEG-6000, PEG-8000, PEG-10000, and PEG-20000 can be used, and most preferably PEG4000 can be used.

[0091] The composition of the present invention can be provided as a pharmaceutical composition or a cosmetic composition.

[0092] The above pharmaceutical composition can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods. Carriers, excipients, and diluents that can be included in the above pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulated, it is prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrating agents, and surfactants that are commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and these solid preparations are prepared by mixing the composition of the present invention with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending agents can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can include witepsol, macrogol, Tween 61, cocoa butter, laurin, and glycerogelatin.

[0093] The dosage of the pharmaceutical composition of the present invention will vary depending on the age, sex, and body weight of the subject to be treated, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the prescriber's judgment. Determining the dosage based on these factors is within the skill of those skilled in the art, and the dosage typically ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferred dosage is 1 mg / kg / day to 500 mg / kg / day. Administration may be administered once a day or divided into several doses. The above dosage does not limit the scope of the present invention in any way.

[0094] The pharmaceutical composition of the present invention can be administered to mammals such as rats, livestock, and humans via various routes. Any route of administration is conceivable, including oral, rectal, intravenous, intramuscular, subcutaneous, intrauterine, or intracerebrovascular injections.

[0095] The cosmetic composition of the present invention may be manufactured in any formulation commonly manufactured in the art, and may be provided in the form of essence, lotion, emulsion, pack, hand cream, foot cream, lip balm, lipstick, eye shadow, eyeliner, eyebrow pencil, blusher, highlighter, general toner, skin, cream, serum, cosmetic soap, emollient toner, medicated toner, body cleanser, cleansing foam, cleansing lotion, gel, cleansing oil, cleansing cream, shampoo, rinse, hair treatment, hair lotion, cleansing tissue, and cleansing water.

[0096] The above cosmetic composition may contain any conventional excipient composition. Preferably, it may contain a fat component, an emollient, a surfactant, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, bactericides, antioxidants, pH adjusters, alcohols, pigments, fragrances, blood circulation promoters, cooling agents, antiperspirants, purified water, and the like.

[0097] In addition, the cosmetic composition may include another active ingredient, and preferably, it may include an anti-wrinkle agent, an anti-aging agent, a skin whitening agent, an antioxidant, an anti-inflammatory agent, a moisturizer, or a hair growth agent.

[0098] As the above-mentioned wrinkle-improving agent, substances that inhibit MMP-1, an extracellular matrix (ECM) protein-decomposing enzyme, can be selected, such as silicic acid, N-methyl-L-serine, isoflavonoids, dehydroepiendrosterone, and paoniflorin. Benzastatins and coenzyme Q10 can be used as substances that prevent skin aging by removing active oxygen that promotes the breakdown of ECM. In addition, adenosine, ascorbyl glucoside, kinetin, auxin, peptide, retinol, retinyl palmitate, and other substances known to have various wrinkle-improving effects can be used. Polyethoxylated retinamide, alpha hydroxyl acid, etc. are available.

[0099] The above whitening agent may include arbutin, niacinamide, ascorbic acid, magnesium ascorbyl phosphate, ascorbyl acid-2-glucoside, mulberry extract, ethyl ascorbyl ether, and soluble licorice extract.

[0100] As the above-mentioned moisturizer, one or more selected from the group consisting of hydroxyproline, glycerin, glycerol, urea, amino acid, lactate, and pyroglutamic acid can be used.

[0101] When the above lipid nanoparticles are applied as a pharmaceutical composition or cosmetic composition, 1 x 10 per mL 2 1 x 10 12 It can be used with dog particles included.

[0102]

[0103] The present invention relates to a composition for improving or treating hair loss, wherein a hair loss drug is loaded onto lipid nanoparticles. The lipid nanoparticles can be loaded with a hydrophobic drug, such as finasteride or dupasteride, at a concentration of 0.30 mol% or more and an encapsulation rate of 90.0% or more relative to the lipid nanoparticles, thereby enhancing transdermal delivery while maintaining long-term water dispersibility, thereby more effectively improving male pattern baldness symptoms on the scalp where localized hair loss or thinning of the scalp is occurring.

[0104]

[0105] Figure 1 is a schematic diagram of nanoparticles manufactured in Example 1 of the present invention.

[0106] Figure 2 is a graph showing the change in size of nanoparticles in the composition manufactured in Example 1 of the present invention over 8 weeks.

[0107] Figure 3 is a graph showing changes in the size distribution of nanoparticles in the composition manufactured in Example 1 of the present invention over 8 weeks.

[0108] Figure 4 is a graph showing the change in the encapsulation rate of nanoparticles in the composition manufactured in Example 1 of the present invention over 8 weeks.

[0109] Figure 5 is a result confirming that the composition prepared in Example 1 of the present invention has a cell protection function by inhibiting hair follicle cell death induced by 5-α-reductase activity.

[0110] Figure 6 shows the results confirming that the concentration of DHT in skin tissue decreased after applying the composition prepared in Example 1 of the present invention to rat skin.

[0111]

[0112] Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the contents introduced herein are provided to ensure thoroughness and completeness, and to sufficiently convey the spirit of the present invention to those skilled in the art.

[0113]

[0114] [Experimental Example 1. Preparation of Lipid Nanoparticles]

[0115] The lipid nanoparticles of the present invention were prepared as follows using a thin film hydration method through freeze-thaw. t-Butyl alcohol was used as the organic solvent.

[0116] As of Example 1, three types of lipids, including 36.9 mM (1900 mg) of DPPG Na as anionic lipid, 39.1 mM (1900 mg) of DSPC as neutral lipid, 24 mM (400 mg) of palmitate, and 1250 mg of dutasteride, were added to 1042 mL of t-butyl alcohol. The types and total content ratios of each lipid and dutasteride for compositions other than Example 1 are as shown in Tables 1 and 2. For reference, the amount of dutasteride added was the same in all cases, but in Example 2, 490 mg, which is twice as much, was added.

[0117] After each raw material was dissolved, the t-butyl alcohol solvent was evaporated to form a lipid film. Next, the lipid film was hydrated with 1042 mL of phosphate buffered saline (PBS) (pH 7.2). After sonicating the hydrated liquid, 245 g of PEG400 was added as a cryopreservative, and the freeze-thaw procedure of freezing and thawing the hydrated lipid film using liquid nitrogen was repeated five times, and sonicated again to hydrate, thereby obtaining a liquid in a dispersion state. The composition in the dispersion state was homogenized twice at 60°C and 27,000 psi using a high pressure homogenizer, if necessary.

[0118] For each lipid nanoparticle manufactured in this way, transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis were measured, and the particle size, dispersion, and dutasteride encapsulation rate are shown in Table 2.

[0119] Classification Lipid Nanoparticle Manufacturing Conditions Note Lipid Properties Raw Material MW Lipid Unit Input (mg) mol Mol % Example 1 Anionic Lipid DPPG Na 79 0.16 10.76 0.000 96 239.1 Neutral Lipid DSPC 744.95 0.76 0.001 02 036.9 Fatty Acid Palmitic Acid 256.4 3 00.16 0.000 62 4 24.0 Total Lipid Content 1.68 0.002 610 Example 2 Anionic Lipid DPPG Na 79 0.16 10.76 0.000 96 239.1 Neutral Lipid DSPC 744.95 0.76 0.001 02 036.9 Fatty Acid Palmitic Acid 256.4 3 00.16 0.000 62 4 24.0 Total Lipid Content 1.680.002610Comparative Example 1Anionic lipidDPPG Na790.1610.760.00096244.4Palmitic acid 1 / 2 decreaseNeutral lipidDSPC744.9500.760.00102041.9Fatty acidPalmitic acid256.4300.080.00031213.6Total lipid content 1.600.002290Comparative Example 2Anionic lipidDPPG Na790.1610.760.00096251.5Palmitic acid excludedNeutral lipidDSPC744.9500.760.00102048.5Fatty acidPalmitic acid256.4300.000.0000000.0Total lipid Content 1.520.001980Comparative Example 3Anionic lipid DPPG Na790.1610.000.00000062.0Excluding DSPCNeutral lipid DSPC744.9500.760.0010200.0Fatty acid Palmitic acid256.4300.160.00062438.0Total lipid content 0.920.001640Comparative Example 4Anionic lipid DPPG Na790.1610.760.0009620.0Excluding DPPGNeutral lipid DSPC744.9500.000.00000060.6Fatty acid Palmitic acid256.4300.160.00062439.4Total lipid content 0.920.001590

[0120]

[0121] ClassificationDutasteride injection conditionsRemarksDrug injection amountMWmolInjection molar ratioExample 10.5 mg528.530.0009460.363018Example 21.0 mg528.530.0018920.726037Drug doubleComparative example 10.5 mg528.530.0009460.412379Comparative example 20.5 mg528.530.0009460.477288Comparative example 30.5 mg528.530.0009460.575372Comparative example 40.5 mg528.530.0009460.596551

[0122]

[0123] For each lipid nanoparticle manufactured in this way, transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis were measured, and the particle size, dispersion, and dutasteride loading rate are shown in Tables 3 and 4.

[0124] Encapsulation rate (%) = (molar ratio of drug loaded into lipid nanoparticles / molar ratio of drug injected into lipid nanoparticles) x 100

[0125]

[0126] Classification Manufacturing Results Dutasteride loading per 1 mg lipid Dutasteride content in unit lipid Actual loading (mol) in lipid nanoparticles Molar drug ratio per unit lipid Encapsulation rate Example 10.29 mg 0.48 72 mg / 1.68 mg 0.000 92 2 0.35 37 25 / 1.68 mg 97.4 % Example 20.58 mg 0.97 44 mg / 1.68 mg 0.001 84 4 0.70 74 5 0 / 1.68 mg 97.4 % Comparative Example 10.22 mg 0.35 20 mg / 1.60 mg 0.000 66 6 0.29 0 32 1 / 1.60 mg 7 0.4 % Comparative Example 20.02 mg 0.03 0 4 mg / 1.52 mg 0.000 58 0.02 90 20 / 1.52 mg6.08 %Comparative Example 30.48 mg0.4416mg / 0.92 mg0.0008360.508179 / 0.92 mg88.3 %Comparative Example 40.09 mg0.0828mg / 0.92 mg0.0001570.098791 / 0.92 mg16.5 %

[0127]

[0128] Classification Manufacturing Results SizePDI Example 1347.0 nm 0.234 Example 2330.0 nm 0.274 Comparative Example 1974.1 nm 0.949 Comparative Example 2690.7 nm 1.000 Comparative Example 3377.7 nm 0.333 Comparative Example 4974.1 nm 0.949

[0129]

[0130] As a result, it was confirmed that lipid nanoparticle compositions containing about 98% of dutasteride were produced in the lipid nanoparticles of the present invention in Examples 1 and 2. The number of nanoparticles produced in the compositions of Examples 1 and 2 was confirmed in an aqueous dispersion state, and was approximately 1.0 x 10 12 6 x 10 12 It was found that the particles / ml.

[0131] Meanwhile, lipid nanoparticles were manufactured under the same conditions as in Examples 3 to 12 by varying the type of lipid, and it was confirmed that lipid nanoparticles were manufactured under conditions almost similar to those in Example 1. At this time, the drug was found to be loaded at 0.35 to 0.75 mol% relative to the lipid nanoparticles.

[0132] Lipid nanoparticle manufacturing conditions (same as Dutasteride input amount of 0.5 mg) Results Lipid Classification Mol % Size PDI Encapsulation rate Example 3 Anionic lipid DOPS Na 39.12 35.1 nm 0.21 598 % Neutral lipid DOPE 36.9 Fatty acid Palmitic acid 24.0 Example 4 Anionic lipid DTPA Na 39.13 69.2 nm 0.24 697 % Neutral lipid DOPC 36.9 Fatty acid Linoleic acid 24.0 Example 5 Anionic lipid POPS 39.12 0 4.3 nm 0.25 0 96 % Neutral lipid DPPC 36.9 Fatty acid oleic acid 24.0 Example 6 Anionic lipid DOPS 39.13 30.4 nm 0.22 297 % Neutral lipid DSPC 36.9 Fatty acid docosapentaenoic acid 24.0 Example 7 Anionic lipid DMPS 39.135 3.1 nm 0.26496 % Neutral lipid HSPC 36.9 Fatty acid Palmitic acid 24.0 Example 8 Anionic lipid DPPS Na 39.124 5.4 nm 0.23697 % Neutral lipid POPE 36.9 Fatty acid Palmitic acid 24.0 Example 9 Anionic lipid DOPA Na 39.126 6.2 nm 0.22097 % Neutral lipid DOPE 36.9 Fatty acid Palmitic acid 24.0 Example 10 Anionic lipid DPPG Na 34.33 33.4 nm 0.25498 % Neutral lipid DSPC 50.5 Fatty acid Palmitic acid 15.2 Example 11 Anionic lipid DPPG Na 42.324 5.4 nm 0.26898 % Neutral lipid DSPC 30.3 Fatty acid Palmitic acid 27.4 Example 12 Anionic lipid DPPG Na 25.3 35 1.2 nm 0.2 3 798 % Neutral lipid DSPC 40.5 Fatty acid Palmitic acid 34.2

[0133] Likewise, under the conditions of the comparative example, lipid nanoparticles were manufactured as follows by varying the lipid content, and not only did most of them deviate from 0.30 to 0.75 mol% compared to lipid nanoparticles, but the encapsulation rate was very low as shown in Table 6 below, making them unsuitable for use as drug delivery vehicles. In addition, the particle size was confirmed to be very non-uniform, indicating that the drug stability was poor.

[0134] Lipid nanoparticle manufacturing conditions (same as Dutasteride input amount of 0.5 mg) Results Lipid Classification Mol % Size PDI Encapsulation rate Comparative example 5 Anionic lipid DOPS Na 50 1 1 4 2.6 nm 0.62 3 3 3 % Neutral lipid DOPE 2 5 Fatty acid Palmitic acid 2 5 Comparative example 6 Anionic lipid DTPA Na 70 4 6 0.4 nm 0.89 2 1 4 % Neutral lipid DOPC 2 0 Fatty acid Palmitic acid 1 0 Comparative example 7 Anionic lipid POPS 50 6 2 1.3 nm 0.56 2 1 1 % Neutral lipid DPPC 2 5 Fatty acid Palmitic acid 2 5 Comparative example 8 Anionic lipid DOPS 70 6 6 4.0 nm 1.13 6 1 4 % Neutral lipid DSPC 2 0 Fatty acid Palmitic acid 1 0 Comparative example 9 Anionic lipid DMPS 50 5 3 5.4 nm 0.94 5 1 2 % Neutral lipid HSPC 25 Fatty acid Palmitic acid 25 Comparative example 10 Anionic lipid DPPS Na 70 8 12.3 nm 1.16 3 21 % Neutral lipid POPE 20 Fatty acid Palmitic acid 10

[0135]

[0136] [Experimental Example 2. Characteristics of lipid nanoparticles loaded with high-content hair loss medication]

[0137] The lipid nanoparticles of Example 1 of the present invention were selected as representative and the particle status was confirmed for 8 weeks, and the status is disclosed in FIGS. 2 to 4. For other lipid nanoparticles with low initial encapsulation rates or low particle stability, separate long-term particle status confirmation tests were not conducted because their subsequent usability itself is meaningless.

[0138] As a result, the lipid nanoparticles of Example 1 were maintained in a stable state with almost no change for 8 weeks, and the drug loading rate was also confirmed to be unchanged even at room temperature of 25°C or under harsh conditions of 40°C.

[0139] In addition, during the 4-week period between the 8 weeks, a high-pressure dispersion process was performed, but no significant aggregation phenomenon was observed, and the dispersion form was still in a stable state.

[0140] Therefore, through these results, it can be proven that the lipid nanoparticles of Example 1 were manufactured in an excellent state for carrying a hair loss drug.

[0141]

[0142] [Experimental Example 3. Cytotoxicity Confirmation]

[0143] The cells for hair loss treatment were prepared using a 5 alpha reductase 2 overexpressing DP (Dermal papilla) cell line (O / E-SRD5a2-DPC) (Korean Patent No. 10-1918250). The cells were cultured at 37°C under 5% CO2 in DPC medium (Gibco) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin.

[0144] To measure the effect on cell viability, 96-well plates (5 x 10 3 O / E-SRD5a2-DPC cells were seeded in a well (cell / well) one day before material treatment, and then treated with lipid nanoparticles (LNPs) loaded with dutasteride of Example 1 manufactured in the present invention. At this time, the drug concentration loaded in the lipid nanoparticles was calculated so that the final concentration of dutasteride in the cell medium was 0 to 1000 nM.

[0145] After 24 hours of culture, the presence or absence of changes in cell morphology was observed under an optical microscope, and cell viability (%) was determined using a Cell counting kit / WST-8 assy.

[0146] To this end, WST-8 reagent (Dojindo) was added to the medium at 37°C for 1 hour, and then the absorbance was measured at 450 nm using a microplate reader (BioTek), and the cell viability (Viability, %) was calculated compared to the lipid nanoparticle-untreated group (Negative control).

[0147] As a result of the experiment, the cell viability of the control group, dutasteride, and dutasteride-LNP treated groups showed almost no change in the result value of approximately 95.0-105.0%, and there was no cytotoxicity.

[0148]

[0149] [Experimental Example 4. Confirmation of Cell Activity]

[0150] After confirming cell viability (toxicity), the cell activity according to dutasteride-LNP treatment was measured to measure the inhibitory effect on the decrease in viability of the cell line for hair loss treatment (O / E-SRD5a2-DPC).

[0151] First, for in vitro activity measurement, testosterone (0.2 M) was treated in SRD5a2-O / E-DPC) cells for 24 hours, and a decrease in cell viability (approximately 30%) due to DHT (dihydrotestosterone) converted by the SRD5a enzyme in the cells was confirmed.

[0152] Subsequently, SRD5a enzyme cells were treated with 0.2 M testosterone and 0-1000 nM dutasteride or dutasteride-LNP for 24 hours, and the activity to inhibit the decrease in cell viability compared to the testosterone-only treatment group was measured using the CCK-8 assay. The experimental results confirmed that when 1 mM dutasteride-LNP was added together with 0.2 M testosterone, cell viability was maintained without decrease.

[0153]

[0154] [Experimental Example 5. Confirmation of 5α-dihydrotestosterone (5α-DHT, 5α-dihydrotestosterone) concentration in skin tissue after application to rat skin]

[0155] Dutasteride-loaded lipid nanoparticles were applied to rat skin, and the reduction in DHT by the active ingredient was compared with the oral administration group. The oral administration group was administered 0.1 mg of dutasteride (once daily) for 7 days, and dutasteride-loaded lipid nanoparticles were applied at a dose of 0.05 mg based on dutasteride (once daily) for 7 days to an area of ​​3 cm in diameter. Before drug treatment, a skin biopsy was performed to measure the DHT concentration in the tissue using ELISA, and this was used as a baseline, and the DHT concentration in the treated skin was measured and compared.

[0156] As a result, as shown in FIG. 6, it can be confirmed that the composition of Example 1 of the present invention reduces the DHT concentration in skin tissue to a level equivalent to that of the oral administration group.

[0157] These results demonstrate that the composition of the present invention significantly enhances the drug delivery properties of a hydrophobic hair loss drug.

Claims

A composition for improving or treating hair loss, wherein a hair loss drug is loaded into lipid nanoparticles, The above hair loss medication is selected from at least one of dutasteride and finasteride, A composition for improving or treating hair loss, characterized in that the lipid nanoparticles comprise anionic lipids, neutral lipids, and fatty acids having 10 to 22 carbon atoms. In the first paragraph, The above drug is loaded at 0.30 mol% or more compared to the above lipid nanoparticles, A composition for improving or treating hair loss, characterized in that it has an encapsulation rate of 90.0% or more according to the following [Formula 1]. [Formula 1] Encapsulation rate (%) = (molar ratio of drug loaded into lipid nanoparticles / molar ratio of drug injected into lipid nanoparticles) x 100 In the first paragraph, The above anionic lipids are SPG (1,2-distearoyl-sn-glycero-3-phosphatidylglycerol), DPPG (dipalmitoylphosphatidylglycerol), DOPG (dioleoylphosphatidylglycerol), SOPG (1-stearoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol), Soy PG (L-α-phosphatidylglycerol (Soy)), Egg PG (L-α-phosphatidylglycerol (Egg, Chicken)), DLPG (1,2-dilauroyl-sn-glycero-3-phosphatidylglycerol), Cardiolipin (1',3'-bis[1,2-dioleoyl-sn-glycero-3-phospho]-glycerol), DMPG (dimyristoylphosphatidylglycerol), POPG (palmitoyloleoylphosphatidylglycerol), DOPS (dioleoylphosphatidylserine), DTPA (diethylenetriamine pentaacetic acid), DPTGA (1,4-dipalmitoyl-tartarate-2,3-diglutaric acid), DSTSA (1,4-disteroyl-tartarate-2,3-disuccinic acid), CHHDA (2-carboxyheptadecanoyl heptadecylamide), DMPS (dimyristoylphosphatidylserine), DPPS (dipalmitoylphosphatidylserine), POPS (palmitoyloleoylphosphatidylserine), DMPA (dimyristoylphosphatidic acid), DPPA (dipalmitoylphosphatidic acid), DOPA (dioleoylphosphatidic acid), POPA (palmitoyl-oleoylphosphatidic acid), CetylP(Cetyl phosphate) and A composition for improving or treating hair loss, characterized in that it is a lipid containing at least one selected from the group consisting of CHEMS (cholesterol hemisuccinate) or a salt thereof. In the first paragraph, The above neutral lipid is HSPC (hydrogenated soy phosphatidylcholine), DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine), DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine), POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine), DSPE (1,2-distearoyl-sn-glycero-3-phosphoethanolamine), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine), POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine), DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine), SM (N-palmitoyl-D-erythro-sphingosylphosphorylcholine), DLPE (1,2-dilauroyl-sn-glycero-3-phosphoethanolamine), DPPE (1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine), phosphatidyl choline, Dimyristoylglycerol, succinoyl-diacylglycerol, phosphatidyl ethanolamine, tetraether lipid, sphingolipid, diacryl glycerol and A composition for improving or treating hair loss, characterized in that it comprises at least one selected from the group consisting of glycerides or a salt thereof. In the first paragraph, The fatty acids having 10 to 22 carbon atoms include both saturated fatty acids and unsaturated fatty acids, A composition for improving or treating hair loss, characterized in that it comprises at least one fatty acid or a salt thereof selected from the group consisting of capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, γ-linolenic acid, α-linolenic acid, docosahexaenoic acid, stearidonic acid, eicosapentaenoic acid, docosapentaenoic acid, and eicosatetraenoic acid. In the first paragraph, A composition for improving or treating hair loss, characterized in that the lipid nanoparticles maintain a polydispersity index (PDI) of 0.30 or less for 8 weeks or longer in an aqueous solution state having a solid content of 0.1 to 2.0 wt%. In the first paragraph, A composition for improving or treating hair loss, characterized in that the lipid nanoparticles have a 5α-dihydrotestosterone (5α-DHT) reducing effect. In the first paragraph, A composition for improving or treating hair loss, characterized in that the size of the lipid nanoparticles is 10 to 600 nm.

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