Formulations with improved absorptivity of low-solubility and / or low-membrane-permeability drugs

Abstract

The present invention provides a pharmaceutical composition comprising a low solubility and / or low membrane permeability drug and an ionic liquid, characterized in that the ionic liquid comprises an anionic component and a cationic component, the anionic component is oleic acid or isostearic acid, and the cationic component is oleic acid or isostearic acid. The cationic component is selected from cationic lipids; the transmucosal absorption enhancer or digestive tract absorption enhancer contains an ionic liquid, and is characterized in that the ionic liquid contains an anionic component and a cationic component, the anionic component is oleic acid or isostearic acid, and the cationic component is selected from cationic lipids.

Description

Technical Field

[0001] The present invention relates to a preparation for improving the absorption of a low-solubility and / or low-membrane permeability drug, and a transmucosal absorption enhancer and a digestive tract absorption enhancer for improving the transmucosal absorption and digestive tract absorption of a low-solubility and / or low-membrane permeability drug. In detail, the present invention relates to a preparation containing a low-solubility and / or low-membrane permeability drug and an ionic liquid, which significantly improves the absorption of the low-solubility and / or low-membrane permeability drug, and a transmucosal absorption enhancer and a digestive tract absorption enhancer containing an ionic liquid, which significantly improves the transmucosal absorption and digestive tract absorption of a low-solubility and / or low-membrane permeability drug. Background Art

[0002] When developing a drug as an oral preparation, if sufficient oral absorbability cannot be obtained, it will lead to the incompleteness of the efficacy evaluation project, etc., so that the development of the drug is delayed or terminated. In recent years, in order to evaluate such risks, BCS (Biopharmaceutics Classification System) has been used as a method to infer the main reason for limiting oral absorbability based on the characteristics of the drug raw materials. In BCS, compounds are classified into four levels based on the solubility and membrane permeability of the drug, and attempts have been made to clarify the digestive tract absorption characteristics of each and use them in formulation development.

[0003] Specifically, in BCS, the solubility in 250 mL buffer at pH 1.0 to 7.5 is combined with the permeability of the cell membrane of Caco-2, MDCK cells, etc. from the gastrointestinal epithelial cells, or the membrane absorption using the permeability of artificial membranes, and the compound group showing high solubility (High Solubility; HS) and high membrane permeability (High Permeability; HP) is classified into Class 1 (HS, HP), the compound group showing low solubility (Low Solubility; LS) and high membrane permeability (HP) is classified into Class 2 (LS, HP), the compound group showing high solubility (HS) and low membrane permeability (Low Permeability; LP) is classified into Class 3 (HS, LP), and the compound group showing low solubility (LS) and low membrane absorption (LP) is classified into Class 4 (LS, LP) (Non-Patent Document 1). In particular, compounds with low solubility and / or low membrane absorption that show low absorption through the mucosa, digestive tract, etc. are difficult to use as oral preparations, so the dosage forms that these compounds can be used in are limited.

[0004] On the other hand, in clinical settings, oral preparations that are less invasive and more convenient for patients are desired. However, there is currently no known method for developing oral preparations with improved absorbability of these compounds.

[0005] In recent years, as an attempt to improve the absorbability of compounds showing low solubility and / or low membrane permeability, absorption enhancers for such compounds have been studied. Absorption enhancers are compounds that promote drug absorption by directly acting on mucous membranes to change their structures. So far, for various compounds including surfactants, bile salts, bacterial toxins, chelating agents or medium-chain fatty acids, in vitro and in vivo tests, the effectiveness of improving the absorption of molecules with low biomembrane permeability is being studied. As absorption enhancers that are practical in oral preparations, there are sodium caprate (C10) and 8-(2-hydroxybenzamide) sodium caprylate (SNAC), etc., but they are mainly used in protein preparations such as GLP-1, and their effect cannot be said to be sufficient.

[0006] Prior art literature

[0007] Patent Literature

[0008] Patent Document 1: Japanese Patent Application Publication No. 2020-73533

[0009] Patent Document 2: U.S. Patent No. 10278923

[0010] Non-patent literature

[0011] Non-patent literature 1: A theoretical basis for a biopharmaceutic drug classification: the correlation of in vitro drug product dissolution and invivo bioavailability, Pharm Res. 1995Mar; 12(3): 413-20. Summary of the invention

[0012] Problems to be solved by the invention

[0013] So far, the present inventors have developed various transdermal absorption preparations that use ionic liquids (ILs), which are salts formed of organic anions and organic cations and are liquid at room temperature, to efficiently percutaneously absorb compounds such as low-molecular drugs, medium-molecular nucleic acids, and low-molecular peptides, and have obtained the knowledge that it is possible to design ionic liquids that are optimal for drugs with different characteristics. However, it is believed that the absorption mechanism of drugs for percutaneous absorption is greatly different from that for digestive tract absorption and intestinal absorption (transmucosal absorption), and even if ionic liquids are used directly in transdermal absorption preparations, improvement in absorbability cannot be expected.

[0014] Therefore, the object of the present invention is to design an ionic liquid that promotes the transmucosal absorption and digestive tract absorption of a drug having low solubility and / or low membrane permeability that is usually extremely difficult to use by oral administration, and to provide an oral preparation containing such a drug and an ionic liquid and having significantly improved drug absorbability, as well as a transmucosal absorption enhancer and a digestive tract absorption enhancer of a drug having low solubility and / or low membrane permeability and having low absorbability through the mucosa and the digestive tract.

[0015] Solutions to Solve Problems

[0016] The present inventors have studied new ionic liquids for promoting intestinal absorption and digestive tract absorption of low-solubility and / or low-membrane permeability drugs. In the study of ionic liquids, PROTAC (e.g., BI-3663, VZ185) was used as a model compound for low-solubility and / or low-membrane permeability drugs.

[0017] The present inventors prepared various ionic liquids capable of dissolving low-solubility and / or low-membrane permeability drugs, and evaluated the permeability of preparations containing these ionic liquids and drugs using a membrane permeability evaluation method, such as an artificial membrane permeability evaluation method (PAMPA method) and a membrane permeability evaluation method using Caco2 cells, which is a cell line derived from human colon cancer and a cell model that reproduces the intestinal function of the small intestine. As a result, it was found that the absorption of low-solubility and / or low-membrane permeability drugs through the mucosa and the digestive tract was improved by using specific ionic liquids, thereby completing the present invention.

[0018] That is, the present invention provides the following aspects.

[0019] [Item 1] A pharmaceutical composition comprising a low-solubility and / or low-membrane-permeability drug and an ionic liquid, wherein the ionic liquid comprises an anionic component and a cationic component, the anionic component is oleic acid or isostearic acid, and the cationic component is selected from cationic lipids.

[0020] [Item 2] The pharmaceutical composition according to Item 1, characterized in that the cationic lipid is DODAP, DODMA or DOTMA.

[0021] [Item 3] The pharmaceutical composition according to Item 1 or Item 2, characterized in that the anionic component is oleic acid and the cationic component is DODAP or DODMA.

[0022] [Item 4] The pharmaceutical composition according to any one of Items 1 to 3, wherein the low solubility and / or low membrane permeability drug is PROTAC.

[0023] [Item 5] An oral preparation comprising the pharmaceutical composition according to any one of Items 1 to 4.

[0024] [Item 6] A transmucosal absorption enhancer (which does not contain a drug), which comprises an ionic liquid, characterized in that the ionic liquid comprises an anionic component and a cationic component, the anionic component is oleic acid or isostearic acid, and the cationic component is selected from cationic lipids.

[0025] [Item 7] The transmucosal absorption enhancer according to Item 6, wherein the cationic lipid is DODAP, DODMA or DOTMA.

[0026] [Item 8] The transmucosal absorption enhancer according to Item 6 or Item 7, wherein the anionic component is oleic acid and the cationic component is DODAP or DODMA.

[0027] [Item 9] A preparation comprising a low-solubility and / or low-membrane-permeability drug and the transmucosal absorption enhancer according to any one of Items 6 to 8.

[0028] [Item 10] The preparation according to Item 9, wherein the low solubility and / or low membrane permeability drug is PROTAC.

[0029] [Item 11] A gastrointestinal absorption enhancer (which does not contain a drug), which comprises an ionic liquid, characterized in that the ionic liquid comprises an anionic component and a cationic component, the anionic component is oleic acid or isostearic acid, and the cationic component is selected from cationic lipids.

[0030] [Item 12] The gastrointestinal absorption enhancer according to Item 11, characterized in that the cationic lipid is DODAP, DODMA or DOTMA.

[0031] [Item 13] The digestive tract absorption enhancer according to Item 11 or Item 12, characterized in that the anion component is oleic acid and the cationic component is DODAP or DODMA.

[0032] [Item 14] An oral preparation comprising a low-solubility and / or low-membrane-permeability drug and the digestive tract absorption enhancer according to any one of Items 11 to 13.

[0033] [Item 15] The oral preparation according to Item 14, wherein the low solubility and / or low membrane permeability drug is PROTAC.

[0034] Effects of the Invention

[0035] According to the present invention, even low solubility and / or low membrane permeability drugs that are difficult to use as oral preparations can be prepared as oral preparations by significantly improving their absorbability. In addition, the transmucosal absorption enhancer or digestive tract absorption enhancer containing ionic liquids according to the present invention can promote the transmucosal or digestive tract absorption of low solubility and / or low membrane permeability drugs.

[0036] Brief description of the attached figure

[0037] Figure 1 The absorption enhancement effect of various ionic liquids on VZ185 based on the PAMPA method is shown. PBS was used as a control.

[0038] Figure 2 The absorption enhancement effect of various ionic liquids on BI-3663 based on the PAMPA method is shown. PBS was used as a control.

[0039] Figure 3 The absorption enhancement effect of various ionic liquids on bifonazole based on the PAMPA method is shown. PBS was used as a control.

[0040] Figure 4 The absorption enhancement effect (permeability coefficient) of various ionic liquids on VZ185 based on the Caco-2 cell membrane permeability test is shown. Transport Buffer (A) was used as a control.

[0041] Figure 5 The absorption enhancement effect (permeability coefficient) of various ionic liquids on BI-3663 based on the Caco-2 cell membrane permeability test is shown. Transport Buffer (A) was used as a control.

[0042] Figure 6 The absorption enhancement effect (permeability coefficient) of ionic liquid (oleic acid-DODMA) on bifonazole based on Caco-2 cell membrane permeability test is shown. Transport Buffer (A) was used as a control.

[0043] Figure 7 The figure shows the absorption enhancement effect (permeability coefficient) of ionic liquid (isostearic acid-DODMA) on ritonavir based on the Caco-2 cell membrane permeability test. Transport Buffer (A) was used as a control.

[0044] Figure 8 The absorption enhancement effect (permeability coefficient) of various ionic liquids on furosemide based on the Caco-2 cell membrane permeability test is shown. The absorption enhancement effect is expressed as the permeability coefficient. Transport Buffer (A) was used as the control. DETAILED DESCRIPTION

[0045] The present invention relates to a preparation containing a low-solubility and / or low-membrane-permeability drug and an ionic liquid, wherein the absorbability of the low-solubility and / or low-membrane-permeability drug is significantly improved, and a transmucosal absorption enhancer and a digestive tract absorption enhancer containing an ionic liquid and significantly improving the transmucosal absorbability and digestive tract absorbability of the low-solubility and / or low-membrane-permeability drug.

[0046] In the present invention, "low solubility drugs" refer to drugs that show a solubility in 250 mL of a buffer solution at pH 1.0 to 7.5 that is less than the maximum drug content contained in one unit preparation, including, for example, drugs equivalent to low solubility (Low Solubility (LS)) defined in BCS. For example, it also includes water-insoluble drugs that show a solubility of less than 100 μg for 1 mL of water, drugs that are difficult to use as injections due to low solubility, and drugs that have low solubility or are completely insoluble in buffer solutions or intestinal fluids or in the small intestine. It should be noted that as a result of its low solubility, "low solubility drugs" have a low absorption through the mucosa and the digestive tract, and therefore sometimes have the property of being a "low membrane permeability drug". Therefore, in the present invention, "low solubility drugs" can also be high membrane permeability as long as they are drugs with low absorption through the mucosa and the digestive tract.

[0047] In the present invention, "low membrane permeability drugs" refer to drugs with low biological membrane permeability, but examples include drugs that are difficult to absorb from the digestive tract due to the poor permeability of the mucus layer present on the gastrointestinal mucosa, drugs that are difficult to be absorbed through the interaction with substances present in the mucus layer, or drugs that are difficult to absorb from the digestive tract due to poor permeability in the digestive tract mucosa, drugs that are difficult to be orally absorbed due to the interaction between the digestive tract mucus layer and the mucosa, or drugs that show poor absorption by forming insoluble complexes with bile acids. Generally speaking, it refers to drugs that are difficult to deliver by oral administration. It should be noted that in the present invention, "low membrane permeability drugs" can also be highly soluble as long as they are drugs with low digestive tract absorption.

[0048] In addition, for example, compounds with low bioavailability, specifically, compounds with an extent of bioavailability of 50% or less are also included. The bioavailability can be calculated by the following formula.

[0049] Bioavailability (%) = 100 × (amount reaching blood via oral administration) / (amount reaching blood via intravenous administration)

[0050] Here, the "amount reaching the blood" is determined as the area of ​​the portion surrounded by the blood drug concentration and the horizontal axis (time axis) (area under the blood drug concentration-time curve: AUC).

[0051] In addition, compounds judged to have low permeability by the monolayer membrane method of cultured cells (Caco-2, MDCK, HT-29, LLC-PK1, etc.), the Immobilized Artificial Membrane chromatography method, the method using the distribution coefficient, the ribosome membrane method, and the PAMPA (Parallel Artificial Membrane Permeation Assay) method are also included.

[0052] In the present invention, "low solubility and / or low membrane permeability drugs" refers to drugs having the properties of both the above-mentioned "low solubility drugs" and "low membrane permeability drugs", for example, compounds classified as Class 4 of BCS (such as PROTAC) can be cited. It should be noted that there are also "low solubility drugs" with low solubility in the small intestine, which are difficult to absorb through the mucosa and from the digestive tract, and there are also "low membrane permeability drugs" with low digestive tract absorption even if they are highly soluble, so "low solubility drugs" or "low membrane permeability drugs" are also included in "low solubility and / or low membrane permeability drugs". As examples thereof, BI-3663, VZ185, ACBI1, bifonazole, hydrochlorothiazide, furosemide, acetazolamide, sulfamethoxazole, chlorothiazide, enalaprilat, mebendazole, bifonazole, methotrexate, amphotericin B, ritonavir, etc. can be cited, but are not limited to these substances.

[0053] In the present invention, the oral absorbability of low-solubility and / or low-membrane permeability drugs should not be particularly limited. Drugs that have low absorbability through the mucosa or in the digestive tract and thus cannot obtain sufficient pharmacological effects in oral administration can obtain the expected pharmacological effects by improving oral absorbability. In addition, drugs that have low absorbability through the mucosa or in the digestive tract but can obtain sufficient pharmacological effects in oral administration can reduce the amount of drug administered and reduce side effects by further improving oral absorbability.

[0054] In the present invention, "ionic liquid" refers to a salt that is a viscous liquid at room temperature formed by an anion component and a cationic component, and is a substance with a melting point of 100°C or less, or a "hydrated ionic liquid". "Hydrated ionic liquid" refers to an ionic liquid formed by an ionic liquid and a small amount of water molecules. The ionic liquid can be prepared by mixing an anion component and a cationic component in an equimolar amount or in excess at room temperature or under heating. The excess of the anion component and the cationic component is preferably within 10 times the molar amount.

[0055] In the present invention, "anion component" refers to an organic anion, for example, organic acids such as lactic acid, citric acid, tartaric acid, succinic acid, malic acid, benzoic acid, levulinic acid, aspartic acid, glutamic acid, caprylic acid, capric acid, lauric acid, myristic acid, stearic acid, behenic acid, isostearic acid, oleic acid, arachidonic acid, docosahexaenoic acid, eicosapentaenoic acid, linoleic acid, linolenic acid, adipic acid, and sebacic acid can be cited. In the present invention, the anion component is preferably a long-chain fatty acid such as oleic acid and isostearic acid.

[0056] In the present invention, "cationic component" refers to an organic cation, and examples thereof include cationic lipids, diethanolamine, triethanolamine, arginine, lysine, meglumine, organic amines such as tromethamine, organic quaternary ammonium cations, basic amino acids, and amino sugars. In the present invention, the cationic component is preferably a cationic lipid.

[0057] Cationic lipids are generally composed of a hydrophobic part and a hydrophilic part, the hydrophobic part is a hydrophobic group such as a fatty acid residue or a sterol residue, and the hydrophilic part is a cationic group such as an amino group or an ammonium group. In particular, a structure (two-chain cationic lipid) containing two hydrophobic groups in the hydrophobic part and one cationic group such as an amino group or an ammonium group in the hydrophilic part is widely known.

[0058] In the present invention, cationic lipids include DODAP (1,2-dioleoyl-3-dimethylaminopropane), DODMA (1,2-dioleoyl-3-dimethylaminopropane), DOTMA (1,2-di-O-octadecenyl-3-trimethylammoniumpropane), DLinDAP (1,2-dilinoleoyl-3-dimethylaminopropane) or salts thereof.

[0059] The ionic liquid of the present invention comprises a combination of the above-mentioned anion component and cation component, but can also be used as a mixed ionic liquid of two or more kinds (including ionic liquids in which only the anion component or the cation component is different). In addition, the ionic liquid can also contain water.

[0060] In the present invention, "transmucosal absorption enhancer" refers to an agent that improves the transmucosal absorption of drugs with low solubility and / or low membrane permeability, such as drugs with low absorption from mucous membranes other than the digestive tract, such as the oral mucosa, rectum, vagina, nasal mucosa, lungs, etc.

[0061] In the present invention, the "digestive tract absorption enhancer" refers to a drug that improves the digestive tract absorption of a drug that has low digestive tract absorption and is not effective when orally administered, such as a low-solubility and / or low-membrane-permeability drug.

[0062] The preparation of the present invention may be contained in combination with a pharmaceutically acceptable carrier. Examples of carriers include excipients, film-forming agents, adhesives, extenders, disintegrants, surfactants, lubricants, diluents, dispersants, buffers, osmotic pressure regulators, pH regulators, emulsifiers, preservatives, stabilizers, antioxidants, colorants, ultraviolet absorbers, humectants, thickeners, activity enhancers, anti-inflammatory agents, bactericides, flavoring agents, and deodorants.

[0063] These carriers may be used alone or in combination of two or more in appropriate amounts.

[0064] In addition, the capsule can be formed into a capsule shape with a water-soluble surrounding agent. The water-soluble surrounding agent is a substance that becomes the capsule membrane, and the content liquid is sealed with the water-soluble surrounding agent to prevent the content liquid from being oxidized and maintain its quality, and can be formed into a capsule.

[0065] When forming the capsule, water-soluble polymers such as hydroxypropyl cellulose, hydroxypropyl methyl cellulose and polyvinyl pyrrolidone may be used.

[0066] Example

[0067] Examples are shown below. These examples are provided for better understanding of the present invention and do not limit the scope of the present invention.

[0068] The physical properties and structures of BI-3663 and VZ185, which are low-solubility and / or low-membrane-permeability drugs used in the following examples, are shown in the following table, but the drugs to be targeted by the present invention are not limited to these compounds.

[0069] [Table 1]

[0070]

[0071] In addition, the drugs used in the following examples, bifonazole, ritonavir, and furosemide were purchased from Tokyo Chemical Industry Co., Ltd.

[0072] Example 1: Evaluation of drug membrane permeability based on Caco-2 cell membrane permeability test (1)

[0073] In order to study the ionic liquids that promote the intestinal absorption effect of low solubility and / or low membrane permeability drugs, various ionic liquids were prepared, and the membrane permeability of the drug was evaluated in the composition containing these ionic liquids and low solubility and / or low membrane permeability drugs. Specifically, as a model drug of low solubility and / or low membrane permeability, VZ185 as PROTAC was used, and for the composition containing various ionic liquids, the membrane permeability of the drug was evaluated by Caco-2 membrane permeability test according to the following steps. As a control, EDTA, SNAC, sodium octanoate, sodium oleate, CAGE and lactic acid reported to promote mucosal absorption of insulin were used instead of ionic liquids.

[0074] (1) Preparation of Transport Buffer

[0075] Sodium hydroxide (0.4 g) was dissolved in water (100 mL) to prepare a 0.1 N NaOH solution. MES (2-morpholineethanesulfonic acid, 1.493 g) and D-glucose (2.453 g) were dissolved in HBSS (+) (without phenol red) (600 mL), and the pH was adjusted to 6.5 with 0.1 N NaOH solution, and then HBSS (+) (without phenol red) was added to make 700 mL. Filter sterilize with a 0.22 μm membrane filter and use it as Transport Buffer (A).

[0076] Next, HEPES (1.192 g), D-glucose (1.752 g) and BSA (1.25 g) were dissolved in HBSS (+) (phenol red free) (400 mL), adjusted to pH 7.4 using 0.1 N NaOH solution, and then HBSS (+) (phenol red free) was added to make 500 mL. The mixture was sterilized by filtration using a 0.22 μm membrane filter and used as Transport Buffer (B).

[0077] (2) Preparation of test solution

[0078] VZ185 (1.5 mg) was mixed with 1% TWEEN80 (1.5 mL) to prepare a 1 mg / mL VZ185 solution. Each organic acid (0.05 mmol) as an anionic component and each cationic lipid (0.05 mmol) as a cationic component were mixed with Transport Buffer (A) (4.98 mL) to prepare an ionic liquid composition containing 10 mM of each component. In addition, as a control, EDTA, SNAC, sodium octanoate, sodium oleate, CAGE (choline geranyl acid) and lactic acid were each mixed with Transport Buffer (A) (4.98 mL) in an amount of 0.05 mmol to prepare a 10 mM control composition.

[0079] A 1 mg / mL VZ185 solution (25 μL) and an ionic liquid composition or a control composition (475 μL) were mixed to prepare a 50 μg / mL drug-ionic liquid composition or a drug-control composition.

[0080] Next, after Caco-2 cells were cultured on a 24-well Transwell for 3 weeks, 50 μg / mL of the drug-ionic liquid composition (100 μL) was added to the apical side of each well, and Transport Buffer (B) (600 μL) was added to the basolateral side, and then incubated in a 37°C, 5% CO2 incubator. After 15, 30, 60, 120, and 240 minutes, 200 μL of samples were collected from the basolateral side of each well and used as the test solution. Transport Buffer (B) (200 μL) was added to each well after the sample was collected, and then incubated again in a 37°C, 5% CO2 incubator.

[0081] In addition, a control solution was prepared in the same manner using a drug-control composition instead of a drug-ionic liquid composition.

[0082] (3) Evaluation of cell membrane integrity

[0083] The integrity of the Caco-2 cell membrane was confirmed using transepithelial electrical resistance (TEER) and Lucifer Yellow.

[0084] (3-1) Before and after the following Caco-2 membrane permeability evaluation, the transepithelial electrical resistance (TEER) was measured. Specifically, after adding Transport Buffer (A) (100 μL) to the apical side of each well and Transport Buffer (B) (600 μL) to the basolateral side, the TEER value of each well was measured using Millicell ERS-2 (Merck Millipore Corporation, Darmstadt, Germany).

[0085] (3-2) After the following Caco-2 membrane permeability evaluation operation was completed, the permeability of fluorescein was measured. Specifically, fluorescein CH dipotassium salt (5.2 mg) was dissolved in Transport Buffer (A) (100 mL) to prepare a 100 μM fluorescein solution. Then, a 100 μM fluorescein solution (100 μL) was added to the top side of each well and Transport Buffer (B) (600 μL) was added to the bottom side. The mixture was incubated in a 37°C, 5% CO2 incubator. After 60 minutes, 200 μL of the sample was collected from the bottom side of each well and used as a sample for fluorescence intensity measurement. The sample for fluorescence intensity measurement (100 μL) and acetonitrile (400 μL) were mixed and centrifuged at 4°C and 3000×g for 5 minutes. The fluorescein concentration in the supernatant was measured using HPLC-FL under the conditions in the following table.

[0086] [Table 2]

[0087] HPLC System LC-2010C HT (Shimadzu Corporation) Fluorescence detector RF-10A XL (Shimadzu Corporation) Injection volume 10μL Flow rate 1mL / min Excitation wavelength (Ex) 428nm Fluorescence wavelength (Em) 538nm Analysis time 1.5 minutes Eluent <![CDATA[H2O / acetonitrile = 1:1]]>

[0088] (4) Membrane permeability evaluation

[0089] Donepezil hydrochloride (10 mg) was dissolved in methanol (100 mL) to prepare a 100 μg / mL donepezil hydrochloride solution. Accurately measure 1 mL of the 100 μg / mL donepezil hydrochloride solution, add methanol and accurately make up to 100 mL, which is used as the IS standard stock solution. Accurately measure 2 mL of the IS standard stock solution, add acetonitrile and accurately make up to 200 mL, which is used as the IS-acetonitrile solution (10 ng / mL donepezil / acetonitrile solution).

[0090] The test solution or control solution (100 μL) and IS-acetonitrile solution (400 μL) were mixed and centrifuged at 4°C and 3000×g for 10 minutes. The drug concentration in the supernatant was measured using LC-MS / MS under the conditions shown in the table below.

[0091] [Table 3]

[0092]

[0093] [Table 4]

[0094] Time program

[0095] Time (minutes) Liquid A(%) Liquid B (%) 0 97 3 1 97 3 3.5 70 30 3.6 10 90 6.5 10 90 6.6 97 3

[0096] [Table 5]

[0097] Multiple reaction monitoring (MRM) setting conditions

[0098] Precursor ions Product ions 996.15 309.15 919.00 442.95

[0099] The results are shown in Table 6. The TEER value was set at >1000Ω and the fluorescent yellow value was set at <1% as the reference value, which were used as indicators for evaluating membrane integrity. Regarding membrane permeability, the membrane permeability coefficient (P app )(x10 6 cm / s).

[0100] (Calculation formula)

[0101] Membrane permeability coefficient (P app )=(dQ / dt)×{1 / (A×C0)}

[0102] dQ / dt: Cumulative permeation on the substrate side

[0103] A: Transwell membrane area (0.33 cm 2 )

[0104] C0: initial concentration on the top side

[0105] [Table 6]

[0106]

[0107] 1: An ionic liquid composition containing each component in an amount of 5 mM which is half the amount.

[0108] 2: Ionic liquid composition comprising 10 mM oleic acid and 20 mM meglumine.

[0109] The results of this experiment suggest that although the TEER value of the ionic liquid group was higher than that of the control group and the permeability of Luciffer Yellow was lower, P app There is a high tendency, especially for the ionic liquids of capric acid-DODAP, oleic acid-DODAP, and isostearic acid-DODAP, the drug permeability coefficient is significantly high, and the ionic liquids of the combination of long-chain fatty acids and cationic lipids have a significant permeation enhancement effect.

[0110] Example 2: Evaluation of drug membrane permeability based on Caco-2 cell membrane permeability test (2)

[0111] The results of Example 1 suggest that the permeation promoting effect of ionic liquids in which long-chain fatty acids and cationic lipids are combined is significant, so various ionic liquids in which DHA (C22), EPA (C20), arachidonic acid (C20), isostearic acid (C18), linoleic acid (C18), linolenic acid (C18) as anionic components and DODAP, DODMA and DOTMA as cationic components are combined were evaluated for Caco-2 membrane permeability and cell membrane integrity. Specifically, the membrane permeability of the drug was evaluated in the same manner as in Example 1. As a control, Transport Buffer (A) prepared in Example 1 was used instead of the ionic liquid.

[0112] The results are shown in Table 7. Regarding membrane integrity, for TEER values, >1000Ω is the expected value, and for fluorescent yellow values, <1% is the expected value, so these values ​​are used as reference values, and the case exceeding the reference value is considered good. Regarding membrane permeability, the membrane permeability coefficient (P app )(x10 6 cm / s).

[0113] [Table 7]

[0114]

[0115] * :Isostearic acid (branched chain),** :Isostearic acid (straight chain)

[0116] The test results showed that all ionic liquids had higher permeability coefficients than the control and promoted membrane permeation, and the combination of isostearic acid and cationic lipids was particularly significant.

[0117] Example 3: Evaluation of drug membrane permeability based on artificial membrane permeability assessment (PAMPA) method (1)

[0118] For various ionic liquid formulations, the membrane permeability of low solubility and / or low membrane permeability drugs was evaluated by the PAMPA method. In this experiment, VZ185 and BI-3663 were used as low solubility and / or low membrane permeability drugs, and isostearic acid-DODMA, isostearic acid-DODAP and oleic acid-DODMA were used as ionic liquids. As a control, PBS was used.

[0119] Specifically, the membrane permeability of the drug was evaluated according to the following procedure using Parallel Artificial Membrane Permeability Assay-Skin Kit (manufactured by BioAssay Systems).

[0120] (1) Preparation of phosphate buffered saline (PBS)

[0121] PBS was prepared by dissolving 1 Phosphate Buffered Saline Tablet (Sigma) in 200 mL of water.

[0122] (2) Preparation of test solution

[0123] VZ185 (1.5 mg) was mixed with 1% TWEEN80 (1.5 mL) to prepare a 1 mg / mL VZ185 solution. Each organic acid (0.025 mmol) as an anionic component and a cationic lipid (0.025 mmol) as a cationic component were mixed with PBS (4.98 mL) prepared in (1) to prepare an ionic liquid composition containing each component in an amount of 5 mM. A 1 mg / mL VZ185 solution (25 μL) and an ionic liquid composition (475 μL) were mixed to prepare a drug-ionic liquid composition of 50 μg / mL. In addition, as a control, a 1 mg / mL VZ185 solution (25 μL) was mixed with PBS (475 μL) to prepare a 50 μg / mL drug-control solution.

[0124] Next, add 4% lecithin / dodecane solution (5 μL) to each donor well and let it stand. Add PBS (300 μL) to each receptor well, and add 50 μg / mL of drug-ionic liquid composition (200 μL) to each donor well, and incubate in a thermostatic bath at 37°C. After 30 minutes, 240 minutes and 1440 minutes, sample 300 μL from each receptor well and use it as a test solution. After adding PBS (300 μL) to each receptor well after sampling, incubate again in a thermostatic bath at 37°C. It should be noted that the test solution for BI-3663 was prepared in the same way.

[0125] In addition, a control solution was prepared in the same manner using a drug-control solution instead of a drug-ionic liquid composition.

[0126] (3) Test methods

[0127] Dissolve donepezil hydrochloride (10 mg) in methanol (100 mL) to prepare a 100 μg / mL IS solution. Accurately measure 1 mL of the 100 μg / mL IS solution, add methanol and accurately dilute to 100 mL, which is used as the IS standard stock solution. Accurately measure 2 mL of the IS standard stock solution, add acetonitrile and accurately dilute to 200 mL, which is used as the IS-acetonitrile solution (10 ng / mL donepezil / acetonitrile solution).

[0128] The test solution or control solution (100 μL) and IS-acetonitrile solution (400 μL) were mixed and centrifuged at 4°C and 3000×g for 10 minutes. The drug concentration in the supernatant was measured using LC-MS / MS under the conditions shown in the table below.

[0129] [Table 8]

[0130]

[0131] [Table 9]

[0132] Time program

[0133] Time (minutes) Liquid A(%) Liquid B (%) 0 97 3 1 97 3 3.5 70 30 3.6 10 90 6.5 10 90 6.6 97 3

[0134] [Table 10]

[0135] Multiple reaction monitoring (MRM) setting conditions

[0136] drug Precursor ions Product ions VZ185 996.15 309.15 BI-3663 919.00 442.95

[0137] The results are as follows Figure 1 and Figure 2 According to the test results, it is shown that the absorbency of VZ185 and BI-3663 is significantly improved by using ionic liquids.

[0138] Example 4: Evaluation of drug membrane permeability based on artificial membrane permeability assessment (PAMPA) method (2)

[0139] For various ionic liquid preparations, the membrane permeability of low-solubility and / or low-membrane permeability drugs was evaluated by the PAMPA method. In this experiment, bifonazole was used as a low-solubility and / or low-membrane permeability drug, and isostearic acid-DODMA, isostearic acid-DODAP, and oleic acid-DODMA were used as ionic liquids. As a control, PBS was used.

[0140] Specifically, the membrane permeability of bifonazole was evaluated in the same manner as in Example 3 using Parallel Artificial Membrane Permeability Assay-Skin Kit (manufactured by BioAssay Systems).

[0141] The results are as follows Figure 3 According to the test results, it is shown that the absorbability of bifonazole is significantly improved by using ionic liquid.

[0142] Example 5: Evaluation of drug membrane permeability based on Caco-2 cell membrane permeability test (3)

[0143] For various ionic liquid formulations, the membrane permeability of low-solubility and / or low-membrane permeability drugs was evaluated by Caco-2 cell membrane permeability test. In this experiment, VZ185 was used as a low-solubility and / or low-membrane permeability drug, and oleic acid-DODMA and oleic acid-DODAP were used as ionic liquids. As a control, Transport Buffer (A) was used.

[0144] Specifically, according to the method of Example 1, the membrane permeability of VZ185 was evaluated.

[0145] The results are as follows Figure 4 According to the results of this experiment, oleic acid-DODMA and oleic acid-DODAP have a significant effect on promoting digestive tract absorption, and the absorbability of VZ185 is significantly improved by using these ionic liquids.

[0146] Example 6: Evaluation of drug membrane permeability based on Caco-2 cell membrane permeability test (4)

[0147] For various ionic liquid preparations, the membrane permeability of low-solubility and / or low-membrane permeability drugs was evaluated by Caco-2 cell membrane permeability test. In this experiment, BI-3663 was used as a low-solubility and / or low-membrane permeability drug, and isostearic acid-DODMA, oleic acid-DODMA and oleic acid-DODAP were used as ionic liquids. As a control, Transport Buffer (A) was used.

[0148] Specifically, according to the method of Example 1, the membrane permeability of BI-3663 was evaluated.

[0149] The results are as follows Figure 5 According to the test results, isostearic acid-DODMA, oleic acid-DODMA and oleic acid-DODAP have a significant effect on promoting digestive tract absorption, and the absorbability of BI-3663 is significantly improved by using these ionic liquids.

[0150] Example 7: Evaluation of drug membrane permeability based on Caco-2 cell membrane permeability test (5)

[0151] For various ionic liquid formulations, the membrane permeability of low-solubility and / or low-membrane permeability drugs was evaluated by Caco-2 cell membrane permeability test. In this experiment, bifonazole was used as a low-solubility and / or low-membrane permeability drug, and oleic acid-DODMA was used as an ionic liquid. As a control, Transport Buffer (A) was used.

[0152] Specifically, according to the method of Example 1, the membrane permeability of bifonazole was evaluated.

[0153] The results are as follows Figure 6 According to the test results, oleic acid-DODMA has a significant effect on promoting digestive tract absorption, and the absorption of bifonazole is significantly improved by using this ionic liquid.

[0154] Example 8: Evaluation of drug membrane permeability based on Caco-2 cell membrane permeability test (6)

[0155] For various ionic liquid preparations, the membrane permeability of low solubility and / or low membrane permeability drugs was evaluated by Caco-2 cell membrane permeability test. In this experiment, ritonavir (a low solubility drug that is almost insoluble in water) was used as a low solubility and / or low membrane permeability drug, and isostearic acid-DODMA was used as an ionic liquid. As a control, Transport Buffer (A) was used.

[0156] Specifically, according to the method of Example 1, the membrane permeability of ritonavir was evaluated.

[0157] The results are as follows Figure 7 According to the test results, isostearic acid-DODMA has a significant effect on promoting digestive tract absorption, and the absorption of ritonavir is significantly improved by using the ionic liquid.

[0158] Example 9: Evaluation of drug membrane permeability based on Caco-2 cell membrane permeability test (7)

[0159] For various ionic liquid preparations, the membrane permeability of low-solubility and / or low-membrane permeability drugs was evaluated by Caco-2 cell membrane permeability test. In this experiment, furosemide (a low-solubility drug that is almost insoluble in water) was used as a low-solubility and / or low-membrane permeability drug, and isostearic acid-DODAP and oleic acid-DODAP were used as ionic liquids. As a control, Transport Buffer (A) was used.

[0160] Specifically, according to the method of Example 1, the membrane permeability of furosemide was evaluated.

[0161] The results are as follows Figure 8 According to the results of this experiment, isostearic acid-DODAP and oleic acid-DODAP have a significant effect on promoting digestive tract absorption, and the absorption of furosemide is significantly improved by using these ionic liquids.

[0162] According to the above test results, it is shown that the absorption of low-solubility and / or low-membrane-permeability drugs is significantly improved by using ionic liquids.

[0163] Industrial Applicability

[0164] According to the present invention, even low solubility and / or low membrane permeability drugs that are difficult to use as oral preparations can be prepared as oral preparations by significantly improving their absorbability. In addition, the transmucosal absorption enhancer or digestive tract absorption enhancer containing ionic liquids according to the present invention can promote the transmucosal or digestive tract absorption of low solubility and / or low membrane permeability drugs.

Claims

1. A pharmaceutical composition comprising a low solubility and / or low membrane permeability drug and an ionic liquid, characterized in that: The ionic liquid comprises an anion component and a cation component, wherein the anion component is oleic acid or isostearic acid, and the cation component is selected from cationic lipids.

2. The composition according to claim 1, characterized in that The cationic lipid is DODAP, DODMA or DOTMA.

3. The pharmaceutical composition according to claim 1 or 2, characterized in that The anionic component is oleic acid, and the cationic component is DODAP or DODMA.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein Drugs with low solubility and / or low membrane permeability are PROTACs.

5. An oral preparation comprising the pharmaceutical composition according to any one of claims 1 to 4.

6. A transmucosal absorption enhancer comprising an ionic liquid but no drug, characterized in that: The ionic liquid comprises an anion component and a cation component, wherein the anion component is oleic acid or isostearic acid, and the cation component is selected from cationic lipids.

7. A preparation comprising a low-solubility and / or low-membrane-permeability drug and the transmucosal absorption enhancer according to claim 6.

8. A digestive tract absorption enhancer comprising an ionic liquid but no drug, characterized in that: The ionic liquid comprises an anion component and a cation component, wherein the anion component is oleic acid or isostearic acid, and the cation component is selected from cationic lipids.

9. The digestive tract absorption enhancer according to claim 8, characterized in that: The cationic lipid is DODAP, DODMA or DOTMA.

10. The digestive tract absorption enhancer according to claim 8 or 9, characterized in that: The anionic component is oleic acid, and the cationic component is DODAP or DODMA.

11. An oral preparation comprising a low-solubility and / or low-membrane-permeability drug and the digestive tract absorption enhancer according to any one of claims 8 to 10.

12. The oral preparation according to claim 11, wherein Drugs with low solubility and / or low membrane permeability are PROTACs.

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