Soluble microneedle compositions for transdermal delivery of polypeptides and uses thereof

Abstract

The application discloses a soluble microneedle composition for transdermal delivery of polypeptides and application thereof. The soluble microneedle composition comprises a first composition and a second composition; wherein the first composition comprises the following components in mass percentage: 20-60% polypeptides, 0.5-20% capryol 98, 10-40% first biocompatible polymer material and 0.5-20% first surfactant; the second composition comprises the following components in mass percentage: 98-99.75% second biocompatible polymer material and 0.25-2% second surfactant. The microneedle composition is used in the microneedle, and the problems of low transdermal efficiency and poor stability of polypeptides are solved.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field. More specifically, it relates to a soluble microneedle composition for transdermal delivery of polypeptides and its application. Background Technology

[0002] Peptides have demonstrated unique advantages in disease treatment, and due to their high specificity, potency, and good safety profile, they have been widely used in various therapeutic areas such as weight loss, metabolic diseases, and cardiovascular diseases. However, existing delivery methods each have their drawbacks: while injections offer high bioavailability, long-term patient compliance is poor; oral administration is limited by severe gastrointestinal enzyme degradation and the first-pass effect in the liver, resulting in extremely low bioavailability. Microneedle technology, as an innovative transdermal delivery platform, can effectively circumvent these problems and achieve painless drug delivery, but its application is limited by two core challenges: first, the natural barrier of the stratum corneum greatly hinders the transdermal penetration efficiency of large peptide molecules; second, peptides are prone to aggregation or degradation during microneedle preparation and storage, affecting formulation stability and efficacy. Summary of the Invention

[0003] The purpose of this invention is to provide a soluble microneedle composition for transdermal delivery of peptides and its application, so as to at least solve the problems of low transdermal peptide efficiency and poor stability.

[0004] To achieve the above objectives, the present invention adopts the following technical solution:

[0005] In a first aspect, the present invention provides a soluble microneedle composition for transdermal delivery of polypeptides, wherein the soluble microneedle composition comprises a first composition and a second composition; wherein,

[0006] The first composition contains the following components in weight percentages:

[0007] 20-60% polypeptides, 0.5-20% octanoic acid / capric acid / polyethylene glycol glycerol esters, 10-40% primary biocompatible polymer materials and 0.5-20% primary surfactants;

[0008] The second composition contains the following components in weight percentages:

[0009] 98-99.75% of the second biocompatible polymer material and 0.25-2% of the second surfactant.

[0010] In this technical solution, PEGylated caprylic / capric acid glyceride possesses a unique amphiphilic structure, which significantly enhances the transdermal penetration of peptides. This unique amphiphilic structure also allows for physical binding with peptide molecules, forming a stable complex that improves microneedle stability. This binding effect significantly enhances transdermal penetration by altering the interfacial properties of the peptide; furthermore, it effectively inhibits peptide aggregation and degradation during formulation by forming a protective microenvironment on the peptide molecule surface, thereby greatly improving the long-term stability of the formulation. By synergistically combining this functional component with polymer matrices, surfactants, and other components, a microneedle delivery system with both high permeability enhancement and excellent stability can be constructed, providing a more ideal solution for the clinical application of peptides.

[0011] Furthermore, the mass ratio of PEGylated caprylic / capric acid to the polypeptide is 1:1 to 1:20. When the mass ratio of PEGylated caprylic / capric acid to the polypeptide is less than 1:20, there is insufficient penetration enhancer to form a stable complex, resulting in ineffective binding of the polypeptide and an unsatisfactory transdermal rate. When the mass ratio is greater than 1:1, excessive penetration enhancer will lead to a decrease in microneedle stiffness, affecting puncture performance.

[0012] Furthermore, the mass ratio of the octanoic acid-capric acid-decalcium glycol glyceride to the polypeptide is 1:5-1:10. Under these conditions, the aforementioned effects are even better.

[0013] For example, by mass percentage, the content of polyethylene glycol glycerol octanoate in the first component includes, but is not limited to, 1-20%, 1-15%, 3-15%, etc.

[0014] For example, the content of polypeptides in the first component, by mass percentage, includes, but is not limited to, 30-60%, 40-60%, 45-60%, etc.

[0015] Furthermore, the polypeptide is selected from one or more of smegglutide, liraglutide, telpoglutide, retaglutide, dulaglutide, loxenatide, lixisenatide, and mascaratide.

[0016] For example, by mass percentage, the content of the first surfactant in the first component includes, but is not limited to, 1-20%, 1-15%, 3-15%, etc.

[0017] Furthermore, the first surfactant and the second surfactant are each independently selected from one or more of Tween, polyethylene glycol, poloxamer 188, polyoxyethylene 40 hydrogenated castor oil, and 15-hydroxystearic acid polyethylene glycol ester.

[0018] For example, by mass percentage, the content of the first biocompatible polymer material in the first component includes, but is not limited to, 20-40%, 30-40%, etc.

[0019] Furthermore, the first biocompatible polymer material is selected from one or more of sodium hyaluronate, dextran, and trehalose.

[0020] Furthermore, the second biocompatible polymer material is selected from a mixture of two or more of sodium hyaluronate, dextran, cyclodextrin derivatives, and sucrose.

[0021] Furthermore, the weight-average molecular weight of the sodium hyaluronate is 100,000 to 250,000.

[0022] Furthermore, the weight-average molecular weight of the dextran is 40,000 to 70,000.

[0023] Furthermore, the cyclodextrin derivative is selected from one or more of sodium sulfobutylcyclodextrin, methylcyclodextrin, and hydroxypropylcyclodextrin.

[0024] Furthermore, the second biocompatible polymer material is a mixture of sodium hyaluronate and one selected from dextran, cyclodextrin derivatives, and sucrose in a mass ratio of 2:1 to 1:3. Under these conditions, the microneedles maintain a smooth appearance and possess suitable mechanical properties. While preserving structural integrity and preventing breakage, it also avoids affecting drug release efficiency due to excessive toughness.

[0025] Furthermore, the second biocompatible polymer material is a mixture of sodium hyaluronate and two of the following selected from dextran, cyclodextrin derivatives, and sucrose in a mass ratio of 2:1 to 1:2. Under these conditions, the microneedles can be better ensured to have a smooth appearance and possess moderate mechanical properties. While maintaining structural integrity and being less prone to breakage, it also avoids affecting drug release efficiency due to excessive toughness.

[0026] Further, the second biocompatible polymer material is a mixture of the following components 1) and 2) in a mass ratio of 2:1 to 1:2:

[0027] 1) Sodium hyaluronate,

[0028] 2) A mixture of sucrose and one of dextran and cyclodextrin derivatives.

[0029] in,

[0030] The mass ratio of dextran or cyclodextrin derivatives to sucrose is 1:1 to 4:1.

[0031] Secondly, the present invention provides a microneedle prepared from a raw material comprising the soluble microneedle composition described above.

[0032] Furthermore, the microneedles consist of a substrate and needles arranged in an array on the substrate.

[0033] In the soluble microneedle composition, the first composition is used to prepare the needle body; the second composition is used to prepare the substrate.

[0034] Furthermore, the raw materials forming the needle also contain water, and the content of the first composition in the raw materials forming the needle is 2-10 wt%, 2-6 wt%, etc.

[0035] Furthermore, the raw materials forming the substrate also contain water, and the content of the second composition in the raw materials forming the substrate is 20-40 wt%, 20-30 wt%, etc.

[0036] Thirdly, the present invention provides a method for preparing the microneedles as described above, comprising the following steps:

[0037] The first composition is dissolved in a solvent to obtain a needle solution;

[0038] The needle body solution is placed in a mold and dried to obtain the needle body layer;

[0039] The second composition was dissolved in water to obtain a base solution;

[0040] The base solution is placed in a mold containing the needle layer and dried to obtain the microneedles.

[0041] Furthermore, the solvent is one or more of water, histidine buffer, and Tris buffer.

[0042] Fourthly, the present invention provides a microneedle patch comprising microneedles as described above and a backing bonded to the microneedles.

[0043] Fifthly, the present invention provides the use of the microneedle composition described above in the preparation of pharmaceutical formulations for treating type II diabetes or obesity.

[0044] Furthermore, the formulation is a microneedle.

[0045] The beneficial effects of this invention are as follows:

[0046] The microneedle composition of this invention introduces polyethylene glycol glyceride (PEG) as a penetration enhancer and stabilizer to form a stable complex with peptides. This effectively overcomes the barrier effect of the stratum corneum on peptide transdermal absorption, significantly improves the transdermal efficiency and bioavailability of peptides, and simultaneously inhibits the aggregation and degradation of peptides during preparation and storage, thereby enhancing the stability of the formulation. Attached Figure Description

[0047] The specific embodiments of the present invention will be described in further detail below with reference to the accompanying drawings.

[0048] Figure 1 The diagram shows the preparation of the needle tip solution in Example 1. The left side shows the solution of water + polyethylene glycol glycerol (EP), the middle side shows the solution of water + polyethylene glycol glycerol (EP) + smegglutinin (SM), and the right side shows the solution of all needle body materials.

[0049] Figure 2 The in vitro transdermal curves of microneedles with and without different penetration enhancers are shown in Examples 2 and Comparative Examples 1-3.

[0050] Figure 3 The in vitro transdermal curves of microneedles containing the penetration enhancer polyethylene glycol glycerol caprylate and caprylic acid, as shown in Examples 4-6, are obtained for different polypeptide model drugs.

[0051] Figure 4 The results of the long-term stability study of the microneedles in each embodiment and Comparative Example 3 are shown in the figure.

[0052] Figure 5 The appearance and folded appearance of the microneedle formulations of Examples 7-8 and Comparative Examples 4-6 are shown. Detailed Implementation

[0053] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments and accompanying drawings, further explains the invention. Similar components in the drawings are indicated by the same reference numerals. Those skilled in the art should understand that the specific description below is illustrative rather than restrictive and should not be construed as limiting the scope of protection of the present invention.

[0054] Example 1

[0055] A soluble microneedle composition for transdermal delivery of peptides comprises a first composition (needle material) and a second composition (base material), the specific formulation of which is shown in Table 1 below.

[0056] A microneedle is prepared from a raw material comprising the soluble microneedle composition described above, specifically including the following steps:

[0057] (1) Preparation of microneedle matrix solution:

[0058] Preparation of needle solution:

[0059] Prepare an aqueous solution of the needle material according to the needle material formula in Table 1, wherein the total concentration of the needle material in the aqueous solution is 5.2 wt%.

[0060] The preparation method of the above-mentioned injection solution specifically includes: weighing the prescribed amount of caprylic / capric acid glyceride (PEG-Glyceryl ester) into a centrifuge tube, adding the prescribed amount of water for injection, stirring to dissolve, and recording the solution phenomena; then adding the prescribed amount of smegglutinin into a centrifuge tube, stirring to dissolve, and recording the solution phenomena; then adding the prescribed amounts of Tween 80, trehalose, and sodium hyaluronate into a centrifuge tube, stirring to dissolve, and recording the solution phenomena to obtain the injection solution. The solution phenomena after each step of dissolution are as follows: Figure 1 As shown.

[0061] Preparation of base solution:

[0062] Prepare an aqueous solution of the substrate material according to the substrate material formulation in Table 1, wherein the total mass content of the substrate material in the aqueous solution is 28.2 wt%.

[0063] The preparation method of the above-mentioned base solution specifically includes: weighing the prescribed amount of injection water into a centrifuge tube, adding the prescribed amounts of sucrose, sodium sulfobutylbenzyl dextrin and sodium hyaluronate, stirring until completely dissolved, centrifuging at 5000 rpm for 15 minutes; and then sterilizing by moist heat at 121°C for 15 minutes to obtain the base solution.

[0064] (2) Preparation of microneedles:

[0065] Transfer 60 μL of the prepared injection solution to a 1 cm volume. 2 Spread the coating evenly on a mold of a certain size, apply negative pressure to the mold for about 15 minutes, transfer the mold to a dry place to dry for 1.5 hours, remove excess residue from the mold surface to obtain the needle layer;

[0066] Then, 80 μL of the base solution was dropped onto the unit mold containing the needle layer and spread evenly. After applying negative pressure to the mold for 30 minutes, the mold was transferred to a dry place to dry for 5 hours to form the base layer and obtain the microneedles. The microneedles were then demolded after drying.

[0067] By combining microneedles with a backing, a microneedle patch is obtained.

[0068] Table 1 Formulation of soluble microneedle composition

[0069]

[0070] Example 2

[0071] The soluble microneedle composition for transdermal delivery of peptides is shown in Table 1.

[0072] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the total mass content of needle material in the needle solution is 3.56 wt%.

[0073] Example 3

[0074] The soluble microneedle composition for transdermal delivery of peptides is shown in Table 1.

[0075] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the total mass content of needle material in the needle solution is 2.94 wt%.

[0076] Comparative Example 1

[0077] The soluble microneedle composition for transdermal delivery of peptides is shown in Table 1.

[0078] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the selection of raw materials for the needle body material is different, and the total mass content of the needle body material in the needle body solution is 3.56 wt%.

[0079] Comparative Example 2

[0080] The soluble microneedle composition for transdermal delivery of peptides is shown in Table 1.

[0081] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the selection of raw materials for the needle body material is different, and the total mass content of the needle body material in the needle body solution is 3.56 wt%.

[0082] Comparative Example 3

[0083] The soluble microneedle composition for transdermal delivery of peptides is shown in Table 1.

[0084] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the selection of raw materials for the needle body material is different, and the total mass content of the needle body material in the needle body solution is 3.26 wt%.

[0085] Examples 4-6

[0086] The soluble microneedle composition for transdermal delivery of peptides is shown in Table 2.

[0087] The preparation method of microneedles and microneedle patches is the same as in Example 1, except that the selection of raw materials for the needle body material is different, and the total mass content of the needle body material in the needle body solution is 5.04 wt%.

[0088] Table 2 Formulation of Soluble Microneedle Compositions

[0089]

[0090] In vitro transdermal test:

[0091] In vitro transdermal assays were performed using the smegglutinin dissolving microneedles prepared in Examples 2, 4-6 and Comparative Examples 1-3. The receiving chamber capacity was 12 ml; 3 ml of sample solution was collected each time, the remaining sample solution was discarded, and then 12 ml of fresh medium was added. The transdermal surface area was 1 cm². 2 Fresh pigskin stored at -20℃ was thawed naturally at room temperature. Skin tissue of uniform size and thickness was then cut and stored in PBS phosphate buffer at 32℃ for 30 minutes. The stratum corneum was then dried with a lint-free cloth. Microneedles were pressed onto the stratum corneum side of the skin for approximately 20 seconds, and the skin with microneedles was fixed between the drug delivery and receiving pools of the diffusion chamber. The temperature of the PBS buffer in the receiving pool was maintained at 32±0.2℃ using instrument heating, and the magnetic stirrer at the bottom of the receiving pool was set to 600 rpm. An automated sampling device was used to collect samples at 1, 2, 4, 6, 8, 12, 18, and 24 hours, with 12 ml of blank receiving solution added simultaneously. Samples from the receiving solution at each time point were filtered through a 0.22 μm PVDF membrane. 3 ml of the filtrate was discarded, and the remaining filtrate was collected for content determination using HPLC.

[0092] Cumulative transdermal penetration curve as follows Figure 2 and Figure 3 As shown in the diagram. According to the in vitro transdermal curve results, the PEGylated caprylic / capric acid ester system of Example 2 exhibited the best permeation-enhancing performance, with a cumulative permeation rate of approximately 70% within 24 hours, demonstrating rapid and efficient permeation. In contrast, the hyaluronidase system of Comparative Example 1 had a cumulative permeation rate of approximately 64% within 24 hours, showing a slightly weaker permeation-enhancing effect than Example 2. The glycine system of Comparative Example 2 and the system without a permeation enhancer of Comparative Example 3 had similar cumulative permeation rates within 24 hours, with the glycine system showing the worst permeation-enhancing effect. In conclusion, the type and combination of permeation enhancers have a decisive influence on the transdermal behavior of microneedle formulations. The PEGylated caprylic / capric acid ester formulation used in the examples was significantly superior to other control regimens in promoting drug penetration.

[0093] The in vitro transdermal curve results show that, although different peptide model drugs were used in Examples 4-6, all three exhibited good transdermal effects under the same penetration enhancer, polyethylene glycol glyceride caprylate-capric acid. This indicates that the penetration enhancer can effectively enhance the penetration of peptides with different structures, demonstrating good versatility.

[0094] Long-term stability assessment:

[0095] To evaluate the stability of the microneedle patch, samples from Examples 1-6 and Comparative Example 3 were packaged in blister packs and aluminum-plastic bags, respectively, and subjected to long-term testing at room temperature. During the 7-month observation period, samples were taken at the end of months 0, 2, 4, and 7, and the content of smegglutinin was determined by high-performance liquid chromatography to evaluate its change over time.

[0096] Stability results were examined as follows Figure 4 As shown, the examples containing PEG-3-caprylate and PEG-3-caprylate exhibited excellent stability, with almost no change in smegglutinin content during the 7-month observation period. This indicates that the penetration enhancer system used (PEG-3-caprylate and PEG-3-caprylate) not only has excellent transdermal effects (due to...) Figure 2 and Figure 3 It is known that this formulation exhibits good compatibility with the main drug component and does not accelerate drug degradation, thus ensuring the quality and efficacy of the formulation during its shelf life. In contrast, Comparative Example 3, which does not contain polyethylene glycol glycerol (PEG), showed a significant decrease in stability.

[0097] Examples 7-8 and Comparative Examples 4-6

[0098] A soluble microneedle composition for transdermal delivery of peptides comprises a first composition (needle material) and a second composition (base material), the specific formulations of which are shown in Table 3 below.

[0099] The preparation method of the microneedles is the same as in Example 1, except that the total mass content of the substrate material in the substrate solution is 28.2 wt% (Example 7), 28.2 wt% (Example 8), 24.2 wt% (Comparative Example 4), 36.2 wt% (Comparative Example 5), and 28.2 wt% (Comparative Example 6), respectively; and in Examples 7, 8, and 4, the drying time for forming the substrate layer is 3-5 hours; in Comparative Example 5, the drying time for forming the substrate layer is 4-6 hours; and in Comparative Example 6, the drying time for forming the substrate layer is 9-10 hours.

[0100] In Example 7, the mass ratio of sodium hyaluronate to (sodium sulfobutylbenzyl dextrin + sucrose) was 1:1, and the ratio of sodium sulfobutylbenzyl dextrin to sucrose was 1.8:1; in Example 8, the mass ratio of sodium hyaluronate to (dextrose + sucrose) was 1:1, and the mass ratio of dextran to sucrose was 2.5:1; in Comparative Example 4, the mass ratio of sodium hyaluronate to (sodium sulfobutylbenzyl dextrin + sucrose) was 3: 1. The mass ratio of sodium sulfobutylbenzyl dextrin to sucrose was 1.8:1; in Comparative Example 5, the mass ratio of sodium hyaluronate to (sodium sulfobutylbenzyl dextrin + sucrose) was 1:3, and the mass ratio of sodium sulfobutylbenzyl dextrin to sucrose was 1.8:1; in Comparative Example 6, the mass ratio of sodium hyaluronate to (sodium sulfobutylbenzyl dextrin + sucrose) was 1:1, and the mass ratio of sodium sulfobutylbenzyl dextrin to sucrose was 0.5:1.

[0101] Table 3 Formulation of Soluble Microneedle Compositions

[0102]

[0103] The microneedles obtained in Examples 7-8 and Comparative Examples 4-6 were photographed and their appearance (flatness and brittleness after folding) were statistically analyzed. The results are as follows: Figure 5 As shown. From Figure 5 As can be seen, the microneedles obtained in Examples 7 and 8 have a smooth appearance, good folding performance, and short drying time. In contrast, the microneedles in Comparative Example 4 are curled, and although they have good folding performance, this affects their puncture performance during use. Although the microneedles in Comparative Example 5 have a smooth appearance, the excessive addition of small molecule excipients makes them brittle, which also affects their puncture performance during use. Comparative Example 6 has a significantly longer drying time, reaching 9-10 hours, which is detrimental to the stability of the peptide. The results of related substances detection for the samples from the examples and comparative examples after drying are shown in Table 4 below. Overall, the microneedle formulations of Examples 7 and 8 have better overall performance than the comparative examples, showing superior practicality and stability.

[0104] Table 4 Results of Related Substance Detection

[0105]

[0106] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. For those skilled in the art, other variations or modifications can be made based on the above description. It is impossible to exhaustively list all the implementation methods here. All obvious variations or modifications derived from the technical solutions of the present invention are still within the protection scope of the present invention.

Claims

1. A microneedle, characterized in that, The microneedles consist of a substrate and needles arranged in an array on the substrate; The microneedles are prepared from raw materials comprising a soluble microneedle composition for transdermal delivery of peptides, wherein the soluble microneedle composition comprises a first composition and a second composition, the first composition being used to prepare the needle body; the second composition being used to prepare the substrate; wherein... The first composition contains the following components in weight percentages: 20-60% polypeptides, 0.5-20% octanoic acid / capric acid / polyethylene glycol glycerol esters, 10-40% primary biocompatible polymer materials and 0.5-20% primary surfactants; The second composition contains the following components in weight percentages: 98-99.75% of a second biocompatible polymer material and 0.25-2% of a second surfactant; The mass ratio of the octanoic acid-capric acid-decanoic acid polyethylene glycol glyceride to the polypeptide is 1:1-1:20; The first biocompatible polymer material is selected from one or more of sodium hyaluronate, dextran, and trehalose; The second biocompatible polymer material is a mixture of two or more selected from sodium hyaluronate, dextran, cyclodextrin derivatives and sucrose; The first surfactant and the second surfactant are each independently selected from one or more of Tween, polyethylene glycol, poloxamer 188, polyoxyethylene 40 hydrogenated castor oil and 15-hydroxystearic acid polyethylene glycol ester; The polypeptide is selected from one or more of smegglutide, liraglutide, telpoglutide, retaglutide, dulaglutide, loxenatide, lixisenatide, and mascaratide.

2. The microneedle according to claim 1, characterized in that, The mass ratio of the octanoic acid-capric acid-decalcium glyceride to the polypeptide is 1:5-1:

10.

3. The microneedle according to claim 1, characterized in that, The second biocompatible polymer is a mixture of sodium hyaluronate and one selected from dextran, cyclodextrin derivatives, and sucrose in a mass ratio of 2:1 to 1:3; or The second biocompatible polymer material is a mixture of sodium hyaluronate and two of the following selected from dextran, cyclodextrin derivatives and sucrose in a mass ratio of 2:1 to 1:

2.

4. The microneedle according to claim 3, characterized in that, The second biocompatible polymer material is a mixture of the following components 1) and 2) in a mass ratio of 2:1 to 1:2: 1) Sodium hyaluronate, 2) A mixture of sucrose and one of dextran and cyclodextrin derivatives. in, The mass ratio of dextran or cyclodextrin derivatives to sucrose is 1:1 to 4:

1.

5. The method for preparing microneedles according to any one of claims 1-4, characterized in that, Includes the following steps: The first composition is dissolved in a solvent to obtain a needle solution; The needle body solution is placed in a mold and dried to obtain the needle body layer; The second composition was dissolved in water to obtain a base solution; The base solution is placed in a mold containing the needle layer and dried to obtain the microneedles.

6. The use of the microneedles as described in any one of claims 1-4 in the preparation of pharmaceutical formulations for treating type II diabetes or obesity.

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