TW201620508A - Microneedle - Google Patents

Abstract

Provided is a microneedle containing a bioactive substance, one or more amino acids selected from the group consisting of arginine and histidine, and an acid having a melting point of 40 DEG C or higher.

Description

Microneedle

The present invention relates to microneedle needles.

The transdermal administration preparation of the physiologically active substance can be administered even if the user has difficulty swallowing, and does not require water. Method. In addition, the method of administering a physiologically active substance by percutaneous administration is easier for the user to perform the administration than the injection, even if the medical practitioner is not surrounded. Easy method of administration. In such a transdermal administration preparation, for example, a patch or a microneedle device can be cited.

The conventional microneedle device is manufactured by coating a microprotrusion with a coating solution containing a physiologically active substance in a microneedle device having microprotrusions (Patent Document 1).

In recent years, methods for reducing skin irritation and waste by using microscopic protrusions as self-dissolving materials have been developed. Examples of the self-dissolving microneedle include a microneedle using a mixture of maltose and dextran as a main component, or a water-soluble high such as sodium carboxymethylcellulose. Microneedle needle composed of molecules (Patent Documents 2 and 3).

[Previous Technical Literature] [Patent Document]

[Patent Document 1] Japanese Patent Laid-Open Publication No. 2008-528509

[Patent Document 2] Japanese Patent Laid-Open Publication No. 2005-152180

[Patent Document 3] Japanese Patent No. 4829497

However, in the microneedle needle described in Patent Document 2, after mixing maltose with a physiologically active substance, it is necessary to heat the melting point of maltose or more to form a lancet shape, and it is difficult to use a physiologically active substance having poor thermal stability. . Further, in the case where the physiologically active substance is a protein, since the physiologically active substance and the maltose undergo a Maillard reaction, the preservation stability of the physiologically active substance is lowered.

On the other hand, in the microneedle needle described in Patent Document 3, the water-soluble polymer (carboxymethylcellulose sodium) which is a material of the microneedle is deformed in the drying step, and it may be difficult to stably manufacture the microneedle.

The present invention has been made in view of the above circumstances to provide a self-dissolving type microneedle which is excellent in moldability and strength.

As a result of intensive review, the inventors of the present invention found a microneedle system containing a physiologically active substance, one or more amino acids selected from the group consisting of arginine and histidine, and an acid having a melting point of 40 ° C or higher. It is excellent in moldability and strength, and has self-solubility, and has completed the present invention.

Preferably, the acid is one or more selected from the group consisting of citric acid, lactic acid, tartaric acid, succinic acid, phosphoric acid and aspartic acid, more preferably selected from the group consisting of citric acid, tartaric acid and aspartic acid. One or more of the acids. The above amino acid is preferably arginine.

The present invention also provides a method of producing the above microneedle.

Further, the present invention provides a microneedle device comprising a microneedle on a substrate, the microneedle comprising a physiologically active substance, an amino acid selected from the group consisting of arginine and histidine, and a melting point It is an acid of 40 ° C or more.

Furthermore, the present invention also provides a method of using a microneedle device comprising applying the above microneedle device to a skin of a subject to be administered a physiologically active substance contained in the microneedle.

The microneedle according to the present invention is excellent in moldability and strength, and has self-solubility, and is excellent in storage stability of a physiologically active substance. In addition, since the microneedle needle of the present invention is a self-dissolving microneedle, it can contain a physiologically active substance inside the microneedle, and is reliable in terms of skin permeability of a physiologically active substance compared to a conventional microneedle. Higher, and remains in the skin even if the microneedle is broken, such as during use In this case, it will gradually dissolve, thus reducing the physical burden on the user. Further, according to the microneedle of the present invention, the microneedle is rapidly dissolved after being punctured to the skin, so that the application time can be shortened compared to the conventional microneedle.

1‧‧‧Microneedle device

2‧‧‧Substrate

3‧‧‧Microneedle

Fig. 1 is a photograph showing the moldability of a microneedle according to an embodiment of the present invention.

Fig. 2 is a schematic view showing a microneedle device according to an embodiment of the present invention.

Figure 3 is a graph showing the results of a skin permeability test.

Figure 4 is a graph showing the results of the strength test (2).

Fig. 5 is a graph showing the results of the stability test of physiologically active substances.

Figure 6 is a graph showing the results of the dissolution test.

The microneedle according to an embodiment of the present invention contains a physiologically active substance, an amino acid selected from the group consisting of arginine and histidine, and an acid having a melting point of 40 ° C or higher.

In the present specification, among the components constituting the microneedle, the components other than the physiologically active substance are collectively referred to as "the material of the microneedle."

As the physiologically active substance which can be used in the present embodiment, For example, it may be not only a low molecular compound but also a polymer compound such as a peptide, a protein, a DNA, or an RNA, a vaccine, or the like. Examples of the physiologically active substance include naltrexone, cetrorelix acetate, taltirelin, nafarelin acetate, and prostaglandin A1. Alprostadil, alpha-interferon, zolmitriptan, beta-interferon, erythropoietin, follicle-stimulating beta, follicle-stimulating alpha, G-CSF, GM-CSF, human villi Gonadal stimulating hormone, leutinizing hormone, salmon calcitonin, glycoside, GNRH antagonist, insulin, human growth hormone, filgrastim, heparin, low molecular weight heparin, somatotropin Incretin, GLP-1 derivatives, and the like. In the microneedle needle according to the present embodiment, the physiologically active substance may be used singly or in combination of two or more.

The content of the physiologically active substance may be 0.1% by mass or more, or may be 1% by mass or more, or may be 10% by mass or more based on the total mass of the microneedle. In addition, the content of the physiologically active substance may be 90% by mass or less, or may be 75% by mass or less, or may be 50% by mass or less based on the total mass of the microneedle.

Examples of the amino acid which can be used in the group selected from the group consisting of arginine and histidine in the present embodiment include L-arginine, D-arginine, and L-histamine. D-histidine. A preferred amino acid is L-arginine or L-histamine. In the microneedle needle according to the embodiment, the amino acid may be used singly or in combination of two or more.

Further, the above-mentioned amino acid may be used as a free form, or a known acid addition salt may be used, or a salt-formed state may be used after desalting by a method known to those skilled in the art. Commercially available products can be used as the above-mentioned amino acid, and those obtained by chemical synthesis can also be used.

The content of the amino acid or more selected from the group consisting of arginine and histidine in the present embodiment may be 1 to 99% by mass, preferably 5, based on the total mass of the microneedle. ~95% by mass, more preferably 10 to 90% by mass. Further, when two or more kinds of amino acids are used in combination, the above content corresponds to the total amount of the amino acids used in the microneedle.

The acid which can be used in the present embodiment, wherein the melting point is 40° C. or higher, may be, for example, one or more selected from the group consisting of citric acid, lactic acid, tartaric acid, succinic acid, phosphoric acid, and aspartic acid. Specific examples of such acids are citric acid, L-lactic acid, D-lactic acid, L-tartaric acid, D-tartaric acid, succinic acid, phosphoric acid, L-aspartic acid, and D-aspartic acid. In the microneedle needle according to the embodiment, the acid may be used singly or in combination of two or more. The acid having a melting point of 40 ° C or higher in the present embodiment may be an acid having a melting point of 50 ° C or higher, or an acid having a melting point of 150 ° C or higher. Further, the measurement of the melting point can be carried out, for example, in accordance with the description of the melting point measurement method of the 15th revised Japanese Pharmacopoeia general test method. When the microneedle is prepared by mixing an acid having a melting point of 40 ° C or higher with arginine or histidine, the moldability of the microneedle is excellent.

The content of the acid having a melting point of 40 ° C or higher in the present embodiment can be 1 to 99% by mass based on the total mass of the microneedle needle. Preferably, it is 5 to 95% by mass, more preferably 10 to 90% by mass. Further, when two or more kinds of acids are used in combination, the above content corresponds to the total amount of the acid used in the microneedle.

In the microneedle using arginine as the amino acid, the preferred molar ratio of acid to arginine differs depending on the type of acid. In the case where arginine is used as the amino acid, as the acid having a melting point of 40 ° C or higher, lactic acid, tartaric acid, citric acid, succinic acid, phosphoric acid or aspartic acid is preferably used. In the case of such a combination, a microneedle needle having more excellent moldability can be obtained. In the case where the acid is lactic acid, the molar number of arginine relative to the molar number of the acid is preferably from 0.5 to 4, more preferably from 0.5 to 3. In the case where the acid is tartaric acid, the molar number of arginine relative to the molar number of the acid is preferably from 0.5 to 6, more preferably from 0.5 to 5. In the case where the acid is citric acid, the number of moles of arginine relative to the number of moles of the acid is preferably from 0.5 to 9, more preferably from 0.5 to 8. In the case where the acid is succinic acid, the molar number of arginine relative to the molar number of the acid is preferably from 0.5 to 6, more preferably from 1 to 5. In the case where the acid is phosphoric acid, it is preferably from 0.5 to 9, more preferably from 2 to 5. In the case where the acid is aspartic acid, the molar number of arginine is preferably from 1 to 4 with respect to the molar number of the acid. When the number of moles of arginine is within the above range with respect to the molar number of the acid, the microneedle is more excellent in moldability and strength.

In the present specification, the excellent microneedle forming property means a microneedle which maintains a transparent amorphous shape in the case of observing the appearance of the formed microneedle by a microscope.

In the microneedle using histamine as the amino acid, the preferred molar ratio of acid to histidine differs depending on the type of acid. In making In the case where histidine is used as the amino acid, as the acid having a melting point of 40 ° C or higher, tartaric acid, citric acid or aspartic acid is preferably used. In the case of such a combination, a microneedle needle having more excellent moldability can be obtained. In the case where the acid is tartaric acid, the molar number of histidine relative to the molar number of the acid is preferably from 0.5 to 3, more preferably from 0.5 to 2. In the case where the acid is citric acid, the number of moles of histidine relative to the number of moles of the acid is preferably from 0.5 to 4, more preferably from 0.5 to 2. In the case where the acid is aspartic acid, the molar number of histidine relative to the molar number of the acid is preferably 0.5 to 1. When the number of moles of histidine is in the above range with respect to the number of moles of acid, the moldability and strength of the microneedle are more excellent.

The shape of the microneedle needle according to the present embodiment may be, for example, a polygonal pyramid shape such as a triangular pyramid or a quadrangular pyramid; or a polygonal pyramidal shape such as a triangular pyramid or a quadrangular pyramid; a conical shape; and a truncated cone shape. It is preferably conical in terms of ease of puncture and reduction in pain at the time of application. Further, the microneedle needle according to the embodiment is preferably amorphous. When the microneedle is amorphous, the strength tends to be excellent, and the solubility tends to be improved.

In the microneedle needle according to the present embodiment, the length in the direction in which the puncture is performed is preferably 10 μm to 2 mm, more preferably 50 μm to 1 mm. When the length in the direction in which the puncture is performed is 10 μm or more, the reliability at the time of puncture is further improved. Further, in the microneedle needle according to the present embodiment, the area of the surface in contact with the substrate is preferably 100 to 10000 μm ² , more preferably 200 to 5000 μm ² . When the area of the surface in contact with the substrate is 10000 μm ² or less, the pain can be further reduced.

<Method for Producing Microneedle Needle According to the Present Embodiment>

The microneedle needle of the present embodiment can be produced, for example, by the following method.

(1) A mold for making a microneedle is produced by a method well known to those skilled in the art.

(2) a physiologically active substance, an amino acid selected from the group consisting of arginine and histidine, and an acid having a melting point of 40 ° C or higher are dissolved in an arbitrary amount of water, and added dropwise to the above mold. After that, it was dried overnight at 37 ° C to prepare a microneedle of the present embodiment. Further, it is preferable that the mold is immersed in water, degassed under reduced pressure, and then taken out and used to remove air bubbles of the mold.

When the microneedle needle of the present embodiment is produced, the mold can be produced by a method known to those skilled in the art. Further, the material of the above mold may be, for example, a metal or a non-metal. As a non-metal mold, a polyoxy oxy rubber is mentioned, for example.

Further, the microneedle of the present embodiment may be a mixture of a physiologically active substance, an amino acid selected from the group consisting of arginine and histidine, and an acid having a melting point of 40 ° C or higher, and heated to A liquid having a suitable temperature and ensuring fluidity flows into the mold, or is cast and gradually cooled.

As shown in FIG. 2, the microneedle device 1 according to an embodiment of the present invention has a plurality of the microneedle pins 3 disposed on the substrate 2. Regarding the number of microneedle needles 3, those skilled in the art can arbitrarily set them.

As the substrate 2 according to the present embodiment, a material well known to those skilled in the art can be used. The substrate 2 and the microneedle 3 can also be used In this case, the substrate 2 and the microneedle 3 are made of the same material.

Further, the microneedle device according to the embodiment may further include an applicator. As the applicator, those skilled in the art can be used.

[Examples]

The microneedle needles of the examples, comparative examples and reference examples were produced in the following manner. Further, in the present embodiment, as long as there is no special note, as citric acid, lactic acid, tartaric acid, succinic acid, phosphoric acid, hydrochloric acid, glacial acetic acid, and aspartic acid, anhydrous citric acid, L-lactic acid, and L-tartaric acid are used, respectively. , succinic acid, anhydrous phosphoric acid, concentrated hydrochloric acid (37% by mass aqueous hydrogen chloride solution), glacial acetic acid and L-aspartic acid. Further, as arginine and histidine, L-arginine and L-histamine are used, respectively.

The mold made of polyoxyxene rubber was immersed in water, deaerated under reduced pressure, and taken out from the water. Next, an aqueous solution of an amino acid and an acid (additive concentration: 15 w/v%) was added dropwise to the above-mentioned mold, and dried at 37 ° C overnight to obtain a microneedle. The obtained microneedle needle was evaluated using the following test.

1. Formability test (1)

The microneedle needle was prepared by a combination of arginine and citric acid, a combination of arginine and hydrochloric acid, or only arginine or only citric acid, and the appearance of the obtained microneedle was compared.

A photograph of a microneedle prepared by a combination of arginine and citric acid, a combination of arginine and hydrochloric acid, or only arginine or only citric acid is shown in Fig. 1. The microneedle needle made of a combination of arginine and citric acid (Fig. 1 (a)) is in a transparent amorphous state, and has good moldability and maintains a conical shape. On the other hand, the combination of arginine and hydrochloric acid (Fig. 1 (b)), arginine alone (Fig. 1 (c)), citric acid alone (Fig. 1 (d)), the microneedle needles are After drying, precipitation of crystals was confirmed, and moldability was poor, and the shape of a conical shape could not be maintained.

2. Formability test (2)

Using one or more amino acids selected from the group consisting of arginine and histidine, and a group selected from the group consisting of citric acid, lactic acid, tartaric acid, succinic acid, phosphoric acid, hydrochloric acid, glacial acetic acid, and aspartic acid One or more kinds of acids are used to produce microneedle needles by the above-described production method. Next, using a microscope, the state of the microneedle obtained was observed, and whether or not the crystal of the salt was precipitated and whether the microneedle was molded was evaluated.

In the case of using arginine, and lactic acid, tartaric acid, citric acid, succinic acid, phosphoric acid or aspartic acid, a transparent amorphous microneedle can be produced, whereas arginine, hydrochloric acid or ice is used. In the case of acetic acid, microneedle needles cannot be obtained. Further, in the case of using histidine, tartaric acid, citric acid or aspartic acid, a transparent amorphous microneedle can be produced, whereas in the case of using histidine, hydrochloric acid or glacial acetic acid, Obtain a microneedle.

3. Skin permeability test

In this test, a microneedle device having 100 microneedle needles prepared by mixing edible red No. 40 (also referred to as R40 or Allura Red AC), arginine and citric acid in the ratio shown in Table 1 was used. (Reference example 1). In addition, the numerical value of Table 1 shows the mass ratio of each component used. In addition, Edible Red No. 40 was used as a substitute for physiologically active substances.

The skin permeability test was carried out in accordance with the following method.

The microneedle device (Reference Example 1) was applied to humans to take out the skin using an applicator. Next, the skin to which the microneedle device was applied was fixed in a FRANZ-type tank, and the receiving liquid after 24 hours was collected, and the concentration of the edible red No. 40 was measured by HPLC to measure the amount of permeation. Further, phosphate buffered physiological saline (PBS) was used in the receiving solution of the test.

In the case where the applicator was used to apply the microneedle to the skin ("applicator application" in Fig. 3), the cumulative permeation amount of the physiologically active substance after 24 hours from the application was about 5 μg. It was confirmed that the individual microneedle needles were quickly dissolved, and the physiologically active substance (food red No. 40) contained in the microneedle needle was released, showing sufficient skin permeability. On the other hand, in the absence of an applicator, only the microneedle is exposed to the skin. (In Fig. 3, "unpressed"), the microneedle did not penetrate the epidermis, and the cumulative permeation amount of the physiologically active substance after 24 hours from the application was 0 μg.

4. Shape stability test

Observe a mixture of arginine and citric acid, maltose, polyvinylpyrrolidone (PVP-K12), polyvinylpyrrolidone (PVP-K90), amylopectin or gelatin in a 15% aqueous solution at 40 ° C. The change in shape.

In the case of a microneedle prepared by polyvinylpyrrolidone (PVP-K90), amylopectin or gelatin, the shape of the microneedle is changed when it is dried overnight at 40 ° C. In the case of a microneedle needle prepared by a mixture of arginine and citric acid, maltose or polyvinylpyrrolidone (PVP-K12), the shape of the microneedle does not change.

5. Strength test (1)

The amino acid was mixed with an acid at a ratio described in Table 2 to produce a microneedle. A urethane sheet (manufactured by Exseal Corporation, trade name: HyperGel sheet hardness 50, Shore-C hardness: 50) was coated with an aluminum foil, and applied using an applicator having the obtained microneedle, and the microneedle was evaluated. Whether it runs through the aluminum foil.

The results are shown in Table 2. The microneedles of Reference Examples 2 to 11 all had strength throughout the aluminum foil.

6. Strength test (2)

The microneedles of Reference Examples 12 to 14 were produced in accordance with Table 3, and the maximum stress at the time of breakage of the obtained microneedle was measured using a texture analyzer (TA.XT plus texture analyzer, manufactured by Stable Micro Systems).

The results are shown in Table 3 and Figure 4. The microneedle needle prepared by the combination of arginine and citric acid is inferior to the microneedle prepared by maltose, but is equivalent to the microneedle prepared by polyvinylpyrrolidone (PVP-K12). , showing the strength required for puncture.

7. Stability test of physiologically active substances

The microneedle needles of Examples 1 to 6 and Comparative Examples 1 to 6 were produced in accordance with Tables 4 and 5. In addition, the numerical values shown in Table 4 and Table 5 mean the mass part. The mass of the physiologically active substance contained in the microneedle needle immediately after the production is a fully automatic enzyme immunoassay device (AIA-360, manufactured by Tosoh Co., Ltd.) in the case where the physiologically active substance is insulin or human growth hormone. It is calculated that when the physiologically active substance is lixisenatide, it is calculated by HPLC. The value obtained by dividing the mass of the obtained physiologically active substance by the mass of the physiologically active substance used for production is recorded as "the content (%) of the initial physiologically active substance". Next, after the obtained microneedle needle was stored at 50 ° C for one week, the mass of the physiologically active substance contained in the microneedle needle was calculated in the same manner. The value obtained by dividing the mass of the physiologically active substance obtained by the mass of the physiologically active substance used for the production was recorded as "the content (%) of the physiologically active substance (50 ° C, 1 week)".

The results are shown in Table 6 and Figure 5. The microneedle system of Example 1 had a high initial insulin content, and the value of the insulin content did not decrease even after storage at 50 ° C for one week. On the other hand, in the microneedle of Comparative Example 1, although the initial insulin content was high, the insulin content after storage at 50 ° C for one week was lowered to 65%. Further, the microneedle of Comparative Example 2 had an initial insulin content of 50%, and the insulin content after storage for one week at 50 ° C was 17%. Since the microneedle of Example 1 showed a high insulin content even after storage at 50 ° C for one week, it was confirmed that the stability of the physiologically active substance was high. On the other hand, the microneedle needles of Examples 1 to 6 have the strength required for puncture, and the stability of the physiologically active substance can be highly maintained. When the microneedles of Comparative Examples 3 and 4 were stored at 50 ° C for one week, the stability of the physiologically active substance was excellent, but the content of the physiologically active substance was remarkably lowered during the production of the microneedle.

8. Dissolution test

According to Table 7, the concentration of the edible red No. 40 (R40) was adjusted to 1% by mass, and the composition of the material of the microneedle was 15% aqueous solution, and the microneedle of Reference Examples 15 to 17 was produced. In addition, R40 is used as a substitute for a physiologically active substance. The obtained microneedle needle was immersed in PBS according to the following method, and the cumulative elution amount of R40 after a certain period of time was measured.

The results are shown in Fig. 6. The microneedles of Reference Examples 15 and 16 reached a constant state after a lapse of 3 minutes after immersion, and the value was about 17 μg/mL. On the other hand, the microneedle of Reference Example 17 passes after immersion 7 At the point of the minute, a constant state was reached with a value of about 15 μg/mL. Further, when all of R40 was eluted from the microneedle, the concentration thereof was 17 μg/mL. The microneedles of Reference Examples 15 and 16 can dissolve physiologically active substances more rapidly.

9. Formability test (3)

The spurs of Examples 7 and 8 were prepared by mixing naltrexone, arginine, histidine, and citric acid at a mass ratio shown in Table 8. The appearances of the microneedles of Examples 1 to 4 and 7 to 8 were compared.

In the same manner as in Fig. 1(a), the microneedle needles of Examples 1 to 4 and 7 to 8 were in a transparent amorphous state, and the moldability was good, and the shape of the conical shape was maintained.

10. Strength test (3)

A urethane sheet (manufactured by Exseal Corporation, trade name: HyperGel sheet hardness 50, Shore-C hardness: 50) having a thickness of 0.5 mm coated with an aluminum foil having a thickness of 12 μm was prepared. 100 microneedle needles of Examples 1 to 8 will be used in the use of an applicator (impact rate of 7 m/sec) The microneedle device is applied to the surface of the above aluminum foil to evaluate whether the microneedle needle penetrates the aluminum foil. Calculate the number of roots that penetrate the aluminum foil among the 100 microneedle needles.

In the case of using the microneedles of Examples 1 to 4 and 7 to 8, all of the microneedle needles showed strength throughout the aluminum foil.

1‧‧‧Microneedle device

2‧‧‧Substrate

3‧‧‧Microneedle

Claims (5)

A microneedle comprising: a physiologically active substance, an amino acid selected from the group consisting of arginine and histidine, and an acid having a melting point of 40 ° C or higher. The microneedle of claim 1, wherein the acid is one or more selected from the group consisting of citric acid, lactic acid, tartaric acid, succinic acid, phosphoric acid, and aspartic acid. The microneedle of claim 1 or 2, wherein the aforementioned amino acid is arginine. The microneedle according to any one of claims 1 to 3, wherein the acid is one or more selected from the group consisting of citric acid, tartaric acid and aspartic acid. A microneedle device comprising the microneedle according to any one of claims 1 to 4 on a substrate.