Microneedle composition containing semaglutide, and preparation method therefor and use thereof

Abstract

The present invention belongs to the technical field of medicine. Disclosed are a microneedle composition containing semaglutide, and a preparation method therefor and the use thereof. The microneedle composition containing semaglutide consists of a needle body and a substrate, wherein the components of the needle body comprise semaglutide, a cosolvent, and an excipient; the cosolvent is selected from one or more of Tween, Span, propylene glycol, glycerol, and lecithin; and the excipient is selected from one or more of dextran, polyvinylpyrrolidone, hyaluronic acid, and water-soluble cyclodextrin. A soluble microneedle is prepared from semaglutide and the microneedle excipient, which has sufficient mechanical strength to pierce the skin, thereby achieving transdermal delivery of a macromolecular polypeptide drug. Moreover, in a pharmacokinetic experiment, the microneedle composition can simultaneously achieve a maximum plasma concentration with an injection dosage form, and the relative bioavailability can be improved by 40-60 times compared with that of an oral dosage form. The microneedle composition is present in a solid form, thus significantly improving the stability of semaglutide and substantially reducing the cost requirements for storage.

Description

A microneedle composition containing semaglutide and a preparation method and application thereof TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and specifically relates to a microneedle composition containing semaglutide and a preparation method and application thereof. BACKGROUND

[0002] Glucagon-like peptide 1 (GLP-1) is a hormone with broad pharmacological potential, which is secreted by human intestinal epithelial L cells after food stimulation. It is the most effective intestinal insulin hormone known, and its insulin secretion capacity accounts for about 50-70% of the total insulin secretion. Under the mediation of GLP-1 receptors, it has multiple effects on glucose. Semaglutide (SM, also known as semaglutide, semaglutide) is one of the GLP-1 receptor agonists, which has 94% homology with GLP-1, and is used for the treatment of type II diabetes and obesity. It reduces blood sugar by stimulating insulin secretion and reducing glucagon secretion, both of which are in a glucose-dependent manner, so that hypoglycemia is less likely to occur when used, and the safety is higher than that of insulin.

[0003] At present, polypeptide drugs on the market are mostly injections. Although this method has high utilization degree, daily or weekly invasive subcutaneous injection can cause inconvenience and psychological stress in patients. Therefore, some non-invasive drug delivery routes are being studied. Among them, oral and transdermal administration are two directions widely studied in recent years. Novo Nordisk has the first oral semaglutide product, which is in the form of tablets, and the absolute bioavailability is only 0.4-1%, which is relatively low. Compared with semaglutide subcutaneous injection once a week, the oral dosage form needs a relatively large dose and a higher administration frequency (once a day), which may reduce patient compliance and increase treatment costs.

[0004] As a new transdermal delivery technology, microneedle technology can solve many problems associated with subcutaneous injection, such as needle stick injury, needle phobia, sharp waste and transmission of blood-borne pathogens. Microneedle administration is painless, non-invasive and self-administered, which does not require professional management and improves patient compliance. In addition, microneedles bypass liver first-pass metabolism and avoid enzymatic degradation, improving drug bioavailability. The solid form of microneedles greatly improves the stability of some drugs, eliminating the need for cold chain storage and transportation, reducing cost requirements. However, the dissolution rate of the high molecular material used in the dissolving microneedle varies greatly, which may have little effect on small molecule drugs, but has a greater impact on the delivery rate of macromolecular drugs such as semaglutide (molecular weight 4113 Da). If polypeptide drugs cannot be quickly delivered into the blood, the therapeutic effect of the drug will be affected. Therefore, it is necessary to develop a semaglutide microneedle patch to deliver the drug into the body as quickly as possible to achieve the onset concentration of the drug, and to store it at room temperature to maintain the stability of macromolecular drugs. SUMMARY

[0005] The purpose of the present application is to provide a microneedle composition containing semaglutide and its preparation method and application. By preparing a microneedle patch with semaglutide as the active ingredient and controlling the microneedle components, semaglutide is quickly delivered to maintain therapeutic effect, and can be stored at room temperature to maintain the stability of semaglutide. Compared with oral dosage forms, the bioavailability of semaglutide is improved.

[0006] To achieve the above-mentioned purpose, the present application provides the following technical solutions:

[0007] One of the technical solutions of the present application: a microneedle composition containing semaglutide is provided, which is composed of a needle body and a substrate; the components of the needle body include semaglutide, a cosolvent and an excipient;

[0008] The cosolvent is selected from one or more of Tween, Span, propylene glycol, glycerol and lecithin;

[0009] The excipient is selected from one or more of dextran, polyvinylpyrrolidone, hyaluronic acid and water-soluble cyclodextrin.

[0010] The excipient selected in the present application is a high molecular material with fast drug delivery rate and not easy to form film. Using this raw material as an excipient can make the drug deposit in the needle body part, rather than form a film on the surface of the mold used in the preparation process of the microneedle composition. More preferably, the excipient is one or more of dextran-40, dextran-70, polyvinylpyrrolidone K90, hyaluronic acid with a weight average molecular weight of 20,000-80,000 and sulfobutyl betadex sodium cyclodextrin.

[0011] The selected cosolvent can stabilize the polypeptide and help to store the microneedle composition for a long time.

[0012] Preferably, the components of the needle body further comprise a protective agent; the protective agent is zinc sulfate, zinc chloride or zinc gluconate.

[0013] More preferably, the mass ratio of the protective agent to the semaglutide is 1:10-100.

[0014] The protective agent added in the needle body of the application is also beneficial to maintain the stability of semaglutide and prolong the storage period of the microneedle composition.

[0015] Preferably, the components of the substrate comprise polyvinyl alcohol.

[0016] More preferably, the components of the substrate comprise polyvinyl alcohol and one or more selected from polyvinylpyrrolidone K90 and hyaluronic acid; the combination of these polymers can better connect with the needle body part, make the needle body of the microneedle completely separate from the mold, and maintain the flatness, toughness and mechanical strength of the surface of the microneedle; meanwhile, the selection of these polymer combinations can also reduce the diffusion of drugs in the needle body into the substrate.

[0017] Preferably, the mass ratio of the semaglutide to the excipient is 1:20-4:1; more preferably, 1:10-2:1.

[0018] The second technical scheme of the application provides a preparation method of the microneedle composition containing semaglutide, comprising the following steps:

[0019] The components in the needle body are dissolved in water to obtain a needle body aqueous solution, which is then added to a mold, dried to obtain a needle body layer; then a prepared substrate aqueous solution is added to the mold, dried to obtain the microneedle composition containing semaglutide.

[0020] Preferably, the mass percentage content of the cosolvent in the needle body aqueous solution is 0.05-2%.

[0021] Preferably, the solid content of the substrate aqueous solution is 20-60%; more preferably, 25-40%.

[0022] The third technical scheme of the application provides an application of the microneedle composition containing semaglutide in preparing a drug preparation for treating type 2 diabetes or obesity.

[0023] The beneficial technical effects of the application are as follows:

[0024] The somatostatin and the microneedle excipient are prepared into soluble microneedles, the microneedles have sufficient mechanical strength to pierce the skin, and realize transdermal delivery of macromolecular polypeptide drugs. In the pharmacokinetic experiment, the microneedle composition can reach the maximum blood drug concentration at the same time as the injection form, and the relative bioavailability can be increased by 40-60 times compared with oral administration. The microneedle composition exists in a solid form, so that the stability of somatostatin is greatly improved, and the cost requirement for storage is greatly reduced. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 is an in-vitro cumulative release curve of the needle body prepared by different microneedle excipients in Examples 1-7 and Comparative Examples 1-6.

[0026] Figure 2 is a body microscope graph of the somatostatin microneedle composition prepared in Example 9.

[0027] Figure 3 is a body microscope graph of the fluorescein isothiocyanate grafted somatostatin microneedle composition prepared in Example 19.

[0028] Figure 4 is a body microscope graph of the fluorescein isothiocyanate grafted somatostatin microneedle composition prepared in Comparative Example 11.

[0029] Figure 5 is a long-term stability test result graph of two batches of somatostatin layered dissolving microneedles prepared in Example 24, wherein a is the long-term stability test result of the first batch of products, and b is the long-term stability test result of the second batch of products.

[0030] Figure 6 is a mechanical property test result of the somatostatin layered dissolving microneedles prepared in Example 17.

[0031] Figure 7 is a pharmacokinetic curve of the somatostatin layered dissolving microneedles and somatostatin injection prepared in Examples 9-10. DETAILED DESCRIPTION

[0032] The detailed description set forth below is intended as a description of various example embodiments of the present application and is not intended to represent the only embodiments in which the present application can be practiced. It is also not intended to represent that the present application cannot be practiced except as set forth in the following description or except as otherwise

[0033] In addition, for numerical ranges of the present application, it is to be understood that every intervening value, to the upper and lower limits, is also specifically disclosed. Each smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within the application. The upper and lower limits of these smaller ranges can independently be included or excluded in the range.

[0034] All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains unless clearly indicated otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, the preferred methods and materials are described.

[0035] As used herein, the terms "comprise", "comprising", "include", "including", "have" and "having" or the like are used synonymously to denote or describe a combination including the stated elements or integers or the like that do not, through explicit reference or implication, foreclose the addition of one or more other elements or integers.

[0036] Examples 1-8 and Comparative Examples 1-6

[0037] Different microneedle excipients were selected to prepare integrated microneedles loaded with semaglutide, and high molecular materials with fast drug release rate and good drug stability were screened as needle body materials.

[0038] (1) Preparation of microneedle matrix solution: according to Table 1, a certain amount of water was weighed in a centrifuge tube, the weighed semaglutide (SM) powder was added, stirred and dissolved, then the corresponding microneedle excipient was added, stirred and dissolved, and centrifuged at 4000 rpm and 4°C for 5 min to obtain the microneedle preparation solution;

[0039] (2) Preparation of microneedles: according to Table 1, the corresponding volume of microneedle preparation solution was removed with a liquid adding gun and added to the PDMS microneedle mold. After vacuumizing the mold for 10 min, the microneedles were dried at room temperature and demolded. The theoretical drug content of the whole microneedles was 3 mg.

[0040] (3) In vitro release test: the microneedle patch was fixed on the inner wall of the reciprocating cylinder near the bottom end with double-sided tape, and the reciprocating cylinder was placed in the dissolution cup. The test was carried out under the temperature condition of 32±0.5°C according to the release condition. Samples were taken at 5, 10, 15, 20, 30, 45 and 60 min, respectively, and filtered with a 0.22 μm filter. The content of semaglutide in the sample solution was determined by high performance liquid chromatography.

[0041] (4) Influence factor test: the microneedle patch containing hydrocolloid was placed in a culture dish and stored in a 60°C environment for 10 days. The content of semaglutide in the integrated microneedle was determined by high performance liquid chromatography at 0th, 5th and 10th day.

[0042] The microneedle excipients used by the inventors to prepare the one-piece dissolving microneedles of Examples 1-8 and Comparative Examples 1-6 are listed in Table 1. The microneedle excipients of the Examples include dextran-40 (DT-40), dextran-70 (DT-70), sulfobutyl ether betacyclodextrin (SEB-β-CD), polyvinylpyrrolidone (PVP K90), sodium hyaluronate 20,000 molecular weight (HA-2), sodium hyaluronate 80,000 molecular weight (HA-8), methyl betacyclodextrin (M-β-CD), polyvinyl alcohol (PVA); the microneedle excipients of the Comparative Examples include sodium carboxymethylcellulose 35 cp (CMC-35 cp), sodium carboxymethylcellulose 117 cp (CMC-117 cp), sodium carboxymethylcellulose 800 cp (CMC-800 cp), polyvinylpyrrolidone (PVP K30), sodium hyaluronate 240,000 molecular weight (HA-24), sodium hyaluronate 800,000 molecular weight (HA-80). The results of the influence factor experiments are shown in Table 1.

[0043] Table 1 Microneedle formulations and results of influence factor experiments of Examples 1-8 and Comparative Examples 1-6

[0044] Note: Residual content = 0 or 5 or 10 day content / 0 day content x 100%

[0045] The above results show that PVP K30 has the worst effect on the stability of SM, and the other microneedle excipients have little difference in their effects on the stability of SM.

[0046] The results of the in vitro release experiments are shown in Figure 1 (release medium: PBS solution, pH = 7.4). The microneedle excipients of M-β-CD, SEB-β-CD, DT-40, DT-70, HA-2, PVP K30, HA-8, and PVP K90 can make SM release faster; the microneedle excipients of PVA, HA-24, HA-80, CMC-35 cp, CMC-800 cp, and CMC-117 cp can make SM release slower. Although these high molecular materials make the drug release rate slower, they have good stability and can be used as microneedle excipients without drugs.

[0047] Examples 9-12

[0048] The raw and auxiliary material ratios of the layered dissolving microneedle compositions of SM in Examples 9-12 are shown in Table 2.

[0049] Table 2 Raw and auxiliary material ratios of SM microneedle compositions

[0050] According to the raw and auxiliary material ratios of Table 2, SM dissolving microneedles were prepared as follows:

[0051] (1) Preparation of the matrix solution of microneedle:

[0052] Preparation of the needle body solution: precisely weigh the amount of propylene glycol in a centrifuge tube, then use a pipette to transfer the amount of ultrapure water into the centrifuge tube, stir and dissolve, add the amount of somatuline peptide into the centrifuge tube, stir and dissolve, then add the amount of DT-40 and HA-2 into the centrifuge tube, stir and dissolve, centrifuge at 3000 rpm for 3 min, and obtain the needle body solution.

[0053] Preparation of the base solution: weigh the amount of ultrapure water in a centrifuge tube, add the amount of polyvinyl alcohol (PVA) into the centrifuge tube, place it in a 90°C vacuum drying oven for swelling, then stir until completely dissolved, centrifuge at 5000 rpm for 10 min. After moist heat sterilization at 121°C, the base solution is obtained.

[0054] (2) Preparation of microneedles: use a liquid adding gun to transfer 25 uL of the prepared needle body solution onto the unit mold, evenly spread it, and perform negative pressure on the mold at room temperature until no solution exists on the surface of the mold; then use a liquid adding gun to transfer 60 uL of the base solution onto the unit mold, evenly spread it, perform negative pressure on the mold for 15 min, and then naturally dry the microneedles at room temperature before demolding. The theoretical drug content of the dissolving microneedles obtained in Examples 9-12 is 0.0625 mg / cm 2 , 0.25 mg / cm 2 , 1 mg / cm 2 , and 0.125 mg / cm 2 , respectively.

[0055] (3) Determination of the content of microneedles and residual patches: apply the dissolving microneedles to the abdominal skin of mice, press for 30 s, wrap the rat abdomen with medical tape, and after 2 h, remove the residual patches from the skin and collect them in a centrifuge tube. Add an appropriate amount of PBS solution to the centrifuge tube, shake for 1 h, filter with a 0.22 μm filter membrane, analyze the drug content of the residual microneedles using high performance liquid chromatography, and calculate the drug utilization rate; drug utilization rate = (microneedle drug content - residual microneedle drug content) / microneedle drug content x 100%. The calculation results are shown in Table 3.

[0056] Table 3 Experimental results of microneedle compositions in Examples 9-12

[0057] As can be seen from Table 3, the drug utilization rates of the two prescription microneedle compositions after being applied to living rats for 2 h are almost above 50%.

[0058] Examples 13-16 and Comparative Examples 7-8

[0059] Investigation of excipients in needle body materials:

[0060] The polymer materials used by the inventors to prepare the layered dissolving microneedles in Examples 13-16 and Comparative Examples 7-8 are listed in Table 4. Microneedle excipients with high delivery efficiency and not prone to film formation were used in the examples, and microneedle excipients with slow drug delivery rate were used in the comparative examples. The cosolvent in the needle body material was all Tween 80.

[0061] Table 4 Raw and auxiliary material ratio of somatostatin microneedle composition

[0062] According to the ratio in Table 4, somatostatin dissolving microneedles were prepared according to the following steps:

[0063] (1) Preparation of microneedle matrix solution:

[0064] Preparation of needle body solution: Precisely weigh the amount of Tween 80 in the centrifuge tube, then use a pipette to remove the amount of ultrapure water in the centrifuge tube, stir and dissolve, then add the amount of somatostatin in the centrifuge tube, stir and dissolve, then add the amount of polymer material in the centrifuge tube, stir and dissolve, centrifuge at 3000 rpm for 3 min, and obtain the needle body solution.

[0065] Preparation of base solution: Weigh the amount of ultrapure water in the centrifuge tube, add the amount of PVP K90 in the centrifuge tube, stir until completely dissolved, then add the amount of PVA in the centrifuge tube, place it in a 90°C vacuum drying oven to swell, then stir until completely dissolved, then add the amount of HA-24 in the centrifuge tube, stir until completely dissolved, centrifuge at 5000 rpm for 10 min. After moist heat sterilization at 121°C, the base solution is obtained.

[0066] (2) Preparation of microneedles: 25uL of the prepared needle body solution was added to the unit mold using a liquid addition gun, evenly spread, and the mold was subjected to negative pressure at room temperature until no solution was present on the surface of the mold; then 80uL of the base solution was added to the unit mold using a liquid addition gun, evenly spread, and the mold was subjected to negative pressure for 30 min, then the microneedles were naturally dried at room temperature and demolded. The theoretical drug content of the layered microneedles obtained was 0.5mg.

[0067] (3) Determination of drug content in microneedle needle body and base: The prepared microneedle sample was cut off with a scalpel, collected in a centrifuge tube, and the corresponding base was placed in another centrifuge tube, labeled. Add an appropriate amount of PBS solution to the centrifuge tube, shake for 1h, filter with a 0.22μm filter, and analyze the corresponding drug content of the needle body and base using high performance liquid chromatography, and calculate the proportion of drug in the needle body.

[0068] (4) Proportion of drug in needle body = drug content in needle body / (drug content in needle body + drug content in base) x 100%.

[0069] Drug utilization rate was calculated by the same formula as in Example 9, where the dissolving microneedles were applied to the human arm for 1 h.

[0070] Table 5 Experimental results of microneedle compositions of Examples 13-16 and Comparative Examples 7-8

[0071] As can be seen from Table 5, the drug content ratio of the needle body of Examples 13-16 is 1.7-2.1 times higher than that of the comparative examples; after applying to the human arm for 1 h, the microneedle drug utilization rate of the examples is twice that of the comparative examples by analyzing the content of the residual patch. This shows that the needle body material selected from the high molecular material which is not easy to form a film can more easily deposit the drug in the needle body, and the delivery rate is faster, which can correspondingly improve the drug utilization rate.

[0072] Examples 17-18 and Comparative Examples 9-10

[0073] The base material was prepared into somatostatin layered dissolving microneedles using high molecular materials with different solid contents, and the ratio is shown in Table 6.

[0074] Table 6 Raw and auxiliary material ratio of somatostatin microneedle composition

[0075] According to the ratio in Table 6, somatostatin layered dissolving microneedles were prepared according to the following steps:

[0076] (1) Preparation of microneedle matrix solution:

[0077] Preparation of needle body solution: accurately weigh the amount of Tween 80 in the centrifuge tube, then use a pipette to transfer the amount of ultrapure water in the centrifuge tube, shake to dissolve, then add the amount of somatostatin in the centrifuge tube, shake to dissolve, then add the amount of high molecular material in the centrifuge tube, shake to dissolve, and stand to obtain the needle body solution.

[0078] Preparation of base solution 1 and 4: weigh the amount of ultrapure water in the centrifuge tube, add the amount of PVPK90 in the centrifuge tube, stir until completely dissolved, then add the amount of PVA in the centrifuge tube, place in a 90°C vacuum drying oven for swelling, then stir until completely dissolved, then add the amount of HA-24 in the centrifuge tube, stir until completely dissolved, centrifuge at 6000 rpm for 15 min. After moist heat sterilization at 121°C, base solution 1 and base solution 4 are obtained.

[0079] Preparation of base solution 2: weigh the amount of ultrapure water in the centrifuge tube, first add the amount of PVPK90, stir until completely dissolved, then add the amount of PVA in the centrifuge tube, place in a 90°C vacuum drying oven for swelling, then stir until completely dissolved, centrifuge at 6000 rpm for 15 min. After moist heat sterilization at 121°C, base solution 2 is obtained.

[0080] The base solution 3 was prepared: the formula amount of ultrapure water was weighed in a centrifuge tube, the formula amount of Tween 80 was first added to the centrifuge tube, stirred and dissolved, then the formula amount of DT-40 was added to the centrifuge tube, stirred until completely dissolved, and finally the formula amount of HA-24 and HA-80 was added to the centrifuge tube, stirred until completely dissolved. Centrifugation was performed at 6000 rpm for 10 min. After moist heat sterilization at 121°C, the base solution 3 was obtained.

[0081] (2) Preparation of microneedles: 50 uL of the prepared needle body solution was added to the unit mold using a liquid adding gun, and evenly spread. The mold was subjected to negative pressure at room temperature until no solution was present on the surface of the mold. Then 80 uL of the base solution was added to the unit mold using a liquid adding gun, and evenly spread. The mold was subjected to negative pressure for 15 min, and then the microneedles were naturally dried at room temperature and demolded.

[0082] (3) Determination of microneedle content and residual patch content: the dissolved microneedles were applied to the isolated pig skin, pressed for 30 s, and after 2 h the residual patch was removed from the skin and collected in a centrifuge tube. An appropriate amount of PBS solution was added to the centrifuge tube, shaken for 1 h, filtered with a 0.22 μm filter membrane, and the residual microneedle drug content was analyzed using high performance liquid chromatography, and the drug utilization rate was calculated, in the same manner as in Example 9.

[0083] Table 7 Experimental results of microneedle compositions of Examples 17-18 and Comparative Examples 9-10

[0084] As can be seen from Table 7, the drug utilization rate of the microneedles of Examples 17-18 is about 50%, while the drug utilization rate of the microneedles of Comparative Examples 9-10 is extremely low. This indicates that when the solid content of the base solution of the microneedles is high, the diffusion of the drug in the needle tip solution into the base solution is relatively small. When the solid content of the base solution of the microneedles is low, the diffusion of the drug in the needle tip solution into the base solution is relatively large, resulting in that when the microneedles are used, the drug contained in the needle body is extremely small, and the drug utilization rate is extremely low.

[0085] Example 19 and Comparative Example 11

[0086] A preparation method of a dissolving microneedle loaded with fluorescein isothiocyanate grafted somatostatin:

[0087] (1) The needle tip solution was a 10 mg / ml fluorescein isothiocyanate grafted somatostatin aqueous solution containing 1% HA-8;

[0088] (2) The base solution 1 was the same as that used in Example 17;

[0089] (3) The base solution 2 was the same as that used in Comparative Example 9;

[0090] Preparation of microneedles of Example 19

[0091] The prepared needle body liquid was added dropwise to the unit mold by using a liquid adding gun, and evenly spread. The mold was subjected to negative pressure until no solution existed on the surface of the mold under room temperature conditions. Then, 80 uL of base solution 1 was added dropwise to the unit mold by using a liquid adding gun, and evenly spread. The mold was subjected to negative pressure for 15 min, and then naturally dried under room temperature conditions. After the microneedle was demolded, a dissolved microneedle loaded with fluorescently labeled somatuline was obtained, as shown in FIG. 3.

[0092] Preparation of microneedles of Comparative Example 11

[0093] The prepared needle body liquid was added dropwise to the unit mold by using a liquid adding gun, and evenly spread. The mold was subjected to negative pressure until no solution existed on the surface of the mold under room temperature conditions. Then, 80 uL of base solution 2 was added dropwise to the unit mold by using a liquid adding gun, and evenly spread. The mold was subjected to negative pressure for 15 min, and then naturally dried under room temperature conditions. After the microneedle was demolded, a dissolved microneedle loaded with fluorescently labeled somatuline was obtained, as shown in FIG. 4.

[0094] As can also be seen from Example 19 and Comparative Example 11, when the solid content of the microneedle base solution is high, the fluorescently labeled drug is more concentrated in the needle body, and only a small amount diffuses upward. When the solid content of the microneedle base solution is low, the drug in the needle body basically diffuses to the outermost edge of the microneedle, and the needle body contains very little drug, which cannot well deliver the drug into the body.

[0095] Examples 20-23

[0096] Somatuline layered dissolved microneedles were prepared by using different liquid adding amounts of the needle body liquid, and the solid content after drying was consistent. The ratio is shown in Table 8.

[0097] Table 8 Raw material ratio of somatuline microneedle composition

[0098] According to the raw material ratio in Table 8, somatuline layered dissolved microneedles were prepared by referring to the preparation method of the dissolved microneedles in Example 9. According to the needle body drug ratio formula in Example 13, the drug content of the needle body and the base was determined to calculate the needle body drug ratio.

[0099] Table 9 Experimental results of microneedle composition of Examples 20-23

[0100] As can be seen from Table 9, the needle body drug ratio of Examples 20-23 can all reach more than 70%. When the total weight after drying is constant, the needle body drug ratio gradually decreases with the increase of the liquid adding amount of the needle body, which indicates that when the microneedle is loaded with the same drug loading amount, the liquid adding amount of the needle body liquid should be as small as possible, and the prepared solid content should be relatively high.

[0101] Examples 24-25

[0102] The raw and auxiliary material ratio of the somatostatin layered dissolving microneedle composition is shown in Table 10.

[0103] Table 10 Raw and auxiliary material ratio of somatostatin microneedle composition

[0104] According to the raw and auxiliary material ratio in Table 10, the somatostatin layered dissolving microneedle was prepared according to the preparation method of the dissolving microneedle in Example 9. The microneedle was placed in a 60°C environment for 10 days. The somatostatin content in the one-piece microneedle was determined by high performance liquid chromatography on the 0th, 5th, and 10th days. The results showed that the addition of a small amount of zinc ions in the somatostatin microneedle preparation can have a certain stabilizing effect on somatostatin.

[0105] Long-term stability test

[0106] The two batches of somatostatin microneedles prepared in Example 24 were used for long-term stability test. The drug content was detected on the 0th, 1st, 2nd, and 3rd months: the microneedle patch adhered to the hydrogel was packaged with a bubble cap and an aluminum plastic bag and stored in-20°C, 4°C, 25°C, and 40°C environments for 3 months. The somatostatin content in the layered microneedle on the 0th, 1st, 2nd, and 3rd months was determined by high performance liquid chromatography.

[0107] The stability results are shown in Figure 5, where a is the long-term stability test results of the first batch of products, and b is the long-term stability test results of the second batch of products. After storage at-20°C, 4°C, 25°C, and 40°C for 3 months, the somatostatin content in the two batches of layered dissolving microneedles of Example 24 had no significant difference and could reach 96%. Compared with the 0th month, the drug content did not decrease significantly after 3 months of storage, which indicated that the stability of the preparation was good.

[0108] Microneedle mechanical property test

[0109] ​Mechanical performance test was performed using the solubility microneedle of semaglutide prepared in Example 17. The solubility microneedle of semaglutide was fixed on the test platform under the force-travel tester with pressure-sensitive adhesive, with the needle tip upward and perpendicular to the mechanical sensor probe above. The sensor probe was 2 mm x 2 mm in size. When the test started, the sensor probe moved vertically downward at a longitudinal speed of 1.1 mm / s to apply a longitudinal force to the microneedle array, with the upper limit of the force being 10 N. The computer recorded the curve of the longitudinal force of the microneedle and the moving distance of the probe simultaneously. The obtained data were plotted into a mechanical performance curve, as shown in FIG. 6. When the displacement reached 0.25 mm, no obvious breakpoint appeared on the mechanical performance curve of the microneedle, indicating that the microneedle did not break during the process, and the compression force of a single needle of the microneedle was greater than 0.058 N, which indicated that the solubility microneedle of semaglutide had sufficient mechanical strength to penetrate the stratum corneum to achieve drug delivery.

[0110] Pharmacokinetic test

[0111] The solubility microneedle of semaglutide prepared in Examples 9-10 and the injection solution were selected for pharmacokinetic study.

[0112] The pharmacokinetic experiment used SD male rats. Twenty-five SD male rats were randomly divided into five groups, with five rats in each group, and were labeled respectively. On the day before the experiment, all the rats were shaved on the abdomen and treated with depilatory cream. Two microneedle administration groups: after the rats were anesthetized with isoflurane, one microneedle patch was pressed on the rat's abdominal skin with the thumb for 30 s, and the rat's abdomen was wrapped with medical tape. After 2 h, the residual patch was removed from the skin and collected into a centrifuge tube, and the drug in the residual patch was extracted with PBS solution. The drug content was analyzed by high performance liquid chromatography. Subcutaneous injection administration group: after the rats were anesthetized with isoflurane, 0.25 mL of SM solution (250 μg / mL) was drawn into a 1 mL syringe and injected subcutaneously into each rat's abdomen. The administration dose was 62.5 μg per rat. At 1, 2, 3, 4, 6, 8, 12, 24, 36, and 48 h after administration, blood was collected from the tail vein or the orbit of each rat, and each rat was anesthetized with isoflurane for a short period. The whole blood was collected in an EDTAK2 centrifuge tube, shaken, and centrifuged at 1600 x g for 10 min. The supernatant was taken, and the semaglutide content in the blood sample was analyzed by liquid chromatography-mass spectrometry to draw the pharmacokinetic curve.

[0113] Pharmacokinetic test results

[0114] Compared with SC administration, the pharmacokinetics of semaglutide was studied after the solubility microneedle of semaglutide was implanted into normal rats. The pharmacokinetic parameters and the pharmacokinetic curve after administration are shown in Table 11 and FIG. 7.

[0115] Table 11 Pharmacokinetic parameters of semaglutide microneedle composition and injection solution

[0116] As can be seen from Table 11, the microneedle high-dose group administered at a dose of 264 μg (129 μg delivered to the body) can have the same pharmacokinetic behavior as the injection group (dose of 62.5 μg), with maximum blood drug concentrations of 655 ng / mL and 667 ng / mL, respectively. The peak level was reached 8 h after administration in the microneedle high-dose, low-dose, and injection groups. With subcutaneous injection as the reference formulation, the relative bioavailability of the microneedle high-dose group was 49.0% based on the intake dose and the calculated AUC value, which indicates that the somatropin released from the microneedle is quickly introduced into the circulating system.

[0117] The above-described embodiments are merely intended to describe the preferred modes of the present application, and are not intended to limit the scope of the present application, and various modifications and improvements to the technical solutions of the present application made by those skilled in the art without departing from the design spirit of the present application shall fall within the scope of protection of the present application as defined by the claims.

Claims

1. A microneedle composition comprising semaglutide, characterized in that, The microneedle composition consists of a needle body and a substrate; the components of the needle body include somatuline, a cosolvent and an excipient; The cosolvent is selected from one or more of Tween, Span, propylene glycol, glycerol and lecithin; The excipient is selected from one or more of dextran, polyvinylpyrrolidone, hyaluronic acid and water-soluble cyclodextrin.

2. The sumatriptan containing microneedle composition of claim 1, wherein, The components of the needle body further include a protective agent; the protective agent is zinc sulfate, zinc chloride or zinc gluconate.

3. The sumatriptan containing microneedle composition of claim 2, wherein, The mass ratio of the protective agent to the somatuline is 1:10-100.

4. The microneedle composition containing semaglutide according to claim 1, characterized in that, The components of the substrate include polyvinyl alcohol.

5. The sumatriptan containing microneedle composition of claim 1, wherein the sumatriptan is present in an amount of about 0.1 mg to about 1 mg. The mass ratio of the somatuline to the excipient is 1:20-4:

1.

6. A process for the preparation of the microneedle composition containing semaglutide according to any one of claims 1 to 5, characterized in that, The method comprises the following steps: The components in the needle body are dissolved in water to obtain a needle body aqueous solution, which is then added to a mold, dried to obtain a needle body layer; then a prepared substrate aqueous solution is added to the mold, dried to obtain the microneedle composition containing somatuline.

7. The production method according to claim 6, characterized by, The mass percentage of the cosolvent in the needle body aqueous solution is 0.05-2%.

8. The preparation method according to claim 6, characterized in that, The solid content of the substrate aqueous solution is 20-60%.

9. Use of the microneedle composition containing somatuline according to any one of claims 1-5 in the preparation of a pharmaceutical preparation for treating type 2 diabetes or obesity.

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