Sustained-release injection composition comprising exenatide and method for preparing same

Abstract

The present invention relates to a sustained-release injection composition comprising exenatide and a method for preparing same. The sustained-release injection composition enables exenatide to be continuously released in the body for 1-2 months with a single injection, thus making it possible to reduce the number of administrations and improve the convenience of administration. In addition, since the sustained-release injection composition contains exenatide in microparticles having a uniform particle size, long-term sustained release of exenatide can be achieved, and thus the bio-absorption rate of exenatide can be increased compared to conventional injectable formulations comprising exenatide.

Description

Sustained-release injectable composition containing exenatide and method for preparing the same

[0001] The present invention relates to a sustained-release injectable composition comprising exenatide and a method for preparing the same.

[0002] Exenatide is a synthetic peptide of Exendin-4, a glucagon-like peptide 1 (GLP-1) analogue, and is a drug used to lower blood sugar levels by promoting insulin secretion in patients with type 2 diabetes.

[0003] It shares approximately 53% homology with natural GLP-1, acting as an agonist for the GLP-1 receptor but resistant to the protease DPP-IV, with a half-life of 2 to 4 hours. The initial dose is 5 mcg subcutaneously twice daily, with a maximum dose of 10 mcg twice daily. However, its use in type 1 diabetes is not yet approved.

[0004] Several studies on exenatide have consistently shown effectiveness in reducing fasting and postprandial blood glucose, glycated hemoglobin, and body weight; however, efforts are ongoing to improve its short half-life.

[0005] Therefore, there is a need to develop a formulation containing highly effective exenatide to improve patient convenience, such as reducing the number of administrations, and to reduce manufacturing costs.

[0006] [Prior Art Literature]

[0007] [Patent Document]

[0008] KR 10-2018-0129825 A1

[0009] The purpose of the present invention is to provide a sustained-release injectable composition containing exenatide and a method for preparing the same.

[0010] Another object of the present invention is to provide a sustained-release injectable composition that can continuously release exenatide in the body for 1 to 2 months with a single injection, thereby reducing the number of administrations and increasing the convenience of administration.

[0011] Another object of the present invention is to provide a method for producing a sustained-release injectable composition comprising exenatide, which comprises exenatide in microparticles having a uniform particle size, and can exhibit a sustained release effect of exenatide for a long period of time, thereby increasing the bioabsorption rate of exenatide compared to conventional injectable formulations of exenatide.

[0012] To achieve the above object, the present invention relates to a sustained-release injectable composition comprising microparticles, wherein the microparticles comprise exenatide and a biodegradable polymer, and the sustained-release injectable composition comprises exenatide having an area under the plasma level-time curve (AUC) of 40,000 pg·d / mL to 100,000 pg·d / mL, according to the release of exenatide over 63 days, when exenatide is administered to SD rats at 8 mg / kg.

[0013] In addition, the above-mentioned sustained-release injection composition exhibits an initial rapid exenatide release effect when exenatide is administered to SD rats at a dose of 8 mg / kg, and the concentration of exenatide in the blood can then be maintained at a steady state.

[0014] Additionally, the above-mentioned western-type injection composition can continuously release exenatide for 1 to 2 months.

[0015] Additionally, the microparticles may contain exenatide and a biodegradable polymer in a weight ratio of 1:3 to 1:7.

[0016] Additionally, the microparticles may have an average diameter of 40 μm to 80 μm.

[0017] Additionally, the biodegradable polymer may be selected from the group consisting of polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA), polyphosphazine, polyiminocarbonate, polyphosphoester, polyanhydride, polyorthoester, polycaprolactone, polyhydroxyvalerate, polyhydroxybutyrate, polyamino acid, and combinations thereof.

[0018] Additionally, the injectable composition may include a suspending solvent.

[0019] According to another embodiment of the present invention, a method for preparing a sustained-release injectable composition comprising exenatide comprises the steps of: 1) preparing an oily solution by mixing exenatide and a biodegradable polymer in an organic solvent; 2) preparing an aqueous solution by dissolving a surfactant in purified water; 3) injecting the oily solution and the aqueous solution into each microchannel and allowing them to flow to generate microparticles at the intersection of the microchannels; 4) collecting the microparticles in a tank containing the aqueous solution; 5) removing the organic solvent present in the collected microparticles; 6) washing and drying the microparticles from which the organic solvent has been removed with purified water; And 7) a step of preparing a sustained-release injection composition by mixing the dried microparticles with a suspending solvent, wherein the sustained-release injection composition is administered to SD rats at 8 mg / kg of exenatide, and the area under the plasma level-time curve (AUC) according to the release of exenatide for 63 days can be 40,000 pg·d / mL to 100,000 pg·d / mL.

[0020] Additionally, the above oily solution can be prepared by dissolving exenatide in a co-solvent and then mixing a biodegradable polymer and an organic solvent.

[0021] The above co-solvent may be selected from the group consisting of acetic acid, dimethyl sulfoxide, dimethyl formamide, tetrahydrofuran, ethyl acetate, acetonitrile, and mixtures thereof.

[0022] Additionally, the co-solvent may be included in a weight ratio of 1:1 to 1:2 with the organic solvent.

[0023] In addition, when the above oil solution is injected into the microchannel, the pressure can be increased at a condition of 1 to 5 mbar / min after injection under a pressure condition of 500 to 700 mbar.

[0024] Additionally, the above-mentioned aqueous solution can be injected into the microchannel under pressure conditions of 2,500 mbar to 3,000 mbar.

[0025] In addition, the above step 5) may be stirred at a speed of 200 to 400 rpm for 4 to 6 hours at 40°C to 50°C.

[0026] The present invention utilizes a sustained-release injection composition to continuously release exenatide in the body for 1 to 2 months with a single injection, thereby reducing the number of administrations and increasing the convenience of administration.

[0027] In addition, since exenatide is contained in microparticles having a uniform particle size, it can exhibit a long-term, sustained release effect of exenatide, and thus, the bioabsorption rate of exenatide can be increased compared to conventional injectable formulations of exenatide.

[0028] Figure 1 is a SEM measurement photograph of microparticles according to one embodiment of the present invention.

[0029] Figure 2 is an SEM measurement photograph of microparticles according to one embodiment of the present invention.

[0030] Figure 3 is an SEM measurement photograph of microparticles according to one embodiment of the present invention.

[0031] Figure 4 is an SEM measurement photograph of microparticles according to one embodiment of the present invention.

[0032] Figure 5 shows the PK measurement results for an injection composition according to one embodiment of the present invention.

[0033] Figure 6 shows the PK measurement results for an injection composition according to one embodiment of the present invention.

[0034] The present invention relates to a sustained-release injectable composition comprising microparticles, wherein the microparticles comprise exenatide and a biodegradable polymer, and the sustained-release injectable composition comprises exenatide having an area under the plasma level-time curve (AUC) of 40,000 pg·d / mL to 100,000 pg·d / mL, based on the release of exenatide over 63 days, when the sustained-release injectable composition is administered to SD rats at 8 mg / kg.

[0035] Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement them. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

[0036] Exenatide is a 39-amino acid peptide that is a potent GLP-1 receptor agonist and an insulin secretagogue with glycemic control effects. It is a Byetta ® (Astra-Zeneca) is a commercially available peptide widely used in the treatment of type 2 diabetes. This peptide has a short in vivo half-life of 2.5 hours, so it is injected twice daily.

[0037] There is a strong desire to extend the half-life of exenatide and related GLP-1 agonist peptides to improve their efficacy, reduce side effects, and ease the burden of treatment on patients.

[0038] Peptide half-life is traditionally extended by one or a combination of several methods: (i) chemical modification of the peptide to delay metabolism; (ii) encapsulation to provide a sustained-release depot formulation; and (iii) conjugation with macromolecules to slow body clearance. [References include, e.g., Cai, et al., Drug Design, Development, and Therapy (2013) 7:963-970].

[0039] Chemical modification of peptides to increase their half-life has resulted in once-daily GLP-1 agonists, such as lixisenatide (Lyxumia®) and liraglutide (Victoza®). Encapsulation of the peptides within PLGA (polylactic-co-glycolic acid) microparticles has been used to prepare a sustained-release formulation, marketed as Bydureon® (Astra-Zeneca), allowing for weekly subcutaneous injections.

[0040] In the case of the above Bydureon®, although it releases exenatide for 10 weeks with a single administration, there is a problem in that the body is not exposed to exenatide to a sufficient extent to exhibit the type 2 diabetes treatment effect due to the release of exenatide. Therefore, it is used for the treatment of type 2 diabetes by subcutaneous injection once a week.

[0041] However, in the case of the method of subcutaneous injection once a week as described above, there is a cumbersome problem of having to administer subcutaneous injection every week, so there is a need to develop a formulation that can exhibit the release effect of exenatide for a longer period of time through a single administration.

[0042] The present invention is intended to solve the above-described problem, and relates to a sustained-release injection composition including microparticles that can release exenatide for 1 to 2 months through a single injection, thereby improving the convenience of administration.

[0043] Specifically, the microparticles include exenatide and a biodegradable polymer, and when the sustained-release injection composition is administered to SD rats at 8 mg / kg of exenatide, the area under the plasma level-time curve (AUC) according to the release of exenatide for 63 days may be 40,000 pg·d / mL to 100,000 pg·d / mL.

[0044] The above AUC is related to the bioabsorption rate according to the release of exenatide, and a higher AUC value indicates a better bioabsorption rate.

[0045] The sustained-release injection composition of the present invention, when exenatide is administered to SD rats at 8 mg / kg, can have an area under the plasma level-time curve (AUC) of 40,000 pg·d / mL to 100,000 pg·d / mL, 45,000 pg·d / mL to 60,000 pg·d / mL, 50,000 pg·d / mL to 60,000 pg·d / mL, and 55,000 pg·d / mL to 580,000 pg·d / mL according to the release of exenatide for 63 days. The AUC value of the sustained-release injection composition of the present invention shows a large difference compared to the above-described Bydureon®.

[0046] That is, even when treated with the same concentration of exenatide as Bydureon® administered once a week, a large difference in AUC value is observed, and thus, the sustained-release injection composition containing exenatide of the present invention can exhibit excellent bioabsorption rate.

[0047] The AUC value as described above is due to the characteristics of the microparticles included in the above-described western-type injection composition. The microparticles of the present invention are particles having a very uniform average diameter, with a smooth spherical shape and an average diameter of 40 µm to 80 µm, 60 µm to 80 µm, and 70 µm to 80 µm, and a standard deviation (SD) of 7 to 8 for the average diameter of the particles.

[0048] On the other hand, the average diameter of the microparticles of the Bydureon® is 76.51㎛, which is not much different from the microparticles of the present invention, but the surface of the particles is not uniform, so there is a big difference in shape, and the standard deviation (SD) for the average diameter of the particles is 24.99, so there is a big difference from the microparticles of the present invention in that the particles do not have a uniform diameter.

[0049] Due to the difference in particle characteristics as described above, the sustained-release injection composition of the present invention can exhibit a sustained release effect of exenatide for 1 to 2 months with a single administration.

[0050] In addition, the sustained-release injection composition can exhibit an initial rapid exenatide release effect and then maintain a steady state when exenatide is administered to SD rats at a dose of 8 mg / kg. That is, when the sustained-release injection composition of the present invention is administered to SD rats, it is characterized in that after an initial rapid release of exenatide is exhibited, the concentration of exenatide in the blood can be continuously maintained above a certain level.

[0051] In the case of conventional microparticles having a uniform diameter, the drug contained within the microparticles is characterized by continuously releasing the drug without initial rapid drug release. However, the microparticles contained in the sustained-release injection composition of the present invention are different in that they exhibit a rapid exenatide release pattern immediately after injection and then continuously release exenatide at a certain level or higher.

[0052] When the sustained-release injection composition of the present invention is administered to SD rats in the above-described manner and the release pattern of exenatide in the blood is confirmed, it can be confirmed that the blood concentration value of exenatide is maintained for a long time after the initial rapid release of exenatide. In contrast, when Bydureon® is administered to SD rats at the same dose, unlike the release pattern of the present invention, a continuous release pattern is shown without the initial rapid release of exenatide. However, from the time of the first injection until 21 days ago, the blood concentration value of exenatide is too low, making it difficult for exenatide to exert its effect, and after 42 days, it can be confirmed that the blood concentration decreases rapidly, confirming that the same effect as the present invention cannot be exerted with a single administration.

[0053] The above microparticles are characterized by containing exenatide and a biodegradable polymer in a weight ratio of 1:3 to 1:7, preferably 1:5, but are not limited to the above example, and when containing exenatide and a biodegradable polymer within the above range, an excellent sustained release effect of exenatide can be exhibited.

[0054] The biodegradable polymer may be selected from the group consisting of polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA), polyphosphazine, polyiminocarbonate, polyphosphoester, polyanhydride, polyorthoester, polycaprolactone, polyhydroxyvalerate, polyhydroxybutyrate, polyamino acid, and combinations thereof, and preferably may be selected from the group consisting of polylactic acid, polylactide-co-glycolide (PLGA), and mixtures thereof, but is not limited to the above examples, and any biodegradable polymer that can be decomposed in vivo to continuously release exenatide may be used without limitation.

[0055] Specifically, the biodegradable polymer may be PLGA, and may include a mixture of two types of PLGA having a viscosity of 0.16 to 0.24 dl / g and PLGA having a viscosity of 0.35 to 0.45 dl / g. When two types of PLGA having different viscosities are included, the release pattern of exenatide unique to the present invention may be exhibited, and continuous release of exenatide may be exhibited for 1 to 2 months, and effects according to the release of exenatide may be exhibited.

[0056] The above-mentioned injectable composition may include a suspending agent. The suspending agent includes a tonicity agent, a suspending agent, and a solvent.

[0057] More specifically, the isotonic agent may be selected from the group consisting of D-Mannitol, Maltitol, Sorbitol, Lactitol, Xylitol, Sodium chloride and mixtures thereof, preferably D-Mannitol, but is not limited to the above examples.

[0058] The above suspending agent is composed of sodium carboxymethylcellulose, polysorbate 80, starch, starch derivatives, polyhydric alcohols, chitosan, chitosan derivatives, cellulose, cellulose derivatives, collagen, gelatin, hyaluronic acid (HA), alginic acid, algin, pectin, carrageenan, chondroitin, chondroitin sulfate, dextran, dextran sulfate, polylysine, titin, fibrin, agarose, fluran, xanthan gum, and mixtures thereof. Selected from the group, preferably sodium carboxymethylcellulose and polysorbate 80, but not limited to the above examples.

[0059] The above solvent can be used as injection water, and any solvent that can be used as injection water can be used without limitation.

[0060] According to another embodiment of the present invention, a method for manufacturing a sustained-release injectable composition comprising exenatide comprises the steps of: 1) preparing an oily solution by mixing exenatide and a biodegradable polymer in an organic solvent; 2) preparing an aqueous solution by dissolving a surfactant in purified water; 3) injecting the oily solution and the aqueous solution into each microchannel and allowing them to flow to generate microparticles at the intersection of the microchannels; 4) collecting the microparticles in a tank containing the aqueous solution; 5) removing the organic solvent present in the collected microparticles; 6) washing and drying the microparticles from which the organic solvent has been removed with purified water; And 7) a step of preparing a sustained-release injection composition by mixing the dried microparticles with a suspending solvent, wherein the sustained-release injection composition is administered to SD rats at 8 mg / kg of exenatide, and the area under the plasma level-time curve (AUC) according to the release of exenatide for 63 days can be 40,000 pg·d / mL to 100,000 pg·d / mL.

[0061] The above step 1) is a step for preparing an oily solution, which is a step for preparing an oily solution by dissolving exenatide and a biodegradable polymer in an organic solvent.

[0062] However, in order to prepare the above oily solution, exenatide may be dissolved in a co-solvent, and then prepared by mixing a biodegradable polymer and an organic solvent.

[0063] The above common solvent may be selected from the group consisting of acetic acid, dimethyl sulfoxide, dimethyl formamide, tetrahydrofuran ethyl acetate, acetonitrile and mixtures thereof, and preferably may be acetic acid and dimethyl sulfoxide.

[0064] As described above, when exenatide is dissolved using a cosolvent and then manufactured into an oily solution, not only can the dissolution of exenatide be assisted, but compared to the case where no cosolvent is used, when manufactured into microparticles and administered into the body, an initial rapid release of exenatide can be induced. In addition, compared to the case where a single cosolvent is used, when acetic acid and dimethyl sulfoxide are used as described above, not only can the initial rapid release of exenatide from the microparticles be induced, but as described above, the bioabsorption rate of exenatide in the body can be increased, and a long-term sustained release effect can be exhibited.

[0065] When dissolving exenatide using the above co-solvent, a first co-solvent and a second co-solvent can be used, and the first co-solvent can be acetic acid, and the second co-solvent can be dimethyl sulfoxide. The first co-solvent and the second co-solvent can be mixed in a weight ratio of 1:1 to 1:5, mixed in a weight ratio of 1:2 to 1:4, or mixed in a weight ratio of 1:3. When the first co-solvent and the second co-solvent are mixed and used within the above range, not only does it facilitate the dissolution of exenatide, but even after being manufactured into microparticles, it can induce an initial rapid release of exenatide from the microparticles, improve the bioabsorption rate, and exhibit the effect of long-term sustained release of exenatide.

[0066] As described above, an oily solution can be prepared by dissolving exenatide using a co-solvent and then mixing a biodegradable polymer and an organic solvent.

[0067] At this time, the weight ratio of the total co-solvent mixed in the oily solution to the weight ratio of the organic solvent may be included in a weight ratio of 1:1 to 1:2. When mixed and used within the above range, exenatide and the biodegradable polymer can be completely dissolved in the organic solvent, so that the oily solution can be manufactured, and the microparticles with a smooth and uniform surface can be manufactured through the manufacturing process described below.

[0068] The biodegradable polymer is selected from the group consisting of polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA), polyphosphazine, polyiminocarbonate, polyphosphoester, polyanhydride, polyorthoester, polycaprolactone, polyhydroxyvalerate, polyhydroxybutyrate, polyamino acid and combinations thereof, preferably polylactide-co-glycolide (PLGA) and / or polylactide (PLA), but is not limited to the above examples.

[0069] In addition, the organic solvent is one that does not mix with water, for example, at least one selected from the group consisting of chloroform, chloroethane, dichloroethane, trichloroethane, and mixtures thereof, preferably dichloromethane, but is not limited to the example, and any organic solvent that can dissolve the biodegradable polymer and exenatide, and is not limited to the example, and any organic solvent that can be easily selected by a person skilled in the art can be used.

[0070] The above oily solution may have a weight ratio of exenatide and biodegradable polymer of 1:3 to 1:7, 1:4 to 1:6, or 1:5. When mixed and used within the above range, exenatide can be continuously released for a long period of time by decomposition of the biodegradable polymer.

[0071] When the weight ratio of the exenatide and the biodegradable polymer is less than 1:3, that is, when the biodegradable polymer is included in an amount less than the weight ratio, the weight ratio of the biodegradable polymer is small compared to the weight of exenatide, and thus it is difficult to manufacture sustained-release particles in which exenatide is evenly distributed and included in spherical biodegradable polymer particles. In addition, when the weight ratio of the biodegradable polymer and exenatide exceeds 1:7, that is, when the biodegradable polymer is included in an amount greater than the weight ratio, the content of exenatide in the sustained-release particles is small, and thus a problem may arise that a large amount of sustained-release particles must be administered to administer the drug at a desired concentration.

[0072] More specifically, the biodegradable polymer in the oil solution is included in an amount of 5 to 15 wt%, 6 to 12 wt%, 7 to 10 wt%, and 9.3 wt%, but is not limited to the above examples.

[0073] The above step 2) is a step for preparing an aqueous solution, and the aqueous solution is prepared by dissolving a surfactant in purified water. The surfactant may be used without limitation as long as it can help the biodegradable polymer solution form a stable emulsion. Specifically, it is at least one selected from the group consisting of nonionic surfactants, anionic surfactants, cationic surfactants, and mixtures thereof, and more specifically, it is at least one selected from the group consisting of methylcellulose, polyvinylpyrrolidone, lecithin, gelatin, polyvinyl alcohol, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene castor oil derivatives, sodium lauryl sulfate, sodium stearate, ester amines, linear diamines, patty amines, and mixtures thereof, and is preferably polyvinyl alcohol, but is not limited thereto.

[0074] The surfactant contained in the above oil solution may be contained in an amount of 0.1 to 1.0 wt%, 0.3 to 0.7 wt%, or 0.5 wt%. The remainder is purified water.

[0075] Step 3) above is a step of injecting and flowing an oil solution and an aqueous solution into a microchannel formed on a wafer.

[0076] More specifically, the microchannel can be formed in a material selected from the group consisting of a silicon wafer or a polymer film, but examples of the material are not limited to the examples above, and any material capable of forming a microchannel can be used.

[0077] The polymer film may be selected from the group consisting of polyimide, polyethylene, fluorinated ethylene propylene, polypropylene, polyethylene terephthalate, polyethylene naphthalate, polysulfone, and mixtures thereof, but is not limited to the above examples.

[0078] As an example, aluminum is deposited on a silicon wafer using an e-beam evaporator, and photoresist is patterned on the aluminum using photolithography. The aluminum is then etched using the photoresist as a mask, the photoresist is removed, and the silicon is etched using DRIE (deep ion reactive etching) using the aluminum as a mask. After the aluminum is removed, glass is anodic bonded to the wafer to seal it, thereby fabricating the microchannels described above.

[0079] The microchannel for manufacturing the microparticles of the present invention can use a 90-channel chip. In the case of the microchip, the average diameter of the microchannel for injecting the oil phase solution and the aqueous solution is 300 μm to 500 μm, and the oil phase solution and the aqueous solution can pass through the resistance channel after moving through each channel. The average diameter of the resistance channel is 10 μm to 50 μm. After passing through the resistance channel, the oil phase solution and the aqueous solution pass through a junction channel where they intersect, and the diameter of the junction channel can be 100 μm to 400 μm. After the oil phase solution and the aqueous solution intersect within the junction channel to form an emulsion, they pass through a microchannel having a diameter of 150 μm to 200 μm, and then pass through a microchannel having a diameter of 200 μm to 300 μm.

[0080] However, the average diameter of the microchannel may vary depending on the range of injection pressure. Furthermore, the average diameter of the microchannel is closely related to the average diameter of the particles, but is also closely related to the injection pressures of the oily and aqueous solutions.

[0081] In addition, the step 3) above is to flow the oil solution and the water solution into the first microchannel and the second microchannel where an intersection is formed under the injection pressure conditions.

[0082] That is, the oily solution flows along the first microchannel, and the aqueous solution flows along the second microchannel shaped to form an intersection with the first microchannel, where it meets the flow of the oily solution.

[0083] More specifically, when the oil solution is injected into the first microchannel, the pressure can be increased under conditions of 1 to 5 mbar / min after injection under conditions of 600 to 700 mbar.

[0084] In addition, when the above-mentioned water solution is injected into the second microchannel, it can be injected under pressure conditions of 2,500 mbar to 3,000 mbar, pressure conditions of 2,600 mbar to 2,900 mbar, or pressure conditions of 2,800 mbar.

[0085] Specifically, in the manufacturing method using the above microchannel, when the flow rate of the oily solution and the water solution flowing inside the microchannel was set to a constant value using a flow meter and the pressure was measured through feedback control, it was confirmed that the pressure required to make the oily solution flow through the microchannel at a constant flow rate gradually increases over time.

[0086] Therefore, by using a method of constantly increasing the pressure applied to the oily solution, the variability of the flow rate can be minimized, and the problem of uneven distribution of microparticles or channel blockage can be prevented due to the oily solution gradually hardening inside the microchannel, and the manufacturing yield of the target microparticles can be increased.

[0087] In addition, the pressure conditions when injecting the oil solution and the water solution into the microchannel are intended to control the average diameter of the manufactured microparticles, and if the above range is not specifically satisfied, the size of the manufactured particles may not be uniform, or a problem may occur in which the average diameter range of the microparticles of the present invention is not satisfied.

[0088] That is, in order to cause the flow of the aqueous solution forming an intersection with the flow of the oily solution injected into the microchannel to flow at a faster flow rate than the flow of the oily solution, the aqueous solution is caused to flow under higher pressure conditions.

[0089] As described above, by making the flow rates of the oil phase solution and the aqueous solution different and making the flow rate of the aqueous solution faster than that of the oil phase solution, the aqueous solution having a relatively faster flow rate compresses the oil phase solution at the point where the flow of the oil phase solution and the flow of the aqueous solution meet, and at this time, due to the repulsive force between the oil phase solution and the aqueous solution, the biodegradable polymer and exenatide in the oil phase solution form spherical microparticles, and more specifically, microparticles in which exenatide is evenly distributed in the spherical biodegradable polymer are formed.

[0090] Step 4) above is a step of collecting microparticles, which collects microparticles in a tank containing a second mixture, thereby preventing aggregation between initially generated microparticles.

[0091] The above step 4) uses the aqueous solution prepared in the above step 2), i.e., a mixed solution of a surfactant and purified water. After the aqueous solution is prepared in the above step 2), some of it is injected into the microchannel, and the other part is moved to the tank of step 4, thereby preventing the agglomeration phenomenon between the collected microparticles.

[0092] The above step 5) is a step for removing the organic solvent present in the microparticles collected in the tank, and the organic solvent present on the surface of the sustained-release particles is evaporated and removed by stirring under constant temperature conditions and stirring speed. At this time, the stirring conditions may be stirring at a speed of 200 to 400 rpm for 4 to 6 hours at 40°C to 50°C, and specifically stirring at a speed of 250 to 350 rpm for 4.5 to 5.5 hours at 40°C to 45°C.

[0093] As described above, when removing the organic solvent under stirring conditions, the formation of pores on the surface of the microparticles is prevented by removing the residual organic solvent, thereby enabling the production of microparticles with a smooth surface, and minimizing the residual organic solvent.

[0094] The temperature at which the above-mentioned oily solution and aqueous solution flow through the microchannel is also 15 to 20°C, and preferably 18°C. That is, after flowing through the microchannel and forming an intersection to generate microparticles, the temperature is maintained at a constant low temperature of 15 to 20°C until the collected microparticles are stirred for the first time. Only when a low temperature is maintained during the process of manufacturing microparticles can spherical particles be manufactured and maintained. In other words, if the temperature is not low, it is difficult to manufacture particles with a consistent spherical shape.

[0095] Lastly, the above step 6) is a step of washing and drying the microparticles. The microparticles, which have had all organic solvents on their surface removed by stirring, are washed several times with sterilized, filtered purified water to remove surfactants remaining in the microparticles, and then freeze-dried.

[0096] The microparticles finally produced are in the form of microparticles made of spherical biodegradable polymers in which exenatide is evenly distributed, and contain exenatide and the biodegradable polymer in a weight ratio of 1:3 to 1:7.

[0097] The weight ratio of exenatide and biodegradable polymer contained in the above microparticles is the same as the weight ratio in the oily solution, which means that by manufacturing the microparticles and removing all organic solvents by evaporation, microparticles containing exenatide and biodegradable polymer in the same weight ratio as in the oily solution can be manufactured.

[0098] The above-mentioned manufactured microparticles can be prepared into an injectable composition by mixing them with a suspension solvent. The description of the suspension solvent is as described above.

[0099]

[0100] Manufacturing Example 1

[0101] Preparation of microparticles containing exenatide

[0102] An API mixture was prepared by dissolving exenatide in acetic acid and dimethyl sulfoxide. An oily solution was prepared by dissolving PDLG7502A, PDLG7504A, and dichloromethane in the API mixture. The weight ratio of exenatide and the biodegradable polymer in the oily solution was 1:5, and the biodegradable polymer in the oily solution was 9.3 wt%.

[0103] Polyvinyl alcohol, a surfactant, was mixed with purified water to prepare an aqueous solution containing 0.5 wt% of polyvinyl alcohol.

[0104] The above oily solution and water-soluble solution were injected into a microchannel formed on a silicon wafer and allowed to flow.

[0105] At this time, in order to flow the oily solution and the aqueous solution at a constant flow rate, the oily solution was started under a pressure condition of 650 mbar and was allowed to flow under a condition of constantly increasing the pressure at a rate of 2 mbar per minute, and the aqueous solution was allowed to flow under a pressure condition of 2800 mbar. The temperature condition was maintained at 18°C, and the stirring speed was maintained at 200 rpm.

[0106] Microparticles generated at the intersection of the flow of the oily solution and the flow of the aqueous solution were collected in a tank containing the aqueous solution. The microparticles collected in the tank were stirred at 43°C for 5 hours at a speed of 300 rpm.

[0107] The microparticles that had completed stirring were washed several times with sterilized, filtered purified water and freeze-dried to produce microparticles.

[0108]

[0109] Manufacturing Examples 2 to 3

[0110] Microparticles were manufactured in the same manner as in the process for manufacturing microparticles of Manufacturing Example 1 above, and the microparticles were manufactured by varying the type of biodegradable polymer, the mixing ratio of two types of biodegradable polymers, the injection pressure of the oily solution, and the temperature conditions for removing the residual solvent.

[0111]

[0112] Examples 1 to 3

[0113] The microparticles of the above Manufacturing Examples 1 to 3 were added to 0.25 ml of a suspension solvent for each vial, and uniformly suspended to prepare a composition for subcutaneous injection. The weight ratio of exenatide and the biodegradable polymer in the composition for subcutaneous injection was 1:5.

[0114] The above suspension solvent was composed as shown in Table 1 below.

[0115] Content standard, mixing purpose, ingredient name, quantity unit, 0.25 mL, isotonic agent, D-Mannitol 12.5 mg, suspending agent, sodium carboxymethylcellulose 0.625 mg, suspending agent, polysorbate 80 0.25 mg, solvent, water for injection, remainder

[0116] As a comparative example, commercially available Bydureon BCise was purchased and used. The manufacturing conditions of Manufacturing Examples 1 to 3 are as shown in Table 2 below:

[0117] Oily solutionAqueous solution(PVA, %)Oily pressure(mbar)Solvent removalPolymerCo-solventSolventTemperature(℃)Stirring(rpm)Time(h)Manufacturing example 2PDLG7504A(3)PDL02A(1)Acetic acidDMSODCM0.5550(up3 / min)433005Manufacturing example 3PDLG7504AAcetic acidDMSODCM0.5600(up2 / min)43.53005Manufacturing example 1PDLG7502A(1)PDLG7504A(1)Acetic acidDMSODCM0.5650(up2 / min)42.53005

[0118]

[0119] Experimental Example 1

[0120] Examination of the properties of microparticles

[0121] To specifically determine the diameter of the microparticles, analysis was conducted using a Microtrac particle size analyzer.

[0122] The measurement results are shown in Figures 1 to 4 and Table 3 below:

[0123] %Tile Manufacturing Example 1 Manufacturing Example 2 Manufacturing Example 3 Comparative Example 10.0064.8865.1465.2047.7320.0066.9567.3867.4955.6130.0068.9069.4369.5762.7340.0070.7871.4471.6269.6650.0072.7873.4873.7076.5 160.0074.9375.6975.9383.4970.0077.3478.0578.3290.7780.0080.2180.6380.929 8.8290.0083.7683.8484.10109.795.0086.2386.0186.34118.3SD7.717.717.8124.99

[0124] As shown in Table 3 above, the microparticles of Manufacturing Examples 1 to 3 of the present invention have a D50 of 72.78 µm, 73.48 µm, and 73.70 µm, and a standard deviation of 7.71, 7.71, and 7.81, confirming that they have a uniform diameter. In contrast, the comparative example has a D50 of 76.51 µm, which is not much different from the microparticles of the present invention, but has a standard deviation of 24.99, confirming that there is a large difference in diameter between the particles, confirming that the diameter is not uniform.

[0125]

[0126] Experimental Example 2

[0127] Pharmacokinetic characterization

[0128] Pharmacokinetic evaluations for Examples 1 to 3 of the present invention and comparative examples were confirmed.

[0129] ClassificationPrescription Dosage (mg)Attached solventComparative exampleBydureon Bcise (QW_4)2.0MCT OilExample 2API(1) : (5)[PDLG7504A(3) : PDL02A(1)]2.0Aqueous solutionExample 3API(1) : (5)[PDLG7504A]2.0Aqueous solutionExample 1API(1) : (5)[PDLG7502A(1) : PDLG7504A(1)]2.0Aqueous solution

[0130] The evaluation was conducted by administering Examples 1 to 3 and the comparative example to SD rats, collecting blood samples, and measuring the blood concentration (PK) of exenatide. The test results are shown in Figures 5, 6, and Table 5:

[0131] Dose 8mg / kg 8mg / kg 8mg / kg 8mg / kg Day Exenatide SCd Example 2 Example 3 Example 1 Comparative Example 0NDNDNDND0.025097.82510618.1852035.423346.5520.082985.9775100.1421131.44189.030.171121.3201770.526501.06157.9010.33489.333441.056261.02769.4140.5337.369 253.732178.89863.7161512.932409.931784.91885.9462294.111281.292671.312191.194188.902133.9431060.353338.8957108.43997.1623052.336217.64310235.109137.4771286. 705289.20014275.403364.0871305.332316.24617437.362248.734927.035508.866211482.0652163.971575.2031042.042243188.7424821.117821.8391330.108281460.0011746.1778 71.7661419.03835570.695974.335816.4991054.61142389.163743.981958.549879.354492 18.772514.262645.167483.87456213.625297.446417.987BQL63209.278526.694143.874ND

[0132] According to Table 5 above, it can be confirmed that the commercially available comparative example exhibits a pattern of releasing a small amount of exenatide initially after injection, and although the release of exenatide continuously increases thereafter, it can be confirmed that the release of exenatide ends quickly compared to the injection composition of the present invention. In contrast, it can be confirmed that Example 1 exhibits a rapid release of exenatide immediately after administration, and then continuously exhibits the effect of releasing exenatide.

[0133] On the other hand, the comparative example confirmed that the initial release effect of exenatide was minimal, and therefore, it was necessary to achieve a steady state through repeated administration on a weekly basis. In addition, in the case of Examples 2 and 3, although an initial rapid release of exenatide was observed, the subsequent release of exenatide was minimal, making it difficult to see the effect of exenatide.

[0134] For more clarity, the results of comparing the PK data between the comparative example and Example 1 are as shown in Figure 6. According to Figure 6, when the sustained-release injection composition of the present invention is administered, it can be confirmed that, unlike the comparative example, an initial rapid release of exenatide occurs and a continuous release of exenatide occurs. On the other hand, it can be confirmed that the comparative example shows insufficient release of exenatide from the beginning of injection until 21 days prior.

[0135] The results of comparing the PK parameters of Comparative Example and Example 1 are shown in Table 6 below:

[0136] PK ParametersExenatide SC Comparative Example Example 1AUC(last)36916.87155333.372Cmax1419.0383052.336Tmax28.007.00

[0137] * AUC: pg·d / mL, Cmax: pg / mL, Tmax & t 1 / 2 : d

[0138] Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements made by those skilled in the art using the basic concept of the present invention defined in the following claims also fall within the scope of the present invention.

[0139] The present invention relates to a sustained-release injectable composition comprising exenatide and a method for preparing the same.

Claims

1. A western-type injection composition containing microparticles, The above microparticles contain exenatide and a biodegradable polymer, The above-mentioned western-type injection composition was administered to SD rats at 8 mg / kg of exenatide, and the area under the plasma level-time curve (AUC) according to the release of exenatide in the blood was 40,000 pg·d / mL to 100,000 pg·d / mL for 63 days. A sustained-release injectable composition comprising exenatide.

2. In paragraph 1, The above-mentioned western-type injection composition exhibited an initial rapid exenatide release effect when exenatide was administered to SD rats at a dose of 8 mg / kg, and then the concentration of exenatide in the blood was maintained at a steady state. A sustained-release injectable composition comprising exenatide.

3. In paragraph 1, The above-mentioned western-type injectable composition continuously releases exenatide for 1 to 2 months. A sustained-release injectable composition comprising exenatide.

4. In paragraph 1, The above microparticles contain exenatide and a biodegradable polymer in a weight ratio of 1:3 to 1:

7. A sustained-release injectable composition comprising exenatide.

5. In paragraph 1, The above micro particles have an average diameter of 40 μm to 80 μm. A sustained-release injectable composition comprising exenatide.

6. In paragraph 1, The above biodegradable polymer is selected from the group consisting of polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA), polyphosphazine, polyiminocarbonate, polyphosphoester, polyanhydride, polyorthoester, polycaprolactone, polyhydroxyvalerate, polyhydroxybutyrate, polyamino acid, and combinations thereof. A sustained-release injectable composition comprising exenatide.

7. In paragraph 1, The above injectable composition comprises a suspending solvent, A sustained-release injectable composition comprising exenatide. 8.1) A step of preparing an oily solution by mixing exenatide and a biodegradable polymer in an organic solvent; 2) A step of preparing an aqueous solution by dissolving a surfactant in purified water; 3) A step of injecting the oil solution and the water solution into each microchannel and allowing them to flow to generate microparticles at the intersection of the microchannels; 4) A step of collecting the above micro particles in a tank containing the above water solution; 5) A step of removing the organic solvent present in the collected micro particles; 6) A step of washing and drying the micro particles from which the organic solvent has been removed with purified water; and 7) A step of preparing a western-type injection composition by mixing the dried microparticles with a suspending solvent, The above-mentioned western-type injection composition was administered to SD rats at 8 mg / kg of exenatide, and the area under the plasma level-time curve (AUC) according to the release of exenatide in the blood was 40,000 pg·d / mL to 100,000 pg·d / mL for 63 days. A method for preparing a sustained-release injectable composition comprising exenatide.

9. In paragraph 8, The above-mentioned oil solution is prepared by dissolving exenatide in a co-solvent and then mixing a biodegradable polymer and an organic solvent. A method for preparing a sustained-release injectable composition comprising exenatide.

10. In paragraph 9, The above common solvent is selected from the group consisting of acetic acid, dimethyl sulfoxide, dimethyl formamide, tetrahydrofuran, ethyl acetate, acetonitrile and mixtures thereof. A method for preparing a sustained-release injectable composition comprising exenatide.

11. In paragraph 9, The above common solvent is included in a weight ratio of 1:1 to 1:2 with the organic solvent. A method for preparing a sustained-release injectable composition comprising exenatide.

12. In paragraph 8, The above oil solution is injected into the microchannel under a pressure condition of 500 mbar to 700 mbar, and then the pressure is increased under a condition of 1 to 5 mbar / min. A method for preparing a sustained-release injectable composition comprising exenatide.

13. In paragraph 8, The above-mentioned water solution is injected into the microchannel under pressure conditions of 2,500 mbar to 3,000 mbar. A method for preparing a sustained-release injectable composition comprising exenatide.

14. In paragraph 8, The above step 5) is to stir at a speed of 200 to 400 rpm for 4 to 6 hours at 40 to 50°C. A method for preparing a sustained-release injectable composition comprising exenatide.

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