Sustained-release microsphere preparation containing texipatide or pharmaceutically acceptable salt thereof and preparation method of sustained-release microsphere preparation

Abstract

The present invention relates to a pharmaceutical composition which does not undergo rapid initial burst release of a drug by comprising sustained release microspheres consisting of texipatide or a pharmaceutically acceptable salt thereof, an initial burst release inhibitor, and a biodegradable polymer, has a high drug content with respect to particle size, and has high bioavailability, thus, the composition can minimize patient pain and inflammatory response that may occur when administered to a human body, and thus can be effectively used in the prevention or treatment of diabetes, beta cell dysfunction, hypertension, hyperlipidemia, obesity, and non-alcoholic steatohepatitis.

Description

Technical Field

[0001] This invention relates to a pharmaceutical composition comprising sustained-release microspheres containing a high content of tesipatide or a pharmaceutically acceptable salt thereof, and to a method for preparing the sustained-release microspheres. Background Technology

[0002] Tirzepatide, marketed under the brand name Mounjaro, is an antidiabetic drug used to treat type 2 diabetes. Tirzepatide is administered once weekly via subcutaneous injection.

[0003] The most common side effects of tesipatide include nausea, vomiting, diarrhea, decreased appetite, constipation, upper abdominal discomfort, and abdominal pain.

[0004] Glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) are hormones involved in blood glucose regulation. After eating, these hormones are secreted from intestinal cells and trigger insulin secretion. Tesipatide is known to be a GIP analog that improves blood glucose regulation by activating GLP-1 and GIP receptors.

[0005] Tesipatide was approved for medical use in the United States in May 2022, the European Union in September 2022, Canada in November 2022, and Australia in December 2022. The U.S. Food and Drug Administration (FDA) considers tesipatide to be the first drug of its kind.

[0006] To effectively administer tesipatide alone, a technology is needed to ensure long-term pharmacological efficacy. Therefore, the inventors of this invention have devised a technique to encapsulate tesipatide in microspheres formed from biodegradable polymers to achieve its long-term release. However, in this case, due to the low bioavailability of the tesipatide encapsulated in the microspheres, or the low drug content contained in the microspheres, a large number of microspheres are required to exhibit long-term pharmacological efficacy. However, when administering large numbers of microspheres in vivo, a problem arises: patients find it difficult to administer (self-administer) due to the difficulty of subcutaneous injection and the high likelihood of pain and inflammatory reactions at the application site. Summary of the Invention

[0007] Technical issues

[0008] The present invention is designed to solve the above-mentioned problems and aims to provide: a pharmaceutical composition comprising sustained-release microspheres, the sustained-release microspheres containing a high content of tesipatide or a pharmaceutically acceptable salt thereof, a biodegradable polymer and an initial burst release inhibitor, the sustained-release microspheres exhibiting long-term stable drug release; and a method for preparing the sustained-release microspheres.

[0009] Technical solution

[0010] The present invention will now be described in detail.

[0011] As used herein, the term "one or more" refers to a "quantity" corresponding to one or more. In this invention, when one or more configurations are present, one, two or more, three or more, one to three, or one to two configurations may be preferred, but are not limited thereto. In this invention, the term "one or more" may be used interchangeably with the term "at least one."

[0012] In order to achieve the above objectives,

[0013] As one aspect of the invention, a pharmaceutical composition is provided comprising a biodegradable polymer and tesipatide sustained-release microspheres, wherein the amount of tesipatide is 8 wt.% or more based on the total weight of the microspheres, and the amount of an initial burst-release inhibitor is from 5 ppm to 2000 ppm relative to the weight of tesipatide.

[0014] On the other hand, biodegradable polymers can be at least one selected from the group consisting of: polymers selected from the group consisting of block copolymers of: polylactide (PLA), polyglycolic acid (PGA), polylactide-co-glycolic acid (PLGA), polydioxane, polycaprolactone (PCL), polylactide-co-glycolic acid-co-caprolactone (PLGC), polylactide-co-hydroxymethylglycolic acid (PLGMGA), polyalkyl carbonate, polytrimethylene carbonate (PTMC), polylactide-co-trimethylene carbonate (PTMC) PLTMC), polyhydroxybutyrate (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoester, polyanhydride, polyanhydride-co-imide, polypropylene fumarate, pseudo-polyamino acid, polyalkyl cyanoacrylate, polyphosphazene, polyphosphate, polysaccharide, and block copolymers of poly(butylene succinate-lactide) (PBSLA); simple mixtures of two or more of the polymers selected; copolymers of the selected polymers with polyethylene glycol (PEG); and polymer-sugar complexes of the polymers or copolymers combined with sugars.

[0015] On the other hand, pharmaceutically acceptable salts of tesiparatide can be sodium salt, acetate, benzoate, hydroxynaphthyl carbamate, naphthalene disulfonate, or dihydroxynaphthyl carbamate.

[0016] On the other hand, the intrinsic viscosity of biodegradable polymers can be from 0.16 to 1.7 dL / g. More specifically, the intrinsic viscosity of polylactide-co-glycolic acid (PLGA) or polylactide (PLA) in biodegradable polymers can be from 0.16 to 1.7 dL / g.

[0017] On the other hand, the average particle size of microspheres containing tesipatide or its pharmaceutically acceptable salt and an initial burst-release inhibitor can be from 5 μm to 100 μm.

[0018] On the other hand, the span value of microspheres containing tesipatide or its pharmaceutically acceptable salt and an initial burst-release inhibitor can be 1.5 or less.

[0019] On the other hand, the total weight of the microspheres comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst release inhibitor contained in the pharmaceutical composition may be 20 to 1000 mg, 20 to 800 mg, 20 to 600 mg, 20 to 400 mg, 20 to 200 mg, 20 to 100 mg, 30 to 1000 mg, 30 to 800 mg, 30 to 600 mg, 30 to 400 mg, 30 to 200 mg, 30 to 100 mg, 40 to 1000 mg, 40 to 800 mg, 40 to 600 mg, 40 to 400 mg, 40 to 200 mg, 40 to 100 mg, 50 to 1000 mg, 50 to 800 mg, 50 to 600 mg, 50 to 400 mg, 50 to 200 mg, or 50 to 100 mg.

[0020] On the other hand, the microspheres may also contain one or more release controllers selected from the group consisting of: butyric acid, valeric acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, palmitic acid, heptadecanic acid, stearic acid, nonadecanic acid, behenic acid, arachidic acid, isocrotonic acid, oleic acid, transoleic acid, sorbic acid, linoleic acid, arachidonic acid, benzoic acid, hydroxynaphthoic acid, naphthalene disulfonic acid, naphthalene sulfonic acid, and dihydroxynaphthoic acid.

[0021] On the other hand, microspheres can contain 10 to 500 mg / kg of Na.

[0022] On the other hand, microspheres can contain 1 to 100 mg / kg of P.

[0023] On the other hand, the present invention provides a method for preparing sustained-release microspheres comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor.

[0024] On the other hand, the preparation method can use a continuous phase containing an initial burst-release inhibitor.

[0025] On the other hand, any substance that can set the pH of the continuous phase to 7 or higher when dissolved in the continuous phase can be used as an initial burst release inhibitor.

[0026] On the other hand, the initial burst-release inhibitor may use one or more substances selected from the group consisting of: phosphates, hydroxides, phosphides, phosphites, carbonates, bicarbonates, chromates, dichromates, oxides, oxalates, silicates, sulfates, sulfides, sulfites, tartrates, tetraborates, thiosulfates, arsenates, arsenites, citrates, ferricyanides, and nitrides. Alternatively, the initial burst-release inhibitor may be at least one selected from the group consisting of disodium hydrogen phosphate, dipotassium hydrogen phosphate, and diammonium phosphate. Furthermore, the present invention provides a pharmaceutical composition and a method for preparing the same, the pharmaceutical composition comprising a biodegradable polymer and sustained-release microspheres, wherein the content of tesipatide is 8 wt.% or more based on the total weight of the microspheres, and the initial drug burst-release rate is 10% or less.

[0027] Beneficial effects

[0028] According to the preparation examples of the present invention, the sustained-release pharmaceutical composition comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst release inhibitor has the following effects: despite containing a high content of tesipatide, rapid initial burst release is also inhibited by the initial burst release inhibitor, not only reducing the dose of microspheres due to the high content, but also exhibiting high bioavailability and a long-lasting effect of one month or longer when administered in vivo, while reducing the dose of microspheres, thus minimizing possible patient pain and side effects at the application site during administration. Attached Figure Description

[0029] Figure 1A and 1B These are scanning electron microscope images confirming the morphological characteristics of the microspheres manufactured according to Example 1 and Comparative Example 1. Detailed Implementation

[0030] The present invention will now be described in detail.

[0031] This invention comprises tesipatide or a pharmaceutically acceptable salt thereof as the active ingredient.

[0032] Tesipatide is an acylated peptide designed to activate GIP and GLP-1 receptors, which are key mediators of insulin secretion and are also expressed in brain regions that regulate food consumption. Tesipatide is designed for once-weekly subcutaneous administration.

[0033] The structure of tesipatide is shown in the following chemical formula 1.

[0034] [Chemical Formula 1]

[0035]

[0036] Tesipatide can exist in the form of a salt, particularly in the form of a pharmaceutically acceptable salt. Any salt commonly used in the art may be used without limitation. As used herein, the term "pharmaceutically acceptable salt" refers to any organic or inorganic addition salt of the compound that, at concentrations relatively non-toxic and harmless to patients, would not diminish the beneficial effects of the active ingredient due to side effects. Specific examples include, but are not limited to, sodium salts, acetates, benzoates, hydroxynaphthylcarbamates, naphthalene disulfonates, or dihydroxynaphthylcarbamates of tesipatide.

[0037] Tilipatide or a pharmaceutically acceptable salt thereof is the active ingredient of the present invention, which may exist in various forms, such as amorphous or crystalline.

[0038] According to the pharmaceutical composition of the present invention, within 24 hours after administration, the area under the total plasma concentration-time curve (AUC) is [value missing]. 总 The area under the plasma concentration-time curve (AUC) of tesipatide 0-24hr () can be 20% or less, 10% or less, 5% or less, 0.1% to 20%, 1% to 10%, or 1% to 5%.

[0039] According to the pharmaceutical composition of the present invention, the area under the total plasma concentration-time curve (AUC) from the date of administration to the day following administration is [data missing]. 总 The area under the plasma concentration-time curve (AUC) of tesipatide 0-QXM In humans, the percentage can be 70% or less, 65% or less, 60% or less, 20% to 70%, 20% to 65%, 20% to 60%, 25% to 70%, 25% to 65%, 25% to 60%, 30% to 70%, 30% to 65%, or 30% to 60%, and in rats, the percentage can be 95% or less, 93% or less, 90% or less, 50% to 95%, 50% to 93%, 50% to 90%, 55% to 95%, 55% to 93%, 55% to 90%, 60% to 95%, 60% to 93%, or 60% to 90%.

[0040] Furthermore, the pharmaceutical composition according to the invention may be a pharmaceutical composition comprising sustained-release microspheres, wherein the content of tesipatide is 8 wt.% or more relative to the total weight of the microspheres, and the initial burst release of the drug is less than 10% within 24 hours.

[0041] The biodegradable polymer contained in the tesipatide sustained-release microspheres in the pharmaceutical composition according to the present invention is selected from, for example, from the group consisting of: polylactide (PLA), polyglycolic acid (PGA), polylactide-co-glycolic acid (PLGA), polydioxanone, polycaprolactone (PCL), polylactide-co-glycolic acid-co-caprolactone (PLGC), polylactide-co-hydroxymethylglycolic acid (PLGMGA), polyalkyl carbonate, polytrimethylene carbonate (PTMC), polylactide-co-trimethylene... Poly(hydroxybutyric acid) carbonate (PLTMC), polyhydroxybutyrate (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoester, polyanhydride, polyanhydride-co-imide, polypropylene fumarate, pseudo-polyamino acid, polyalkyl cyanoacrylate, polyphosphazene, polyphosphate, polysaccharide, and poly(butylene succinate-lactide) (PBSLA); simple mixtures of two or more of the polymers selected from the polymers; copolymers of the polymers with polyethylene glycol (PEG); and polymer-sugar complexes of the polymers or copolymers combined with sugars, but not limited thereto.

[0042] Furthermore, the biodegradable polymer is a simple mixture of two or more (i.e., a simple mixture comprising two or more of the polymers described herein), and may include two or more different types of polymers illustrated (but not limited to) herein, or may be a combination or blend of polymers of the same type, but may be a combination of polymers with different intrinsic viscosities and / or monomer ratios (e.g., a combination or blend of two or more polylactide-co-glycolic acid polymers with different intrinsic viscosities), or may be polymers of the same type with different terminal groups (e.g., polymers of the same type having ester terminal groups or acid terminal groups).

[0043] In a specific aspect, the pharmaceutical composition according to the invention may include microspheres comprising two or more biodegradable polymers. As another specific embodiment, the pharmaceutical composition according to the invention may include two or more microspheres, each microsphere comprising at least one polymer selected from two or more biodegradable polymers.

[0044] Considering factors such as drug release characteristics and manufacturing processes, biodegradable polymers can be biodegradable polymers with intrinsic viscosities of 0.16 to 1.7 dL / g, 0.2 to 1.3 dL / g, or 0.24 to 1.2 dL / g. Intrinsic viscosity refers to the viscosity measured at 25°C in chloroform at a concentration of 0.1% (w / v) using an Ubbelohde viscometer.

[0045] In the case of polylactide-co-glycoli, the molar ratio of lactide to glycolide in the copolymer can be 40:60 to 90:10, 45:55 to 85:15, or 50:50 to 75:25, for example 45:55, 50:50, 75:25 or 85:15.

[0046] When the intrinsic viscosity of a biodegradable polymer is less than 0.16 dL / g, the polymer's molecular weight is insufficient, making it difficult to exhibit the sustained-release effect of tesiparatide or its pharmaceutically acceptable salts. When the intrinsic viscosity of a biodegradable polymer exceeds 1.7 dL / g, the release of tesiparatide or its pharmaceutically acceptable salts may be excessively delayed. Furthermore, when using polymers with high intrinsic viscosity to manufacture microspheres, excessive amounts of manufacturing solvents are used due to the high viscosity, making it difficult to produce reproducible microspheres. Examples of commercially available polymers with the above-mentioned properties include Evonik's Resomer® series RG502H, RG503H, RG504H, RG502, RG503, RG504, RG653H, RG752H, RG753H, RG752S, RG755S, RG750S, RG757S, RG858S, R202H, R203H, R205H, R202S, R203S, R205S, R206S, and R207S, and Corbion's PDL 02A, PDL 02, PDL 04, PDL 05, PDLG 7502A, PDLG 7502, PDLG 7507, PDLG 5002A, PDLG5002, PDLG 5004A, and PDLG 5004.

[0047] The amount of biodegradable polymer in sustained-release microspheres comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor according to the present invention may, based on the total weight of the microspheres, be selected as an upper limit of 60 wt.% or more, 62 wt.% or more, 65 wt.% or more, 67 wt.% or more, or 70 wt.% or more, and a lower limit of 92 wt.% or less, 91 wt.% or less, 90 wt.% or less, 89 wt.% or less, or 88 wt.% or less, or may be included in a range formed by a combination of the upper and lower limits. For example, the range may be 60 wt.% to 92 wt.%, 65 wt.% to 90 wt.%, or 70 wt.% to 88 wt.%, but is not limited thereto.

[0048] In the sustained-release microspheres of the present invention comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor, the content of tesipatide or a pharmaceutically acceptable salt thereof relative to the total weight of the microspheres is preferably 8 wt.% or more, 12 wt.% or more, 14 wt.% or more, 37 wt.% or less, 35 wt.% or less, or 33 wt.% or less. Based on tesipatide, when the content of tesipatide or a pharmaceutically acceptable salt thereof in the microspheres is less than 8 wt.%, the amount of polymer used based on the drug may be too high, which may lead to a decrease in the bioavailability of tesipatide or a pharmaceutically acceptable salt thereof; when the content is too high, the initial burst-release effect of tesipatide or a pharmaceutically acceptable salt thereof may be enhanced, and therefore may not be ideal.

[0049] The pharmaceutical composition comprising sustained-release microspheres according to the present invention may contain less than a certain amount of initial burst-release inhibitor in the microspheres.

[0050] The initial burst-release inhibitor in this invention is a substance contained in the continuous phase, used to inhibit the rapid release of the active ingredient during microsphere manufacturing, and according to this invention, its content in the microspheres can be below a certain amount. The continuous phase refers to a substance that, when using emulsion and solvent extraction evaporation methods in the microsphere manufacturing process, enables the dispersion of a dispersed phase containing the active ingredient (e.g., tesipatide) and a biodegradable polymer. In this invention, it can be an aqueous solution containing a surfactant, but is not limited thereto. The initial burst-release inhibitor according to this invention is contained in the continuous phase used in the manufacturing method; therefore, after manufacturing, the initial burst-release inhibitor can be contained in a certain amount in the microspheres according to this invention. Matters concerning the continuous phase can be applied in the same manner as those described in the microsphere preparation method according to this invention, as follows.

[0051] Specifically, based on the total weight of the microspheres, the initial burst-release inhibitor can contain 5 ppm or more, 10 ppm or more, 20 ppm or more, 30 ppm or more, 40 ppm or more, 50 ppm or more, 100 ppm or more, 150 ppm or more, 200 ppm or more, 250 ppm or more, 200 ppm or more, 300 ppm or more, 350 ppm or more, 400 ppm or more, 450 ppm or more, 500 ppm or more, 550 ppm or more, 600 ppm or more, 650 ppm or more, 700 ppm or more, 750 ppm or more, 800 ppm or more, 850 ppm or more, 900 ppm or more, 950 ppm or more. The upper limits are 1000 ppm or more, 1100 ppm or more, 1200 ppm or more, 1300 ppm or more, 1400 ppm or more, or 1500 ppm or more, and the lower limits are 2000 ppm or less, 1900 ppm or less, 1800 ppm or less, 1700 ppm or less, 1600 ppm or less, 1500 ppm or less, 1400 ppm or less, 1300 ppm or less, 1200 ppm or less, 1100 ppm or less, 1000 ppm or less, 900 ppm or less, 800 ppm or less, 700 ppm or less, 600 ppm or less, or 500 ppm or less, and may be included within the range formed by the combination of the upper and lower limits. For example, based on the total weight of the microspheres, the initial burst-release inhibitor may be 5 ppm to 2000 ppm, preferably 10 ppm to 1500 ppm, more preferably 20 ppm to 1000 ppm, and most preferably 20 ppm to 500 ppm.

[0052] In a specific sense, any substance that, when dissolved in the continuous phase, enables the pH of the continuous phase to be set to 7 or higher can be used as an initial burst release inhibitor.

[0053] The initial burst inhibitor may be at least one selected from the group consisting of phosphates, hydroxides, phosphides, phosphites, carbonates, bicarbonates, chromates, dichromates, oxides, oxalates, silicates, sulfates, sulfides, sulfites, tartrates, tetraborates, thiosulfates, arsenates, arsenites, citrates, ferricyanides, and nitrides, but is not limited thereto.

[0054] More specifically, the initial burst inhibitor may be selected from at least one of the following groups: phosphates of two or more alkali metals, carbonates of one or more alkali metals, and phosphites of ammonium, but is not limited thereto.

[0055] More specifically, phosphates containing two or more alkali metals refer to phosphates containing two or more alkali metal ions, such as disodium hydrogen phosphate (Na₂HPO₄) or dipotassium hydrogen phosphate (K₂HPO₄); bicarbonates containing one or more alkali metals refer to bicarbonates containing one or more alkali metal ions, such as sodium bicarbonate (NaHCO₃); carbonates containing two or more alkali metals refer to carbonates containing two or more alkali metal ions, such as sodium carbonate (Na₂CO₃); and phosphates containing two or more ammonium ions refer to phosphates containing two or more ammonium ions, such as diammonium hydrogen phosphate ((NH₄)₂HPO₄). These salts can be used alone or in combination of two or more.

[0056] The microspheres comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor according to the present invention preferably have a uniform particle size distribution, with average particle sizes of 5 μm to 100 μm, 5 μm to 90 μm, 5 μm to 80 μm, 10 μm to 90 μm, 10 μm to 80 μm, 15 μm to 100 μm, 15 μm to 90 μm, 15 μm to 80 μm, 70 μm to 100 μm, 70 μm to 90 μm, and 70 μm to 80 μm. The particle sizes are 60 μm to 100 μm, 60 μm to 80 μm, 60 μm to 70 μm, 20 μm to 90 μm, 20 μm to 70 μm, 20 μm to 60 μm, 30 μm to 80 μm, 30 μm to 60 μm, 40 μm to 70 μm, 40 μm to 50 μm, 30 μm to 40 μm, 20 μm to 30 μm, 5 μm to 30 μm, 5 μm to 20 μm, 10 μm to 20 μm, or 5 μm to 10 μm. The term "average particle size" used in this invention refers to the particle size corresponding to 50% of the volume in the particle size distribution curve, which is the median diameter, denoted as D50 or D(v, 0.5).

[0057] When the average particle size of microspheres containing tesiparatide or its pharmaceutically acceptable salts and an initial burst-release inhibitor is less than 5 μm, the release of tesiparatide or its pharmaceutically acceptable salts from the microspheres may be too rapid, and therefore may be undesirable. When the average particle size exceeds 100 μm, the injection needle may become too large for human administration, resulting in pain during injection or potential leakage of the drug from the injection site after injection, and therefore may be undesirable.

[0058] The microspheres of the present invention comprising tesiparatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor preferably have a uniform particle distribution. Compared with non-uniform microspheres, microspheres comprising tesiparatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor exhibit less deviation during injection and can be administered in more accurate amounts. The span value of the microspheres of the present invention comprising tesiparatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor is preferably 1.5 or less. More preferably, the span value is preferably 1.2 or less. More specifically, the span value can be 1.5 or less, 1.2 or less, 0.1 to 1.5, 0.3 to 1.5, 0.5 to 1.5, 0.1 to 1.0, 0.4 to 1.0, 0.6 to 1.0, 0.2 to 0.8, or 0.4 to 0.8. The term "span value" used in this invention is an indicator of microsphere particle size uniformity, referring to the value obtained by the formula "span value = (Dv0.9 - Dv0.1) / Dv0.5". Here, Dv0.1 refers to the particle size corresponding to 10% of the volume in the microsphere particle size distribution curve, Dv0.5 refers to the particle size corresponding to 50% of the volume in the microsphere particle size distribution curve, and Dv0.9 refers to the particle size corresponding to 90% of the volume in the microsphere particle size distribution curve.

[0059] The sustained-release microspheres of the present invention, comprising tesiparatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor, are administered via injection (e.g., subcutaneous injection), and are particularly suitable for self-administration; therefore, it is preferred that tesiparatide is released over a relatively long period of time. Preferably, the sustained-release microspheres in the pharmaceutical compositions according to the present invention can release tesiparatide or a pharmaceutically acceptable salt thereof for 1 month or longer, 2 months or longer, 3 months or longer, 1 to 2 months, 1 to 3 months, 1 to 4 months, 1 to 5 months, 1 to 6 months, 2 to 6 months, 2 to 5 months, 2 to 4 months, 2 to 3 months, 3 to 5 months, or 3 to 4 months, but are not limited thereto. Furthermore, the sustained-release microspheres of the present invention comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor are not particularly limited in their release mode, but preferably the upper limit of the release rate (release amount) of tesipatide or a pharmaceutically acceptable salt thereof after 24 hours of in vivo administration is less than 10%, less than 15%, or less than 20%, or the lower limit of the release rate is 0.001% or more, 0.01% or more, 0.1% or more, or 1%, or the release rate is within a combination of the upper and lower limits.

[0060] Furthermore, the total amount (total weight) of the sustained-release microsphere formulation comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor in the pharmaceutical composition of the present invention may be 30 to 3000 mg, 30 to 2500 mg, 30 to 2000 mg, 30 to 1500 mg, 30 to 1250 mg, 30 to 1000 mg, 60 to 3000 mg, 60 to 2500 mg, 60 to 2000 mg, 60 to 1500 mg, 60 to 1250 mg, 100 to 3000 mg, 100 to 2500 mg, 100 to 2000 mg, 100 to 1500 mg, 200 to 3000 mg, 200 to 2500 mg, 200 to 1500 mg, 400 to 3000 mg, 400 to 2500 mg, 400 to 2000 mg, or 400 to 1500 mg. Since sustained-release microspheres containing tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor are included in the composition within the range described above, the compositions according to the invention have the advantage of minimizing inflammatory response at the application site and allowing patients to administer them themselves.

[0061] The microspheres comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor included in the compositions of the present invention may further comprise a release controller. Examples of substances used as release controllers may be at least one selected from the following: butyric acid, valeric acid, hexanoic acid, heptanoic acid, caprylic acid, nonanoic acid, decanoic acid, undecanoic acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, palmitic acid, heptadecanoic acid, stearic acid, nonadecanoic acid, behenic acid, arachidic acid, isocrotonic acid, oleic acid, transoleic acid, sorbic acid, linoleic acid, arachidonic acid, benzoic acid, hydroxynaphthoic acid, naphthalene disulfonic acid, naphthalene sulfonic acid, and dihydroxynaphthoic acid, but are not limited thereto. Preferably, the release controller may be hydroxynaphthoic acid, naphthalene disulfonic acid, or dihydroxynaphthoic acid, but is not limited thereto.

[0062] The pharmaceutical composition according to the invention, comprising microspheres containing tesiparatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor, can be formulated into various forms of formulations, for example, it can be a known parenteral dosage form. Therefore, in addition to tesiparatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor, the pharmaceutical composition according to the invention may also contain a thickener, stabilizer, isotonic agent, surfactant, excipient, and / or carrier. The isotonic agent may be a water-soluble excipient or sugar, such as mannitol, sucrose, sorbitol, trehalose, lactose, sodium chloride, etc. Examples of thickeners may include sodium carboxymethyl cellulose, sodium carboxymethyl cellulose, povidone, etc. Furthermore, sodium dihydrogen phosphate, anhydrous citric acid, sodium hydroxide, sodium chloride, etc., can be used as stabilizers.

[0063] The pharmaceutical composition according to the invention can be administered in a therapeutically effective amount of tesiparatide, for example, in the treatment of diabetes (especially type 2 diabetes), β-cell dysfunction, hypertension, hyperlipidemia, obesity, non-alcoholic steatohepatitis, or degenerative neurological diseases (such as Alzheimer's disease and Parkinson's disease). A physician can assess the therapeutically effective amount of tesiparatide. According to the invention, the pharmaceutical composition containing tesiparatide can be administered monthly or quarterly. In some specific embodiments, based on tesipatide, the monthly dose of the composition according to the present invention can be 5 mg to 500 mg, 5 mg to 400 mg, 5 mg to 300 mg, 5 mg to 150 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg to 40 mg, 5 mg to 30 mg, 10 mg to 500 mg, 10 mg to 400 mg, 10 mg to 300 mg, 10 mg to 150 mg, 10 mg to 100 mg, 10 mg to 50 mg, 10 mg to 40 mg, or 10 mg to 30 mg. 20 mg to 500 mg, 20 mg to 400 mg, 20 mg to 300 mg, 20 mg to 150 mg, 20 mg to 100 mg, 40 mg to 500 mg, 40 mg to 400 mg, 40 mg to 300 mg, 40 mg to 150 mg, 50 mg to 500 mg, 50 mg to 400 mg, 50 mg to 300 mg, 50 mg to 150 mg, 100 mg to 500 mg, 100 mg to 400 mg, 100 mg to 300 mg, or 100 mg to 150 mg. Pharmaceutical compositions according to the invention comprising microspheres containing tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor can also be administered parenterally, for example, by subcutaneous injection. The pharmaceutical composition according to the present invention can consist of a drug phase and a solvent phase. The drug phase comprises microspheres containing tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor. The solvent phase is used to suspend the microspheres. The pharmaceutical composition can be in the form of a dual-chamber syringe, with one chamber containing the drug phase and the other chamber containing the solvent phase; or it can be in the form of a pre-filled syringe, wherein the drug phase is suspended in the solvent phase. When configured as a pre-filled syringe, the drug phase is suspended in the solvent phase of the pre-filled syringe. The solvent phase used can be an injectable oil, including medium-chain oils, mineral oils, etc.

[0064] In specific embodiments, the sustained-release microspheres comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst release inhibitor, included in pharmaceutical compositions according to the present invention, have a high drug content based on the microsphere content, while inhibiting excessive initial burst release of the drug that may lead to fatal side effects, and exhibit sufficient pharmaceutical efficacy as a GIP analog within the required time due to their high bioavailability. Therefore, they can be used for the prevention or treatment of diabetes (especially type 2 diabetes), β-cell dysfunction, hypertension, hyperlipidemia, obesity, non-alcoholic steatohepatitis, Alzheimer's disease, or Parkinson's disease.

[0065] On the other hand, the present invention provides a method for preparing sustained-release microspheres comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor.

[0066] On the other hand, the present invention provides a method for preparing sustained-release microspheres, which, although containing high amounts of tesipatide or its pharmaceutically acceptable salts, have significantly inhibited initial burst release.

[0067] The preparation method of the sustained-release microsphere injection comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor of the present invention will be described in detail below.

[0068] The sustained-release microsphere injections comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor according to the present invention can be manufactured, for example, by an emulsion solvent extraction and evaporation method, but are not limited thereto.

[0069] As one aspect of the specific preparation method, the present invention provides a method for manufacturing sustained-release microspheres using an oil-in-water (O / W) emulsion, the method comprising the following steps:

[0070] (a) Preparation of an oil phase (O phase), which is a dispersion obtained by dissolving tesipatide or a pharmaceutically acceptable salt thereof and one or more biodegradable polymers in an organic solvent;

[0071] (b) An initial burst release inhibitor is added to an aqueous solution containing a surfactant to prepare a continuous phase (W phase), and the dispersed phase from step (a) is added to obtain a dispersed phase in an emulsion state.

[0072] (c) Extracting the organic solvent from the dispersed phase in the emulsion state obtained in step (b) into the continuous phase (W phase) and evaporating the organic solvent to form microspheres; and

[0073] (d) Recover microspheres.

[0074] On the other hand, the present invention provides a method for preparing sustained-release microspheres using a water-in-oil-in-water (W / O / W) emulsion, the method comprising the following steps:

[0075] (a') A primary W1 / O emulsion is prepared by dissolving tesipatide or a pharmaceutically acceptable salt thereof in an aqueous solution to prepare an aqueous phase (W1 phase) and by dissolving one or more biodegradable polymers in an organic solvent to prepare an oil phase (O phase) as a dispersed phase in which the aqueous phase (W1 phase) and the oil phase (O phase) are mixed.

[0076] (b') An initial burst release inhibitor is added to an aqueous solution containing a surfactant to prepare a continuous phase (W2 phase), and the dispersed phase obtained in step (a') is added to obtain a dispersed phase in a secondary W1 / O / W2 emulsion state.

[0077] (c') Extracting the organic solvent from the dispersed phase of the secondary W1 / O / W2 emulsion obtained in step (b') into the continuous phase (W2 phase) and / or evaporating the organic solvent to form microspheres; and

[0078] (d') Recycle microspheres.

[0079] On the other hand, the pH of the continuous phase used in the preparation method can be 7 or higher.

[0080] In preparing sustained-release microspheres according to the present invention comprising tesiparatide or a pharmaceutically acceptable salt thereof and an initial burst release inhibitor, the sustained-release microspheres contain a high amount of tesiparatide or a pharmaceutically acceptable salt thereof relative to the weight of the microspheres, while excessive initial burst release of tesiparatide or a pharmaceutically acceptable salt thereof is inhibited, resulting in high bioavailability, and can use the following biodegradable polymers to release at a constant concentration over the desired long period of time, such as 1 month or longer, 3 months or longer, 1 to 2 months, 1 to 3 months, 1 to 4 months, 1 to 5 months, 1 to 6 months, 2 to 6 months, 2 to 5 months, 2 to 4 months, 2 to 3 months, 3 to 5 months, or 3 to 4 months, but not limited thereto. Specifically, it is preferred to use one or more polymers selected from the group consisting of: polylactide (PLA), polyglycolic acid (PGA), polylactide-co-glycolic acid (PLGA), polydioxanone, polycaprolactone (PCL), polylactide-co-glycolic acid-co-caprolactone (PLGC), polylactide-co-hydroxymethylglycolic acid (PLGMGA), polyalkyl carbonate, polytrimethylene carbonate (PTMC), polylactide-co-hydroxymethylglycolic acid (PTMC), and polyalkyl carbonate. Methyl carbonate (PLTMC), polyhydroxybutyrate (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoester, polyanhydride, polyanhydride-co-imide, polypropylene glycol fumarate, pseudo-polyamino acid, polyalkyl cyanoacrylate, polyphosphazene, polyphosphate, polysaccharide, and poly(butylene succinate-lactide) (PBSLA); simple mixtures of two or more of the polymers selected from said polymers; copolymers of said polymers with polyethylene glycol (PEG); and polymer-sugar complexes of polymers or copolymers combined with sugars. As a specific embodiment, in the manufacturing method according to the invention, poly(lactide-co-glycolic acid) and / or polylactide (PLA) polymers can be used as biodegradable polymers.

[0081] In this invention, two or more different biodegradable polymers may include two or more polymers having different repeating units constituting the polymer, and two or more polymers having different molar ratios of repeating units when containing two or more repeating units. For example, the microspheres may be a mixture of microspheres containing polylactide-co-glycolic acid and microspheres containing polylactide (PLA) polymer, or microspheres containing both polylactide-co-glycolic acid and polylactide (PLA) polymer.

[0082] Furthermore, as a specific embodiment, when there are two different types of biodegradable polymers, the content ratio of the biodegradable polymers can be 0.5:10 to 10:0.5, 0.5:8 to 8:0.5, 1:10 to 10:1, 1:4 to 4:1, 1:3 to 3:1, or 1:2 to 2:1 by weight, but is not limited thereto.

[0083] Furthermore, unless otherwise defined, the description of the biodegradable polymers described in the microspheres of this invention can be applied directly.

[0084] More specifically, when polylactide-co-glycolic acid is used as two or more biodegradable polymers to manufacture sustained-release microspheres according to the invention comprising tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor, at least one biodegradable polymer having an intrinsic viscosity of 0.16 dL / g to 0.45 dL / g may be included.

[0085] In the case of polylactide-co-glycoli, the molar ratio of lactide to glycolide in the copolymer can be 40:60 to 90:10, 45:55 to 85:15, or 50:50 to 75:25, for example 45:55, 50:50, 75:25, or 85:15.

[0086] Furthermore, unless otherwise defined, the description of the biodegradable polymers described in the microspheres of this invention can be applied in the same manner.

[0087] In step (a) or (a') of this specific preparation method, the organic solvent used to dissolve at least one biodegradable polymer is one or more organic solvents. Alternatively, a mixed organic solvent, consisting of two or more organic solvents, can also be used. Specifically, the mixed solvent can be a mixture of a water-miscible organic solvent and a water-immiscible organic solvent. In this case, it is preferable to use an organic solvent that is water-immiscible, with a concentration of 50% (v / v) or higher, 60% (v / v) or higher, 50% to 99.9% (v / v), 50% to 90% (v / v), 50% to 80% (v / v), 50% to 70% (v / v), 60% to 90% (v / v), or 60% to 80% (v / v). Utilizing the water-immiscibility of the organic solvent, the dispersed phase can be uniformly mixed with a continuous phase, including a surfactant, in subsequent step (b) or (b') to form an emulsion. There are no particular limitations on the type of organic solvent used to dissolve one or more biodegradable polymers, but a mixture of one or more solvents selected from the group consisting of dichloromethane (DCM), chloroform, ethyl acetate, methyl ethyl ketone, acetone, acetonitrile, dimethyl sulfoxide, dimethylformamide, N-methylpyrrolidone, acetic acid, methanol, ethanol, propanol, and benzyl alcohol is preferred. More preferably, a solvent selected from dichloromethane (DCM) and ethyl acetate, and one or more organic solvents selected from dimethyl sulfoxide, N-methylpyrrolidone, methanol, and acetic acid may be used. In one embodiment, a mixture of dichloromethane (DCM) and acetic acid (glacial acetic acid) may be used as the organic solvent.

[0088] In step (b) or (b'), there are no particular limitations on the method for uniformly mixing the continuous phase, including the dispersed phase and the surfactant, but a high-speed stirrer, an in-line mixer, a membrane emulsion method, a microfluidic emulsion method, an ultrasonic mixer, or a static mixer can be used alone or in combination with two or more of these types. When forming an emulsion using a high-speed stirrer, an in-line mixer, an ultrasonic mixer, or a static mixer, it is difficult to obtain a uniform emulsion; therefore, it is preferable to perform an additional sieving process, etc., between subsequent steps (c) and (d) or between steps (c') and (d').

[0089] There are no particular restrictions on the type of surfactant used in step (b) or (b'), and any type that can help form a dispersed phase with stable droplets within the continuous phase can be used. The following surfactants can be used alone or in combination of two or more: polyvinyl alcohol, methylcellulose, polyvinylpyrrolidone, carboxymethylcellulose, lecithin, gelatin, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene castor oil derivatives, etc.

[0090] In step (b) or (b'), based on the total volume of the continuous phase containing the surfactant, the surfactant content in the continuous phase can be from 0.01 w / v% to 20 w / v, preferably from 0.03 w / v% to 18 w / v%, 0.05 w / v% to 15 w / v%, 0.07 w / v% to 10 w / v%, or 0.1 w / v% to 5 w / v. When the surfactant content is less than 0.01 w / v%, droplet-like dispersed phase or emulsion may not form in the continuous phase; when the surfactant content exceeds 20 w / v%, due to the excess surfactant, fine particles may form in the continuous phase, making surfactant removal difficult.

[0091] An aqueous solution containing a surfactant and an initial burst-release inhibitor can be used as the continuous phase in step (b) or (b'). Alternatively, the continuous phase can be used by adding the surfactant to the aqueous solution to achieve the final concentration of the initial burst-release inhibitor.

[0092] Initial burst release inhibitors can be used to control the initial burst release of microspheres encapsulating high drug concentrations, and substances that enable the pH of the continuous phase to be set to 7.0 or higher can be used as initial burst release inhibitors.

[0093] There are no particular restrictions on the type of initial burst inhibitor; any basic salt that is soluble in the continuous phase and can maintain a pH of 7.0 or higher can be used. Initial burst inhibitors can be one or more substances selected from the group consisting of: phosphates, hydroxides, phosphides, phosphites, carbonates, bicarbonates, chromates, dichromates, oxides, oxalates, silicates, sulfates, sulfides, sulfites, tartrates, tetraborates, thiosulfates, arsenates, arsenites, citrates, ferricyanides, and nitrides.

[0094] In a specific respect, the initial burst inhibitor may be selected from at least one of the following groups: phosphates of two or more alkali metals, carbonates of one or more alkali metals, bicarbonates of one or more alkali metals, and phosphates of two or more ammonium metals, but is not limited thereto.

[0095] More specifically, the phosphates of two or more alkali metals are disodium hydrogen phosphate (Na2HPO4) or dipotassium hydrogen phosphate (K2HPO4), the carbonates of one or more alkali metals are sodium bicarbonate (NaHCO3) or sodium carbonate (Na2CO3), and the phosphates of two or more ammonium metals are diammonium hydrogen phosphate ((NH4)2HPO4), and may be used alone or in combination of two or more of them.

[0096] In the continuous phase, the final concentration of the initial burst-release inhibitor can be from 0.1 to 5.0 (w / v)%, preferably from 0.1 to 4.0 (w / v)%, more preferably from 0.2 to 4.0 (w / v)%, even more preferably from 0.3 to 3.5 (w / v)%, particularly preferably from 0.4 to 3.0 (w / v)%, and even more preferably from 0.5 to 2.5 (w / v)%.

[0097] When the concentration of the initial burst release inhibitor exceeds 5.0 w / v%, emulsion droplet bursting may occur during microsphere manufacturing; conversely, when the concentration is below 0.1 w / v%, the initial burst release of microspheres containing high concentrations of tesipatide may not be adequately inhibited.

[0098] When the initial burst release inhibitor is included in the continuous phase during the microsphere manufacturing process, the initial burst release in microspheres containing 12 (w / w)% or more tesipatide can preferably be reduced to 10% or less, more preferably to 8% or less, and most preferably to 5% or less.

[0099] The continuous phase used in step (b) or (b') may also include at least one selected from the group consisting of methanol, ethanol, propanol and ethyl acetate to control the rate of extraction of the organic solvent from the dispersed phase in the emulsion state.

[0100] The initial burst-release inhibitor may be added at the time point in step (b) or (b') during the preparation of the continuous phase, or at any time point during the supply of the continuous phase in step (c) or (c'), and the method of addition is not limited.

[0101] Furthermore, the pH of the continuous phase can be 7.0 or higher, 7.2 or higher, 7.4 or higher, 8.0 or higher, 8.5 or higher, 9.0 or higher, 7.0 to 9.0, or 7.0 to 8.5, but is not limited to these values. When the pH of the continuous phase is controlled within this range, the bioavailability of microspheres containing tesipatide or its pharmaceutically acceptable salts can be further improved.

[0102] In step (c) or (c'), when the emulsion comprising a continuous phase containing droplet-like dispersed phase and a surfactant is held or stirred for a period of time (e.g., 2 hours to 48 hours) below the boiling point of the organic solvent, the organic solvent can be extracted from a solution of tesiparatide or its pharmaceutically acceptable salts and polymers, which is a droplet-like dispersed phase, into the continuous phase. A portion of the organic solvent extracted into the continuous phase may evaporate from its surface. During the extraction and evaporation of the organic solvent from the droplet-like tesiparatide or its pharmaceutically acceptable salts and polymers, the droplet-like dispersed phase can solidify to form microspheres.

[0103] To further and more effectively remove the organic solvent in step (c) or (c'), the continuous phase can be heated for a period of time. The heating temperature is not limited and can be appropriately controlled by those skilled in the art depending on the organic solvent used. For example, when dichloromethane (DCM) is used as the organic solvent, heating can be maintained at 30°C or higher, 40°C or higher, 45°C or higher, 30 to 50°C, 40 to 50°C, or 45°C.

[0104] In step (d) or (d'), the method for recovering microspheres containing tesipatide or a pharmaceutically acceptable salt thereof and an initial burst-release inhibitor can be performed using various known techniques, such as filtration or centrifugation.

[0105] Between steps (c) and (d) or between steps (c') and (d'), residual surfactant can be removed by filtration and washing, and then filtered again to recover the microspheres.

[0106] Water can usually be used for the washing step to remove residual surfactants, and the washing step can be repeated several times.

[0107] In addition, as mentioned above, an additional sieving process can be performed between steps (c) and (d) or between steps (c') and (d') to obtain uniform microspheres. The sieving process can be carried out using known techniques, using sieves of different sizes to filter out small and large particles of microspheres to obtain microspheres of uniform size.

[0108] The manufacturing method of the present invention may, after step (d) or step (d'), or after the filtration and washing steps, use a conventional drying method to dry the obtained microspheres to finally obtain dried microspheres.

[0109] In addition to the above, all items not specifically defined, including tesipatide, initial burst release inhibitors, biodegradable polymers and their amounts, may be applied as defined in the pharmaceutical composition.

[0110] According to the manufacturing method of the present invention, sustained-release microsphere injections comprising tesiparatide or a pharmaceutically acceptable salt thereof can be manufactured, wherein the drug tesiparatide or a pharmaceutically acceptable salt thereof maintains an effective concentration for a desired time without initial burst release of the drug and has high bioavailability. Furthermore, sustained-release microsphere injections comprising tesiparatide or a pharmaceutically acceptable salt thereof and an initial burst release inhibitor, having uniform particle size and good administration capability, can also be prepared.

[0111] The sustained-release microspheres according to the present invention can be blended with microspheres containing two or more of the same drug but with different compositions and manufacturing conditions. In the case of blending microspheres containing two or more drug microspheres, the purpose of such blending can be to optimize the drug release profile, control the release time, etc.

[0112] Specifically, different compositions and manufacturing conditions may be selected from at least one of the following groups: drug dosage, polymer type, particle size distribution (e.g., average particle size), roundness, polymer dosage, type and amount of dispersed phase solvent, type and amount of co-solvent, type and amount of continuous phase, curing temperature and time, theoretical drug content, etc., but not limited to these.

[0113] Microsphere blending can be, for example, mixing drug microspheres having one or more different compositions and manufacturing conditions in a specific ratio.

[0114] Specifically, when blending microspheres, the bioavailability of the active ingredients in each microsphere can be considered to appropriately control the mixing ratio (weight ratio or number of microspheres).

[0115] As a specific embodiment, two or more different drug microspheres may be drug microspheres with different components and composition ratios (hereinafter referred to as drug microspheres with different compositions), and / or drug microspheres with different drug release characteristics (e.g., at least one selected from the group consisting of polymer type, polymer content, drug content, etc.). The difference in polymer type of microspheres may be a difference selected from at least one of the following: repeating units of polymer, terminal groups of polymer, molecular weight of polymer, intrinsic viscosity of polymer, etc.

[0116] Microsphere blending can be achieved either by preparing individual microspheres separately and then mixing them, or by preparing the microspheres in a single preparation process. Specifically, as an example, an O / W emulsion method for preparing microsphere blends in a single preparation process can be a preparation method including steps 1 to 4.

[0117] Two or more dispersed phase solutions are prepared separately by dissolving biodegradable polymers and drugs with different compositions in organic solvents (step 1).

[0118] Each of the two or more dispersed phases prepared in step 1 is injected into an aqueous solution (continuous phase) containing a surfactant to form a two or more dispersed phase emulsion (step 2).

[0119] The organic solvent in the dispersed emulsion prepared in step 2 is extracted into the continuous phase and evaporated to form microspheres (step 3); and

[0120] The microspheres are recovered after step 3 (step 4).

[0121] Specifically, as an example, the W / O / W emulsion method for preparing microsphere blends in a single preparation process may include steps 1 to 4.

[0122] A dispersed phase solution is prepared by dissolving the drug in an aqueous solution to prepare an aqueous phase (W1 phase), by dissolving biodegradable polymers of different compositions in an organic solvent to prepare two or more oil phases (O phases), and by mixing the aqueous phase (W1 phase) and two or more oil phases (O phases) to prepare a dispersed phase solution separately (step 1).

[0123] Each of the two or more dispersed phases prepared in step 1 is injected into an aqueous solution containing a surfactant (W2 phase, continuous phase) to form an emulsion of two or more dispersed phases (step 2).

[0124] The organic solvent in the dispersed emulsion prepared in step 2 is extracted into the continuous phase (W2 phase) and the organic solvent is evaporated to form microspheres (step 3); and

[0125] The microspheres are recovered after step 3 (step 4).

[0126] Invention Embodiments

[0127] [Example]

[0128] The present invention will now be described in more detail through the following manufacturing examples. However, the following manufacturing examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

[0129] Examples 1-1 to 1-3 (O / W method): Preparation of biodegradable polymers containing tesipatide and an initial burst release inhibitor Compound microspheres

[0130] Tesiparatide (manufacturer: Zhejiang Paipai Biotechnology Co., Ltd., China) was used as the drug, and RG653H and RG753H (manufacturer: Evonik, Germany) in a 1:1 weight ratio were used as biodegradable polymers, weighed to ensure batch amounts ranged from 0.5 g to 1.5 g (0.5 g for Examples 1-1 and 1-2, and 1.5 g for Examples 1-3). The drug and polymers were homogeneously dissolved in a mixed solvent of dichloromethane (DCM) and glacial acetic acid as a co-solvent to prepare an oil phase (O phase) as the dispersed phase.

[0131] Na2HPO4 (disodium hydrogen phosphate) was added as an initial burst release inhibitor to a 0.1 (w / v)% aqueous solution of polyvinyl alcohol (viscosity: 4.8 to 5.8 mPa·s) to make a final concentration of 2.0 (w / v)%, and used as the continuous phase (W phase).

[0132] After connecting the continuous phase to an emulsification apparatus equipped with a porous membrane with a diameter of 40 μm, the prepared dispersed phase was injected together with the continuous phase into the porous membrane. An emulsion containing biodegradable polymer droplets containing tesipatide was prepared, and the suspension was placed in a preparation vessel and stirred at 200 to 300 rpm. The temperature of the preparation vessel was maintained at 25°C. After the dispersed phase was injected, the suspension was maintained at 40°C for 3 hours while removing the organic solvent. Subsequently, the suspension was cooled to 25°C, filtered with triple-distilled water to remove residual polyvinyl alcohol, and then freeze-dried.

[0133] Table 1

[0134]

[0135] *T / L: Target load

[0136] Examples 2-1 to 2-9 (O / W method): Preparation of biodegradable polymers containing tesipatide and an initial burst release inhibitor Compound microspheres (with varying types and concentrations of initial burst-release inhibitors)

[0137] Tesiparatide (manufacturer: Chengdu Shengnuo Biopharmaceutical Co., Ltd., China) was used as the drug, and RG653H and RG753H (manufacturer: Evonik, Germany) in a 1:1 weight ratio were used as biodegradable polymers, weighed to ensure a batch weight of 0.5 g (target loading of 20%). The drug and polymers were homogeneously dissolved in a mixed solvent of dichloromethane (DCM) and glacial acetic acid as a co-solvent to prepare an oil phase (O phase) as the dispersed phase.

[0138] The initial burst-release inhibitor was added to a 0.1 (w / v)% aqueous solution of polyvinyl alcohol (viscosity: 4.8 to 5.8 mPa·s) to the final concentration and used as the continuous phase (W phase).

[0139] After connecting the continuous phase to an emulsification apparatus equipped with a porous membrane with a diameter of 40 μm, the prepared dispersed phase was injected together with the continuous phase into the porous membrane. An emulsion containing biodegradable polymer droplets containing tesipatide was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm.

[0140] The preparation container was maintained at 25°C. After the dispersed phase was injected, the suspension was kept at 40°C for 3 hours to remove the organic solvent. The suspension was then cooled to 25°C, filtered with triple-distilled water to remove residual polyvinyl alcohol, and then freeze-dried.

[0141] Table 2

[0142]

[0143] Examples 3-1 to 3-2 (O / W method): Preparation of biodegradable polymers containing tesipatide and an initial burst release inhibitor Composite microspheres (different polymer types)

[0144] Tesiparatide (manufacturer: Chengdu Shengnuo Biopharmaceutical Co., Ltd., China) was used as the drug, and RG503H and RG203H (manufacturer: Evonik, Germany) were used as biodegradable polymers, weighed to ensure a batch weight of 0.5 g (target loading of 20%). The drug and polymers were homogeneously dissolved in a mixed solvent of dichloromethane (DCM) solvent and glacial acetic acid co-solvent to prepare an oil phase (O phase) as the dispersed phase.

[0145] Na2HPO4 (disodium hydrogen phosphate) was added as an initial burst release inhibitor to a 0.1 (w / v)% aqueous solution of polyvinyl alcohol (viscosity: 4.8 to 5.8 mPa·s) to make a final concentration of 2.0 (w / v)%, and used as the continuous phase (W phase).

[0146] After connecting the continuous phase to an emulsification apparatus equipped with a porous membrane with a diameter of 40 μm, the prepared dispersed phase was injected together with the continuous phase into the porous membrane. An emulsion containing biodegradable polymer droplets containing tesipatide was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm.

[0147] The preparation container was maintained at 25°C. After the dispersed phase was injected, the suspension was kept at 40°C for 3 hours to remove organic solvents. The suspension was then cooled to 25°C, filtered with triple-distilled water to remove residual polyvinyl alcohol, and then freeze-dried.

[0148] Table 3

[0149]

[0150] Examples 4-2 to 4-3 (W / O / W method): Preparation of biodegradable formulations containing tesipatide and an initial burst release inhibitor Polymer microspheres (different in terms of drug loading or polymer type)

[0151] Tesiparatide (manufacturer: Chengdu Shengnuo Biopharmaceutical Co., Ltd., China) was dissolved in triple-distilled water to prepare the primary aqueous phase. RG653H and RG753H (manufacturer: Evonik, Germany), as biodegradable polymers, were dissolved in dichloromethane (DCM) at a 1:1 weight ratio, or RG503H (manufacturer: Evonik, Germany) alone, to prepare the oil phase (O phase). The aqueous phase was then dissolved in the oil phase using a homogenizer to prepare a primary W1 / O emulsion (dispersed phase).

[0152] Na2HPO4 (disodium hydrogen phosphate) was added as an initial burst release inhibitor to a 0.1 (w / v)% aqueous solution of polyvinyl alcohol (viscosity: 4.8 to 5.8 mPa·s) to make a final concentration of 2.0 (w / v)%, and used as the continuous phase (W2 phase).

[0153] After connecting the continuous phase to an emulsification apparatus equipped with a porous membrane with a diameter of 40 μm, the prepared dispersed phase was injected together with the continuous phase into the porous membrane. A secondary W1 / O / W2 emulsion containing biodegradable polymer microdroplets of tesipatide was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm.

[0154] The preparation container was maintained at 25°C. After the dispersed phase was injected, the suspension was kept at 40°C for 3 hours to remove the organic solvent. The suspension was then cooled to 25°C, filtered with triple-distilled water to remove residual polyvinyl alcohol, and then freeze-dried.

[0155] Table 4

[0156]

[0157] Comparative Example 1 (O / W Method): Preparation of a biodegradable polymer containing tesipatide but without an initial burst release inhibitor microspheres

[0158] 0.045 g of tesiparatide (manufacturer: Zhejiang Paipai Biotechnology Co., Ltd., China) was used as the drug, and 0.255 g of RG653H and RG753H (manufacturer: Evonik, Germany) were used as biodegradable polymers (weight ratio 1:1), weighed to achieve a batch weight of 0.3 g (target loading 15%). Dichloromethane (DCM) was used as the solvent for preparing the dispersed phase, and glacial acetic acid was used as a co-solvent to ensure homogeneous dissolution.

[0159] A 2,000 ml aqueous solution of 0.1% polyvinyl alcohol (viscosity: 4.8~5.8 mPa·s) was used as the continuous phase.

[0160] After connecting the continuous phase to an emulsification apparatus equipped with a porous membrane with a diameter of 40 μm, the prepared dispersed phase was injected together with the continuous phase into the porous membrane. An emulsion containing biodegradable polymer droplets containing tesipatide was prepared, and the suspension was placed in a preparation vessel and stirred at a speed of 200 to 300 rpm.

[0161] The preparation container was maintained at 25°C. After the dispersed phase was injected, the suspension was kept at 40°C for 3 hours to remove the organic solvent. The suspension was then cooled to 25°C, filtered with triple-distilled water to remove residual polyvinyl alcohol, and then freeze-dried.

[0162] Table 5

[0163]

[0164] Experimental Example 1. Measuring the encapsulation amount of endicipatide in microspheres

[0165] To determine the encapsulation amount of tesipatide in the microspheres manufactured in the examples and comparative examples, 10 mg of microspheres were completely dissolved in DMSO. 20 μL of the diluted solution was injected into the HPLC, and measurements were taken at a detection wavelength of 280 nm. The column used in this experimental example was a ZORBAX 300SB-C18, 5 μm, 4.6 x 150 mm, using a mixture of acetonitrile and an aqueous solution containing 0.1% (w / w) trifluoroacetic acid in a 40:60 (v / v) ratio.

[0166] Table 6

[0167]

[0168] As shown in Table 6

[0169] When observing the results of Examples 1-1 to 1-3 manufactured using the O / W method, it can be confirmed that when the T / L reaches 20%, the encapsulation yield remains at 70% or higher.

[0170] When observing the results of Examples 2-1 to 2-9, which were manufactured using the O / W method but with different initial burst-release inhibitor types or concentrations, it can be confirmed that the encapsulation yield remains between 50% and 70%.

[0171] When observing the results of Examples 3-1 and 3-2, which were manufactured using the O / W method but with different polymer types, it can be confirmed that the encapsulation efficiency remained between 50% and 70%.

[0172] When observing the results of Examples 4-2 and 4-3 with different drug loading or polymer types in the W / O / W method, it can be confirmed that the encapsulation efficiency remains at 70% or higher when T / L reaches 30%.

[0173] Experimental Example 2. In vitro measurement of the initial burst release of tesipatide microspheres

[0174] To confirm the initial release of the drug from the microspheres manufactured in the examples and comparative examples, the following experiments were performed. 10 mg of microspheres were placed in an HDPE wide-mouth bottle filled with 50 mL of release test solution and stored in a 37°C incubator. After 24 hours, 1 mL of the test solution was centrifuged to obtain the supernatant. The content and release rate of tesipatide were analyzed by HPLC under the same analytical conditions as in Example 1. The release test solution used for this measurement was a pH 7.4 PBS solution containing sodium dodecyl sulfate and sodium azide.

[0175] Table 7

[0176]

[0177] As shown in Table 7, it was confirmed that the initial burst release of the drug in microspheres containing tesipatide was significantly reduced when an initial burst release inhibitor was included in the continuous phase during microsphere manufacturing. Meanwhile, in Comparative Example 1, which did not contain an initial burst release inhibitor, the release rate on day 1 was 44.59%, confirming that an initial burst release occurred.

[0178] Experimental Example 3. Morphological Analysis Using Electron Microscopy

[0179] The morphology of the microspheres of this invention was analyzed using electron microscopy. The experimental procedure is as follows: 5 mg of microspheres manufactured in the examples and manufacturing examples were placed on an aluminum sample stage with attached carbon ribbons, and platinum was deposited on them using an ion plating machine (COXEM, Korea). The aluminum sample stage was mounted on a scanning electron microscope (COXEM EM-30, Korea), and the morphology of the microspheres was observed at an accelerating voltage of 10 kV. The results are as follows. Figure 1A As shown in Examples 1-1 and 1B (Comparative Example 1).

[0180] Figure 1A These are scanning electron microscope images confirming the morphological characteristics of the microspheres manufactured in Example 1-1.

[0181] Figure 1B These are scanning electron microscope images confirming the morphological characteristics of the microspheres manufactured in Comparative Example 1.

[0182] Example 4. In vivo pharmacokinetic evaluation using rats.

[0183] To evaluate the in vivo drug release pattern of the microspheres according to embodiments of the present invention, the concentration of tesipatide in the blood was measured after administration to rats.

[0184] The microspheres were measured at 3.6 mg / rat (12.0 mg / kg) of tesipatide, dispersed in 0.5 mL of suspension, and then subcutaneously injected into Sprague-Dawley (SD) rats (300 g). 0.5 mL of blood was collected at predetermined times, and the concentration of tesipatide in the blood was measured using HPLC.

[0185] Table 8

[0186]

[0187] As shown in Table 8, it can be confirmed that the bioavailability of the tesipatide microspheres manufactured in the examples is at a good level.

[0188] Example 5. Pharmacokinetic experiment using SD rats after single-dose subcutaneous administration.

[0189] To evaluate the potential of the microspheres of the present invention as a sustained-release therapeutic agent, the concentration of tesipatide in rat blood was measured using the following method.

[0190] Specifically, the microspheres were measured to achieve a tesipatide dose of 3.6 mg / rat, dispersed in a 0.5 mL suspension, and then subcutaneously injected into SD rats. Blood samples of 0.25 to 0.5 mL were collected at predetermined times, and the concentration of tesipatide in the blood was measured using HPLC.

[0191] Table 9

[0192]

[0193] As shown in Table 9, the microspheres prepared in the examples exhibited excellent sustained-release curves, with no initial burst release by day 1 and continued release until day 49. On the other hand, the microspheres of Comparative Example 1, which were confirmed to not contain an initial burst release inhibitor, exhibited an initial burst release by day 1, thereby releasing most of the drug, and then released at a negligible concentration starting from day 10.

[0194] Example 6. Measurement of the residual amount of the initial burst-release inhibitor in the microspheres.

[0195] To measure the content of the initial burst release inhibitor in the microspheres manufactured in the examples, the residual amounts of sodium (Na) and phosphorus (P) in the microspheres were measured.

[0196] Specifically, 300 mg of microspheres were mixed with 6 mL of nitric acid aqueous solution (mixed with ultrapure water in a 1:1 ratio) and 3 mL of hydrogen peroxide. The mixture was heated to 100 °C or higher, and then acid was added until the gas produced during dissolution changed from yellow to white. The resulting sample was weighed, dissolved in ultrapure water, and then injected into an inductively coupled plasma optical emission spectrometer (ICP-OES) (Thermo Scientific Co., UK, iCAP 6300 Duo) for measurement at a detection wavelength of 598.5 nm.

[0197] Table 10

[0198]

[0199] As shown in Table 10, the residual sodium and phosphorus content in the microspheres containing the initial burst release inhibitor in the continuous phase varies depending on the microsphere production conditions. Therefore, the residual initial burst release inhibitor content in the microspheres is confirmed to be: 50 ppm to 500 ppm based on sodium (Na) and 10 ppm to 100 ppm based on phosphorus (P).

Claims

1. A pharmaceutical composition for preventing or treating diabetes, type 2 diabetes, beta cell dysfunction, obesity, or nonalcoholic steatohepatitis, the pharmaceutical composition comprising: sustained-release microspheres consisting of tesidiposide or a pharmaceutically acceptable salt thereof, an initial burst inhibitor, and a biodegradable polymer, wherein the content of tesidiposide or a pharmaceutically acceptable salt thereof is 8 wt.% or more based on the total weight of the microspheres in terms of tesidiposide, and the content of the initial burst inhibitor is 5 ppm to 2000 ppm. The sustained-release microspheres comprising tesidiposide or a pharmaceutically acceptable salt thereof and the initial burst inhibitor release less than 20% of tesidiposide or a pharmaceutically acceptable salt thereof within 24 hours when administered in vivo. The sustained-release microspheres comprising tesidiposide or a pharmaceutically acceptable salt thereof and the initial burst inhibitor release less than 15% of tesidiposide or a pharmaceutically acceptable salt thereof within 24 hours when administered in vivo. The sustained-release microspheres comprising tesidiposide or a pharmaceutically acceptable salt thereof and the initial burst inhibitor release less than 10% of tesidiposide or a pharmaceutically acceptable salt thereof within 24 hours when administered in vivo.

2. The pharmaceutical composition according to claim 1, wherein, The biodegradable polymer is at least one selected from the group consisting of:

3. The pharmaceutical composition of claim 1, wherein, The polymer is selected from the group consisting of polylactide (PLA), polyglycolide (PGA), poly(lactide-co-glycolide) (PLGA), polydioxanone, polycaprolactone (PCL), poly(lactide-co-glycolide-co-caprolactone) (PLGC), poly(lactide-co-glycolymetacrylate) (PLGMGA), polyalkylcarbonate, polytrimethylene carbonate (PTMC), poly(lactide-co-trimethylene carbonate) (PLTMC), polyhydroxybutyric acid (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoester, polyanhydride, poly(anhydride-co-imide), polypropylene fumarate, pseudo polyamino acid, polyalkylcyanoacrylate, polyphosphazene, polyphosphoester, polysaccharide, and poly(butylene succinate-lactide) (PBSLA); a copolymer or a simple mixture of two or more selected from the polymers; a copolymer of the selected polymer with polyethylene glycol (PEG); a polymer-saccharide complex of the selected polymer or the copolymer with a saccharide.

4. The pharmaceutical composition of claim 1, wherein, The initial burst inhibitor is one or more selected from the group consisting of phosphate, hydroxide, phosphide, phosphite, carbonate, chromate, dichromate, oxide, oxalate, silicate, sulfate, sulfide, sulfite, tartrate, tetraborate, thiosulfate, arsenate, arsenite, citrate, ferricyanide, and nitride salts of alkali metals, alkaline earth metals, and ammonium.

5. The pharmaceutical composition of claim 1, wherein, The average particle size of the microspheres comprising tesidiposide or a pharmaceutically acceptable salt thereof and the initial burst inhibitor is 5 μm to 100 μm. The weight of the microspheres comprising tesidiposide or a pharmaceutically acceptable salt thereof and the initial burst inhibitor is 20 mg to 3000 mg.

6. The pharmaceutical composition of claim 1, wherein, The intrinsic viscosity of the biodegradable polymer is 0.16 dL / g to 1.7 dL / g.

7. The pharmaceutical composition of claim 1, wherein, ​ 8. The pharmaceutical composition of claim 1, wherein, ​ 9. The pharmaceutical composition of claim 1, wherein, ​ 10. The pharmaceutical composition of claim 1, wherein, The microspheres further comprise one or more release controlling agents selected from the group consisting of butyric acid, valeric acid, caproic acid, heptanoic acid, caprylic acid, pelargonic acid, capric acid, undecanoic acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, palmitic acid, margaric acid, stearic acid, nonadecanoic acid, behenic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, benzoic acid, hydroxynaphthoic acid, naphthalene disulfonic acid, naphthalenesulfonic acid, and pamoic acid.

11. A method for preparing microspheres with an oil-in-water (O / W) emulsion comprising telipotide or a pharmaceutically acceptable salt thereof, an initial burst inhibitor, and a biodegradable polymer, the method comprising: (a) preparing an oil phase (O phase) as a dispersed phase by dissolving telipotide or a pharmaceutically acceptable salt thereof and one or more biodegradable polymers in an organic solvent; (b) preparing a continuous phase (W phase) by adding an initial burst inhibitor to an aqueous solution containing a surfactant, and then adding the dispersed phase of step (a) to form an emulsion state of the dispersed phase; (c) extracting the organic solvent in the dispersed phase in the emulsion state obtained in step (b) into the continuous phase (W phase) and evaporating the organic solvent to form microspheres; and (d) recovering the microspheres.

12. A method for preparing microspheres with a water-in-oil-in-water (W / O / W) emulsion comprising telipotide or a pharmaceutically acceptable salt thereof, an initial burst inhibitor, and a biodegradable polymer, the method comprising: (a') preparing a primary water phase (W1 phase) by dissolving telipotide or a pharmaceutically acceptable salt thereof in an aqueous solution, and preparing an oil phase (O phase) by dissolving one or more biodegradable polymers in an organic solvent, to prepare a primary W1 / O emulsion as a dispersed phase in which the water phase (W1 phase) and the oil phase (O phase) are mixed; (b') preparing a continuous phase (W2 phase) by adding an initial burst inhibitor to an aqueous solution containing a surfactant, and then adding the dispersed phase of step (a') to prepare a dispersed phase in a secondary W1 / O / W2 emulsion state; (c') extracting the organic solvent in the dispersed phase in the secondary W1 / O / W2 emulsion state in step (b') into the continuous phase (W2 phase) and / or evaporating the organic solvent to form microspheres; and (d') recovering the microspheres.

13. The method of any one of claims 11 and 12, wherein, The biodegradable polymer is at least one selected from the group consisting of a polymer selected from the group consisting of poly-lactide (PLA), poly-glycolide (PGA), poly-lactide-co-glycolide (PLGA), polydioxanone, polycaprolactone (PCL), poly-lactide-co-glycolide-co-caprolactone (PLGC), poly-lactide-co-glycolide-methylcaprolactone (PLGMGA), polyalkylcarbonate, polytrimethylene carbonate (PTMC), poly-lactide-co-trimethylene carbonate (PLTMC), polyhydroxybutyric acid (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBV), polyorthoester, polyanhydride, poly-anhydride-co-imide, polypropylene fumarate, pseudo polyamino acid, polyalkylcyanoacrylate, polyphosphazene, polyphosphoester, polysaccharide, and poly(butylene succinate-lactide) (PBSLA); a copolymer or a simple mixture of two or more selected from the polymers; a copolymer of the selected polymer with polyethylene glycol (PEG); a polymer-saccharide complex of the selected polymer or the copolymer with a saccharide.

14. The method of any one of claims 11 and 12, wherein, The dispersed phase in step (a) or (a') further comprises one or more release controlling agents selected from the group consisting of butyric acid, valeric acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, palmitic acid, heptadecanoic acid, stearic acid, nonadecanoic acid, arachidic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, hydroxynaphthoic acid, naphthalene disulfonic acid, and pamoic acid.

15. The method of any one of claims 11 and 12, wherein, The organic solvent in step (a) or (a') is one or more organic solvents selected from the group consisting of dichloromethane (DCM), chloroform, ethyl acetate, methyl ethyl ketone, acetone, acetonitrile, dimethyl sulfoxide, dimethylformamide, N-methylpyrrolidone, acetic acid, methanol, ethanol, propanol, and benzyl alcohol.

16. The method of any one of claims 11 and 12, wherein, The surfactant in step (b) or (b') is polyvinyl alcohol.

17. The method of any one of claims 11 and 12, wherein, The continuous phase in step (b) or (b') further comprises at least one selected from the group consisting of methanol, ethanol, propanol, and ethyl acetate.

18. The method of any one of claims 11 and 12, wherein, The continuous phase in step (b) is used by adding the initial burst suppressant to a final concentration of 0.1 to 5.0 (w / v)% to an aqueous solution containing a surfactant.

19. The method of any one of claims 11 and 12, wherein, The initial burst suppressant is at least one selected from the group consisting of a phosphate salt of two or more alkali metals, a carbonate salt of one or more alkali metals, and a phosphate salt of two or more ammoniums.

20. The method of any one of claims 11 and 12, wherein, The pH of the continuous phase in step (b) or (b') is 7 or higher.

21. The method of any one of claims 11 and 12, wherein, When the prepared sustained-release microspheres containing tesevimer or a pharmaceutically acceptable salt thereof and an initial burst suppressant are administered in vivo, the release rate of tesevimer or a pharmaceutically acceptable salt thereof is less than 15% in 24 hours.

22. The method of any one of claims 11 and 12, wherein, When the prepared sustained-release microspheres containing tesevimer or a pharmaceutically acceptable salt thereof and an initial burst suppressant are administered in vivo, the release rate of tesevimer or a pharmaceutically acceptable salt thereof is less than 10% in 24 hours.

Images