KR20080054355A - Plaster composition comprising alendronate as an effective ingredient for treating osteoporosis - Google Patents

Abstract

A composition comprising alendronate is provided to increase bioavailability and reduce side effects of alendronate by administering it percutaneously and improving its permeability to the skin by using a permeation improver, so that the composition is useful for treating osteoporosis. A plaster composition for treating osteoporosis comprises 0.1-10% of alendronate, 5-50% of solvent, 0.1-10% of percutaneous permeation improver, and 30-90% of pressure-sensitive adhesive, wherein the percutaneous permeation improver is selected from lauric acid, oleic acid, linoleic acid, caproic acid, caprylic acid, myristic acid, oleyl alcohol, isopropylmyristate and propyleneglycol laurate. The plaster for treating osteoporosis comprises contains a supporter for inhibiting permeation of drug, a drug layer containing alendronate solubilized in solvent and pressure-sensitive adhesive, and a release layer for treating alendronate with silicon.

Description

Plaster composition comprising alendronate as an effective ingredient for treating osteoporosis}

The present invention relates to a plaster composition for osteoporosis containing the alendronate as an active ingredient.

Reference 1 Keller RK and Fliesler SJ, Biochem . Biophys . Res . Commun ., 266 , pp 560-3, 1999

Document 2 Hardman JG and Limbird LE, Goodman & Gilman's The Pharmacological Basis of Therapeutics , 10 th ed ., pp 1733-1735, 1997

Ansel HC et al., Pharmaceutical Dosage Forms and Drug Delivery Systems , Chapter 10 , pp357-372, 1995

4 Gwak HS et al., Arch . Pharm . Res . , 26 , pp 644-648, 2003

Ptacek, P. et al., J. Chromatogr . B. , 767 , pp 111-116, 2002

6 Gwak HS and Chun IK, Arch . Pharm . Res . , 24 , pp 578-583, 2001

Document 7 Kenneth W. Renton. et al., Original Research . , 18 , pp98-101, 2004

8 Ptacek, P. et al., J. Chromatogr . B. , 767 , pp 111-116, 2002

Alendronate is a bisphosphonate derivative that is widely used for osteoporosis and Paget's disease. Alendronate acts on osteoclasts or their precursors and inhibits bone resorption. Unlike other bisphosphonate drugs, this drug is not metabolized by ATP analogs, but isophores such as geranylgeranyl diphosphate, which is required for the phenylation of various proteins that play an important role in osteoclast function. Inhibits the conversion of mevalonate to isoprenoid lipids and cholesterol (Keller RK and Fliesler SJ, Biochem. Biophys. Res. Commun., 266 , pp560-3, 1999) .

Alendronate has a very low absorption by the gastrointestinal tract (0.6-0.7%) and should be administered at least 30 minutes before taking another food or drug in the first hour, especially because absorption is reduced by up to 60% by other drugs or foods. Hardman JG and Limbird LE, Goodman & Gilman's The Pharmacological Basis of Therapeutics , 10 th ed ., pp 1733-1735, 1997). In addition, to prevent esophagitis, a major side effect, it should be taken with a large amount of water, and at least 30 minutes after sitting in a straight position. Because of the complexity of these medications, their use is limited, especially in the elderly, patients with low medication compliance, who should not consume much water, or who cannot sit straight for a long time. To compensate for this problem, there have been improvements in the usage of 70mg once a week from 10mg once a day. However, despite these improvements, there are still reports of adverse gastrointestinal side effects due to reluctance to take this drug or to take it. Although there are many problems in oral administration, few studies have been conducted on alternative routes of allendronate.

Due to the above weakness, transdermal absorption preparations have attracted attention as an alternative route of oral administration preparations. Transdermal absorption preparations effectively improve the bioavailability by effectively delivering the drugs with gastrointestinal side effects, drugs that are difficult to be absorbed from the gastrointestinal tract, drugs that receive hepatic metabolism, and drugs that are easily degraded in the gastrointestinal tract through the body, through the skin, and improve side effects such as gastrointestinal side effects. It is known to reduce (Ansel HC et al., Pharmaceutical Dosage Forms and Drug Delivery Systems , Chapter 10 , pp357-372, 1995). At the end of 1970, Ciba-Geigy's motion sickness prevention scopolamine patch (Transderm-Scop) was developed as a transdermal absorbent, and a nitroglycerin patch (Transderm-Nitro) was developed for the purpose of preventing angina, 1984 Since launching in 2009 and securing a significant market in the United States, several pharmaceutical companies have spurred the development of new transdermal therapeutic systems (TTS).

Alendronate, the subject of this study, is a drug with very low bioavailability and high gastrointestinal side effects, as mentioned above. Therefore, it is expected to improve bioavailability, minimize side effects, and increase medication compliance according to the development of gastric transdermal absorption agents. However, there is currently no research on transdermal absorption of the drug at home and abroad. This is thought to be due to the low permeability of the drug, but considering that the bioavailability is up to 1% even in the case of oral administration, it is expected that a sufficient effect can be achieved even if only a small amount of permeation is used using an appropriate permeation accelerator. .

Osteoporosis is a disease that is not a big problem if continuous treatment is done, but if it is not properly treated, it can cause other problems such as fracture, which can greatly affect the national medical budget, etc. It is a disease that requires proper medicine. Therefore, if this study can open the possibility of commercialization of percutaneous absorption of alendronate, it can have a great effect on the treatment of osteoporosis, which is expected to be of great help to the country and individual patients.

In the present invention, it is intended to develop an alendronate transdermal absorption system and to compare drug absorption in animals and healthy volunteers with the oral case. In addition, through these pharmacokinetics and clinical results, the ultimate goal is to commercialize transdermal absorbents of Alendronate.

However, none of the above documents discloses or teaches transdermal absorbents containing alendronate.

Accordingly, the present inventors have completed the present invention by confirming the persistence and safety of the alendronate-containing transdermal absorbent as a result of performing the quantitative test of the alendronate and the skin permeation experiment.

      In order to achieve the above object, the present invention provides a plaster composition for the treatment and prevention of osteoporosis containing an alendronate as an active ingredient, containing a solvent, a transdermal absorption accelerator, a pressure-sensitive adhesive.

      The solvent of the present invention isopropyl myristate, propylene glycol monolaurate, propylene glycol monocaprylate, diethylene glycol monoethyl ether ), Propylene glycol or ethanol, preferably propylene glycol (PG).

      The transdermal absorption accelerator of the present invention is lauric acid, oleic acid, linolenic acid, capric acid, caprylic acid, myristic acid, oleyl alcohol, isopropyl myristate or propylene glycol laurate, preferably lauric acid, Oleic acid, linolenic acid, capric acid or caprylic acid, more preferably oleic acid or caprylic acid, most preferably 5-10% oleic acid or 5-10% caprylic acid-containing propylene glycol (PG) The pressure-sensitive adhesive is an acrylic pressure-sensitive adhesive made of an acrylate polymer, a rubber pressure-sensitive adhesive and a water-soluble pressure-sensitive adhesive, preferably an acrylate polymer, to which additives such as petroleum resin, rosin resin, and oil are added to natural or synthetic rubber. Is available.

       In addition, the present invention provides an alendronate-containing plaster composed of a support layer preventing drug permeation, an active ingredient dissolved in a solvent, an alendronate, a drug layer containing a pressure-sensitive adhesive, and a release layer treated with silicon (see FIG. 1).

       Hereinafter, the present invention will be described in detail.

       The plaster according to the present invention contains an effective drug, a solvent, a percutaneous absorption accelerator, a pressure-sensitive adhesive and other commonly used additives.

       In the present invention, as an active ingredient, alendronate contains 0.1 to 10%, preferably 0.5 to 5% in the total composition. This blending ratio provides an alendronate-containing plaster that maintains good skin permeability, sustainability of drug, and drug stability.

In addition, the active drug is optionally a bisphosphonate preparation, preferably pamidronate, etidronate, clodronate, tiludronate, nerdronate ( neridronate, olpadronate, ibandronate, risedronate or zledronate, more preferably pamidronate can be used.

     Ethanol, propylene glycol, propylene glycol monocaprylate, propylene glycol monolaurate, isopropyl myristate used as a solvent in the present invention Or diethylene glycol monoethyl ether is used for the purpose of increasing the permeation rate of the matrix transdermal drug delivery system and dissolving the drug, and 5 to 50%, preferably 10 to 30% of the total composition. It contains.

       In addition, in the present invention, as a transdermal absorption accelerator in addition to a solvent for maintaining a high concentration of the drug in the plaster in order to further promote skin penetration of the alendronate, lauric acid, oleic acid, linoleic acid as lauric acid ), Capric acid, caprylic acid, and the like can be used. Such transdermal absorption accelerators may be used alone or in combination of two or more thereof, and the total amount thereof includes 0.1 to 10% of the total composition of the drug layer, more preferably 0.5 to 5%.

      In addition, the pressure-sensitive adhesive used in the plaster of the present invention uses an acrylic adhesive consisting of an acrylate polymer, preferably Duro-Tak® 87-2196, Duro-Tak® 87-2100 or Duro-Tak® 87- 2510 (Bridgewater, NJ, USA).

      The pressure-sensitive adhesive is used alone or in combination of two or more, the total amount of the total composition comprising the drug layer in the range of 30 to 90%, more preferably containing in the range of 50 to 80%.

      In addition, natural or synthetic rubbers, tackifier resins such as petroleum resins, rosin resins, terpene resins, rubber-based adhesives and water-soluble adhesives to which additives such as oils and antioxidants are added may be used.

      In the present invention, in order to improve the physical properties of the formulation in addition to the above composition, additives such as essential oil components such as 1-menthol and peppermint oil, antioxidants, preservatives and inorganic fillers can be blended, and the amount of the composition is used as the total composition of the drug layer. 0.1 to 10%, preferably 0.2 to 5%.

       In the alendronate-containing plaster of the present invention, all components in the formulation having the above composition are uniformly dispersed without aggregation, and high skin permeability is stably maintained to maintain the effect of the alendronate for a long time, and stability and adhesion of the preparation great.

       The present invention includes a plaster formulated using the composition as described above, for example, the structure of the plaster according to the present invention can be prepared as shown in Figure 1 below.

       The alendronate-containing plaster shown in FIG. 1 is composed of three components so that the drug can be effectively transdermally administered during skin application, that is, a support (1) which prevents the penetration of the drug, an alendronate which is an active ingredient dissolved in the above-mentioned solvent, and reduced pressure. It consists of the drug layer 2 containing the sex adhesive, and the peeling layer 3 processed by silicone.

       The support 1 is a material having no interaction with drugs and other compositions in the drug layer 2, preferably made of a material having good moisture permeability and elasticity. Examples of the support that can be used in the present invention include synthetic resin films such as polyethylene, polypropylene, polyester, and polyurethane, sheets, sheet-like foams, woven or nonwoven fabrics, and their repellents, and preferably polyester films. Used.

       The drug layer (2) is mainly composed of an alendronate, a pressure-sensitive adhesive, a solvent and a transdermal absorption accelerator, etc., may be added to the skin irritant or plasticizer, if necessary. The drug layer containing the alendronate and the other composition has a single layer or a multilayer structure of 2-3 layers, and the thickness of the entire adhesive layer is suitably 0.01 to 1 mm, preferably 0.1 to 0.5 mm.

        The release layer 3 may be a polyester film coated with silicon, a fluorine-coated polyester film, polyethylene film or paper, and the like, and may be preferably silicon.

        Plaster of the present invention can be modified and modified to a shape or shape well known in the art as required, such as round, square, oval.

  In addition, the order of mix | blending each composition in the said manufacturing method is only describing the example, The manufacturing method of the additive of this invention is not limited to the method of this mixing procedure.

 As mentioned above, the osteoporosis plaster containing the alendronate of the present invention as an active ingredient can be usefully used as a plaster that has superior durability, excellent safety and no side effects compared to conventional oral preparations.

      Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples and Experimental Examples.

Reference Example  1. Preparation and breeding conditions of experimental animals

In vitro experiment ( In Hairless mice (6-8 weeks old, males) and animal experiments ( in vitro ) vivo) respectively for purchase in rats (8-9 weeks old, 280-320g, male) (main) Sam tacos (Osan, Gyeonggi-do) was used within a week.

Reference Example  2. Drugs and reagents

Alendronate sodium (sodium alendronate) was used in Hwanin Pharmaceutical Co., Ltd., was used as the solvent isodium myostate (isopropyl myristate; IPM, Sigma Chemical Co. , St. Louis, MO, USA), propylene glycol monolaurate (PGML, Lauroglycol® 90), propylene glycol monocaprylate (PGMC, Capryol® 90) or diethylene glycol mono Ethylene glycol monoethyl ether (DGME, Transcutol® P) is a commercially available reagent grade propylene glycol (PG), ethanol, and calcium chloride from Gattefoss, Gennevilliers Cedex, France. , Disodium hydrogen phosphate, sodium hydroxide, hydrochloric acid and potassium phosphate, sodium acetate e), acetic acid, sodium citrate, and citric acid were purchased from Duksan Pure Chemical Co., Ansan, Gyeonggi-do, Korea.

Acetonitrile and methanol were used as HPLC grade. Water was purified water and passed through an ultrapure water production system (Puris MR-UP9000, MIRRE SCI) was used that was 18 MΩ or more. Lauric acid, oleic acid, linoleic acid, capric acid, caprylic acid and fluorescent derivatives 9-fluorenyl, used as skin permeation accelerators Methyl chloroformate (9-fluorenyl methylchloroformate; FMOC) was purchased from Sigma (St. Louis, MO, USA).

Pressure sensitive adhesives include Duro-Tak® 87-2196 {copolymer: acrylate-vinylacetate, functional group: -COOH, 45% solution of self-crosslinking acrylic copolymer, 3,000 cps, solubility parameter 16}, Duro-Tak® 87-2100 {copolymer: acrylate, functional group: -COOH, 51.5% solution of self-crosslinking acrylic copolymer, 8,500 cps, solubility parameter 16} and Duro-Tak® 87-2510 {copolymer: acrylate, functional group: -OH, 40.5% solution of non-crosslinking acrylic copolymer, 4,200 cps , Solubility parameter} was obtained from National Starch and Chemical Company (Bridgewater, NJ, USA) and used. Other reagents used commercial reagent grade.

Reference Example 3. Equipment and Devices

Liquid chromatography used a pump (Hitachi Pump, L-7100, Japan), a detector (Hitachi FL detector, L-7480, Japan) and a recording device (Hitachi integrator, D-7500, Japan). In addition, experimental devices and apparatuses such as a centrifuge (PLC-02, Cannic. INC. USA) and an adhesive coating machine (Sheen Instruments Ltd., London, UK) were used.

Example 1 Design of Alendronate Transdermal Absorbents

90 mg of sodium alendronate (69.0 mg as alendronate) was dissolved with each solvent (Experimental Example 1, Table 1) for 2 to 3 minutes at room temperature, and with pressure-sensitive adhesive solution (Duro-Tak® 87-2196) After mixing evenly, the siliconized release paper (Gelrofex ALU-PET 100μ-2S DR, 3M, USA) was coated with aluminum on one side of the polyester film, and then the silver release paper with silicon on both sides (weight 140 ± 7 g / m ² , thickness 95-106 μm) was cut and fixed horizontally on the glass plate, and applied to a width of 10 cm x 15 cm using a film casting machine (W100 mm, 0.01 mm, Sheen). .

The coating was left in an oven at 20 ° C. for 20 minutes to volatilize the solvent, and then left to dry at room temperature for 24 hours or more, and then adhered to a support (low allergy-producing flesh-colored polyester thin film, Scotchpak 1109, 3M, USA). Prepared (see FIG. 1).

Experimental Example 1. Preparation and Evaluation of Alendronate Plaster

The development of a matrix transdermal drug delivery system requires the selection of a suitable solvent that can increase the permeation rate and dissolve the drug. Therefore, the influence of single and co-solvent on release and permeation of the alendronate from the acrylic PSA pressure-sensitive adhesive matrix was examined to identify the optimum permeation promoting solvent or accelerator. (Gwak HS et al., Arch. Pharm. Res. , 26 , pp 644-648, 2003)

To dissolve the drug by selecting a solvent consisting of Preparation Example 1 in Table 1, three kinds of Duro-Tak® 87-2100, Duro-Tak® 87-2196 and Duro-Tak® 87-2510 as a skin-applied PSA adhesive The drug-containing adhesive matrix was prepared by varying the prescription conditions.

First, when the adhesive is mixed with the drug-containing solvent and dried, good skin adhesion and peeling force need to be ensured, so that 5 g of the pressure-sensitive adhesive, 2 mL of solvent, and 90 mg of alendronate sodium, which are dissolved in an organic solvent and supplied in a liquid state, are dispersed. The solution was mixed and evenly mixed, and evenly applied to the release paper by a manual applicator at a thickness of 0.45 mm and a width of 10 cm × 15 cm, and then the adhesive solvent was volatilized in an oven at 90 ° C. for 20 minutes, and then dried at room temperature for 24 hours or more. Thereafter, the support was brought into close contact to prepare a transdermal absorption preparation (see Table 1).

Production Example Alendronate Sodium Dose (mg) Vehicles Preparation Example 1 90 Propylene Glycol (PG) Preparation Example 2 90 Propylene Glycol Containing 3% Caprylic Acid Preparation Example 3 90 Propylene Glycol Containing 6% Caprylic Acid Preparation Example 4 90 Propylene Glycol Containing 10% Caprylic Acid Preparation Example 5 90 Propylene Glycol Containing 3% Capric Acid Preparation Example 6 90 Propylene Glycol Containing 6% Capric Acid Preparation Example 7 90 Propylene Glycol Containing 10% Capric Acid Preparation Example 8 90  Propylene Glycol Containing 3% Lauric Acid Preparation Example 9 90  Propylene Glycol Containing 6% Lauric Acid Preparation Example 10 90 Propylene Glycol Containing 10% Lauric Acid Preparation Example 11 90 Propylene Glycol Containing 3% Oleic Acid Preparation Example 12 90 Propylene Glycol Containing 6% Oleic Acid Preparation Example 13 90 Propylene Glycol Containing 10% Oleic Acid Preparation Example 14 90 Propylene Glycol Containing 3% Linoleic Acid Preparation Example 15 90 Propylene Glycol Containing 6% Linoleic Acid Preparation Example 16 90 Propylene Glycol Containing 10% Linoleic Acid Preparation Example 17 90 Ethanol Preparation Example 18 90 Propylene Glycol Monocaprylate (PGMC) Preparation Example 19 90 Propylene Glycol Monolaurite (PGML) Preparation Example 20 90 Isopropyl Myristate (IPM) Preparation Example 21 90 Diethylene glycol monoethyl ether (DGME) Preparation Example 22 70 Propylene Glycol Containing 6% Caprylic Acid Preparation Example 22 is used in clinical trials (Experimental Example 4).

    Among the used pressure-sensitive adhesives, Duro-Tak® 87-2100 and Duro-Tak® 87-2510 did not mix well with drug dispersion and were found to be inadequate for the design of transdermal absorbents. Accordingly, in the present invention, Duro-Tak® 87-2196 was used, and an allandronate transdermal absorption preparation was prepared by mixing drug dispersions in various solvents.

One week after the preparation was left at room temperature, microscopic observation was performed on the pressure-sensitive adhesive layer. As a result, no formation of crystals or precipitates was observed. These results confirmed that when the solubility in the solvent or the initial loading (high loading dose) is high, there will be no problems of emission and permeation reduction due to crystal growth that can occur during time-keeping.

Experimental Example  2. Alendronate Percutaneous Absorption  In vitro ( In in vitro )  Experiment

2-1. Create calibration curve

The standard product of Alendronate was dissolved in water to make 1,000 / mL, and this solution was refrigerated at 4 ° C as a standard stock solution, and the standard stock solution was appropriately diluted with water to obtain final concentrations of 50, 100, 200, 500, 1,000 2,000 ng / mL.

4 mL of water was added to 1 mL of each standard solution, and then mixed with 300 mL of an internal standard solution (150 / mL aqueous solution of pamideronide). 100 μl of 0.1 M calcium chloride was added thereto, and 0.1 M potassium phosphate was mixed. 100 μl of phosphate) was added to the mixture, and then 400 μl of 1 M sodium hydroxide was added and mixed.

The mixture was centrifuged for 5 minutes, the precipitate was taken, dissolved in 500 μl of 0.2 M acetic acid (acetic acid), diluted with 5 mL distilled water, and then mixed with 400 μl of 1 M sodium hydroxide, followed by centrifugation for 5 minutes.

The supernatant was discarded, and the remaining precipitate was dissolved in 1 mL of 0.2 M acetate buffer (pH 6.0), diluted with 2 mL distilled water, and 25 μl of acetic acid was added thereto.

The Bond Elute DEA solid phase extraction cartridge (Varian, USA) washed with distilled water was first washed with 1 mL distilled water, loaded with the mixture, and finally 0.2 M citric acid. Eluted with 1.6 mL of sodium citrate.

100 μl of sodium carbonate buffer (pH 11.9) and 100 μl of FMOC solution (0.25 mg / mL) were added to 270 μl of the eluate, followed by reaction at room temperature for 3 minutes, and 100 μl of 1 M citric acid. Was added to terminate the reaction.

A calibration curve was prepared from the chromatogram obtained by injecting 20 µl of the final reaction solution into HPLC, and the results are shown in Table 2 and FIG. 2 (Ptacek, P. et al., J. Chromatogr . B. , 767 , pp 111-). 116, 2002).

Concentration (ng / mL) Peak Area of Internal Standard (IS) Drug Peak Area (Drug) Rate (Drug / IS) 0 14891210 0 0 50 13497684 467020 0.0346 100 15476141 1201972 0.077666 200 16543980 2309540 0.1396 500 14981188 5405739 0.360835 1000 13073076 9707396 0.742549 2000 14569807 20391902 1.3996

2-2. Skin extraction

The dorsal skin of 6-8 week old hairless mouse males was harvested and used at 1.5 cm × 1.5 cm.

First, the hairless mice were anesthetized with ether in an airtight chamber and fixed on the operating table, and then subcutaneous fat, tissues and blood vessels were carefully removed so as not to damage the stratum corneum. The extracted skin was cut to a size of about 1.5 cm x 1.5 cm and immediately used for permeation test.

2-3. Emission test

     In order to examine the release pattern of the alendronate plaster, the dispersion 2 was prepared by dispersing the drug to a concentration of 90 mg / mL using a transdermal absorbent prepared in Preparation Examples 2, 3, 6, 7 and 11 of Example 1 as a solvent. A drug release test of the alendronate transdermal absorption preparation prepared as shown in Table 1 of Experimental Example 1 by mixing mL and 5 g of Duro-Tak® 87-2196 was performed.

The prepared plasters were cut into three pieces of 2 cm × 2 cm each, and the release paper was removed, and the drug-containing adhesive layer was attached to the Franz Cell Permeation System, and then mounted. 12 mL of pH 7.4 phosphate buffer was added to the test solution, followed by stirring with a magnetic star-head bar for 36 hours while maintaining a constant temperature of 37 ° C. Take 1 mL each at intervals of 3, 6, 9, 12, 24, and 36 hours, add 4 mL of water, mix with 300 μl of internal standard solution (150 / mL aqueous solution of pamideronate), and then follow the same procedure as the calibration curve for Experiment 2-1. After pretreatment of the sample with 20 μl of this solution, the peak area ratio was calculated from the chromatogram obtained by HPLC, and the cumulative emission amount was calculated. The results are shown in FIG. 3 (Gwak HS and Chun IK, Arch . Pharm . Res . , 24 , pp 578-583, 2001). (See Figure 3)

2-4. Skin Permeation Test Method

Ethanol, propylene glycol (PG), propylene glycol monocaprylate (PGMC), propylene glycol monolaurite (PGML), iso as a solvent on the dorsal skin of the hairless mice extracted in Experimental Example 2-2 Example 1 was mixed with 2 mL of a dispersion of sodium allandronate and 90 g of Duro-Tak® 87-2196 dispersed in propyl myristate (IPM) and diethylene glycol monoethyl ether (DGME) to 90 mg / mL. The Alendronate transdermal absorbents prepared as described above were attached in close contact with each other, mounted in a Franz Cell Permeation System maintained at 37 ° C, and then 12 mL of pH 7.4 phosphate buffer was added to the receptor cells. Put in.

1 mL of permeate was taken from the receptor cells every 3, 6, 9, 12, 24 and 36 hours over time and immediately supplemented with the same amount of receptor fluid. Add 1 mL of permeate to 4 mL of water and mix with 300 μl of internal standard solution (150 / mL pamideronate solution) .Pre-treat the sample in the same way as in the calibration curve of Experimental Example 2-1, and then 20 μl is injected into HPLC. The peak area ratio was calculated from the chromatogram obtained by the calculation of the cumulative permeation (Gwak HS et al., Arch . Pharm. Res . , 25 , pp 392-396, 2002).

As a result of the experiment, as shown in Table 3 and Table 4, propylene glycol showed the highest permeability among single solvents, and when fatty acid was added as permeation accelerator, permeation promotion effect was up to 2.9 times compared to propylene glycol alone. Could be confirmed.

Among the fatty acids used, 6% of saturated fatty acid caprylic acid (caprylic acid), 6% of unsaturated fatty acid oleic acid, and 10%, it was confirmed that a very high permeate flux (flux).

 Solution Permeation Parameters J _s (ng / ㎠ / hour) T _L (hours) Ethanol 29.19 ± 10.09 1.0 ± 0.2 Polypropylene Glycol (PG) 67.25 ± 5.77 0.5 ± 0.4 Propylene Glycol Monocaprylate (PGMC) 15.10 ± 7.42 NA Propylene Glycol Monolaurite (PGML) 32.90 ± 8.51 NA Isopropyl Myristate (IPM) 31.09 ± 8.20 1.3 ± 0.4 Diethylene glycol monoethyl ether (DGME)  15.82 ± 8.31 1.1 ± 0.4 Values are expressed as mean ± standard deviation (n = 3). -J _s : steady state flux, T _L : lag time-NA: not available

Solution Permeation Parameters J _s (ng / ㎠ / hour) T _L (hours) -(No enhancer) 67.25 ± 5.77  0.5 ± 0.4 Caprylic Acid (3%) 86.46 ± 1.34  3.2 ± 0.03 Caprylic Acid (6%)  195.68 ± 26.60 0.6 ± 0.3 Caprylic Acid (10%)  55.05 ± 6.80 0.2 ± 0.1 Capric acid (3%)  74.44 ± 11.47  1.3 ± 0.5 Capric acid (6%)  30.07 ± 23.25 1.2 ± 0.9 Capric acid (10%)  22.18 ± 8.42  0.9 ± 0.5 Lauric acid (3%) 62.51 ± 3.47  NA Lauric acid (6%) 32.75 ± 2.54  NA Lauric acid (10%) 51.61 ± 4.04  NA Oleic acid (3%)  46.28 ± 1.89  0.04 ± 0.04 Oleic acid (6%) 128.22 ± 10.28  0.4 ± 0.3 Oleic acid (10%) 142.63 ± 2.75  0.5 ± 0.3 Linoleic acid (3%)  41.06 ± 4.17 0.1 ± 0.1 Linoleic acid (6%) 74.29 ± 4.17 0.3 ± 0.1 Linoleic acid (10%)  79.78 ± 8.14 0.1 ± 0.1 Values are expressed as mean ± standard deviation (n = 3). -J _s : steady state flux, T _L : lag time-NA: not available

2-5. HPLC By law Of alendronate  dose

Analytical conditions for the samples were the Luna 5u C18 100A column (5 μm, 150 × 4.60 mm,) equipped with the SecurityGuard Guard Cartridge KJO-4282 (SecurityGuard Guard Catridges KJO-4282; C18, Phenomenex, USA) as a column. Phenomenex (USA) was used as a mobile phase, and a solution A, a mixture of acetonitrile and methanol (methanol) in a 1: 1 ratio, solution B, a mixture of 25 mM citric acid buffer solution and 25 mM phosphate buffer solution 1: 1, was converted into 28:72. After 13 minutes, A solution, B solution and methanol (methanol) were mixed at 20:28:52 and used for 13 minutes.

The mobile phase was flowed at a flow rate of 1.5 mL / min (min) to obtain a peak area ratio from the chromatogram obtained by detection at an excitation wavelength of 260 nm and a fluorescent wavelength of 310 nm. One example of the results is shown in Table 5 below. (Kenneth W. Renton. Et al., Original Research . , 18 , pp98-101, 2004).

Time (h) Peak Area of Internal Standard (IS) Drug Peak Area (Drug) Rate (Drug / IS) 3 16959764 224712 0.01325

The unknown drug concentration was calculated using the following equation 1 from the calibration curve of FIG. 2 using the peak area ratio (IS) of the internal standard obtained by HPLC analysis and the peak area ratio (Drug) between drugs.

 As a result, it was confirmed that the sample concentration (Concentration) of the third hour of Preparation Example 3 of Table 5 was about 10.357 ng / mL.

Experimental Example 3. Animal of the prepared alendronate transdermal absorption preparation ( In vivo)  Experiment

3-1. Create calibration curve

Alendronate sodium standard was dissolved in water to make 1,000 / mL, and this solution was refrigerated as a standard stock solution. After diluting the standard stock solution properly, it was mixed with urine. 200, 500, 1,000 and 2,000 ng / mL.

300 μl of the internal standard solution (150 / mL aqueous solution of pamideronate) was added thereto, followed by mixing, 100 μl of 0.1 M calcium chloride, and 100 μl of 0.1 M potassium phosphate, followed by mixing. 400 μl M sodium hydroxide was added and mixed.

The mixture was centrifuged for 5 minutes, the precipitate was taken, dissolved in 500 μl of 0.2 M acetic acid (acetic acid), diluted with 5 mL distilled water, and then mixed with 400 μl of 1 M sodium hydroxide, followed by centrifugation for 5 minutes.

The supernatant was discarded, and the remaining precipitate was dissolved in 1 mL of 0.2 M acetate buffer (pH 6.0), diluted with 2 mL distilled water, and 25 μl of acetic acid was added thereto.

The Bond Elute DEA solid phase extraction cartridge (Varian, USA) washed with distilled water was first washed with 1 mL distilled water, loaded with the mixture, and finally 0.2 M citric acid. Eluted with 1.6 mL of sodium (sodium citrate).

100 μl of sodium carbonate buffer (pH 11.9) and 100 μl of FMOC solution (0.25 mg / mL) were added to 270 μl of the eluate, followed by reaction at room temperature for 3 minutes, and 100 μl of 1 M citric acid. Was added to terminate the reaction.

A calibration curve was prepared from the chromatogram obtained by injecting 20 µl of the final reaction solution into HPLC, and the results are shown in Table 6 and FIG. 4 (Ptacek, P. et al., J. Chromatogr . B. , 767 , pp 111-). 116, 2002).

Concentration (ng / mL) Peak Area of Internal Standard Drug Rate (Drug / IS) 0 16629323 0 0 50 18366232 416272 0.02266507 100 17761632 841220 0.04736164 200 17242633 2543918 0.14753652 500 13632934 4509471 0.33077773 1000 13153055 8794506 0.66862839 2000 9584386 13260968 1.3836012

In the HPLC method applied to the permeation test of the present invention, the concentration of the alendronate sodium was changed to 50-2,000 ng / mL to obtain a chromatogram, and a peak area ratio was obtained. As a result, a calibration curve was formed. y = 0.0007x-0.0095, R ² = 0.9999) was obtained.

3-2. Accuracy, Precision and Quantitative Limits of Alendronate Methods

As in Experiment 3-1, the urine concentrations of the alendronate quality standard (alendronate quality control sample) were treated to 50, 100, 200, 500, 1,000 and 2,000 ng / mL, and then the test was performed five times a day. Daily reproducibility was obtained, and the experiment was continuously performed for 5 days to obtain daily reproducibility.

Accuracy was calculated as a percentage of the ratio of the mean value of the concentration quantified by the calibration curve divided by the known concentration. Quantitative limits were determined at concentrations of more than 10 times S _N.

Samples of the Allendronate standard at six concentrations of 50, 100, 200, 500, 1,000 and 2,000 ng / mL in urine were sampled in the following manner with reference to the method described in the literature (Ptacek, P. et al., J. Chromatogr.B. , 767 , pp 111-116, 2002).

The standard deviation of the peak area ratios of the alendronate and the internal standard was calculated as a percentage (%) divided by the average value of the peak area ratios of the alendronate and the internal standard. Intra-day precision (expressed in% CV) was obtained 5 times a day, and daily precision (expressed in% CV) was obtained by repeating the 5-day experiment.

The average value of the concentration quantified by the calibration curve was calculated as the percentage (%) of the ratio divided by the known concentration.

Signal to noise ratio (S _N ratio) on the chromatogram was determined to a concentration satisfying the conditions of more than 10, precision of 20% or less, and accuracy of 80-120%.

As a result, as shown in Table 7 below, the precision CV (%) of this analysis method was 12.4% in daily precision and 7.0% in daily precision at the lowest concentration (50 ng / mL) of the calibration curve. The average daily precision at the lowest concentration of 14.6% and the calibration curve was 13.4%. The average accuracy was 98.9%, the accuracy at the lowest concentration of the calibration curve was 96.8, and the quantitative limit concentration of alendronate was at least 50 ng / mL.

Concentration (ng / mL) Precision (C.V.%) accuracy(%) In days (n = 5) Days (n = 5) 50 7.0 13.4 96.8 100 11.2 14.6 91.5 200 12.4 10.3 98.4 500 10.2 7.5 102.7 1000 5.4 4.0 102.4 2000 6.1 5.4 101.8 -C.V. (%) = Standard deviation (S.D.) / Mean * 100

3-3. Alendronate Percutaneous Absorption  After oral administration Urinary  Drug amount measurement

3-3-1. Oral administration

Male rats weighing 280-320 g were fed for 12 hours under constant conditions, and water was freely supplied before the experiment.

Six healthy rats were made into one group to prepare a 0.1% aqueous solution of alendronate sodium, and then orally administered using a sonde in an amount of 30 mg / kg.

After oral administration, urine was collected for 12, 24, 36 and 48 hours. Chromatography obtained by mixing 300 μl of the internal standard solution (150 / mL pamideronate solution) with 5 mL of the urine solution, and pretreating the sample in the same manner as in the calibration curve of Experimental Example 3-1. The peak area ratio was quantified from grams, and one example of the result was shown in Table 8 (Ptacek, P. et al., J. Chromatogr . B. , 767 , pp 111-116, 2002).

Time (hours) Peak Area of Internal Standard (IS) Drug Rate (Drug / IS) 36 4076301 7347 0.001802

The unknown drug concentration was calculated using the following equation (2) from the calibration curve of FIG. 4 using the peak area ratio (IS) of the internal standard obtained by HPLC analysis and the peak area ratio between drugs.

As a result, it was confirmed that the concentration in the urine (Concentration) of 36 hours after oral administration of Table 8 is about 16.15 ng / mL.

      In addition, after oral administration, the urine excretion with time of the alendronate in the rat is shown in FIG. As a result, the amount of accumulated excretion drug in urine up to 48 hours was 604.9 ng, it was confirmed that a significant portion was detected within 12 hours (see Table 10).

3-3-2. Transdermal Administration

After carefully removing the flanks of male rats weighing 280-320 g with an animal razor, the prepared transdermal absorbents were attached to a size of 3 cm x 4 cm, followed by urination for 12, 24, 48 and 72 hours. It was.

5 μl of urine solution was mixed with 300 μl of the internal standard solution (150 / mL pamideronate solution), and then 20 μl of this solution was injected into HPLC by pretreatment of the sample in the same manner as in the calibration curve of Experimental Example 3-1. The peak area ratio was determined from the chromatogram and quantified. One example of the results is shown in Table 9 (Ptacek, P. et al., J. Chromatogr. B. , 767 , pp 111-116, 2002).

Time (hours) Peak Area of Internal Standard Drug Rate (Drug / IS) 24 13701876 120701 0.010269

The unknown drug concentration was calculated using the following equation (3) from the calibration curve of FIG. 4 using the peak area ratio between IS and the drug obtained by HPLC analysis.

     As a result, as shown in Table 9, the concentration of urine (Concentration) at 24 hours of Preparation Example 3 was confirmed to be about 28.24 ng / mL.

      In addition, after applying the prepared transdermal absorbent preparations (Preparation Examples 1-1 to 1-7 and 1-11 to 1-16 of Experimental Example 1) to the rats, the total amount of excretion in the urine for 72 hours is shown in Table 10 Indicated.

As a result, as shown in Table 10, the total amount of drug detected in urine was the highest as 1042.9 ng when 6% caprylic acid was used as fatty acid, followed by capric acid ) 6% net. In addition, propylene glycol (PG) alone and 3% and 10% caprylic acid, 3% and 6% capric acid, 3% and 10% oleic acid and 6% linoleic acid are added to propylene glycol. In one case, it was confirmed that the total amount of excretion over 72 hours represents more than 200 ng.

In general, for 6% caprylic acid, which showed the highest drug amount, the drug began to be detected after 12 hours, and the detection amount increased over time, whereas for 6% capric acid, the initial high Excretion was decreased until 48 hours and increased again at 72 hours. In the case of oral administration, it showed high absorption up to 12 hours at the beginning, but only 1.8 ng was absorbed at 48 hours, and even in the case of PG alone, the absorption pattern was even during the initial 48 hours (see FIG. 5).

Vehicles Accumulation (ng) Oral 604.9 ± 93.3 Example 1-Alendronate Transdermal Absorbent Prepared Using Polypropylene Glycol (PG) of Preparation Example 1 235.3 ± 37.8  Alendronate Transdermal Absorbents Prepared Using Polypropylene Glycol Containing 3% Caprylic Acid of Example 1-Preparation Example 2 282.4 ± 52.6  Alendronate Transdermal Absorbents Prepared Using Polypropylene Glycol Containing 6% Caprylic Acid of Example 1-Preparation Example 3 1042.9 ± 226.7 Alendronate Transdermal Absorbents Prepared Using Polypropylene Glycol Containing 10% Caprylic Acid of Example 1-Preparation Example 4 209.6 ± 32.2  Alendronate Transdermal Absorbents Prepared Using Polypropylene Glycol Containing 3% Capric Acid of Example 1-Preparation 5 232.9 ± 34.2 Example 1- Alendronate Percutaneous Absorbent Prepared Using Polypropylene Glycol Containing 6% Capric Acid of Preparation Example 6 516.4 ± 177.9 Example 1- Alendronate Transdermal Absorbents Prepared Using Polypropylene Glycol Containing 10% Capric Acid of Preparation Example 7 114.6 ± 68.5  Example 1- Alendronate Transdermal Absorbent Prepared Using Propylene Glycol Containing 3% Oleic Acid of Preparation Example 11 202.4 ± 92.0 Example 1- Alendronate Transdermal Absorbent Prepared Using Propylene Glycol Containing 6% Oleic Acid of Preparation Example 12 73.4 ± 10.2 Example 1-Alendronate Percutaneous Absorbent Prepared Using Propylene Glycol Containing 10% Oleic Acid of Preparation Example 13 219.7 ± 30.6 Example 1-Alendronate Transdermal Absorbents Prepared Using Propylene Glycol Containing 3% Linoleic Acid of Preparation Example 14 82.0 ± 25.4 Example 1-Alendronate Transdermal Absorbents Prepared Using Propylene Glycol Containing 6% Linoleic Acid of Preparation Example 15 219.4 ± 31.2 Example 1-Alendronate Percutaneous Absorbent Prepared Using Propylene Glycol Containing 10% Linoleic Acid of Preparation Example 16 58.5 ± 10.1 Urine was collected at 72 hours except oral (48 hours). The figures are expressed as mean ± standard error (n = 6).

3-4. HPLC By law Of alendronate  dose

Analytical conditions for the samples were obtained by using a CAPCELL PAK C18 UG 120 column (5, 150 × 4.60 mm, Shiseido) equipped with a SecurityGuard Guard Catridges KJO-4282; C18, Phenomenex, USA. As a mobile phase, solution A, which mixed acetonitrile and methanol in a 1: 1 ratio, and solution B, in which a 25 mM citric acid buffer solution and a 25 mM phosphate buffer solution were mixed 1: 1, flowed at 28:72 for 13 minutes, and then 13 minutes A solution, B solution and methanol (methanol) was mixed at 20:28:52 and used again for 13 minutes.

The mobile phase was flowed out at a flow rate of 1.5 mL / min (min) and detected at an excitation wavelength of 260 nm and a fluorescent wavelength of 310 nm.

Experimental Example 4. Clinical Experiment

Three healthy volunteers (female 24, female 28, female 45) were prepared with percutaneous absorbent preparation (53.8 mg as alendronate) and 70 mg (allendronate) containing 70 mg of alendronate sodium in preparation 22 of Example 1 above. After administration of 70 mg), the amount of drug in urine over time was measured.

There was a one-week holiday between the transdermal and oral administrations, and blank urine was collected before application of the drug to determine whether the drug remained. The urine analysis was used mutatis mutandis in the animal ( in vivo) experiment of Experimental Example 3.

In animal experiments, transdermal absorption using 6% caprylic acid, which showed the highest urinary drug content, was applied to humans to measure urine drug content over time.

As a result, as shown in Figure 6, the total amount of excreted drug up to 72 hours oral was 237.20 g, it was confirmed that 112.03 in the case of transdermal absorption preparations. In the case of oral, it is not detected after 24 hours and can be regarded as the total amount absorbed into the body. However, in the case of transdermal absorbents, the total application time up to 72 hours after analysis was limited to 72 hours. Considering that the amount of excretion increases with time, it is considered that more amount can be detected in urine when applied for a longer time. In addition, the amount of alendronate in the oral preparation was 70 mg, whereas the amount of alendronate in the percutaneous absorption preparation was calculated as 53.8 mg, so that the relative bioavailability was expected to increase further.

From the above experiments, the possibility of designing the percutaneous absorbent of alendronate was found, which is optimal when the various concentrations of fatty acids were mixed with alcohols such as propylene glycol. Could be confirmed.

     1 is a cross-sectional view of the alendronate transdermal absorption agent,

     2 is a diagram showing a calibration curve in an aqueous solution of alendronate,

     Figure 3 is a diagram showing the results of the release test for alendronate transdermal absorption agent,

     4 is a diagram showing a calibration curve in urine of alendronate,

     FIG. 5 shows the time-lapse of alendronate when administered an oral formulation and propylene glycol alone, propylene glycol containing 6% caprylic acid and propylene glycol containing 6% capric acid, in rats. Is a diagram showing the amount of excretion in urine,

     FIG. 6 is a diagram showing urinary excretion with time of Alendronate after oral administration of Alendronate and administration of an Alendronate transdermal absorbent prepared using propylene glycol containing 6% caprylic acid in a healthy volunteer.

Claims (6)

 Plaster composition for the prevention and treatment of osteoporosis, which contains alendronate as an active ingredient, and contains a solvent, a transdermal absorption accelerator, and a pressure-sensitive adhesive. Prevention of allendronate-containing osteoporosis, characterized in that the active ingredient contains 0.1 to 10% of allenronate, 5 to 50% of solvent, 0.1 to 10% of transdermal absorption accelerator, and 30 to 90% of pressure-sensitive adhesive. Therapeutic plaster composition. The method of claim 2, wherein the solvent is at least one solvent selected from the group consisting of isopropyl myristate, propylene glycol monolaurate, propylene glycol monocaprylate, diethylene glycol monoethyl ether, propylene glycol or ethanol Alendronate-containing plaster composition. According to claim 2, wherein the percutaneous absorption enhancer is selected from the group consisting of lauric acid, oleic acid, linoleic acid, capric acid, caprylic acid, myristic acid, oleyl alcohol, isopropyl myristate or propylene glycol laurate Alendronate-containing plaster composition, characterized in that the percutaneous absorption accelerator. The pressure-sensitive adhesive of claim 2, wherein the pressure-sensitive adhesive is at least one pressure-sensitive adhesive selected from an acrylic pressure-sensitive adhesive made of an acrylate polymer, a rubber-based pressure-sensitive adhesive in which additives such as petroleum resin, rosin paper, and oils are added to natural or synthetic rubber, and a water-soluble pressure-sensitive adhesive. Alendronate-containing plaster composition, characterized in that.  Alendronate, comprising: a support (1) that prevents drug permeation, an active ingredient dissolved in a solvent, a drug layer (2) containing a pressure-sensitive adhesive, and a release layer (3) treated with an allandronate Plaster for the prevention and treatment of containing osteoporosis.

Images