JP2007056045A - Cyclodextrin composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a soluble cyclosporin composition and also provide a soluble cyclosporin composition having reduced hemolytic property. <P>SOLUTION: The composition comprises cyclosporins or macrolide compounds and β-cyclodextrin derivative. In another embodiment, the composition comprises cyclosporins or macrolide compounds, glucosyl-α-cyclodextrin and/or maltosyl-α-cyclodextrin. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a composition of a cyclosporine or a macrolide compound (hereinafter sometimes referred to as cyclosporine or the like). In particular, it relates to a soluble cyclosporine composition.

  Cyclosporine refers to a known group of cyclic undecapeptides having pharmaceutical activity. For example, in addition to the compound commercially available under the trademark SANDIMMUN (cyclosporin A), dihydrocyclosporin D and cyclosporin G, 2- (O-2-Hydroxyethyl-D-serine) -Cyclosporin A, (S ′ Many compounds such as -keto-Bmt1)-(Val2) -Cyclosporin are known (see JP-A-2-17127). Among these, cyclosporin A is currently widely used clinically for the purpose of inhibiting rejection in organ transplantation and graft-versus-host disease.

  Also, various autoimmune diseases and inflammatory conditions, particularly inflammatory conditions with etiology involving autoimmune elements such as psoriasis, arthritis (eg, rheumatoid arthritis, progressive prevalent arthritis and osteoarthritis), rheumatic diseases and respiratory system Its usefulness against diseases (eg, asthma) has been clinically confirmed.

  Furthermore, an action against malaria, coccidioidomycosis and schistosomiasis, anthelmintic, particularly an antigenic animal action, an action against alopecia, and an action against a multi-drug resistant tumor have been confirmed.

On the other hand, macrolide is a macrocyclic lactone and is a general term for compounds having 12 or more ring members. A lactam macrolide is an interesting compound having a lactam (amide) bond in the ring in addition to a lactone (ester) bond, and includes lactams produced by microorganisms of the genus Streptomyces such as rapamycin, ascomycin, and FK-506. There are abundant types of macrolides and their derivatives and analogs. These lactam macrolides are said to have unique pharmacological properties, and in particular, immunosuppression and anti-inflammatory effects are attracting attention.

Rapamycin is a lactam macrolide with immunosuppressive action and is produced by Streptomyces hygroscopicus . The structure of rapamycin is described in Kesseler, H. et al. Et al. In 1993 (Hely. Chim. Acta; 76: 117). As the derivatives of rapamycin, many types such as 40-O-alkylated derivatives such as 40-O- (2-hydroxy) ethyl-rapamicin disclosed in WO94 / 09010 pamphlet have been synthesized. Rapamycin and its structurally similar compounds and derivatives are collectively referred to as rapamycins.

  Ascomycins are different classes of lactam macrolides, and typical compounds include FK-506 and ascomycin. Many of these have strong immunosuppressive and anti-inflammatory effects. The structural formula of FK-506 is described as an A5 item in Appendix of Merck Index, 11th edition (1989). Ascomycin is described in US Pat. No. 3,244,592. Many derivatives of ascomycin and FK-506 have been synthesized, such as halogenated derivatives such as 33-epi-chloro-33-desoxy-asmycin published in European Patent Application Publication No. 427 680. Ascomycin, FK-506, and structurally similar compounds and derivatives thereof are collectively referred to as ascomycins.

  Since the solubility of cyclosporine or macrolide compounds in water is extremely low, it is currently prepared as an oily solution preparation containing ethanol.

  Such a preparation easily precipitates crystals such as cyclosporine upon contact with water, and is not a preferred composition for oral preparations and mucosal application agents.

  In addition, a surfactant, that is, a polyoxyethylene castor oil derivative is blended in the injection as a solubilizer to prevent the precipitation of cyclosporine and the like. However, polyoxyethylene castor oil derivatives are not always preferable for safety because they may cause serious hypersensitivity reactions such as shock.

  Accordingly, a composition that contains a therapeutically effective amount of cyclosporine and the like and is sufficiently soluble without using a surfactant or the like has been desired.

  In the prior art, it is known that α-cyclodextrin is effective in improving the solubility of cyclosporines, but the effect of β-cyclodextrins has not been recognized in this application.

  Japanese Patent Application Laid-Open No. 64-85921 discloses a preparation comprising cyclosporine and α-cyclodextrin and / or a derivative thereof.

  However, α-cyclodextrin has a relatively high hemolytic property and cannot be applied externally, particularly to mucous membranes. Therefore, a cyclosporine composition that has low hemolysis and is soluble has been desired.

  Contrary to the teaching of Japanese Patent Application Laid-Open No. 64-85951, the inventors of the present application dramatically increased the solubility of cyclosporine and the like in water by mixing β-cyclodextrins and cyclosporine and the like. -It was found to be very good compared to the effects known for the mixture of cyclodextrin and cyclosporine. That is, the present invention provides a composition comprising cyclosporine or the like and a β-cyclodextrin derivative.

  The present inventors have also found that specific α-cyclodextrin derivatives have relatively low hemolytic properties and significantly improve the solubility of cyclosporines in water. That is, the present invention provides a composition comprising cyclosporine and glucosyl-α-cyclodextrin and / or maltosyl-α-cyclodextrin (hereinafter sometimes referred to as α-cyclodextrin derivative).

  As the cyclosporins used in the present invention, the aforementioned cyclosporin A is preferable.

  The present invention also provides a composition comprising a macrolide compound and a β-cyclodextrin derivative or glucosyl-α-cyclodextrin and / or maltosyl-α-cyclodextrin.

  Examples of β-cyclodextrin derivatives used in the present invention include methyl-β-cyclodextrin, dimethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, glucosyl-β-cyclodextrin, maltosyl-β-cyclodextrin, Sulfonated-β-cyclodextrin, sulfonated alkyl ether-β-cyclodextrin (for example, C1-4 alkyl) and the like, and methyl-β-cyclodextrin and dimethyl-β-cyclodextrin are preferable, and dimethyl-β-cyclodextrin Dextrin is particularly preferred. Further, β-cyclodextrins in which further substituents are introduced into these compounds can also be used in the present invention.

  The composition of the present invention comprises a powdery cyclosporine and a β-cyclodextrin derivative or glucosyl-α-cyclodextrin and / or maltosyl-α-cyclodextrin in an appropriate amount of water or a mixture of water and alcohol. It can be produced by adding a solvent and kneading, and can also be produced by spray drying, freeze drying or the like.

  The composition of the present invention can be formulated into any formulation known in the art such as powders, granules, tablets, injection preparations, suppositories, solutions, inhalants, dry syrups and capsules.

  The compounding ratio of cyclosporine and the like to the β-cyclodextrin derivative is 0.1: 99.9 to 90:10, preferably 0.5: 99.5 to 50:50, more preferably 1:99 to 20:80. It is a range.

  The compounding ratio of cyclosporine or the like and the α-cyclodextrin derivative is 0.1: 99.9 to 90:10, preferably 0.5: 99.5 to 70:30, more preferably 1:99 to 50:50. It is a range.

  In the composition of the present invention, in addition to the above components, excipients, binders, disintegrants, lubricants, isotonic agents, pH adjusters, preservatives, moisturizing agents generally added to pharmaceutical preparations A coloring agent, a coloring agent, and the like can be appropriately blended.

  The composition of the present invention can be applied to all diseases to which conventional cyclosporine or macrolide compounds have been applied.

  The composition of the present invention containing a β-cyclodextrin derivative can be administered by a method such as oral, intravenous injection, subcutaneous injection, instillation, eye drop, topical application, inhalation, etc., and oral administration is particularly preferable.

  When the composition of the present invention is used, the absorption of the active ingredient into the body is improved as compared with any conventionally known cyclosporine preparations. Therefore, the dose of the active ingredient can be expected to be reduced as compared with the conventional preparations (about 40 -About 80%).

  Further, the composition of the present invention containing an α-cyclodextrin derivative can be administered by a method such as oral, intravenous injection, subcutaneous injection, drip, inhalation, instillation, instillation, topical application, etc., but inhalation, instillation, instillation, etc. Is preferred.

  Since the composition of the present invention is soluble and has a low hemolytic property, it can be administered by inhalation or nasal administration, which was impossible with the conventionally known cyclosporine / α-cyclodextrin. .

  The appropriate amount of active ingredient or formulation to be administered therapeutically is determined by various factors such as dosing conditions, dosing interval, release rate of the active ingredient and the like. For example, the amount of the active ingredient in the preparation necessary for maintaining a therapeutically effective blood concentration, or the release rate of the active ingredient is determined from the results of known in-vitro and in-vivo experiments.

  The oral maintenance dose for transplant patients is about 3 to about 10 mg (potency) / kg daily and is administered by capsules containing 25 to 100 mg of cyclosporine.

  Although the mechanism of the intermolecular interaction of the present invention has not yet been fully elucidated, in the combination of the present invention, the drug is considered to have some binding state with cyclodextrins.

  Hereinafter, the present invention will be described in more detail with reference to examples.

Reference example 1
Table 1 shows the solubilizing effects of various cyclodextrins such as cyclosporine.

Reference example 2
A comparison of the solubilizing effect of dimethyl-β-cyclodextrin and α-cyclodextrin on cyclosporin A at 37 ° C. is shown in FIG. Further, 2- (O-2-Hydroxyethyl-D-serine) -Cyclosporin A, (S′-keto-Bmt1)-(Val2) -Cyclosporin, 33-epi-chloro-33-desoxy of dimethyl-β-cyclodextrin. The solubilizing effect at 37 ° C. on -ascomycin is shown in FIG. 2, FIG. 3, and FIG. 4, respectively.

Example 1
Capsules containing cyclosporin A and cyclodextrins were prepared as follows.

  Cyclosporin A 0.3 g and dimethyl-β-cyclodextrin 11.7 g were mixed for 10 minutes, and 12 ml of water was added and kneaded for 3 hours. The obtained kneaded material was dried at 40 ° C. for 2 hours and further dried under reduced pressure for 24 hours. Thereafter, the mixture was sieved with a 60 mesh sieve to produce a powder agent (agent 1).

Example 2
Agent 2 was produced by repeating the method of Example 1 except that 12 ml of a mixed solvent of water / ethanol = 1/1 was used instead of 12 ml of water.

  In the above-described embodiments, cyclosporin A can be replaced with 40-O- (2-hydroxy) ethyl-rapamycin, 33-epi-chloro-33-desoxy-ascomycin, or the like. In this case, a significant improvement in solubility was observed.

Comparative Example 1
Agent 3 was prepared by repeating the method of Example 1 except that α-cyclodextrin was used instead of dimethyl-β-cyclodextrin.

Test 1 (In vitro test)
540 mg of Agents 1 to 3 were filled into a No. 0 hard gelatin capsule to give a capsule preparation. These were subjected to elution test (eluate volume: 500 ml) at 37 ° C. and 100 rpm using JP I liquid (pH = 1.2) in the paddle method, and quantified using the HPLC method. The results are shown in FIG.

  As is clear from FIG. 5, the capsule containing the composition of the present invention is superior in dissolution property as compared with the capsule of Comparative Example 1 using α-cyclodextrin.

Test 2 (in vivo test)
No. 1 hard gelatin capsule filled with Agent 1 and Sandy Mün capsule (trademark, manufactured by Sand Yakuhin) are administered to male beagle dogs (7 months old) after fasting for 24 hours, the same amount of drug will be administered The blood concentration of cyclosporine was measured by HPLC immediately before administration and 0.5, 1, 2, 4, 6, 8, and 24 hours after administration. The results are shown in FIG.

  As is apparent from FIG. 6, when the capsule containing the composition of the present invention is used, the blood concentration of cyclosporine is higher than when Sandimune Capsule (trademark) is used.

Formulation Example 1 (tablet)
Cyclosporin A 75 g and dimethyl-β-cyclodextrin 1500 g were mixed, and 350 ml of purified water and 30 g of methylcellulose were added and kneaded. This was granulated with an extrusion granulator, dried and then sized. To this, 90 g of corn starch and 70 g of carboxymethylcellulose were added and mixed. Further, 8 g of magnesium stearate was mixed, and the mixture was tableted to obtain tablets.

Formulation Example 2 (granule)
Cyclosporin A 75 g and dimethyl-β-cyclodextrin 1500 g were mixed, and 350 ml of purified water and 30 g of methylcellulose were added and kneaded. This was granulated with an extrusion granulator, dried and then sized. Further, sieving was performed to obtain granules.

Formulation Example 3 (Liquid)
10 g of cyclosporin A and 300 g of dimethyl-β-cyclodextrin were dissolved in 1 l of purified water, and 2 g of methyl paraoxybenzoate and 0.2 g of propyl paraoxybenzoate were further added to obtain a solution.

Reference example 3
The hemolytic properties of various cyclodextrins are shown in FIG.

Reference example 4
The hemolytic properties of cyclodextrins under cyclosporine saturation concentration are shown in FIGS.

Reference Example 5
Table 2 shows the solubilizing effect of various cyclodextrins such as cyclosporin A.

Example 3
Capsules containing cyclosporin A and cyclodextrins were prepared as follows.

  0.6 g of cyclosporin A and 11.4 g of glucosyl-α-cyclodextrin were mixed for 10 minutes, and 12 ml of water was added and kneaded for 3 hours. The obtained kneaded material was dried at 40 ° C. for 2 hours and further dried under reduced pressure for 24 hours. Thereafter, the mixture was sieved with a 60-mesh sieve and filled into a No. 0 hard gelatin capsule to produce a capsule preparation (agent 4).

  In this embodiment, cyclosporin A can be replaced with 40-O- (2-hydroxy) ethyl-rapamycin, 33-epi-chloro-33-desoxy-ascomycin, or the like.

Comparative Example 2
Agent 5 was produced by repeating the method of Example 3 except that α-cyclodextrin was used instead of glucosyl-α-cyclodextrin.

Test 3 (in vivo test)
Agents 4 and 5 were orally administered to male beagle dogs (7 months old) after fasting for 24 hours so that the amount of drug to be administered would be the same, immediately before administration, 0.5, 1, 2, The blood concentration of cyclosporine after 4, 6, 8, and 24 hours was measured by HPLC. The results are shown in FIG.

  As is clear from FIG. 11, when the capsule containing the composition of the present invention is used, the blood concentration of cyclosporine is higher than when the capsule of Comparative Example 2 using α-cyclodextrin is used.

Formulation Example 4 (tablet)
Cyclosporin A 75 g and glucosyl-α-cyclodextrin 1500 g were mixed, and 350 ml of purified water and 30 g of methylcellulose were added and kneaded. This was granulated with an extrusion granulator, dried and then sized. To this, 90 g of corn starch and 70 g of carboxymethylcellulose were added and mixed. This was mixed with 8 g of magnesium stearate, and this mixture was tableted to obtain tablets.

Formulation Example 5 (granule)
Cyclosporin A 75 g and glucosyl-α-cyclodextrin 1500 g were mixed, and 350 ml of purified water and 30 g of methylcellulose were added and kneaded. This was granulated with an extrusion granulator, dried and then sized. Further, sieving was performed to obtain granules.

Formulation Example 6 (Inhalant)
25 g of cyclosporin A and 75 g of glucosyl-α-cyclodextrin were dissolved in 10 l of a mixed solvent of purified water / isopropyl alcohol = 1/1, and spray-dried with a spray dryer to form fine particles. This was filled into a hard gelatin capsule to obtain a powder inhalant.

Formulation Example 7 (eye drops)
Cyclosporine A 24 mg, glucosyl-α-cyclodextrin 10 g, sodium dihydrogen phosphate 0.56 g, sodium monohydrogen phosphate 0.28 g and benzalkonium chloride 10 mg were dissolved in 100 ml of sterile purified water. Sodium chloride was added thereto to make it isotonic to obtain an eye drop.

Formulation Example 8 (Nasal solution)
Cyclosporine A 24 mg, glucosyl-α-cyclodextrin 10 g, sodium dihydrogen phosphate 0.56 g, sodium monohydrogen phosphate 0.28 g and benzalkonium chloride 10 mg were dissolved in 100 ml of sterile purified water. Glycerol was added thereto to make it isotonic to obtain a nasal solution.

It is a graph of the solubilization effect in 37 degreeC with respect to cyclosporin A of a dimethyl-beta-cyclodextrin and an alpha cyclodextrin. Cyclodextrin and an excess amount of cyclosporine were added to 10 ml of water and stirred at 37 ° C. for 3 hours. Thereafter, centrifugation was performed, and the supernatant was analyzed by HPLC. It is a graph of the solubilization effect in 37 degreeC with respect to 2- (O-2-hydroxyethyl-D-serine) -CyclosporinA of dimethyl- (beta) -cyclodextrin. Cyclodextrin and an excess amount of cyclosporine were added to 10 ml of water and stirred at 37 ° C. for 3 hours. Thereafter, centrifugation was performed, and the supernatant was analyzed by HPLC. It is a graph of the solubilization effect in 37 degreeC with respect to (S'-keto-Bmt1)-(Val2) -Cyclosporin of a dimethyl-beta-cyclodextrin. Cyclodextrin and an excess amount of cyclosporine were added to 10 ml of water and stirred at 37 ° C. for 3 hours. Thereafter, centrifugation was performed, and the supernatant was analyzed by HPLC. It is a graph of the solubilization effect in 37 degreeC with respect to 33-epi-chloro-33-desoxy-ascomycin of a dimethyl-beta-cyclodextrin. Cyclodextrin and an excess amount of ascomycin were added to 10 ml of water and stirred at 37 ° C. for 3 hours. Thereafter, centrifugation was performed, and the supernatant was analyzed by HPLC. It is a graph of the in vitro dissolution test of the capsule containing various compositions. It is a graph of the blood concentration of cyclosporine with respect to the elapsed time after administration of various capsules. It is a graph which shows the hemolytic property of various cyclodextrins / cyclosporin A aqueous solution. Cyclodextrin was dissolved in 5 ml of 0.85% saline, 0.05 ml of rabbit blood was added, incubated at 37 ° C. for 30 minutes, centrifuged (5 minutes), and the absorbance of the supernatant was measured at 540 nm. 100% hemolysis was calculated with 0.1% saline. It is a graph which shows the hemolytic property of cyclodextrins under cyclosporine saturation concentration. Cyclodextrin is dissolved in 5 ml of 0.85% saline, cyclosporine is added to saturation, then 0.05 ml of rabbit blood is added to the test solution, incubated at 37 ° C. for 30 minutes, and centrifuged (5 minutes ) Thereafter, the absorbance of the supernatant at 540 nm was measured. 100% hemolysis was calculated with 0.1% saline. It is a graph which shows the hemolytic property of cyclodextrins under 2- (O-2-Hydroxyethyl-D-serine) -Cyclosporin A saturation concentration. Cyclodextrin is dissolved in 5 ml of 0.85% saline, cyclosporine is added to saturation, then 0.05 ml of rabbit blood is added to the test solution, incubated at 37 ° C. for 30 minutes, and centrifuged (5 minutes ) Thereafter, the absorbance of the supernatant at 540 nm was measured. 100% hemolysis was calculated with 0.1% saline. It is a graph which shows the hemolytic property of cyclodextrins under the (S'-keto-Bmt1)-(Val2) -Cyclosporin saturation concentration. Cyclodextrin is dissolved in 5 ml of 0.85% saline, cyclosporine is added to saturation, then 0.05 ml of rabbit blood is added to the test solution, incubated at 37 ° C. for 30 minutes, and centrifuged (5 minutes ) Thereafter, the absorbance of the supernatant at 540 nm was measured. 100% hemolysis was calculated with 0.1% saline. It is a graph which shows the cyclosporin blood dynamics after administration of various capsules.

Claims (3)

  A composition comprising cyclosporines and glucosyl-α-cyclodextrin and / or maltosyl-α-cyclodextrin.   The composition according to claim 1, wherein the cyclosporine is cyclosporin A.   The composition according to claim 1 or 2, which is in the form of an inhalant.

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