Composition for liraglutide-containing microneedle with enhanced stability and use thereof

Abstract

The present invention provides: a composition for a liraglutide-containing microneedle, which contains liraglutide as an active ingredient, does not cause the denaturation of the active ingredient during manufacturing, and retains stability even after manufacturing; and a liraglutide-containing microneedle which is manufactured using said composition and has enhanced stability. The composition for a microneedle according to the present invention uses a specific combination of a biodegradable polymer, a stabilizer, and an antioxidant, and thus the denaturation of liraglutide, which is an active ingredient, is prevented during the manufacture of the microneedle and the stability of liraglutide is maintained even after the microneedle is manufactured.

Description

Composition for liraglutide-containing microneedles with enhanced stability and use thereof

[0001] The present invention relates to a composition for liraglutide-containing microneedles with enhanced stability and to the use thereof, and more particularly, to a composition for liraglutide-containing microneedles containing liraglutide as an active ingredient, in which the active ingredient is not denatured during manufacture and stability is maintained after manufacture, and to liraglutide-containing microneedles with enhanced stability manufactured using the composition.

[0002] Liraglutide is a human glucagon-like peptide (GLP)-1 analogue that binds to and activates the GLP-1 receptor, increasing glucose-dependent insulin secretion and suppressing inappropriate glucagon secretion. It is used to treat type 2 diabetes and obesity. Liraglutide is sold under the brand names Victoza for type 2 diabetes and Saxenda for obesity.

[0003] Currently, liraglutide is only used as a subcutaneous injection, which requires the hassle of disinfecting and injecting the injection site at a set time every day, as well as the risk of secondary infection. In addition, repeated injections can cause pain, bleeding, rash, and swelling at the injection site, and in the case of self-injection, patients must inject the drug themselves, which can lead to low compliance due to fear and reluctance, and thus, a decrease in the therapeutic effect.

[0004] To improve such problems, some attempts have been made to formulate the drug using microneedles, but a method for manufacturing microneedles containing liraglutide that does not bounce off the skin surface layer, does not break or bend, and shortens the drug manufacturing time, making it easy to mass-produce (Korean Patent Publication No. 10-2022-0151995) has been disclosed.

[0005] Meanwhile, liraglutide has low stability and is easily denatured by various factors during the manufacturing process in the form of microneedle formulations, and its activity may also decrease due to denaturation during storage after manufacturing.

[0006] Therefore, there is a need for the development of liraglutide-containing microneedles that prevent denaturation of liraglutide during manufacturing with microneedles, thereby retaining its activity, and that prevent denaturation during storage after manufacturing, thereby enhancing stability.

[0007] Accordingly, the inventors of the present invention have conducted continuous research to meet the needs of the prior art, and as a result, they have surprisingly confirmed that liraglutide-containing microneedles manufactured using a specific combination of a biodegradable polymer, a stabilizer, and an antioxidant are prevented from denaturing during manufacture, so that the activity is retained, and that denaturation is prevented during storage after manufacture, so that the stability is enhanced, thereby completing the present invention.

[0008] Accordingly, an object of the present invention is to provide a composition for liraglutide-containing microneedles with enhanced stability.

[0009] Another object of the present invention is to provide liraglutide-containing microneedles with enhanced stability manufactured from the above composition.

[0010] Another object of the present invention is to provide a microneedle patch for transdermal delivery of liraglutide comprising the microneedles.

[0011] In order to achieve the above object of the present invention, a composition for liraglutide-containing microneedles with enhanced stability is provided.

[0012] The composition for liraglutide-containing microneedles with enhanced stability of the present invention comprises liraglutide, a biodegradable polymer, a stabilizer, an antioxidant, and a solvent.

[0013] Liraglutide has the structure of chemical formula 1 and is a human GLP (glucagon-like peptide)-1 analogue that binds to and activates the GLP-1 receptor, thereby increasing glucose-dependent insulin secretion and suppressing inappropriate glucagon secretion, and is used to treat type 2 diabetes and obesity:

[0014]

[0015] (Chemical formula 1)

[0016] In the composition of the present invention, the biodegradable polymer is capable of being naturally biodegraded in the body and thus excreted from the body, and at least one selected from the group consisting of hyaluronic acid or a salt thereof and carboxymethyl cellulose (CMC) may be used, most preferably hyaluronic acid or a salt thereof. Compared to other types of biodegradable polymers, the composition for microneedles containing these biodegradable polymers can prevent denaturation of liraglutide during the manufacture and storage of microneedles, thereby enhancing stability.

[0017] In the composition of the present invention, the biodegradable polymer may be included in an amount of 10% to 20% by weight based on the total composition (100% by weight).

[0018] In the composition of the present invention, the stabilizer plays a role in helping to ensure stability during the manufacture of microneedles, and at least one selected from the group consisting of xylitol and trehalose may be used. Compared to other types of stabilizers, the composition for microneedles containing the stabilizer can ensure the stable manufacture of microneedles while preventing denaturation of liraglutide during the manufacture and storage of microneedles, thereby enhancing stability.

[0019] In the composition of the present invention, the stabilizer may be included in an amount of 3% to 10% by weight relative to the entire composition.

[0020] In the composition of the present invention, at least one antioxidant selected from the group consisting of DPG (Dipotassium glycyrrhizinate) and EDTA may be used. Compared to other types of antioxidants, the composition for microneedles containing the antioxidant can prevent denaturation of liraglutide during microneedle manufacturing and storage, thereby enhancing stability.

[0021] In the composition of the present invention, the antioxidant may be included in an amount relative to the weight of liraglutide, that is, a weight ratio of liraglutide: antioxidant of 1:0.3 to 5.

[0022] In the composition of the present invention, liraglutide may be included in an amount of 0.1% to 2.0% by weight relative to the entire composition.

[0023] In the composition of the present invention, the solvent is water, preferably deionized water, or potassium phosphate buffer (PPB).

[0024] The composition of the present invention may additionally include a pH adjuster, if necessary. The pH adjuster may be any of those commonly used in the manufacture of microneedles, such as NaOH. The composition according to the present invention has a pH of 7.5 to 8.7, preferably 7.8 to 8.4.

[0025] In addition, the composition of the present invention may additionally include a plasticizer, surfactant, preservative, etc. that can be commonly used in the manufacture of microneedles, as needed.

[0026] According to another object of the present invention, the present invention provides a liraglutide-containing microneedle with enhanced stability manufactured from the composition.

[0027] The microneedle of the present invention may include a needle portion protruding in one direction and a mattress layer supporting the needle portion. The needle portion has a shape that is easy to penetrate the skin. The shape of the needle portion of the microneedle according to the present invention may be a cone, a pyramid, a sphere, a short-headed wedge, a blade shape, etc., and these should all have a shape that can commonly penetrate the skin. The length of the needle portion is 500 to 1000 μm, preferably 750 μm. The mattress layer has a thickness of 0.1 to 1 mm, preferably 0.1 to 0.3 mm. The needle portion of the microneedle of the present invention may be manufactured so as to be separated from the mattress layer when inserted into the skin, if necessary. The composition according to the present invention may be used in the manufacture of the needle portion and the mattress layer of the microneedle, and the mattress layer may be manufactured from a different material, if necessary. Therefore, the microneedle of the present invention can have liraglutide evenly distributed in the needle portion and the mattress layer, or liraglutide can be distributed only in the needle portion.

[0028] The microneedle of the present invention is a soluble microneedle that releases liraglutide when decomposed in vivo.

[0029] The microneedles of the present invention can be manufactured using a mold.

[0030] According to another object of the present invention, a microneedle patch for transdermal delivery of liraglutide comprising the above microneedles is provided.

[0031] The microneedle patch of the present invention has an adhesive layer laminated on one side of the mattress layer, so that the microneedle patch can be used by attaching it to the skin. The microneedle patch may also include a protective film on the adhesive layer.

[0032] The microneedle patch for transdermal delivery of liraglutide of the present invention can be used for the treatment and improvement of obesity and the treatment and improvement of type 2 diabetes.

[0033] The composition for microneedles according to the present invention uses a specific combination of a biodegradable polymer, a stabilizer, and an antioxidant to prevent denaturation of liraglutide, an active ingredient, during the manufacture of microneedles and to maintain the stability of liraglutide even after the manufacture of microneedles.

[0034] The microneedle according to the present invention is a liraglutide-containing microneedle in which denaturation of liraglutide is prevented during manufacturing, so that activity is maintained, and denaturation is prevented even during storage after manufacturing, so that stability is enhanced.

[0035] The microneedle patch for transdermal delivery according to the present invention is useful for treating and improving obesity and type 2 diabetes because the stability of the active ingredient liraglutide is improved.

[0036] Figure 1 is a graph showing the results of stability analysis according to biodegradable polymers.

[0037] Figure 2 is a graph showing the results of stability analysis according to the stabilizer.

[0038] Figure 3 is a photograph of a microneedle according to the present invention.

[0039] Figure 4 is a graph showing the results of stability analysis of microneedles according to the present invention.

[0040] Hereinafter, to aid understanding of the present invention, the composition and effects of the present invention will be described in more detail through specific examples. However, the following examples are provided solely to illustrate the present invention more clearly, and the scope of the present invention is not limited by the examples.

[0041] Manufacturing Example 1: Manufacturing of a composition for microneedles using biodegradable polymers

[0042] In order to select a biodegradable polymer suitable for manufacturing liraglutide-containing microneedles with enhanced stability, a composition for microneedles for each biodegradable polymer was prepared with the composition shown in Table 1, and the composition was manufactured into a film form (thin film) to test the stability.

[0043] Content (mg) Composition 1 Composition 2 Composition 3 Composition 4 Composition 4 Liraglutide (Lira) 10.0 10.0 10.0 10.0 10.0 Sodium Alginate (Alginate)-30.0---Sodium Hyaluronate (HA)--30.0--Carboxy methyl cellulose (CMC)---30.0-Polyvinyl pyrrolidone (PVP)----30.0

[0044] Specifically, liraglutide and biodegradable polymers were weighed as shown in Table 1 in 1 ml of purified water (deionized water; DW), placed in a cup tube, dissolved for 5 minutes using a shaking mixer, and dried at 25°C for 24 hours using a dryer. Then, the solvent was completely evaporated using an evaporator under nitrogen to produce a film (thickness 50–100 μm) capable of simulating microneedles. The produced films were subjected to stability analysis immediately after production and after storage for one week in a constant temperature and humidity stability chamber at 25°C and 60% acceleration conditions.

[0045] Specifically, for stability analysis, the entire amount of each film sample was added to 5 ml of a dilution solvent (1:1 mixture of phosphate buffer solution pH 3.5: 78% Acetonitrile), mixed by shaking, and then quantitative analysis of liraglutide was performed using the HPLC method under the conditions shown in Table 2 below. The results are shown in Table 3 and Figure 1.

[0046] Device: Thermo U3000, Column C8, 4.6 x 100 mm, 3.5 μm, Wavelength 215 nm, Column temperature 45 °C, Flow rate 1.0 ml / min, Injection volume 10 μl, Operating time 40 min, Fluid phase A: Phosphate buffer solution (pH 3.5), B: 78% Acetonitrile, Gradient 0-40 min: 56-64% B

[0047] Content (%) Composition 1 (Lira) Composition 2 (Lira + Alginate) Composition 3 (Lira + HA) Composition 4 (Lira + CMC) Composition 5 (Lira + PVP) Immediately after manufacturing 99.5 ± 0.3 98.6 ± 2.8 104.3 ± 1.2 102.4 ± 3.3 98.2 ± 4.7 After 1 week of storage 94.2 ± 3.7 84.2 ± 9.6 101.5 ± 3.8 98.4 ± 2.4 92.7 ± 3.8

[0048] As shown in Table 3 and Fig. 1, when hyaluronic acid and CMC were used as biodegradable polymers, liraglutide denaturation was prevented immediately after film production and after one week of storage, thereby enhancing stability. When alginic acid or PVP were used as biodegradable polymers, liraglutide denaturation appeared to have progressed somewhat immediately after production and after one week of storage. Therefore, hyaluronic acid and CMC are suitable biodegradable polymers for liraglutide-containing microneedles, and hyaluronic acid is particularly suitable.

[0049] Manufacturing Example 2: Manufacturing of a composition for microneedles by stabilizer

[0050] In order to select a stabilizer suitable for manufacturing liraglutide-containing microneedles with enhanced stability, a composition for microneedles according to the stabilizer composition shown in Table 4 was manufactured and manufactured into a film form (thin film) in the same manner as in Manufacturing Example 1, and a stability analysis was performed in the same manner as in Manufacturing Example 1.

[0051] Content (mg) Composition 1 (Lira) Composition 6 Lira + Glycerin Composition 7 Lira + Trehalose Composition 8 Lira + xylitol Composition 9 Lira + GH815 Liraglutide (Lira) 10.0 10.0 10.0 10.0 10.0 Glycerin-30.0---Trehalose--30.0--Xylitol---30.0-GH-815 (Propanediol, Caprylyl Glycol, Ethylhexylglycerin)----10.0

[0052] The results are shown in Table 5 and Figure 2.

[0053] Content (%) Composition 1 (Lira) Composition 6 Lira + Glycerin Composition 7 Lira + Trehalose Composition 8 Lira + xylitol Composition 9 Lira + GH8 After preparation 99.5 ± 0.3 10 2.0 ± 1.5 10 4.2 ± 1.9 9 8.2 ± 4.7 10 2.4 ± 3.0 1 After storage 94.2 ± 3.7 8 4.6 ± 5.3 10 0.9 ± 0.7 9 9.7 ± 3.8 8 0.3 ± 5.8

[0054] As shown in Table 5 and Fig. 2, when trehalose and xylitol were used as stabilizers, liraglutide denaturation was prevented immediately after film production and after one week of storage, thereby enhancing stability. When glycerin or GH-815 (a mixture of propanediol, caprylyl glycol, and ethylhexylglycerin) was used as stabilizers, liraglutide denaturation appeared to have progressed after one week of storage. Therefore, it can be confirmed that trehalose and xylitol are suitable stabilizers for liraglutide-containing microneedles.

[0055] Manufacturing Example 3: Manufacturing of a composition for microneedles by antioxidant

[0056] In order to select an antioxidant suitable for manufacturing liraglutide-containing microneedles with enhanced stability, hyaluronic acid was used as a biodegradable polymer, and compositions for microneedles for each antioxidant were manufactured with the compositions shown in Table 6, manufactured in the form of a film (thin film) in the same manner as in Manufacturing Example 1, and stability analysis was performed in the same manner as in Manufacturing Example 1.

[0057] Content (mg) Composition 10 Lira+HA Composition 11 Lira+HA+BHA Composition 12 Lira+HA+BHT Composition 13 Lira+HA+EDTA Composition 14 Lira+HA+DPGLiraglutide(Lira)3.003.003.003.003.003.00HA2.822.822.822.822.82BHA (Butylated hydroxyanisole)-0.3---BHT (Butylated hydroxytoluene)--0.3--EDTA (Disodium Dihydrogen Ethylenediaminetetraacetate Dihydrate)---0.3-DPG (Dipotassium glycyrrhizinate)----0.3

[0058] The results are shown in Table 7.

[0059] Content (%) Composition 10 Lira+HA Composition 11 Lira+HA+BHA Composition 12 Lira+HA+BHT Composition 13 Lira+HA+EDTA Composition 14 Lira+HA+DPG Immediately after manufacturing 86.8±2.4 79.8±7.3 76.4±5.3 89.3±1.2 88.3±2.1 After 1 week of storage 82.1±2.4 69.8±2.1 54.2±1.3 90.5±3.4 87.1±0.8

[0060] As shown in Table 7, when EDTA and DPG (Dipotassium glycyrrhizinate) were used as antioxidants, liraglutide denaturation was prevented and stability was enhanced immediately after film production and after one week of storage. When BHA (Butylated hydroxyanisole) or BHT (Butylated hydroxytoluene) were used as antioxidants, liraglutide denaturation appeared to have progressed somewhat immediately after production and after one week of storage. Therefore, EDTA and DPG are confirmed to be suitable antioxidants for liraglutide-containing microneedles.

[0061] Manufacturing Example 4: Manufacturing of liraglutide-containing microneedles

[0062] <Manufacturing of a composition for microneedles>

[0063] A composition for microneedles was prepared with the composition shown in Table 8:

[0064] Content (mg) Example 1 Example 2 Example 3 Example 4 Liraglutide 0.26 0.26 0.26 0.26 0.26 HA 39.29 39.29 39.29 39.29 Xylitol 13.10- 13.10 Trehalose -13.08- 13.10 DPG 0.26 0.26 0.26 EDTA--- 0.26 0.1M NaOH aqueous solution 41.65 41.65 41.06 5 41.65 DW remainder remainder 0.005M PPB remainder remainder Total (mg) 200 200 200 200

[0065] Specifically, the compositions of Examples 1 to 4 were prepared by adding DPG or EDTA to a conical tube, mixing purified water or potassium phosphate buffer (PPB) and 0.1 M NaOH aqueous solution, and vortexing for 10 minutes. The resulting solution was then added to the tube and vortexed for 10 minutes. Then, xylitol or trehalose was added and vortexed for 5 minutes, liraglutide was added and vortexed for 5 minutes, and sodium hyaluronate as hyaluronic acid (HA) was added and vortexed for 30 minutes to complete the composition. The pH of the prepared composition was about 7.8 to 8.4.

[0066] Microneedle manufacturing

[0067] Each of the compositions manufactured above was loaded into a silicone negative mold engraved with a microneedle shape. After loading, the mold was placed in a desiccator, decompressed to -0.04 MPa, maintained for 5 minutes, ventilated, and any resulting air bubbles removed using an air gun. The silicone mold was then placed in a hot air dryer and dried at 30°C for 2 hours. The completed microneedles were then separated from the silicone mold using tweezers to produce microneedles. A photograph of the manufactured microneedles is shown in Figure 3.

[0068] The recovered microneedles can be made more usable in the form of a patch by attaching a colloid band or other device to the back, as needed.

[0069] As shown in Fig. 3, it can be confirmed that microneedles are well formed when the compositions of Examples 1 to 4 are used.

[0070] Microneedles manufactured with the compositions of Examples 1 to 4 also had sufficient strength to penetrate the skin.

[0071] Test Example 1: Drug Stability Analysis of Microneedles

[0072] For each of the microneedles manufactured in Examples 1 to 4 in Manufacturing Example 4, a stability analysis was performed in the same manner as in Manufacturing Example 1, and the results are shown in Table 9 and Figure 4.

[0073] Content (%)Example 1Example 2Example 3Example 4Immediately after manufacture101.86±0.39103.41±2.03102.80±1.2699.55±1.57After storage for 1 week104.36±1.38108.54±0.78105.64±2.6599.93±1.68

[0074] As shown in Table 9 and Fig. 4, the microneedles of Examples 1 to 3, which were manufactured with a composition containing hyaluronic acid as a biodegradable polymer, xylitol or trehalose as a stabilizer, and DPG as an antioxidant, exhibited excellent stability by preventing liraglutide denaturation immediately after manufacture, and maintained excellent stability even after one week of storage. In addition, the microneedles of Example 4, which were manufactured with a composition containing EDTA as an antioxidant, maintained excellent stability by preventing liraglutide denaturation after one week of storage. Therefore, it can be confirmed that the stability of the liraglutide-containing microneedles is enhanced only when a specific combination of the biodegradable polymer, stabilizer, and antioxidant according to the present invention is included.

Claims

1. A composition for liraglutide-containing microneedles with enhanced stability, The composition comprises liraglutide, a biodegradable polymer, a stabilizer, an antioxidant and a solvent. The above biodegradable polymer is at least one selected from the group consisting of hyaluronic acid or a salt thereof and carboxymethylcellulose (CMC), The above stabilizer is at least one selected from the group consisting of xylitol and trehalose, A composition for microneedles, wherein the antioxidant is at least one selected from the group consisting of DPG (Dipotassium glycyrrhizinate) and EDTA.

2. A composition for liraglutide-containing microneedles with enhanced stability, wherein the biodegradable polymer in claim 1 is hyaluronic acid or a salt thereof.

3. A composition for liraglutide-containing microneedles with enhanced stability, wherein the stabilizer in claim 1 is xylitol.

4. A composition for liraglutide-containing microneedles with enhanced stability, wherein the antioxidant in claim 1 is DPG.

5. A composition for liraglutide-containing microneedles with enhanced stability, wherein the biodegradable polymer in claim 1 is hyaluronic acid or a salt thereof, the stabilizer is xylitol, and the antioxidant is DPG.

6. A composition for liraglutide-containing microneedles with enhanced stability, wherein the biodegradable polymer in claim 1 is hyaluronic acid or a salt thereof, the stabilizer is trehalose, and the antioxidant is DPG.

7. A composition for liraglutide-containing microneedles with enhanced stability, wherein the biodegradable polymer is included in an amount of 10 to 20 wt% based on the total composition in the first paragraph.

8. A composition for liraglutide-containing microneedles with enhanced stability, wherein the stabilizer is included in an amount of 3 to 10 wt% based on the total composition in the first paragraph.

9. A composition for liraglutide-containing microneedles with enhanced stability, wherein the antioxidant is included in the first paragraph so that the weight ratio of liraglutide:antioxidant is 1:0.3 to 5.

10. A liraglutide-containing microneedle with enhanced stability manufactured using the composition according to paragraph 1, A microneedle comprising a needle portion protruding in one direction and a mattress layer supporting the needle portion.

11. A liraglutide-containing microneedle with enhanced stability, wherein the needle portion can be separated from the mattress layer when inserted into the skin in the 10th paragraph.

12. In the 10th paragraph, the microneedle is manufactured using a mold, a liraglutide-containing microneedle with enhanced stability.

13. A microneedle patch for transdermal delivery of liraglutide comprising microneedles according to Article 10.

14. A microneedle patch for transdermal delivery of liraglutide, wherein the microneedle patch is for the treatment and improvement of obesity in accordance with claim 13.

15. In claim 13, the microneedle patch is a microneedle patch for transdermal delivery of liraglutide for the treatment and improvement of type 2 diabetes.

Images