KR20160110245A

KR20160110245A - A method to improve montelukast's bio-availability

Info

Publication number: KR20160110245A
Application number: KR1020160029470A
Authority: KR
Inventors: 이영주; 오주희
Original assignee: 경희대학교 산학협력단
Priority date: 2015-03-13
Filing date: 2016-03-11
Publication date: 2016-09-21
Also published as: WO2016148455A3; KR101892340B1; WO2016148455A2

Abstract

The present invention relates to a method for increasing the bioavailability of montelukast, and a method for administering a solubilizing agent such as an endogenous surfactant (bile acid) or a polymeric surfactant according to the present invention in combination with a montelukast salt is a montelukast or a pharmacologically acceptable salt thereof Since the reprecipitation of the montelukast that occurs when an acceptable salt is administered alone is effectively prevented, the absorption rate of montelukast in vivo increases, the absorption rate is increased, and an excellent bioavailability is exhibited.

Description

[0001] The present invention relates to a method for improving the bioavailability of montelukast,

The present invention relates to a method for increasing the bioavailability of montelukast.

Cholesterol, an organic acid derived from cholesterol, is a natural ionic detergent that plays a decisive role in the absorption, transport and secretion of lipids. In bile acid chemistry, the steroid nucleus of the bile acid salt has a perhydrocyclopentanophenanthrene nucleus common to all the perhydrosteroids. The salient features of the bile acid salt include saturated 19-carbon sterol nucleus, beta-oriented hydrogen in position 5, branched saturated 5-carbon side chain of carboxylic acid end, and alpha-oriented hydroxy group in position 3 . The only substituent present in most natural bile acids is the hydroxy group. In most mammals, the hydroxy group is at position 3, 6, 7 or 12.

Typical bile acids differ mainly in the number and orientation of the hydroxy groups in the sterol ring. The term primary bile acid refers to those newly synthesized by the liver. In humans, the primary bile acid is the cholanic acid (3 alpha, 7 alpha, 12 alpha -trihydroxy-5 beta -cholanic acid) ("CA") and the chenodeoxycholic acid (3 alpha, 7 alpha -dihydroxy- "CDCA"). When these bile acids are dehydroxylated by enteric bacteria, the more hydrophobic secondary bile acids, namely deoxycholic acid (3α, 12α-dihydroxy-5β-cholanic acid) ("DCA") and lithocholic acid -Hydroxy-5? -Cholanic acid) ("LCA"). These four bile acids CA, CDCA, DCA and LCA generally constitute more than 99 percent of the bile salt pool in the human body. Secondary bile acids metabolized by the liver are sometimes referred to as tertiary bile acids.

Keto-bile acid is produced secondarily in the body as a result of oxidation of bile acid hydroxy groups, especially 7-hydroxy groups, by the colon bacteria. However, keto-bile acids are rapidly reduced to the corresponding? Or? -Hydroxy bile acids by the liver. For example, the corresponding keto bile acid of CDCA is 7-ketolitholic acid, and one of the reduction products by its corresponding? -Hydroxyl bile acid is the ursodeoxycholic acid (3? -7-dihydroxy-5? -Cholanic acid) ("DCA").

Typically, more than 99 percent of the natural bile acid secreted in human bile is conjugated. The conjugate is a bile acid in which a second organic substituent (e.g., glycine, taurine, glucuronate, sulfate, etc.) is attached to the side chain carboxylic acid or to one of the ring hydroxy groups via an ester, ether, or amide linkage . Thus, the ionization properties of conjugated bile acids with glycine or taurine are determined by the acidity of glycine or taurine substituents.

The free and non-conjugated bile acid monomers have a pKa value of about 5.0. However, the pKa value of the glycine-conjugated bile acids averages about 3.9 and the pKa value of the taurine conjugated bile acids is less than 1.0. Thus, the effect of conjugation is to reduce the pKa value of bile acid so that most are ionized at any given pH. Since ionized salts are more water soluble than protonated, the conjugate increases solubility at low pH. While free bile salts precipitate from aqueous solutions of pH 6.5 to 7, precipitation of glycine conjugated bile acids occurs only at a pH of less than 5. Taurine conjugated bile acids remain in aqueous solution only under extremely acidic conditions (pH less than 1).

Montelukast, on the other hand, is an antagonist that inhibits the systeinyl leukotriene type 1 (CysLT1) receptor and has been used for the prevention and treatment of leukotriene-mediated diseases and disorders. In particular, montelukast is known to be effective for conjunctivitis including allergic rhinitis, atopic dermatitis, chronic urticaria, sinusitis, nonpolyposis, chronic obstructive pulmonary disease, conjunctivitis, migraine, cystic fibrosis and viral bronchiolitis (SE Dahlen, Eur . J. Pharmacol., 533 (1-3), 40-56 (2006)). In addition, Singulair (MSD) using montelukast sodium salt is currently approved for the treatment of asthma in adults and pediatric patients over 2 years old.

Montelukast is used in the form of sodium salt with increased solubility. The montelukast salt is freely dissolved (0.1-1 g / mL) in aqueous solution, but once precipitated by rapid re-precipitation, Is known to increase with the passage of time. This reprecipitation is likely to cause a decrease in the absorption of montelukast and inter-individual variation after oral administration.

Accordingly, the inventors of the present invention have found that, when studying a method for improving the bioavailability of montelukast, when a solubilizing agent such as an endogenous surfactant (bile acid) or a polymer surfactant is administered in combination with a montelukast salt, the reprecipitation of the montelukast salt is effectively prevented And that the absorption rate of montelukast in vivo is increased to show excellent bioavailability, and the present invention has been completed.

It is an object of the present invention to provide a method for increasing the bioavailability of montelukast using a solubilizing agent.

Another object of the present invention is to provide a montelukast or a pharmaceutically acceptable salt thereof; And

Solubilizing agent, and a solubilizing agent.

In order to achieve the above object, the present invention provides a method for increasing the bioavailability of montelukast using a solubilizing agent.

The present invention also relates to montelukast or a pharmaceutically acceptable salt thereof; And

A solubilizing agent,

Wherein the solubilizing agent is at least one selected from the group consisting of bile acids, poloxamers, tweens, clemophytes and glycols.

The method for coadministering a solubilizing agent such as an endogenous surfactant (bile acid) or a polymeric surfactant according to the present invention with a montelukast salt is a method in which montelukast or a pharmacologically acceptable salt thereof is administered alone, The absorption rate of montelukast in vivo is increased, and thus an excellent bioavailability is exhibited.

1 is a microscope image showing reprecipitation of a montelukast sodium salt aqueous solution over time.
FIG. 2 is an image showing a general photograph of Monte Carast re-precipitation formation observed over time. FIG.
FIG. 3 is a micrograph showing the formation of the montetracer precipitate over time. FIG. 3 is an image showing the state of the solubilizer before the addition of the montelukast. FIG.
FIG. 4 is a micrograph showing the formation of a monteurocast precipitate over time. FIG. 4 is an image showing the state immediately after the addition of montelukast (0 min).
5 is an image showing a state after the addition of montelukast to the solubilizing agent (60 minutes) by observing the formation of the montetracer precipitate over time.
6 is an image showing the solubility change of montelukast over time.
7 and 8 are images showing the solubility of montelukast when the solubilizing agent is 0.1, 1, 10, or 100 parts by weight based on 1 part by weight of montelukast expressed as a concentration ratio (%) at 60 minutes relative to the concentration at 0 minutes.
9 is an image schematically representing solubility after 60 minutes by adding montelukast to the solubilizing agent.
FIG. 10 is a graph showing the solubility of montelukast when the solubilizing agent is 1 part by weight based on 1 part by weight of montelukast in an acidic environment, expressed as a concentration ratio (%) at 60 minutes relative to the concentration at 0 minutes.
11 is an image of a system of a Hydrophilic-Lipophilic Balance (HLB) system.
Fig. 12 is an image showing changes in the body of montelukast and somatic taurocholate combined treatment and monterocaster alone treatment group.

Hereinafter, the present invention will be described in detail.

The present invention provides a method for increasing the bioavailability of montelukast using a solubilizing agent. Specifically, the method is characterized by increasing bioavailability by preventing re-precipitation of montelukast in vivo.

Herein, the solubilizing agent acts to prevent reprecipitation of montelukast in vivo, and bile acids, poloxamers, twins, clemophytes, glycols and the like can be used, and bile acids are preferably used .

Specifically, the bile acids may be sodium taurocholate, sodium deoxycholate, sodium glycocholate, sodium cholate (CA), or the like; It is preferred to use sodium taurocholate or sodium deoxycholate (DCA); It is most preferred to use sodium taurocholate.

The poloxamer may be selected from the group consisting of poloxamer 124, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, and the like. have.

Further, the twin types are tween 20, tween 21, tween 40, tween 60, tween 61, tween 65, tween 80, (Tween 80), Tween 81 (Tween 81), Tween 85 (Tween 85) and the like.

In addition, the crested females may be selected from the group consisting of Cremophor A6, Cremophor A20, Cremophor A25, Cremophor EL, Cremophor ELP Cremophor RH40, Cremophor RH60, Cremophor RH410, Cremophor WO7, and the like can be used.

Further, the glycols may be polyethylene glycol having a weight-average molecular weight (Mw) of 100 to 8,000.

A solubilizing agent,

Specifically, the bile acids may be sodium taurocholate, sodium deoxycholate, sodium glycocholate (GCA), sodium cholate, or the like; It is preferred to use sodium taurocholate (TCA) or sodium deoxycholate; It is most preferred to use sodium taurocholate.

The formulation may be an enteric agent, but is not limited thereto.

The formulation may be administered by oral administration, oral administration, mucosal administration, intranasal administration, intraperitoneal administration, subcutaneous injection, intramuscular injection, transdermal administration, or intravenous injection, preferably oral administration have.

The solubilizing agent is preferably used in an amount of 0.1 to 100 parts by weight, more preferably 0.3 to 70 parts by weight, and more preferably 0.5 to 50 parts by weight, based on 1 part by weight of the montelukast or a pharmaceutically acceptable salt thereof Is most preferable.

If the amount of the solubilizing agent is less than 0.1 parts by weight based on 1 part by weight of the montelukast or its pharmaceutically acceptable salt, the reprecipitation of the montelukast can not be effectively prevented. When the solubilizing agent is used in an amount exceeding 100 parts by weight, There is a problem that an excessive amount of the solubilizing agent is contained in the formulation to cause side effects in vivo or the weight of the preparation becomes excessively large.

In the case of a surfactant, it has a hydrophilic group and a hydrophobic group in one molecule and dissolves in a liquid to significantly lower the surface (interfacial) tension. Surfactants have the effect of inhibiting or slowing the rate of formation of the supersaturated solution depending on the type and concentration. Surfactants can be classified by the Hydrophilic-Lipophilic Balance (HLB) system. The HLB of the surfactant used as a solubilizing agent is generally 15-18 (see FIG. 11).

11 is an image of a system of a Hydrophilic-Lipophilic Balance (HLB) system.

Specifically, the montelukast sodium salt was dissolved in deionized water (DDW), phosphate buffered saline (PBS), polyethylene glycol monomethyl ether solution 400, precipitated as white precipitate in sodium cholate. On the other hand, very fine precipitates were observed in the surfactants Tween 80 and Poloxamer 407, and in particular Poloxamer 407 showed no turbidity until 60 minutes, but a small amount of precipitate was observed at 120 minutes after the last measurement. In contrast, the bile acid sodium deoxycholate and sodium taurocholate showed almost no sedimentation in 120 minutes, while sodium glycocholate showed a small amount of sedimentation in 120 minutes, but the amount was very small compared to other experimental groups 1-5 of Experimental Example 1).

In addition, experiments were carried out to evaluate the solubility changes of montelukast sodium salt with the solubilizing agent added to montelukast sodium salt. As a result, in deionized water, phosphate buffer and polyethylene glycol 400, Respectively. On the other hand, it was shown that the addition of the polymeric surfactant Tween 80 and Poloxamer 407, and sodium cholate, sodium deoxycholate, sodium glycocholate, and sodium taurocholate, which are endogenous surfactants (bile acid) 6 of Experimental Example 2).

Further, 0.1, 1, 10, and 100 parts by weight of solubilizing agent were added to 1 part by weight of the montelukast sodium salt to evaluate the solubility change of the montelukast sodium salt. As a result, twin 40, tween 60, poloxamer 407, The solubility of Mopor EL and Cremophor RH40 after 60 minutes showed a remarkable effect of preventing precipitation with a solubility of 90% or more as compared to 0 minutes even at a weight ratio of montelukast and solubilizer of 1: 1. Tween 20, Tween 80, Twin 85, Cremopore A25, and Cremophor RH60 have low solubility at a weight ratio of 1: 1, but at a ratio of 1:10 weight, Effect. In addition, it was confirmed that re-precipitation of montelukast could be prevented when bile acid was added at a weight ratio of 1: 100 (see FIG. 7, FIG. 8 and FIG. 9 of Experimental Example 3).

In order to increase the bioavailability during oral administration of montelukast, the montelukast must remain dissolved in the acidic environment of the stomach. Therefore, in order to evaluate the montelukast solubility, it was found that, in acidic conditions, (From FIG. 10 of Experimental Example 4). It was found that the Klemmofoo fish is most preferable in terms of prevention of the precipitation of montelukast depending on the stomach acid.

Considering that the montelukast is easily precipitated under the acidic condition of the stomach, the effect of increasing the bioavailability of montelukast can be expected as an enteric preparation containing the above additives, and it is expected that the effect of decreasing the variation of bioavailability by preventing precipitation .

The HLB value of the polymer surfactant used in Experimental Example 1-3 was 11-16.7 in the twin class, 18-23 in the poloxamer series, and 15-17 in the crime mold class. Therefore, the HLB value of the polymer surfactant applicable to the solubility improvement of montelukast can be estimated to be about 11-23 from the experimental results of Experimental Examples 1-3.

Bile acid acts as an endogenous surfactant to help digest fat in the digestive tract. Although the HLB of bile acid, sodium cholate, was 18 and the HLB of sodium deoxycholate was 16, it was within the usable range of the above polymer surfactant. However, since the molecular properties of bile acid and polymer surfactant are different, There is a dot.

In addition, when the montelukast sodium salt and sodium taurocholate were administered concomitantly, the concentration of montelukast in plasma was measured to evaluate the absorption rate in the body. As a result, in the case of montelukast and sodium taurocholate combination treatment group, The AUC and C _max were increased about 2-3 times and the T _max was reduced by one third compared with the treated group, and the absorption was observed to be rapid. Therefore, it can be expected that the solubilizing agent such as sodium taurocholate greatly affects the absorption of montelukast into the body (see FIG. 12 of Experimental Example 5).

The above results suggest that the solubility of montelukast in combination with the solubilizing agent can be improved in the actual body, and it is expected that the montelukast or the solubility of the drug will be improved Can be expected to be applicable. In particular, when 240 mL of the human intestinal volume and 10 mg of montelukast were used, it was found that coexistence of the solubilizer effectively inhibited the re-precipitation of montelukast, .

In case of taurocholate, there was an effect of preventing precipitation in 100 parts by weight or more of 1 part by weight of montelukast, but bioavailability was also increased when it was administered to rats at a weight ratio of "montelukast: taurocholate = 1: 1.65". Thus, in vivo, it is possible to improve the bioavailability of montelukast at a lower dose ratio than in vitro, which is due to a separate mechanism other than the prevention of re-precipitation.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.

However, the following examples and experimental examples are illustrative of the present invention, and the present invention is not limited thereto.

Experimental Example 1 Evaluation of reprecipitation of montelukast sodium salt

Montelukast dissolves well in water, but there is a problem of re-precipitation after a certain period of time. Thus, the following experiment was carried out in order to observe the re-precipitation of the montelukast sodium salt after the lapse of a certain time by adding the solubilizing agent to the montelukast sodium salt.

<1-1> Preparation of Solubilizing Agent

Tween 80, TW80, polyethylene glycol 400 and PEG400 and poloxamer 407 and PLX407 were dissolved in phosphate buffered saline (pH 7.4) at a concentration of 0.5%.

Sodium cholate, sodium deoxycholate, sodium glycocholate, and sodium taurocholate were added to a phosphate buffered saline (pH 7.4) at a concentration of 10 mM &Lt; / RTI >

In general, in the case of solubilizing agents such as twin types, polyethylene glycols and poloxamers, they are prepared in a weight or volume percentage and set to 0.5% in order to use as little as possible. In the case of sodium cholate, sodium deoxycholate, sodium glycocholate, and sodium taurocholate, bile acids were prepared with reference to the bile acid concentration in the gastrointestinal tract (Reference: Concentration of BAs in the intestinal lumen, but usually high, estimated in the medium millimolar range. Marin, Intestianl even acid physiology and pathophysiology, World J Gastroenterol (2008) 14 (37): 5630-5640).

<1-2> Preparation of montelukast sodium salt

The montelukast sodium salt was prepared to be 10 mg / mL as montelukast using deionized water.

<1-3> Montelukast reprecipitation observation

Tween 80, polyethylene glycol 400, poloxamer 407 and phosphate buffer (pH 7.4) dissolved in 0.5% concentration in deionized water, phosphate buffered saline (pH 7.4) and phosphate buffer (pH 7.4) mM sodium cholate, sodium deoxycholate, sodium glycocholate, and sodium taurocholate were dissolved in a 12-well plate in an amount of 1.8 mL each. 0.2 mL of montelukast sodium salt was added to the plate, diluted to 1 mg / mL, and then photographed or photographed at regular intervals. The details of the used microscope are as follows:

1. Microscope: OLYMPUS JP / CKX41SF2

2. Microscope lens: 10X / 0.25 PhP / - / FN22

3. Camera: moticam 2000 2.0M Pixel USB 2.0

4. Program: Motic Image Plus 2.0

The results are shown in Figs. 1-5.

FIG. 1 is a microphotograph showing reprecipitation of a montelukast sodium salt aqueous solution over time. FIG.

Fig. 2 is a photograph of the reprecipitation of montelukast observed over time.

FIG. 3 is a microscope photograph showing the formation of a precipitate of montelukast over time. FIG. 3 shows the state of the solubilizer before the addition of montelukast.

FIG. 4 is a photograph of the formation of a precipitate of montelukast over time and is a photograph immediately after the addition of montelukast (0 min).

FIG. 5 is a photograph of a microscope photograph showing the formation of a precipitate of montelukast over time and a photograph taken at 60 minutes after the addition of montelukast to the solubilizing agent.

As shown in Fig. 1, montelukast is dissolved well in water, but re-precipitation occurs after a certain time. As shown in FIG. 2, when the solubilizing agent was added to the aqueous solution in which the montelukast was dissolved, the reprecipitation formation of the montelukast was observed over time. As shown in FIG. 2, when the solubilizing agent was added, The degree of precipitation inhibition was controlled.

More specifically, in order to observe whether the montelukast reprecipitation was observed, it was observed through a microscope. As shown in FIG. 3 and FIG. 4, in the state of the solubilizer before addition of montelukast and the state immediately after dissolving montelukast (0 min) No precipitation was observed. On the other hand, montelukast sodium salt precipitated a white precipitate in deionized water, phosphate buffer (pH 7.4), polyethylene glycol 400 and sodium cholate in 60 minutes after addition of montelukast. On the other hand, very fine precipitates were observed in the surfactants Tween 80 and Poloxamer 407, and in particular Poloxamer 407 showed no turbidity until 60 minutes, but a small amount of precipitate was observed at 120 minutes after the last measurement. On the other hand, in the case of sodium glycocholate, a small amount of precipitation was observed in 120 minutes, but the amount of sodium glycolokolate was extremely small compared with other experimental groups. In the case of sodium deoxycholate and sodium taurocholate, precipitation was not observed even in 120 minutes.

<Experimental Example 2> Evaluation of solubility of montelukast sodium salt

The following experiment was carried out in order to evaluate the solubility change of the montelukast sodium salt with the addition of the solubilizing agent to the montelukast sodium salt.

<2-1> Preparation of Solubilizing Agent

Tween 80, polyethylene glycol 400 and poloxamer 407 were dissolved in phosphate buffered saline (pH 7.4) at a concentration of 0.5%.

<2-2> Preparation of montelukast sodium salt

The montelukast sodium salt was prepared to be 0.4 mg / mL as montelukast using deionized water.

<2-3> Evaluation of solubility

Tween 80, polyethylene glycol 400, poloxamer 407 and phosphate buffer (pH 7.4) dissolved in 0.5% concentration in deionized water, phosphate buffered saline (pH 7.4) and phosphate buffer (pH 7.4) mM sodium cholate, sodium deoxycholate, sodium glycocholate, and sodium taurocholate were dissolved in 9 mL of each tube. To the tube was added 1 mL of montelukast sodium salt, diluted to a final concentration of 40 μg / mL, and shaken. 1 mL of the sample was taken at 0, 15, and 60 minutes, filtered through a 0.5-μm syringe filter, immediately diluted 1/10 with high-performance liquid chromatography (HPLC) mobile phase, Respectively. The details of the HPLC used are as follows:

1. HPLC system: Agilent 1200 serises;

2. Mobile phase: 60% acetonitrile containing 0.2% trifluoroacetic acid, 10% methanol and 30% deionized water mixture;

3. Flow rate: 1 mL / min;

4. Column: Phenomenex Gemini (250 × 4.5 mm, 5 μm, 100 Å, PN 00G-4435-E0, SN 290750-12);

5. Injection volume: 20 μL; And

6. UV wavelength: 389 nm.

The results are shown in Fig.

6 is an image showing the solubility change of the montelukast sodium salt with time.

As shown in Figure 6, rapid depletion was observed over time in deionized water, phosphate buffer (pH 7.4) and polyethylene glycol 400. On the other hand, it was shown that the addition of polymeric surfactants Tween 80 and Poloxamer 407, sodium cholate, sodium deoxycholate, sodium glycocholate, and sodium taurocholate, which are endogenous surfactants (bile acid)

Therefore, in the case of the human intestinal volume of 240 mL and the montelukast clinical dose of 10 mg, when the solubilizer coexists, the solubility of the montelukast is maintained higher than that of the montelukast alone, .

<Experimental Example 3> Evaluation of the weight ratio of solubilizing agent to montelukast sodium salt

In order to evaluate the solubility changes of the montelukast sodium salt by adding 0.1, 1, 10, and 100 parts by weight of solubilizing agent to 1 part by weight of the montelukast sodium salt, the following experiment was conducted.

<3-1> Preparation of Solubilizing Agent

Tween 20, Tween 40, Tween 60, Tween 80, Tween 85, Tween 85, and Polyethylene Glycol 400 were added to a phosphate buffered saline (pH 7.4) polyethylene glycol 400, poloxamer 188, poloxamer 407, Cremophor A25, Cremophor EL, Cremophor RH 40, Sodium cholate, sodium deoxycholate, sodium glycocholate and sodium taurocholate were added to 0.00444 mg / mL (twin 40, Tween 60, Poloxamer 407, Cremophor EL, and Cremophor RH40 only), 0.0444 mg / mL, 0.444 mg / mL, and 4.44 mg / mL.

<3-2> Preparation of montelukast sodium salt

<3-3> Evaluation of solubility

Deionized water, phosphate buffered saline (pH 7.4), and the solubilizing agent prepared in <3-1> were put into a tube in an amount of 1.8 mL each. To the tube was added 0.2 mL of montelukast sodium salt, diluted to a final concentration of 40 μg / mL, and shaken (each solubilizer was added in amounts of 1, 10, and 100 parts by weight with respect to 1 part by weight of montelukast). At 0 and 60 minutes, 0.5 mL of the sample was filtered through a 0.5 μm syringe filter and immediately diluted 1/10 with high-performance liquid chromatography (HPLC) mobile phase, and analyzed by HPLC to determine the concentration. The details of the HPLC used are as follows:

1. HPLC system: Agilent 1200 serises;

3. Flow rate: 1 mL / min;

5. Injection volume: 20 μL; And

6. UV wavelength: 389 nm.

The results are shown in Figs. 7, 8 and 9. Fig.

7 and 8 are images showing the solubility of montelukast when the solubilizing agent is 0.1, 1, 10 or 100 parts by weight based on 1 part by weight of the montelukast in terms of the concentration ratio (%) at 60 minutes to the concentration at 0 minutes The concentration ratio (%) represents the concentration ratio (%) of the dissolved montelukast, that is, the concentration ratio at the elapse of 60 minutes with respect to the concentration at 0 minutes after mixing.

9 is an image schematically showing the solubility of the solubilizing agent after 60 minutes of addition of the montelukast.

As shown in FIGS. 7, 8 and 9, the solubility of Tween 40, Tween 60, Poloxamer 407, Cremophor EL, and Cremophor RH40 after 60 minutes in the 1: 1 weight ratio of montelukast and solubilizer The solubility of 90% or more showed very good sedimentation prevention effect. Tween 20, Tween 80, Twin 85, Cremopore A25, and Cremophor RH60 have low solubility at a weight ratio of 1: 1, but at a ratio of 1:10 weight, Effect. In addition, it was confirmed that the addition of bile acid at a weight ratio of 1: 100 prevents the reprecipitation of montelukast.

<Experimental Example 4> Evaluation of solubility of montelukast by solubilizing agent under acidic condition

In order to increase the bioavailability of montelukast, the montelukast should remain dissolved in the acidic environment of gastrointestinal tract.

7-8 of Experimental Example 3, solubilizing agents (Tween 40, Twin 60, Poloxamer 407, Cremophor EL, Kreflex), which were found to increase the solubility of montelukast in neutral condition (pH 7.4) Mopore RH40).

&Lt; 4-1 > Preparation of Solubilizing Agent

Tween 40, Tween 60, Poloxamer 407, Cremophor EL, or Cremophor RH40 were dissolved in artificial gastric juice (the first liquid of the 11th revision of Korea Pharmacopoeia, pH 1.2) at a concentration of 0.0444 mg / mL.

<4-2> Monterukast Preparation of sodium salt

<4-3> Evaluation of solubility

Deionized water, artificial gastric juice (pH 1.2), and the solubilizing agent prepared in the above <4-1> were each put into a tube in an amount of 1.8 mL. To the tube was added 0.2 mL of the montelukast sodium salt prepared in <4-2>, and the diluted solution was diluted to a final concentration of 40 μg / mL, followed by shaking. Each of the solubilizing agents is 1 part by weight based on 1 part by weight of the montelukast.

Immediately after the addition of the montelukast sodium salt (0 min) and after 60 min, 0.5 mL of sample was filtered through a 0.5 μm syringe filter and immediately diluted 1/10 with high-performance liquid chromatography (HPLC) mobile phase And then analyzed by HPLC to determine the concentration. The details of the HPLC used are as follows.

1. HPLC system: Agilent 1200 serises;

3. Flow rate: 1 mL / min;

4. Column: Phenomenex Luna (250 × 4.6 mm, 5 μm, 100 Å, PN 00G-4252-E0, SN 290750-12);

5. Injection volume: 20 μL; And

6. UV wavelength: 389 nm.

The results are shown in Fig.

FIG. 10 is a graph showing the solubility of montelukast when the solubilizing agent is 1 part by weight based on 1 part by weight of montelukast in an acidic environment, expressed as a concentration ratio (%) at 60 minutes relative to the concentration at 0 minutes.

As shown in Fig. 10, montelukast rapidly precipitated even at 0 min under the condition of only artificial gastric juice (pH 1.2) without addition of solubilizer, and the concentration of the solution portion except for the filtrate, mL).

In the case of adding the solubilizing agent, it was confirmed that the precipitation of montelukast was inhibited compared with the case where only the artificial gastric juice was added without adding the solubilizing agent, and the solubility of montelukast in acidic conditions .

From this, it can be seen that Klempo fish is most preferable in terms of prevention of the precipitation of montelukast depending on the stomach acid.

&Lt; Experimental Example 5 >

Montelukast sodium salt and sodium taurocholate were administered in combination, and the concentration of montelukast in plasma was measured to evaluate the absorption rate in the body. The following experiment was conducted.

8-week-old male Sprague-Dawley (SD) rats were anesthetized by inhalation of ether and intubated into the right carotid artery (PE60). Subsequently, the montelukast sodium salt (5 mg / kg as montelukast) and sodium taurocholate (8.25 mg / kg as taurocholate) in powder form were orally administered using a mini capsule for rats and sonde. As a control group, montelukast sodium salt (montelukast 5 mg / kg) was orally administered orally. 300 μL of blood was sampled over 0, 0.08, 0.17, 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8 and 24 hours and the same amount of heparin salt water was added. Centrifugation was carried out at 12,000 rpm for 3 minutes, and the plasma of the upper layer was stored at -80 ° C. The plasma was shaken by adding 4 times the amount of acetonitrile containing the internal standard, centrifuged, and the upper layer was analyzed by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Details of the LC-MS / MS used are as follows.

1. HPLC conditions

- System: Flexar FX15 UPLC-UVD (Perkin Elmer);

- mobile phase: 80% acetonitrile containing 0.1% formic acid;

Flow rate: 0.4 mL / min;

- Column: SepaxGP-C18, 2.1 x 5.0 mm, 3 m; And

- Injection volume: 10 μL.

2. MS / MS conditions

System: Waters Quattro micro ^TM API mass spectrometer; And

- ES positive mode: 586.4 - 422.2 for montelukast, 592.4 - 427.2 for internal standard.

The results are shown in Fig.

FIG. 12 is an image showing changes in the body dynamics of the monotherapeutic group treated with montelukast and sodium taurocholate and monotherapy alone;

As shown in Fig. 12, in the case of montelukast and sodium taurocholate-treated group, the AUC and C _max were increased about 2-3 times and the T _max was reduced by one-third when compared with the montelukast alone treatment group, . Therefore, it can be expected that solubilizing agents such as sodium taurocholate have a great effect on the absorption of montelukast into the body.

Claims

A method of improving the bioavailability of montelukast using a solubilizing agent.

The method according to claim 1,
Wherein the method comprises preventing reprecipitation of montelukast in vivo to increase bioavailability.

The method according to claim 1,
Wherein the method comprises preventing repretation of montelukast in vivo and thereby accelerating bioavailability.

The method according to claim 1,
Wherein said method prevents re-precipitation of montelukast in vivo, thereby reducing bioavailability variation.

The method according to claim 1,
Wherein the solubilizing agent is at least one selected from the group consisting of bile acids, poloxamers, twins, crmaphos, and glycols.

The method according to claim 1,
Wherein the solubilizing agent is a bile acid.

The method according to claim 5 or 6,
Wherein the bile acid is at least one selected from the group consisting of sodium taurocholate, sodium deoxycholate, sodium glycocholate and sodium cholate. .

The method according to claim 5 or 6,
Wherein the bile acid is sodium taurocholate or sodium deoxycholate.

6. The method of claim 5,
The poloxamers are selected from the group consisting of poloxamer 124, poloxamer 188, poloxamer 237, poloxamer 338, and poloxamer 407. Or more.

6. The method of claim 5,
The twin classes are Tween 20, Tween 21, Tween 40, Tween 60, Tween 61, Tween 65, Tween 80, Tween 80, 80), Tween 81 (Tween 81), and Tween 85 (Tween 85).

6. The method of claim 5,
The crested females are selected from the group consisting of Cremophor A6, Cremophor A20, Cremophor A25, Cremophor EL, Cremophor ELP, And at least one selected from the group consisting of Cremophor RH40, Cremophor RH60, Cremophor RH410 and Cremophor WO7. Way.

6. The method of claim 5,
Wherein the glycols are polyethylene glycols having a weight-average molecular weight (Mw) of 100 to 8000.

Montelukast or a pharmaceutically acceptable salt thereof; And
A solubilizing agent,
Wherein the solubilizing agent is at least one selected from the group consisting of bile acids, poloxamers, tweens, crimp fish and glycols.

14. The method of claim 13,
Wherein the formulation is an enteric preparation.

14. The method of claim 13,
Wherein the formulation is administered by oral administration, oral administration, mucosal administration, intranasal administration, intraperitoneal administration, subcutaneous injection, intramuscular injection, transdermal administration or intravenous injection.