Semaglutide depot system and its uses

Abstract

The present invention provides a parenteral pharmaceutical composition comprising a therapeutically effective amount of semaglutide or a pharma- ceutical acceptable salt thereof, the parenteral pharmaceutical composition being formulated in a depot form to provide a low burst release and sustained release profile.The present invention further provides methods of use of the parenteral pharmaceutical composition for treating type 2 diabetes, obesity, and Parkinson's disease.

Description

[Technical field]

[0001] The present invention relates to parenteral sustained release pharmaceutical compositions comprising semaglutide and their use in the treatment of type 2 diabetes, obesity and Parkinson's disease. [Background technology]

[0002] Type II or type 2 diabetes (previously called non-insulin-dependent diabetes mellitus (NIDDM) or adult-onset diabetes) is a disorder characterized by high blood glucose levels associated with insulin resistance and relative insulin deficiency. It is often initially managed with increased exercise and dietary modifications, but medications are typically required as the disease progresses.

[0003] Despite advances in the treatment of type 2 diabetes, optimal glycemic control is often not achieved. The hypoglycemia and weight gain associated with many antidiabetic drugs can prevent intensive treatment and long-term application. Current treatments focus on increasing insulin availability (either through direct insulin administration or through agents that enhance insulin secretion), improving insulin sensitivity, delaying carbohydrate delivery and absorption from the gastrointestinal tract, or increasing urinary glucose excretion.

[0004] Glucagon-like peptide-1 (GLP-1) is a naturally occurring peptide of 30 amino acid residues secreted by intestinal cells after a meal. It stimulates glucose-dependent insulin release and inhibits postprandial glucagon secretion. GLP-1 is less effective when administered as a therapeutic agent due to its short pharmacokinetic half-life, i.e., about 2-5 min, mainly due to its susceptibility to enzymatic degradation by the dipeptidyl peptidase enzyme (DPP-4). To extend the half-life of GLP-1, metabolically stable GLP-1 analogs containing chemical modifications and amino acid substitutions have been developed.

[0005] Semaglutide was designed to have a long circulating human half-life through increased albumin binding with the aim of providing a release profile suitable for once-weekly dosing. The sequence of liraglutide (a modified acylated version of the human GLP-1 peptide) was derivatized at position 8 with a 2-aminoisobutyric acid (Aib) group, retaining the arginine at position 34 and the glycine at position 37. The conjugation chemistry to the fatty acid moiety and Lys at position 26 were key features to provide the desired increase in albumin binding affinity. Substitution of Lys26 (γGlu-C16 acid) in liraglutide with extended Lys26 (γGlu-2xOEG-C18 acid) provided semaglutide ([Aib8,Lys26(γGlu-2xOEG-C18 acid),Arg34]-hGLP-1-(7-37)-OH) with a binding affinity of 0.38 nM and a functional potency of 6.2 pM at the human GLP-1 receptor. Incorporation of the Aib amino acid residue at position 8 stabilized semaglutide through protection from DPP-4 enzymatic degradation and, combined with increased albumin affinity, the plasma half-life was extended to 46.1 hours after IV administration and the mean residence time after SC administration was 63.6 hours in minipigs. In a db / db mouse model (a hyperglycemic, hyperinsulinemic obese model of type 2 diabetes), semaglutide reduced ED 50 It produced sustained dose-dependent efficacy at <2 nmol / kg (Suzuki et al., J. Med. Chem., 2020, 13; 63(3): 905-927; doi: 10.1021 / acs.jmedchem.9b00835).

[0006] Semaglutide is indicated as an adjunct to diet and exercise to improve glycemic control in adults with type 2 diabetes and to reduce the risk of major adverse cardiovascular events in adults with type 2 diabetes and established cardiovascular disease. Recently, the US Food and Drug Administration (FDA) approved the use of weekly injections of semaglutide for chronic weight management in adults with obesity or overweight status and at least one weight-related condition (e.g., hypertension, type 2 diabetes, and high cholesterol).

[0007] Yu et al. (Adv. Drug Deli. Rev., 130, p. 113-130) describes various half-life extension strategies employed for GLP-1 receptor agonists, including sequential modification, attachment of fatty acids to peptides, fusion with human serum albumin, fusion with fragment crystallizable (Fc) regions of monoclonal antibodies, sustained drug delivery systems, and PEGylation, and the manner in which these strategies affect the pharmacokinetics, pharmacodynamics, safety, patient utility, and ultimately the commercial success of GLP-1 receptor agonist products. Yu et al. disclose that one of the disadvantages of the PLGA microparticles in the Bydureon® (exenatide) formulation is that they cannot deliver the GLP-1 receptor agonist at a steady rate.

[0008] WO 2020 / 210764 describes a depot that includes a treatment region that includes a therapeutic agent and a control region that includes a bioabsorbable polymer and a release agent mixed with the polymer. The release agent may be configured to dissolve when the depot is placed in vivo to form a diffusion opening in the control region. The depot may be configured to be implanted in vivo at a treatment site and to release the therapeutic agent at the treatment site over an extended period of time while implanted.

[0009] WO 2018 / 136909 describes a method for producing fine particles of an active ingredient via an in-line recirculating mixing system, the in-line recirculating mixing system comprising a mixer and a conduit coupled to the mixer.

[0010] US Patent Application Publication No. 2020 / 0298196 describes methods for producing biodegradable microspheres with improved safety and storage stability, as well as methods for producing the same.

[0011] WO 2020 / 028907 describes compositions comprising GLP-1 or an analogue thereof, such as exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, semaglutide, or taspoglutide, encapsulated or incorporated into polymer particles.

[0012] WO 2021 / 020885 describes a pharmaceutical composition for the treatment or prevention of levodopa-induced dyskinesia. When administered in combination with levodopa, a GLP-1 receptor agonist or a controlled release formulation thereof exhibits the effect of reducing the severe side effects caused by long-term administration of levodopa and alleviating levodopa-induced involuntary dyskinesia.

[0013] China Patent No. 110101846 describes low sudden release rate semaglutide microspheres and the preparation method thereof. The preparation is a long-acting injection prepared from the active ingredient semaglutide, whose weight is 1-20% of the weight of the microsphere, a biocompatible polymer matrix, whose weight is 60-99% of the weight of the microsphere, and other pharma- ceutically acceptable auxiliary materials, whose weight is 0-20% of the weight of the microsphere.

[0014] China Patent No. 113018277 describes a sustained release preparation for injection and its preparation method, which is used to prepare sustained release microspheres of protein polypeptide pharmaceuticals based on water-in-oil-in-water (W1 / O / W2) double emulsion.

[0015] US Patent Application Publication No. 2019 / 0133952 describes a method for preparing sustained-release microspheres, which is characterized by comprising the following steps: 1) preparing a solid dispersion of a water-soluble drug and a biodegradable and biocompatible poorly water-soluble polymer; 2) dissolving the solid dispersion prepared in step 1) in an organic solvent C to form a solid dispersion emulsion, where the organic solvent C is an organic solvent that is not capable of dissolving the water-soluble drug but is capable of dissolving the poorly water-soluble polymer, has a boiling point lower than that of water, and is insoluble or poorly soluble in water; 3) adding the solid dispersion emulsion obtained in step 2) to a surfactant-containing aqueous solution to form a uniform emulsion; and 4) solidifying the microparticles in the emulsion by solvent evaporation or solvent extraction, collecting the microparticles, washing them with ultrapure water several times to remove the surfactant attached to the surface of the microparticles, and drying to obtain sustained-release microparticles.

[0016] There remains an unmet need for improved long-acting semaglutide formulations that can provide safe and effective release of the active ingredient over an extended period of time. Summary of the Invention

[0017] The present invention provides a parenteral pharmaceutical composition comprising a therapeutically effective amount of a GLP-1 receptor agonist, in particular semaglutide or a pharma- ceutically acceptable salt thereof, the parenteral pharmaceutical composition being formulated in a depot form.The present invention further provides a method for treating type 2 diabetes, obesity and Parkinson's disease, comprising administering to a subject in need of treatment a parenteral pharmaceutical composition of the present invention.

[0018] The present invention is based in part on the surprising discovery that a depot formulation of semaglutide and a biodegradable carrier, for example comprising polylactide, polyglycolide, and / or polycaprolactone, provides a long-acting therapeutically effective plasma concentration of semaglutide active ingredient for at least 6 weeks after a single administration.The formulation of the present invention is effective in reducing HbA1c and plasma glucose levels, and provides therapeutic efficacy equivalent to or superior to that of a once-weekly injectable dosage form of semaglutide, with reduced incidence and / or reduced severity of side effects.Compared to formulations derived from other sustained release drug delivery technologies, the semaglutide sustained release formulation of the present invention provides superior release kinetics, with minimal burst, minimal lag time after administration, increased duration of drug release with less frequent injections, continuous plasma levels that are substantially identical to those obtained by administration of a commercially available semaglutide immediate release formulation, and improved local tissue tolerance due to reduced injection volume and small particle size.

[0019] Accordingly, in one aspect, the invention provides a long-acting parenteral pharmaceutical composition comprising dry microparticles, wherein the long-acting parenteral pharmaceutical composition is a long-acting depot composition suitable for administration to a medically acceptable location in a subject in need thereof at a frequency of from once every four weeks to once every six months, wherein the dry microparticles comprise dry microparticles formed by drying water-in-oil-in-water (w / o / w) double emulsion droplets comprising an internal aqueous phase comprising a therapeutically effective amount of semaglutide or a pharma- ceutically acceptable salt thereof, a water-immiscible polymer phase comprising a biodegradable carrier selected from the group consisting of polylactide, polyglycolide, polycaprolactone, and combinations thereof, and an external aqueous phase, wherein the dry microparticles are characterized by a median particle size in the range of about 5 to about 20 μm, including each value within the specified range.

[0020] In one embodiment, the dry particulates are characterized by a median particle size in the range of about 7 to about 17 μm, inclusive, hi another embodiment, the dry particulates are characterized by a median particle size in the range of about 10 to about 15 μm, inclusive, hi another embodiment, the dry particulates are characterized by a median particle size in the range of about 10 to about 15 μm, inclusive.

[0021] In some embodiments, the composition provides a 24-hour semaglutide burst release of less than 20% of the administered dose after administration. In other embodiments, the composition has physical properties such that it releases less than 20% of the semaglutide or a pharma- ceutically acceptable salt thereof over 24 hours in a phosphate buffer at pH 7.4. In yet a further embodiment, the composition has physical properties such that it releases less than 80% of the semaglutide or a pharma- ceutically acceptable salt thereof over 14 days in a phosphate buffer at pH 7.4. In yet another embodiment, the composition has physical properties such that it releases more than 80% of the semaglutide or a pharma- ceutically acceptable salt thereof over 28 days in a phosphate buffer at pH 7.4.

[0022] In various embodiments, the composition provides a mean human steady state plasma concentration (C) of semaglutide of at least about 1 nmol / mL for about 4 weeks to about 6 months following a single dose. ss,avg In additional embodiments, the composition has physical properties such as to provide a mean human steady state plasma concentration (C) of semaglutide of about 1 nmol / mL to about 5 nmol / mL for about 4 weeks to about 6 months following a single dose, including each value within the specified ranges. ss,avg ) and has physical properties that provide

[0023] In another embodiment, the semaglutide is released continuously from the composition.

[0024] In yet another embodiment, the semaglutide is released from the composition in a controlled release sequence selected from zero, first, second and third release sequences, and any pseudo-sequences thereof. Each possibility represents a separate embodiment.

[0025] In certain embodiments, semaglutide is present in the pharmaceutical composition as the only active ingredient.

[0026] In various embodiments, the biodegradable carrier is a biodegradable polymer selected from the group consisting of poly(D,L-lactide-co-glycolide) (PLGA), poly(D,L-lactide) (PLA), polyglycolide (PGA), polycaprolactone (PCL), and combinations thereof. Each possibility represents a separate embodiment. In one currently preferred embodiment, the biodegradable carrier is poly(D,L-lactide-co-glycolide) (PLGA). In another currently preferred embodiment, the biodegradable carrier is poly(D,L-lactide) (PLA). In yet another currently preferred embodiment, the biodegradable carrier is poly(D,L-lactide)-polycaprolactone (PLA-PCL). In an additional currently preferred embodiment, the biodegradable carrier is a blend of poly(D,L-lactide-co-glycolide) and poly(D,L-lactide)-polycaprolactone (PLGA / PLA-PCL).

[0027] In certain embodiments, the biodegradable carrier is poly(D,L-lactide-co-glycolide) (PLGA) having a molecular weight of less than 20 kDa. In other embodiments, the biodegradable carrier is poly(D,L-lactide-co-glycolide) (PLGA) having a molecular weight ranging from about 2 to about 18 kDa, including each value within the specified range.

[0028] In additional embodiments, the water-immiscible polymer phase further comprises a first surfactant comprising a fatty acid or a derivative thereof. In certain embodiments, the first surfactant is lecithin, hydrogenated lecithin, stearic acid, or a mixture or combination thereof. Each possibility represents a separate embodiment. In specific embodiments, the first surfactant comprises hydrogenated lecithin comprising a phosphatidylcholine content of greater than 90% by weight.

[0029] In some embodiments, each of the internal and external aqueous phases independently further comprises a second surfactant. In certain embodiments, the second surfactant is selected from the group consisting of polyvinyl alcohol (PVA), polysorbate, polyethylene oxide-polypropylene oxide block copolymer, polyethylene glycol, and cellulose ester. Each possibility represents a separate embodiment. In a currently preferred embodiment, the second surfactant is PVA. In another currently preferred embodiment, the second surfactant is polyethylene glycol.

[0030] In other embodiments, each of the internal and external aqueous phases independently further comprises a tonicity adjuster. In certain embodiments, the tonicity adjuster is an ionic tonicity adjuster comprising sodium chloride. In further embodiments, the tonicity adjuster is a non-ionic tonicity adjuster comprising a sugar or sugar alcohol. In a specific embodiment, the sugar is sucrose.

[0031] In some embodiments, the ratio of semaglutide or a pharma- ceutically acceptable salt thereof to the biodegradable carrier is in the range of about 1:2 to about 1:30 (w / w), including all iterations of the ratio within the specified range. In other embodiments, the ratio of semaglutide or a pharma- ceutically acceptable salt thereof to the biodegradable carrier is in the range of about 1:5 to about 1:20 (w / w), including all iterations of the ratio within the specified range. In yet other embodiments, the ratio of semaglutide or a pharma- ceutically acceptable salt thereof to the biodegradable carrier is in the range of about 1:5 to about 1:15 (w / w), including all iterations of the ratio within the specified range.

[0032] In various embodiments, the dry microparticles are characterized by a semaglutide drug loading capacity in the range of about 5% to about 15%, inclusive. In other embodiments, the dry microparticles are characterized by a semaglutide drug loading capacity in the range of about 5% to about 12%, inclusive. In additional embodiments, the dry microparticles are characterized by a semaglutide drug loading capacity in the range of about 5% to about 10%, inclusive. In further embodiments, the dry microparticles are characterized by a semaglutide drug loading capacity in the range of about 7% to about 9%, inclusive.

[0033] In certain embodiments, the depot composition of the present invention is prepared by a water-in-oil-in-water (w / o / w) double emulsion process, (i) dispersing an aqueous suspension or solution of semaglutide or a pharma- ceutically acceptable salt thereof in a solution of a biodegradable carrier in a water-immiscible volatile organic solvent, thereby obtaining a water-in-oil emulsion; (ii) dispersing a water-in-oil emulsion in a continuous external aqueous phase to form microparticles comprising water-in-oil-in-water (w / o / w) double emulsion droplets.

[0034] In another aspect, the invention provides a method for preparing a long-acting parenteral pharmaceutical composition comprising semaglutide or a pharma- ceutical acceptable salt suitable for administration to a medically acceptable location in a subject in need thereof at a frequency ranging from once every four weeks to once every six months, the method comprising: (i) dispersing an aqueous suspension or solution of semaglutide or a pharma- ceutically acceptable salt thereof in a solution of a biodegradable carrier in a water-immiscible volatile organic solvent, thereby obtaining a water-in-oil emulsion; (ii) dispersing the water-in-oil emulsion in a continuous external aqueous phase to form microparticles comprising water-in-oil-in-water (w / o / w) double emulsion droplets.

[0035] In some embodiments, the process further comprises (iii) collecting the microparticles so formed by filtration or centrifugation. In other embodiments, the process further comprises (iv) washing the collected microparticles. In one embodiment, washing is performed with purified water, a buffer solution, an external aqueous phase, or a mixture or combination thereof. Each possibility represents a separate embodiment. In another embodiment, washing is performed with an aqueous solution comprising divalent cations. In a further embodiment, the process further comprises drying the collected or washed microparticles. In one embodiment, drying the collected or washed microparticles is performed by lyophilization.

[0036] In some embodiments, the internal aqueous phase has a pH of about 7 to about 9, including each value within the specified range. In other embodiments, the internal aqueous phase has a pH of about 7.5 to about 9.5, including each value within the specified range. In yet other embodiments, the internal aqueous phase has a pH of about 7.8. In various embodiments, the pH is adjusted with an acid or a base. Each possibility represents a separate embodiment. Preferably, the pH is adjusted using sodium hydroxide.

[0037] In additional embodiments, the depot composition of the present invention is in the form of solid particulates, a solution, or a suspension. Each possibility represents a separate embodiment. In a currently preferred embodiment, the composition is in the form of a suspension comprising solid particulates suspended in a physiologically acceptable solvent. In additional embodiments, the suspension comprises dry particulates reconstituted in a physiologically acceptable solvent.

[0038] In a further embodiment, the composition is suitable for a dosing schedule of about once every 4 weeks to about once every 6 months, including values ​​within the specified ranges. In other embodiments, the composition releases the semaglutide active ingredient over a period of about 1 month to about 3 months, including values ​​within the specified ranges. In additional embodiments, the composition releases the semaglutide active ingredient over a period of about 4 weeks to about 6 weeks, including values ​​within the specified ranges. In a currently preferred embodiment, the composition releases the semaglutide active ingredient over a period of about 1 month to about 2 months, including values ​​within the specified ranges.

[0039] In other embodiments, the composition is administered at a dose of about 5 mg to about 100 mg of semaglutide, including each value within the range specified. In some embodiments, the composition is administered intramuscularly. In further embodiments, the composition is administered subcutaneously.

[0040] As contemplated herein, the compositions of the present invention are useful for treating subjects suffering from diabetes, particularly type 2 diabetes.

[0041] Thus, in some embodiments, the present invention provides a method of treating type 2 diabetes, comprising administering to a subject in need of treatment a parenteral pharmaceutical composition disclosed herein at a frequency ranging from once every 4 weeks to once every 6 months, including values ​​within the specified ranges. In one embodiment, the treatment comprises reducing fasting blood glucose levels in the subject by at least about 5%, preferably at least about 10%, more preferably at least about 15%, and most preferably at least about 20%. Each possibility represents a separate embodiment. In another embodiment, the treatment comprises reducing fed blood glucose levels in the subject by at least about 5%, preferably at least about 10%, more preferably at least about 15%, and most preferably at least about 20%. Each possibility represents a separate embodiment. In yet another embodiment, the treatment comprises reducing hemoglobin A1c (HbA1c) levels in the subject by at least about 0.5%, preferably at least about 1%, more preferably at least about 1.5%, and most preferably at least about 2%. Each possibility represents a separate embodiment.

[0042] According to additional embodiments, the compositions of the present invention are useful for treating a subject suffering from Parkinson's disease. Thus, in some embodiments, the present invention provides a method of treating Parkinson's disease, comprising administering to a subject in need of treatment a parenteral pharmaceutical composition disclosed herein at a frequency ranging from once every 4 weeks to once every 6 months, including values ​​within the specified ranges.

[0043] According to further embodiments, the compositions of the present invention are useful for treating obesity. Thus, in some embodiments, the present invention provides a method of treating obesity, comprising administering to a subject in need of treatment a parenteral pharmaceutical composition disclosed herein at a frequency ranging from once every 4 weeks to once every 6 months, including values ​​within the specified ranges.

[0044] According to yet another aspect, the present invention provides a method for treating a patient with a semaglutide-related inflammatory bowel disease, comprising administering to the patient a mean human steady-state plasma concentration (C ss,avg The present invention provides a method for achieving a long-acting parenteral pharmaceutical composition comprising dry microparticles, the dry microparticles being formed by drying water-in-oil-in-water (w / o / w) double emulsion droplets comprising an internal aqueous phase comprising a therapeutically effective amount of semaglutide or a pharma- ceutical acceptable salt thereof, a water-immiscible polymer phase comprising a biodegradable carrier selected from the group consisting of polylactide, polyglycolide, polycaprolactone, and combinations thereof, and an external aqueous phase, comprising administering to a subject in need thereof a long-acting parenteral pharmaceutical composition comprising dry microparticles, the dry microparticles comprising a water-in-oil-in-water (w / o / w) double emulsion droplets comprising an internal aqueous phase comprising a therapeutically effective amount of semaglutide or a pharma- ceutical acceptable salt thereof, a water-immiscible polymer phase comprising a biodegradable carrier selected from the group consisting of polylactide, polyglycolide, polycaprolactone, and combinations thereof, and an external aqueous phase. In various embodiments, the present invention provides a method for achieving a mean human steady-state plasma concentration (C) of semaglutide of about 1 nmol / mL to about 5 nmol / mL for about 4 weeks to about 6 months after a single dose, including each value within the specified ranges. ss,avg ) to achieve this.

[0045] Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description given hereinafter. It should be understood, however, that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. [Brief description of the drawings]

[0046] [Figure 1A] 1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Figure 1B]1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Figure 1C] 1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Figure 1D] 1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Figure 1E] 1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Figure 1F] 1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Figure 1G]1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Figure 1H] 1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Figure 1I] 1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Figure 1J] 1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Figure 1K] 1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Figure 1L]1A and 1B show microscopy images of exemplary microparticles prepared according to the formulations detailed in Example 1. FIG. 1A: MPS-23, FIG. 1B: MPS-25, FIG. 1C: MPS-27M, FIG. 1D: MPS-34HL, FIG. 1E: MPS-40, FIG. 1F: MPS-41, FIG. 1G: MPS-43, FIG. 1H: MPS-44, FIG. 1I: MPS-08, FIG. 1J: MPS-15, FIG. 1K: MPS-45, and FIG. 1L: MPS-46. [Diagram 2] Figure 2 shows the in vitro release of semaglutide from depot formulations MPS-23 (τ), MPS-25 (ν), MPS-27M (Ψ) and MPS-34HL (×). The arrow indicates the time point when porcine pancreas lipase (PPL) was added. [Diagram 3] Figure 2 shows the in vitro release of semaglutide from depot formulations MPS-40 (→), MPS-41 (λ), MPS-43 (+) and MPS-44 (←). The arrow indicates the time point when porcine pancreatic lipase (PPL) was added. [Figure 4A] Figure 4A shows the in vitro release of semaglutide from depot formulation MPS-46 and Figure 4B shows the % semaglutide content in the MPS-46 depot formulation during dissolution testing. [Figure 4B] Figure 4A shows the in vitro release of semaglutide from depot formulation MPS-46 and Figure 4B shows the % semaglutide content in the MPS-46 depot formulation during dissolution testing. [Diagram 5] Ad libitum fed blood glucose levels are shown for Group 1 - naive control (λ), Group 2 - vehicle control (ν), Group 3 - semaglutide 0.06 mg / kg SC daily (Ψ), Group 4 - semaglutide 0.4 mg / kg SC daily (▼), and Group 5 - semaglutide depot (τ). Data are presented as mean + SEM, *p<0.05 vs vehicle. One-way ANOVA followed by Dunnett's test. [Figure 6] Fasting blood glucose levels are shown on days 14 and 28. Data are presented as mean + SEM, *p<0.05 vs. vehicle. One-way ANOVA followed by Dunnett's test. [Figure 7]HbA1c in db / db mice on days 0 and 28. Data are presented as mean + SEM, *p<0.05 vs. vehicle. One-way ANOVA followed by Dunnett's test. [Figure 8] Body weights of db / db mice are shown in Group 1 - naive control (λ), Group 2 - vehicle control (ν), Group 3 - semaglutide 0.06 mg / kg SC daily (Ψ), Group 4 - semaglutide 0.4 mg / kg SC daily (▼), and Group 5 - semaglutide depot (τ). [Figure 9A] Food intake is shown. Figure 9A: Group 1 - naive control (λ and Group 2 - vehicle control (ν), Figure 9B: Group 2 - vehicle control (ν) and Group 3 - semaglutide 0.06 mg / kg SC daily (Ψ), Figure 9C: Group 2 - vehicle control (ν) and Group 4 - semaglutide 0.4 mg / kg SC daily (▼), Figure 9D: Group 2 - vehicle control (ν and Group 5 - semaglutide depot (τ). Data are presented as mean + SEM. [Figure 9B] Food intake is shown. Figure 9A: Group 1 - naive control (λ and Group 2 - vehicle control (ν), Figure 9B: Group 2 - vehicle control (ν) and Group 3 - semaglutide 0.06 mg / kg SC daily (Ψ), Figure 9C: Group 2 - vehicle control (ν) and Group 4 - semaglutide 0.4 mg / kg SC daily (▼), Figure 9D: Group 2 - vehicle control (ν and Group 5 - semaglutide depot (τ). Data are presented as mean + SEM. [Figure 9C] Food intake is shown. Figure 9A: Group 1 - naive control (λ and Group 2 - vehicle control (ν), Figure 9B: Group 2 - vehicle control (ν) and Group 3 - semaglutide 0.06 mg / kg SC daily (Ψ), Figure 9C: Group 2 - vehicle control (ν) and Group 4 - semaglutide 0.4 mg / kg SC daily (▼), Figure 9D: Group 2 - vehicle control (ν and Group 5 - semaglutide depot (τ). Data are presented as mean + SEM. [Figure 9D]Food intake is shown. Figure 9A: Group 1 - naive control (λ and Group 2 - vehicle control (ν), Figure 9B: Group 2 - vehicle control (ν) and Group 3 - semaglutide 0.06 mg / kg SC daily (Ψ), Figure 9C: Group 2 - vehicle control (ν) and Group 4 - semaglutide 0.4 mg / kg SC daily (▼), Figure 9D: Group 2 - vehicle control (ν and Group 5 - semaglutide depot (τ). Data are presented as mean + SEM. [Figure 10A] Semaglutide plasma concentrations in db / db mice are shown. Figure 10A: Group 3 - 0.06 mg / kg semaglutide SC daily, Figure 10B: Group 4 - 0.4 mg / kg semaglutide SC daily, Figure 10C: Group 5 - semaglutide depot. Data are shown as mean + SEM. Figure 10D: Semaglutide depot (Ψ) after a single IM dose versus semaglutide solution administered subcutaneously every 24 hours (λ). [Figure 10B] Semaglutide plasma concentrations in db / db mice are shown. Figure 10A: Group 3 - 0.06 mg / kg semaglutide SC daily, Figure 10B: Group 4 - 0.4 mg / kg semaglutide SC daily, Figure 10C: Group 5 - semaglutide depot. Data are shown as mean + SEM. Figure 10D: Semaglutide depot (Ψ) after a single IM dose versus semaglutide solution administered subcutaneously every 24 hours (λ). [Figure 10C] Semaglutide plasma concentrations in db / db mice are shown. Figure 10A: Group 3 - 0.06 mg / kg semaglutide SC daily, Figure 10B: Group 4 - 0.4 mg / kg semaglutide SC daily, Figure 10C: Group 5 - semaglutide depot. Data are shown as mean + SEM. Figure 10D: Semaglutide depot (Ψ) after a single IM dose versus semaglutide solution administered subcutaneously every 24 hours (λ). [Figure 10D] Semaglutide plasma concentrations in db / db mice are shown. Figure 10A: Group 3 - 0.06 mg / kg semaglutide SC daily, Figure 10B: Group 4 - 0.4 mg / kg semaglutide SC daily, Figure 10C: Group 5 - semaglutide depot. Data are shown as mean + SEM. Figure 10D: Semaglutide depot (Ψ) after a single IM dose versus semaglutide solution administered subcutaneously every 24 hours (λ). [Figure 11] Figure 2 shows mean semaglutide plasma concentrations in minipigs. Semaglutide depot (Ψ) after a single IM administration versus semaglutide solution administered subcutaneously every 72 hours (λ). [Figure 12] Ad libitum feeding blood glucose levels are shown in db / db mice for Group 1 - naive control (λ), Group 2 - vehicle control (ν), Group 3 - semaglutide 0.02 mg / kg SC daily (Ψ), Group 4 - semaglutide depot 2 mg / kg IM once on day 1 (▼), and Group 5 - semaglutide depot 2 mg / kg SC once on day 1 (τ). Data are presented as mean + SEM, *p<0.05 vs vehicle. Two-way ANOVA followed by Bonferroni's multiple comparison test. [Figure 13] Figure 1 shows fasting blood glucose levels in db / db mice on days 14, 28, and 42. Data are presented as mean + SEM, *p<0.05 vs. vehicle. One-way ANOVA followed by Dunnett's test. [Figure 14] HbA1c in db / db mice on days 0, 28 and 42. Data are presented as mean + SEM, *p<0.05 vs. vehicle. One-way ANOVA followed by Dunnett's test. [Figure 15] Shown are body weights of db / db mice in Group 1 - naive control (λ), Group 2 - vehicle control (ν), Group 3 - semaglutide 0.02 mg / kg SC daily (Ψ), Group 4 - semaglutide depot 2 mg / kg IM once on day 1 (▼), and Group 5 - semaglutide depot 2 mg / kg SC once on day 1 (τ). Data are presented as mean + SEM, *p<0.05 vs. vehicle. Two-way ANOVA followed by Bonferroni's multiple comparison test. [Figure 16]Sample intake in db / db mice is shown for Group 1 - naive control (λ), Group 2 - vehicle control (ν), Group 3 - semaglutide 0.02 mg / kg SC daily (Ψ), Group 4 - semaglutide depot 2 mg / kg IM once on day 1 (▼), and Group 5 - semaglutide depot 2 mg / kg SC once on day 1 (τ). Data are presented as mean + SEM, *p<0.05 vs vehicle. Two-way ANOVA followed by Bonferroni's multiple comparison test. [Figure 17A] Figure 17 shows semaglutide plasma concentrations in db / db mice. Figure 17A: Group 3 - semaglutide 0.02 mg / kg SC daily, Figure 17B: Group 5 - semaglutide depot 2 mg / kg SC once on day 1. Data are shown as mean + SEM. [Figure 17B] Figure 17 shows semaglutide plasma concentrations in db / db mice. Figure 17A: Group 3 - semaglutide 0.02 mg / kg SC daily, Figure 17B: Group 5 - semaglutide depot 2 mg / kg SC once on day 1. Data are shown as mean + SEM. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0047] The present invention provides a long-acting pharmaceutical preparation of semaglutide or a pharma- ceutically acceptable salt thereof, which provides therapeutic efficacy equivalent to or superior to that of a once-weekly composition, but is designed for administration at a frequency of once every 4 weeks to once every 6 months, thus providing improved patient compliance. In addition to providing the same or superior therapeutic efficacy, the pharmaceutical preparation of the present invention reduces side effects (local and / or systemic) resulting from frequent injections of semaglutide, including lipoatrophy, lipohypertrophy, local allergic reactions, abscess formation, and scarring. The present invention further provides a low burst release semaglutide depot formulation, which provides a continuous release of the active ingredient, thereby avoiding undesirable release of high concentrations of semaglutide immediately after administration or delayed release of semaglutide after administration, thus providing improved glycemic control and prevention of hypoglycemic and hyperglycemic events within the first 48 hours after administration. The low burst release of semaglutide within the first 48 hours after administration is also advantageous in preventing the adverse events of nausea and vomiting that are often associated with semaglutide administration.

[0048] The long-acting parenteral compositions disclosed herein include microparticles characterized by small median particle size in the range of about 5 to about 20 μm, which is associated with improved physical stability and easy injectability. The microparticles also feature improved drug loading capacity. In accordance with the principles provided herein, effective plasma concentrations within the clinical range are achieved and maintained for at least four weeks following a single administration of the compositions of the present invention. The compositions are biodegradable, resulting in rapid clearance from the injection site.

[0049] According to some aspects and embodiments, the pharmaceutical formulations and dosages of the present invention are conveniently provided in a form suitable for parenteral administration, for example, by injection, implantation, or infusion. Each possibility represents a separate embodiment. The term "parenteral" as used herein refers to a route of administration selected from subcutaneous (SC), intravenous (IV), intramuscular (IM), intradermal (ID), intraperitoneal (IP), and the like. Each possibility represents a separate embodiment. In a currently preferred embodiment, the pharmaceutical composition is administered via the intramuscular (IM) route. In other currently preferred embodiments, the pharmaceutical composition is administered subcutaneously.

[0050] Within the scope of the present invention are sustained release depot formulations. The term "sustained" as used herein refers to a pharmaceutical composition that provides a long-term, extended, or prolonged release of a therapeutically effective amount of semaglutide or any pharma- ceutically acceptable salt thereof to the general circulation of a subject or to a local site of action in a subject. This term may further refer to a pharmaceutical composition that provides a long-term, extended, or prolonged exposure (pharmacokinetics) to a therapeutically effective amount of semaglutide or any pharma-ceutically acceptable salt thereof, and a duration of its (pharmacodynamics) action in a subject. In particular, the sustained release pharmaceutical composition of the present invention provides a dosing regimen of once every 4 weeks, once every month, once every 1.5 months, once every 2 months, once every 3 months, once every 4 months, once every 5 months, or once every 6 months. Each possibility represents a separate embodiment.

[0051] Depending on the duration of action required, each depot or implantable device of the invention is designed to provide release of semaglutide or a pharma- ceutically acceptable salt thereof over a period selected from the group consisting of 4 weeks, 1 month, 1.5 months, 2 months, 2.5 months, 3 months, 3.5 months, 4 months, 4.5 months, 5 months, 5.5 months, and 6 months. Each possibility represents a separate embodiment.

[0052] The depot system of the present invention encompasses a form selected from the group consisting of biodegradable microspheres, non-biodegradable microspheres, implants of any suitable geometric shape, sustained release gels, and erodible matrices. Each possibility represents a separate embodiment. According to certain embodiments, the implants of any suitable geometric shape are selected from the group consisting of implantable capsules, implantable rods, and implantable rings. Each possibility represents a separate embodiment.

[0053] According to some embodiments, suitable forms of parenteral pharmaceutical compositions include, but are not limited to, injectable compositions containing microparticles. The microparticles contain a therapeutically effective amount of an active ingredient encapsulated in a biodegradable or non-biodegradable polymer. Each possibility represents a separate embodiment. In certain embodiments, the microparticles contain semaglutide in an amount ranging from about 30 mg to about 130 mg per gram of microparticles, including each value within the range specified. In other embodiments, the microparticles contain semaglutide in an amount ranging from about 50 mg to about 100 mg per gram of microparticles, including each value within the range specified. In various embodiments, the microparticles are formed by drying water-in-oil-in-water (w / o / w) double emulsion droplets. The double emulsion droplets according to the principles of the present invention include an internal aqueous phase containing a therapeutically effective amount of semaglutide or a pharma- ceutically acceptable salt thereof, a water-immiscible polymer phase containing a biodegradable carrier selected from the group consisting of polylactide, polyglycolide, polycaprolactone, and combinations thereof, and an external aqueous phase.

[0054] According to the principles of the present invention, semaglutide may be present in the composition in the form of a free base or in the form of its salt or mixtures thereof.Each possibility represents a separate embodiment.Representative examples of salts include, but are not limited to, salts with suitable inorganic acids, such as hydrochloric acid, hydrobromic acid, and the like.Each possibility represents a separate embodiment.Representative examples of salts also include, but are not limited to, salts with organic acids, such as formic acid, acetic acid, propionic acid, lactic acid, tartaric acid, ascorbic acid, citric acid, and the like.Each possibility represents a separate embodiment. Representative examples of salts include, but are not limited to, salts containing bases such as triethanolamine, diethylamine, meglumine, arginine, alanine, leucine, diethylethanolamine, olamine, triethylamine, tromethamine, choline, trimethylamine, taurine, benzamine, methylamine, dimethylamine, trimethylamine, methylethanolamine, propylamine, isopropylamine, adenine, guanine, cytosine, thymine, uracil, thymine, xanthine, hypoxanthine, and the like. Each possibility represents a separate embodiment. According to further embodiments, pharma- ceutically acceptable salts include acid addition salts such as those containing sulfate, phosphate, sulfamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclohexylsulfamate, and quinate. Each possibility represents a separate embodiment.

[0055] Salts according to the principles of the present invention can be prepared, for example, by reacting the free acid or free base form with one or more equivalents of the appropriate base or acid, respectively, in a solvent or medium in which the salt is insoluble, or in a solvent such as water which is then removed in vacuo, by lyophilization, or by exchanging an ion of an existing salt for another ion on a suitable ion exchange resin. Each possibility represents a separate embodiment.

[0056] According to various aspects and embodiments, semaglutide or a salt thereof is present in the parenteral composition disclosed herein as the only active ingredient. Typically, semaglutide or a semaglutide salt is present in the parenteral composition of the present invention in a therapeutically effective amount. As used herein, the term "therapeutically effective amount" is intended to encompass an amount of semaglutide or a semaglutide salt that provides the following response after administration of the parenteral composition: stimulation of glucose-dependent insulin release and / or suppression of postprandial glucagon secretion in patients with type 2 diabetes. In some embodiments, semaglutide or a semaglutide salt is present in the parenteral composition disclosed herein in a dose of about 5 mg to about 100 mg, including each value within the specified range. Exemplary doses within the scope of the present invention include, but are not limited to, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, or about 100 mg, with each possibility representing a separate embodiment.

[0057] The internal aqueous phase may further comprise a surfactant and / or a tonicity adjuster according to the principles of the present invention. Suitable surfactants in the aqueous phase include, but are not limited to, polyvinyl alcohol (PVA), polysorbates, polyethylene oxide-polypropylene oxide block copolymers, polyethylene glycol, and cellulose esters. Each possibility represents a separate embodiment. Currently preferred embodiments include the use of PVA and / or PEG. Suitable tonicity adjusters include, but are not limited to, ionic and non-ionic tonicity adjusters, such as sodium chloride, sugars (e.g., sucrose), or sugar alcohols (e.g., mannitol, sorbitol). Each possibility represents a separate embodiment. In one embodiment, the internal aqueous phase consists essentially of semaglutide or a pharma- ceutically acceptable salt thereof and water. In another embodiment, the internal aqueous phase is devoid of polyethylene glycol, gelatin, glycerin, mannitol, sucrose, trehalose, lactose, glucose, propylene glycol, sorbitol, zinc chloride, zinc sulfate, zinc acetate, and human serum proteins.

[0058] In certain embodiments, the dosage forms include, but are not limited to, biodegradable injectable depot systems, such as PLGA-based injectable depot systems, non-PLGA-based injectable depot systems, and injectable biodegradable gels or dispersions. Each possibility represents a separate embodiment. The term "biodegradable" as used herein refers to a component that erodes or degrades at its surface over time, at least in part, due to contact with materials found in the surrounding tissue fluids or by cellular action. In some embodiments, the biodegradable polymer has an average molecular weight of about 1,000 to about 200,000 daltons, including each value within the specified range.

[0059] Suitable biodegradable or non-biodegradable depot systems within the scope of the present invention include at least one of the following polymers: polyanhydrides, poly(sebacic acid) (SA), poly(ricinoleic acid) (RA), poly(fumaric acid) (FA), poly(fatty acid dimer) (FAD), poly(fatty acid dimmer) (FAD), poly(terephthalic acid) (TA), poly(isophthalic acid ... acid), poly(p-{carboxyphenoxy}methane), CPM, poly(p-{carboxyphenoxy}methane), CPP, poly(p-{carboxyphenoxy}propane), CPH, polyamine, polyurethane, polyesteramide, polyorthoester {CHDM: cis / trans cyclohexyldimethanol, HD: 1,6-hexanediol, DETOU: (3,9-diethylidene-2,4,8,10-Tetraoxaspirondecane), polydioxanone, polyhydroxybutyrate, polyalkylene oxalate, polyamide, polyesteramide, polyacetal, polyketal, polycarbonate, polyorthocarbonate, polysiloxane, polyphosphazene, succinate, hyaluronic acid, poly(malic acid), poly(amino acid), polyhydroxyvalerate, polyalkylene succinate, polyvinylpyrrolidone, polystyrene, synthetic cellulose ester, polyacrylic acid, polybutyric acid, triblock copolymer (PEG-PLGA), ... Examples of suitable systems include, but are not limited to, systems comprising poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymers (PEO-PPO-PEO), polyvaleric acid, polyethylene glycol, polyhydroxyalkylcellulose, chitin, chitosan, polyorthoesters and copolymers, terpolymers, cholesterol, lipids such as lecithin, poly(glutamic acid-co-ethyl glutamate) and the like, or mixtures thereof. Each possibility represents a separate embodiment.

[0060] Additional depot systems within the scope of the present invention include, but are not limited to, systems comprising at least one of the following polymers: poly(D,L-lactide-co-glycolide) (PLGA), poly(D,L-lactide) (PLA), polyglycolide (PGA), polycaprolactone (PCL), polyhydroxybutyrate, polyorthoesters, polyalkane anhydrides, gelatin, collagen, oxidized cellulose, polyphosphazene, and any combination thereof. Each possibility represents a separate embodiment.

[0061] In particular, biodegradable polymers include, but are not limited to, polylactides, e.g., poly(D,L-lactide), i.e., lactic acid-based polymers such as PLA, polyglycolide (PGA), e.g., glycolic acid-based polymers such as Lactel® from Durect, poly(D,L-lactide-co-glycolide), i.e., PLGA, polycaprolactones such as (Resomer® RG-504, Resomer RG-502, Resomer® RG-504H, Resomer® RG-502H, Resomer® RG-504S, Resomer® RG-502S from Boehringer, Lactel® from Durect), and poly(ε-caprolactone), i.e., PCL (Lactel® from Durect). Each possibility represents a separate embodiment.

[0062] A currently preferred biodegradable polymer is a lactic acid based polymer, more preferably polylactide, or poly(D,L-lactide-co-glycolide), i.e., PLGA. Another currently preferred biodegradable polymer is polycaprolactone (PCL). Yet another currently preferred biodegradable polymer is polylactic acid (PLA). A further currently preferred biodegradable polymer is PLA-PCL. An additional currently preferred biodegradable polymer is a mixture of PLGA with PLA-PCL. In one embodiment, the weight percent ratio of PLGA to PLA-PCL is in the range of 9:1 to 1:9, including all iterations of the ratio within the specified range. In another embodiment, the weight percent ratio of PLGA to PLA-PCL is in the range of 9:1 to 7:3, including all iterations of the ratio within the specified range. In yet another embodiment, the weight percent ratio of PLGA to PLA-PCL is 8:2. Typically, the biodegradable polymer is present in an amount of about 10% to about 98% w / w of the solid composition (e.g., microparticles), including each value within the range specified, however, it is understood that the amount of biodegradable polymer is determined by parameters such as duration of use and the like.

[0063] In some embodiments, the lactic acid based polymer PLGA has a monomer ratio of lactic acid to glycolic acid ranging from 100:0 to about 0:100, preferably 100:0 to about 10:90, including all iterations of the ratio within the specified range. In one embodiment, the lactic acid based polymer has a monomer ratio of lactic acid to glycolic acid of 80:20. In another embodiment, the lactic acid based polymer has a monomer ratio of lactic acid to glycolic acid of 75:25. In yet another embodiment, the lactic acid based polymer has a monomer ratio of lactic acid to glycolic acid of 50:50. In various embodiments, the PLGA has a molecular weight of less than 20 kDa, for example, from about 2 to about 18 kDa, including each value within the specified range. Exemplary molecular weights of PLGA used in the depot formulations disclosed herein include, but are not limited to, about 2 kDa, about 3 kDa, about 4 kDa, about 5 kDa, about 6 kDa, about 7 kDa, about 8 kDa, about 9 kDa, about 10 kDa, about 11 kDa, about 12 kDa, about 13 kDa, about 14 kDa, about 15 kDa, about 16 kDa, about 17 kDa, about 18 kDa, about 19 kDa, and up to 20 kDa. Each possibility represents a separate embodiment. Advantageously, the use of low molecular weight PLGA allows for rapid excretion of the biodegradable polymer from the injection site, thereby minimizing local side effects.

[0064] According to certain aspects and embodiments, the polymer phase further comprises a surfactant, which may be a fatty acid or a derivative thereof. Suitable surfactants that may be incorporated into the oil phase include, but are not limited to, lecithin, hydrogenated lecithin, stearic acid, or mixtures or combinations thereof. Each possibility represents a separate embodiment. Within the scope of the present invention are natural or synthetic lecithin surfactants. In some embodiments, the oil phase comprises a phospholipid surfactant. A phospholipid is a phosphorus-containing lipid found in lecithin. Exemplary phospholipids include, but are not limited to, egg yolk phospholipids, soybean phospholipids, hydrogenated phospholipids, phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine, phosphatidic acid, PEGylated phospholipids, refined oils, refined fatty acids and fatty acid salts, cationic lipids, glycerophosphocholine, and mixtures or combinations thereof. Each possibility represents a separate embodiment. Natural phospholipids may be derived, for example, from soybeans, sunflowers, and rapeseed (canola). Each possibility represents a separate embodiment. A currently preferred surfactant in the oil phase is hydrogenated lecithin containing a phosphatidylcholine content of greater than 90% by weight, e.g., from about 91 to about 99% by weight, including each value within the range specified. Without being bound by any theory or mechanism of action, the presence of a surfactant containing a fatty acid or a derivative thereof in the oil phase confers improved properties to the microparticles, including uniform particle size distribution, enhanced drug loading capacity, and improved stability and pharmacokinetic properties.

[0065] In accordance with the principles of the present invention, the ratio of semaglutide or a pharma- ceutically acceptable salt thereof to biodegradable carrier is typically in the range of about 1:2 to about 1:30 (w / w), including all iterations of ratios within the specified range. Exemplary ratios include, but are not limited to, about 1:2 to about 1:25, about 1:2 to about 1:20, about 1:2 to about 1:15, about 1:2 to about 1:10, about 1:2 to about 1:5, about 1:5 to about 1:30, about 1:5 to about 1:25, about 1:5 to about 1:20, about 1:5 to about 1:15, about 1:5 to about 1:10, about 1:10 to about 1:30, about 1:10 to about 1:25, about 1:10 to about 1:20, about 1:10 to about 1:15, about 1:15 to about 1:30, about 1:15 to about 1:25, about 1:15 to about 1:20, about 1:20 to about 1:30, or about 1:25 to about 1:30 (w / w). Each possibility represents a separate embodiment.

[0066] The microparticles of the present invention provide a high amount / loading of semaglutide in the microparticles. Thus, according to the principles of the present invention, the semaglutide drug loading capacity is in the range of about 5% to about 15%, inclusive of each value within the range specified. The term "drug loading capacity" as used herein refers to the weight percentage of drug (i.e., semaglutide or a pharma- ceutically acceptable salt thereof) encapsulated in the dry microparticles. Typically, the drug loading capacity is in the range of about 5% to about 12%, about 5% to about 10%, or about 7% to about 9%, inclusive of each value within the range specified. It is contemplated that the high amount / loading of semaglutide in the microparticles of the present invention results in high efficacy of the formulation with the smallest injection volume administered.

[0067] Without being bound to a particular theory, it is believed that the release of semaglutide from the depot formulation may occur by one of two different mechanisms. The first mechanism involves release by diffusion through aqueous-filled channels created in the polymer matrix, such as by dissolution of the biologically active agent or by voids created by removal of the polymer solvent during preparation of the sustained release composition. Additional channels may be formed using pore-forming agents, such as zinc oxide. The second mechanism involves release of the biologically active agent due to degradation of the polymer. The rate of degradation can be controlled by adjusting the polymer properties that affect its hydration rate. These properties include, for example, the ratio of lactide to glycolide containing polymer, the use of L-isomers of monomers instead of racemic mixtures, and the molecular weight of the polymer. These properties can affect the hydrophilicity and crystallinity, which control the hydration rate of the polymer. The release profile can be controlled by varying the properties of the polymer. For example, increasing the glycolide content of a poly(lactide-co-glycolide) polymer and decreasing the molecular weight of the polymer can enhance hydrolysis of the polymer and therefore provide increased release of the biologically active agent from polymer erosion.

[0068] According to various aspects and embodiments, the release of semaglutide from the composition occurs continuously. The release profile can be a zero-order release profile, a first-order release profile, a second-order release profile, a third-order release profile, or any known pseudo-order. Each possibility represents a separate embodiment. According to certain aspects and embodiments, the composition provides a low burst release, providing a 24-hour semaglutide release of less than 20% of the administered dose after administration. The release of semaglutide from the formulation can also be determined in vitro. In some embodiments, less than 20% of semaglutide is released from the depot formulation in a phosphate buffer at pH 7.4 within 1 day. In other embodiments, less than 80% of semaglutide is released from the depot formulation in a phosphate buffer at pH 7.4 within 14 days. In a further embodiment, more than 80% of semaglutide is released from the depot formulation in a phosphate buffer at pH 7.4 within 28 days.

[0069] The composition of the present invention maintains a mean human steady-state plasma concentration (C ) of semaglutide of at least about 1 nmol / mL for about 4 weeks to about 6 months after a single administration. ss,avg In some embodiments, the compositions of the invention achieve a mean human steady state plasma concentration (C) of semaglutide of about 1 nmol / mL to about 5 nmol / mL for about 4 weeks to about 6 months following a single dose. ss,avg In another embodiment, the compositions of the invention achieve a mean human steady state plasma concentration (C) of semaglutide of about 1 nmol / mL to about 10 nmol / mL for about 4 weeks to about 6 months following a single dose. ss,avg ) to be achieved.

[0070] According to further aspects and embodiments, the emulsion droplets comprise an external aqueous phase, which in currently preferred embodiments further comprises at least one of a surfactant and a tonicity adjuster, as detailed above for the internal aqueous phase.

[0071] According to the principles of the present invention, the water-in-oil-in-water (w / o / w) double emulsion droplets are subsequently dried to provide dry microparticles. The dry microparticles can be administered as is. According to some aspects and embodiments, the dry microparticles are suspended in an inert oil, preferably a vegetable oil such as sesame oil, peanut oil, olive oil, or other acceptable carrier. Each possibility represents a separate embodiment. Preferably, the dry microparticles are suspended in an aqueous carrier, such as an isotonic buffer at a pH of about 3.0 to about 7.0, more preferably about 4.0 to about 6.0, and most preferably about 4.0 to about 5.0, including each value within the specified range. These compositions can be sterilized by conventional sterilization techniques or can be sterile filtered.

[0072] The compositions disclosed herein may further contain pharma- ceutically acceptable auxiliary substances required to approximate physiological conditions, such as pH buffers. Suitable buffers include, but are not limited to, sodium acetate / acetic acid buffers. The desired isotonicity may be achieved using sodium chloride or other pharma- ceutically acceptable agents, such as dextrose, boric acid, sodium tartrate, propylene glycol, polyols (e.g., mannitol and sorbitol), or other inorganic or organic solutes. Each possibility represents a separate embodiment. Sodium chloride is particularly preferred for buffers containing sodium ions.

[0073] According to some embodiments, carriers or excipients can be used to facilitate administration of the dosage of the present invention. Examples of carriers and excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars such as lactose, or various types of starch, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols, and physiologically compatible solvents. Each possibility represents a separate embodiment.

[0074] According to various embodiments, the solutions of the dosage forms may be thickened with a thickening agent, such as, but not limited to, methylcellulose. They may be prepared in either water-in-oil or oil-in-water emulsified form. Each possibility represents a separate embodiment. Any of a wide variety of pharma- ceutically acceptable emulsifiers may be employed, including, for example, acacia powder, non-ionic surfactants (such as Tween), or ionic surfactants (alkali polyether alcohol sulfates or sulfonates, e.g., Triton). Each possibility represents a separate embodiment.

[0075] According to additional embodiments, the pharma- ceutically acceptable carrier is a liquid. According to further embodiments, the liquid is selected from the group consisting of an aqueous or non-aqueous solvent, an emulsion, and a suspension. Each possibility represents a separate embodiment. According to other embodiments, the liquid is an aqueous solvent selected from the group consisting of saline, a dextrose solution, and a glycerol solution. Each possibility represents a separate embodiment.

[0076] The compositions of the present invention may further comprise one or more pharma- ceutically acceptable excipients selected from, but not limited to, co-surfactants / solubilizers, solvents / co-solvents, water-immiscible solvents, water, water-miscible solvents, oily components, hydrophilic solvents, emulsifiers, preservatives, antioxidants, antifoaming agents, stabilizers, buffers or pH adjusters, osmotic agents, pore formers, osmolality adjusters, and the like. Each possibility represents a separate embodiment. Suitable co-surfactants or solubilizers include, but are not limited to, polyethylene glycols, polyoxyethylene-polyoxypropylene block copolymers known as "poloxamers", polyglycerin fatty acid esters such as decaglyceryl monolaurate and decaglyceryl monomyristate, sorbitan fatty acid esters such as sorbitan monostearate, polyoxyethylene sorbitan fatty acid esters such as polyoxyethylene sorbitan monooleate (Tween), polyethylene glycol fatty acid esters such as polyoxyethylene monostearate, polyoxyethylene alkyl ethers such as polyoxyethylene lauryl ether, polyoxyethylene castor oils such as polyoxyethylene hydrogenated castor oils and hydrogenated castor oils, and the like, or mixtures thereof. Each possibility represents a separate embodiment. Suitable solvents / co-solvents include, but are not limited to, alcohols, triacetin, dimethyl isosorbide, glycofurol, propylene carbonate, water, dimethylacetamide, and the like, or mixtures thereof. Each possibility represents a separate embodiment. Suitable antifoaming agents include, but are not limited to, silicone emulsion or sorbitan sesquioleate. Each possibility represents a separate embodiment. Suitable stabilizers for preventing or reducing the deterioration of ingredients in the compositions of the present invention include, but are not limited to, antioxidants such as glycine, alpha-tocopherol or ascorbate, BHA, BHT, and the like, or mixtures thereof. Each possibility represents a separate embodiment. Suitable tonicity adjusting agents include, but are not limited to, mannitol, sodium chloride, and glucose. Each possibility represents a separate embodiment. Suitable buffering agents include, but are not limited to, acetates, phosphates, and citrates with suitable cations.Each possibility represents a separate embodiment.

[0077] The sustained release depot system of the present invention can be prepared by any manner known in the art.Currently preferred is to incorporate semaglutide or a pharma-ceutically acceptable salt thereof into a colloid delivery system, such as a biodegradable microparticle, thus allowing delayed release by diffusion through the polymeric wall of the particle and by polymer degradation in aqueous media or biological fluids in the body.In some embodiments, the biodegradable microparticle lacks any coating layer.

[0078] According to some embodiments, the sustained release microparticles of the present invention are prepared in the form of injectable dry microparticles by a process known as "double emulsion". Briefly, a concentrated aqueous solution or suspension of semaglutide or a pharma- ceutically acceptable salt thereof, optionally containing a surfactant (e.g., polyvinyl alcohol-PVA, polysorbate, polyethylene oxide-polypropylene oxide block copolymer, cellulose ester, and the like) and / or a tonicity adjuster (e.g., sucralose), is prepared. The pH of the aqueous solution is typically adjusted to a range of about 7 to about 9, or about 7.5 to about 9.5, including each value within the range specified. The adjustment of the pH can be performed using any acid or base, for example, sodium hydroxide. The aqueous solution or suspension is then dispersed in a solution of a biodegradable or non-biodegradable polymer in a water-immiscible volatile organic solvent (e.g., methylene chloride, chloroform, water-saturated DCM, and the like), optionally containing a surfactant (e.g., a fatty acid or its derivative, such as hydrogenated lecithin). The "water-in-oil" (w / o) emulsion thus obtained is then dispersed in a continuous external aqueous phase, which typically contains a surfactant (e.g., polyvinyl alcohol-PVA, polysorbates, polyethylene oxide-polypropylene oxide block copolymers, cellulose esters, and the like), and optionally further contains a tonicity adjusting agent (e.g., sodium chloride), to form "water-in-oil-in-water (w / o / w) double emulsion" droplets. After evaporation of the organic solvent, the microparticles solidify and are collected by filtration or centrifugation. The terms "oil phase" and "water-immiscible phase" may be used interchangeably herein. The collected microparticles (MP) are washed (e.g., with purified water, a buffer such as a phosphate buffer, an external aqueous solution, or a mixture thereof, or with an aqueous solution containing a divalent cation, e.g., magnesium, calcium, zinc, and the like, or a mixture thereof) to eliminate most of the surfactant and free peptide, and are centrifuged again. The washed MPs are collected and dried (eg, lyophilized) either without additives or with the addition of a cryoprotectant (mannitol) to facilitate their subsequent reconstitution.

[0079] According to further embodiments, the particle size of the "water-in-oil-in-water (w / o / w) double emulsion" droplets can be controlled by various parameters, including, but not limited to, the amount of force applied, the rate of mixing, the type and concentration of surfactant, etc. After solidification, the microparticles are typically characterized by a particle size ranging from about 1 to about 100 μm, including each value within the specified ranges. For example, the microparticles typically have a size ranging from about 3 to about 50 μm, from about 3 to about 40 μm, or from about 3 to about 30 μm, with each possibility representing a separate embodiment of the present invention.

[0080] According to certain aspects and embodiments, the microparticles have a median particle size within the range of about 5 to about 20 μm, inclusive of each value within the specified range. As used herein, "median" or "d 50 The term "median particle diameter" refers to the particle size value at which the cumulative distribution reaches 50%. In other words, the median particle diameter (or median particle size) represents the value below which half of the particle population has a particle size smaller than this value and above which half of the particle population has a particle size larger than this value. Exemplary median particle diameters include values ​​within the specified range, such as d from about 7 to about 17 μm. 50 or particles having a d of about 10 to about 15 μm 50 Exemplary median particle sizes of the microparticles of the present invention include, but are not limited to, about 5 μm, about 6 μm, about 7 μm, about 8 μm, about 9 μm, about 10 μm, about 11 μm, about 12 μm, about 13 μm, about 14 μm, about 15 μm, about 16 μm, about 17 μm, about 18 μm, about 19 μm, and about 20 μm. Each possibility represents a separate embodiment. It is contemplated that the small particle size of the microparticles disclosed herein provides improved physical stability of suspensions containing the microparticles, as well as superior injectability by using smaller gauge needles while avoiding needle clogging.

[0081] How to use The present invention provides methods for treating or delaying the progression or onset of diabetes, particularly type 2 diabetes, including diabetic complications such as retinopathy, neuropathy, nephropathy and delayed wound healing, and related disorders such as insulin resistance (impaired glucose homeostasis), hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, obesity, hyperlipidemia including hypertriglyceridemia, Syndrome X, atherosclerosis, hypertension, and cardiovascular diseases and events such as coronary heart disease, cerebrovascular disease, peripheral arterial disease, rheumatic heart disease, congenital heart disease, deep vein thrombosis and pulmonary embolism, non-fatal myocardial infarction or non-fatal stroke, and high density lipoprotein levels. The method comprises administering to a medically acceptable location in a subject in need thereof a long-acting parenteral pharmaceutical composition comprising dry microparticles, the long-acting parenteral pharmaceutical composition being a long-acting depot composition suitable for administration to a medically acceptable location in a subject in need thereof at a frequency of from once every four weeks to once every six months, the dry microparticles being formed by drying water-in-oil-in-water (w / o / w) double emulsion droplets comprising an internal aqueous phase comprising a therapeutically effective amount of semaglutide or a pharma- ceutical acceptable salt thereof, a water-immiscible polymer phase comprising a biodegradable carrier selected from the group consisting of polylactide, polyglycolide, polycaprolactone, and combinations thereof, and an external aqueous phase.

[0082] The term "treating" as used herein in relation to type 2 diabetes refers to the suppression or alleviation of short-term and long-term symptoms and complications associated with type 2 diabetes, such as hyperglycemia, and any one of the aforementioned complications. In various embodiments, the compositions disclosed herein reduce fasting blood glucose levels by at least about 5%, preferably at least about 10%, more preferably at least about 15%, and most preferably at least about 20%. In other embodiments, the compositions disclosed herein reduce fed blood glucose levels by at least about 5%, preferably at least about 10%, more preferably at least about 15%, and most preferably at least about 20%. In further embodiments, the compositions disclosed herein reduce hemoglobin A1c (HbA1c) levels in the subject by at least about 0.5%, preferably at least about 1%, more preferably at least about 1.5%, and most preferably at least about 2%. The aforementioned reductions in fasting blood glucose levels, fed blood glucose levels, and hemoglobin A1c (HbA1c) levels are contemplated for at least about four weeks to about six months, or any period therebetween, following a single dose.

[0083] Within the scope of the present invention are fasting blood glucose levels after a single parenteral administration of the composition of the present invention of about 70 to about 400 mg / dL, including each value within the specified range. For example, fasting blood glucose levels after a single parenteral administration may be about 70 mg / dL, about 75 mg / dL, about 80 mg / dL, about 85 mg / dL, about 90 mg / dL, about 95 mg / dL, about 100 mg / dL, about 110 mg / dL, about 120 mg / dL, about 130 mg / dL, about 140 mg / dL, about 150 mg / dL, about 160 mg / dL, about 170 mg / dL, about 180 mg / dL, about 190 mg / dL, about 200 mg / dL, about 210 mg / dL, about 220 mg / dL, and the like. Examples of possible ranges include, but are not limited to, about 230 mg / dL, about 240 mg / dL, about 250 mg / dL, about 260 mg / dL, about 270 mg / dL, about 280 mg / dL, about 290 mg / dL, about 300 mg / dL, about 310 mg / dL, about 320 mg / dL, about 330 mg / dL, about 340 mg / dL, about 350 mg / dL, about 360 mg / dL, about 370 mg / dL, about 380 mg / dL, about 390 mg / dL, and about 400 mg / dL. Each possibility represents a separate embodiment.

[0084] Within the scope of the present invention are fed blood glucose levels after a single parenteral administration of the compositions of the present invention of about 120 to about 650 mg / dL, including each value within the specified range. For example, fed blood glucose levels after a single parenteral administration may be about 120 mg / dL, about 130 mg / dL, about 140 mg / dL, about 150 mg / dL, about 160 mg / dL, about 170 mg / dL, about 180 mg / dL, about 190 mg / dL, about 200 mg / dL, about 210 mg / dL, about 220 mg / dL, about 230 mg / dL, about 240 mg / dL, about 250 mg / dL, about 260 mg / dL, about 270 mg / dL, about 280 mg / dL, about 290 mg / dL, about 300 mg / dL, about 310 mg / dL, about 320 mg / dL, about 330 mg / dL, about 340 mg / dL, about 350 mg / dL, about 360 mg / dL, about 370 mg / dL, about 380 mg / dL, about 390 mg / dL, about 400 mg / dL, about 410 mg / dL, about 420 mg / dL, about 430 mg / dL, about 440 mg / dL, about 450 mg / dL, about 460 mg / dL, about 470 mg / dL, about 480 mg / dL, about 490 mg / dL, about 500 mg / dL, about 510 mg / dL, about 520 mg / dL, about 530 mg / dL, about 540 mg / dL, about 550 mg / dL, about 56 40mg / dL, approx. 250mg / dL, approx. 260mg / dL, approx. 270mg / dL, approx. 280mg / dL, approx. 290mg / dL, approx. 300mg / dL, approx. 310m g / dL, approx. 320 mg / dL, approx. 330 mg / dL, approx. 340 mg / dL, approx. 350 mg / dL, approx. 360 mg / dL, approx. 370 mg / dL, approx. 380 mg / d L, approx. 390 mg / dL, approx. 400 mg / dL, approx. 410 mg / dL, approx. 420 mg / dL, approx. 430 mg / dL, approx. 440 mg / dL, approx. 450 mg / dL, approx. 460mg / dL, approx. 470mg / dL, approx. 480mg / dL, approx. 490mg / dL, approx. 500mg / dL, approx. 510mg / dL, approx. 520mg / dL, approx. 530 Examples of possible ranges include, but are not limited to, about 540 mg / dL, about 550 mg / dL, about 560 mg / dL, about 570 mg / dL, about 580 mg / dL, about 590 mg / dL, about 600 mg / dL, about 610 mg / dL, about 620 mg / dL, about 630 mg / dL, about 640 mg / dL, and about 650 mg / dL. Each possibility represents a separate embodiment.

[0085] Within the scope of the present invention are hemoglobin A1c (HbA1c) levels following a single parenteral administration of a composition of the present invention of from about 4% to about 10.5%, inclusive of each value within the range specified. For example, hemoglobin A1c (HbA1c) levels after a single parenteral administration include about 4.0%, about 4.1%, about 4.2%, about 4.3%, about 4.4%, about 4.5%, about 4.6%, about 4.7%, about 4.8%, about 4.9%, about 5.0%, about 5.1%, about 5.2%, about 5.3%, about 5.4%, about 5.5%, about 5.6%, about 5.7%, about 5.8%, about 5.9%, about 6.0%, about 6.1%, about 6.2%, about 6.3%, about 6.4%, about 6.5%, about 6.6%, about 6.7%, about 6.8%, about 6.9%, about 7.0%, about 7.1%, Examples include, but are not limited to, about 7.2%, about 7.3%, about 7.4%, about 7.5%, about 7.6%, about 7.7%, about 7.8%, about 7.9%, about 8.0%, about 8.1%, about 8.2%, about 8.3%, about 8.4%, about 8.5%, about 8.6%, about 8.7%, about 8.8%, about 8.9%, about 9.0%, about 9.1%, about 9.2%, about 9.3%, about 9.4%, about 9.5%, about 9.6%, about 9.7%, about 9.8%, about 9.9%, about 10.0%, about 10.1%, about 10.2%, about 10.3%, about 10.4%, and about 10.5%. Each possibility represents a separate embodiment.

[0086] In addition, the present invention provides a method of treating obesity, the method comprising the step of administering to a subject in need thereof a long-acting parenteral pharmaceutical composition comprising dry microparticles, wherein the long-acting parenteral pharmaceutical composition is a long-acting depot composition suitable for administration to a medically acceptable location in a subject in need thereof at a frequency of from once every four weeks to once every six months, the dry microparticles being formed by drying water-in-oil-in-water (w / o / w) double emulsion droplets comprising an internal aqueous phase comprising a therapeutically effective amount of semaglutide or a pharma- ceutical acceptable salt thereof, a water-immiscible polymer phase comprising a biodegradable carrier selected from the group consisting of polylactide, polyglycolide, polycaprolactone, and combinations thereof, and an external aqueous phase.

[0087] The term "treating" as used herein in relation to obesity refers to preventing weight gain, promoting weight loss, reducing excess weight, or treating obesity, including morbid obesity, and related diseases and conditions, including, but not limited to, obesity-related inflammation, obesity-related gallbladder disease, and obesity-induced sleep apnea (e.g., by controlling appetite, eating, food intake, caloric intake, and / or energy expenditure).

[0088] In addition, the present invention provides a method of treating Parkinson's disease, the method comprising the step of administering to a subject in need thereof a long-acting parenteral pharmaceutical composition comprising dry microparticles, the long-acting parenteral pharmaceutical composition being a long-acting depot composition suitable for administration to a medically acceptable location in a subject in need thereof at a frequency of from once every four weeks to once every six months, the dry microparticles being formed by drying water-in-oil-in-water (w / o / w) double emulsion droplets comprising an internal aqueous phase comprising a therapeutically effective amount of semaglutide or a pharma- ceutically acceptable salt thereof, a water-immiscible polymer phase comprising a biodegradable carrier selected from the group consisting of polylactide, polyglycolide, polycaprolactone, and combinations thereof, and an external aqueous phase.

[0089] As used herein, the term "treating" with respect to Parkinson's disease refers to reversing, alleviating, ameliorating, inhibiting, slowing and / or halting the progression or severity of at least one adverse effect or symptom of Parkinson's disease, including, for example, those associated with impaired motor function.

[0090] It is understood that the amount of semaglutide administered will be determined by a physician according to various parameters, including the selected route of administration, age, weight, and the severity of the patient's disease and symptoms. The required plasma concentration of semaglutide to provide therapeutic efficacy can be determined, for example, from in vitro and in vivo models, as known in the art. According to some specific exemplary embodiments, the steady-state mean plasma concentration of semaglutide is about 0.001 μg / ml to about 100 μg / ml, including each value within the specified range. According to other embodiments, the steady-state mean plasma concentration of semaglutide is about 0.01 μg / ml to about 100 μg / ml, including each value within the specified range. According to further embodiments, the steady-state mean plasma concentration of semaglutide is about 0.05 μg / ml to about 50 μg / ml, including each value within the specified range. According to further embodiments, the steady state mean plasma concentration of semaglutide is from about 0.05 μg / ml to about 10 μg / ml, inclusive of each value within the range specified. According to other embodiments, the steady state mean plasma concentration of semaglutide is from about 0.1 μg / ml to about 1 μg / ml, inclusive of each value within the range specified. In one embodiment, after the minimum burst release, the mean plasma concentration of semaglutide is substantially constant for at least about 4 weeks after a single dose.

[0091] According to further embodiments, the mean plasma concentration of semaglutide is from about 1 ng / ml to about 100 μg / ml, inclusive. According to other embodiments, the mean plasma concentration of semaglutide is from about 1 ng / ml to about 5,000 ng / ml, inclusive. According to yet other embodiments, the mean plasma concentration of semaglutide is from about 1 ng / ml to about 1,000 ng / ml, inclusive. According to additional embodiments, the mean plasma concentration of semaglutide is from about 1 ng / ml to about 500 ng / ml, inclusive. According to certain embodiments, the mean plasma concentration of semaglutide is from about 1 ng / ml to about 300 ng / ml, inclusive. According to some embodiments, the mean plasma concentration of semaglutide is from about 10 ng / ml to about 250 ng / ml, inclusive.

[0092] According to further embodiments, the compositions of the invention provide therapeutic efficacy that is equivalent to or superior to a weekly injectable dosage form of semaglutide, with a reduced incidence and / or reduced severity of side effects. Each possibility represents a separate embodiment of the present invention.

[0093] According to some embodiments, the parenteral pharmaceutical depot composition of the present invention can be administered in vivo to a subject in need thereof. In some embodiments, the "subject" to which the depot composition is administered is a mammal, preferably, but not limited to, a human.

[0094] As used in this specification and the appended claims, the term "about" refers to ±10%.

[0095] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, reference to "a biodegradable carrier" includes a plurality of such carriers. It should be noted that the term "and" or "or" is generally employed in its sense including "and / or" unless the context clearly dictates otherwise.

[0096] The following examples are presented to more fully illustrate certain embodiments of the present invention. However, they should in no way be construed as limiting the broad scope of the present invention. Those skilled in the art can easily devise many variations and modifications of the principles disclosed herein without departing from the scope of the present invention. EXAMPLES

[0097] Example 1: Preparation of PLGA-based injectable semaglutide microparticles Semaglutide microspheres were prepared using a water / oil / water (w1 / o / w2) double emulsion solvent evaporation method according to the following exemplary procedure.

[0098] Inner aqueous phase: 50 mg of semaglutide was hydrated by mixing with water (350 μl) followed by adjusting the pH to 7.8 with NaOH.

[0099] Organic phase: Methylene chloride was saturated with water at room temperature by mixing 50 ml of dichloromethane (DCM) and 5 ml of water in a tightly closed bottle. Following phase separation after 30 min at room temperature, the bottom layer of water-saturated DCM was used for preparation of the organic phase. PLGA (Resomer® RG 502H poly(D,L-lactide-co-glycolide) 50:50 acid terminated, MW 7-17,000, or Resomer® RG 502 poly(D,L-lactide-co-glycolide) 50:50 ester terminated MW 7-17,000) was dissolved in water-saturated DCM.

[0100] External aqueous phase: Sodium chloride and PVA were dissolved in 50 mL of water to yield a total concentration of 0.25% PVA and 1.75% NaCl.

[0101] The inner aqueous phase was then mixed into the PLGA-containing organic phase to form a w / o inner emulsion using an ultrasonic indenter (20 kHz, 20–30 s, ca. 50 watts, ice-water bath) or a Polytron dispersion agglomerator (12 mm, 20,000–30,000 rpm, 1 min). The inner w / o emulsion was then mixed with the outer aqueous phase to form the final w / o / w double emulsion using a high-shear rotor-stator mixer (Polytron dispersion agglomerator (12 mm, 11,000 rpm, 40 s) or a Caframo A231 straight mixer (1,500 rpm, 2 min)) at different temperatures. The obtained double emulsion was then kept in an open container with continuous stirring to allow evaporation of the DCM and solidification of the peptide-containing microparticles (MPs). Once coagulation was complete, the suspension was centrifuged (3,000-5,000 rpm) and the precipitate was washed with phosphate buffer (pH 7.4), centrifuged again, washed with water, collected using a small amount of pure water, dried on a Petri dish, and protected from light to obtain dry MP. MP were collected and stored in tightly closed vials in the refrigerator.

[0102] Different semaglutide depot formulations and their preparation methods are detailed in Tables 1A-1I below. Microscopic images of representative microparticles are shown in Figures 1A-1L.

[0103] [Table 1]

[0104] [Table 2]

[0105] [Table 3]

[0106] [Table 4]

[0107] [Table 5]

[0108] [Table 6]

[0109] [Table 7]

[0110] [Table 8]

[0111] [Table 9]

[0112] Example 2: In vitro release profile The in vitro release profile was measured for the depot formulation prepared according to Example 1. Due to the instability of semaglutide in the release medium, semaglutide release from the microparticles was carried out using separate tubes for each time point at high MP:release medium ratios.

[0113] 15 mg of MP loaded with semaglutide was placed in 3 ml of 0.1 M phosphate buffer (pH 7.4) containing 0.2% BSA and 0.05% sodium azide. The shaker speed was set to approximately 100 strokes / min. The release experiment was carried out at a ratio of 15 mg of MP per 3 ml of buffer. After 3 weeks of release, the buffer was replaced with the same medium additionally containing 200 μg / ml of porcine pancreatic lipase.

[0114] The release rate was determined by assessing the amount of semaglutide remaining in the MP at a given time point. Semaglutide was extracted from the microparticles and separated from the residue by centrifugation. The clear supernatant was transferred into a spectrophotometer cell and analyzed for semaglutide content using a UV calibration curve for the two-point (293-350 nm) method based on a first derivative calibration curve. Alternatively, samples were tested by HPLC.

[0115] Determination of semaglutide burst in the supernatant after 24 hours release was determined as follows: the release vial was placed in a centrifuge at 3,600 rpm for 12 minutes. 1.5 ml of the supernatant was transferred into a 2 ml Eppendorf tube at 10,000 rpm for 8 minutes. 1 ml of the supernatant was placed in a spectrophotometer cell and analyzed for semaglutide content using a UV calibration curve for the two-point (293-350 nm) method based on the first derivative of the spectral calibration curve.

[0116] The release profiles of several formulations are shown in Figures 2-3. Figure 4A shows the release profile of semaglutide from MPS-46 depot formulations and Figure 4B shows the semaglutide content (%) in the MPs during release. The results are summarized in Table 2.

[0117] [Table 10]

[0118] Example 3: Effect of divalent ions and porcine pancreatic lipase on the release of semaglutide from MP The semaglutide-loaded microparticles were then mixed with divalent ions (Mg 2+ or Zn 2+ ) and the release of semaglutide from the microparticles was determined as detailed in Example 2. The results are summarized in Table 3.

[0119] [Table 11]

[0120] The results show that washing with a solution of divalent cations reduces the initial burst.

[0121] The effect of porcine pancreatic lipase (PPL) on the release of semaglutide from microparticles was tested. The results are summarized in Table 4. The interaction of MP with PPL demonstrated that different formulations have different susceptibility to PPL as follows: the combination of Resomer® 502 and 502H was found to be less susceptible to PPL hydrolysis (MPs-27 and 41), while MPs composed of Resomer® 502H alone degraded faster (MPs-34). Also, degradation in the presence of PPL was independent of the enzyme concentration for pure Resomer® 502H MPs, but may be sensitive to the enzyme concentration in the polymer mixture.

[0122] [Table 12]

[0123] Example 4: Comparison of semaglutide formulations in animal models of diabetes The aim of this study was to examine the pharmacokinetic and pharmacodynamic effects of different formulations of semaglutide in genetically diabetic male db / db mice.

[0124] [Table 13]

[0125] Formulation preparation: SEMA-031120, 25.3 mg / kg: 25.3 mg of semaglutide depot was dissolved in 1.0 ml water for injection and used immediately. Stock solution A: 10.0 mg of semaglutide API was dissolved in 25.0 ml of water for injection, and the solution (referred to as stock solution A) was used for one week and stored in a refrigerator at 2-8°C. S-API: 0.06 mg / kg dose: 1.5 ml of stock solution A was transferred to a 10 ml volumetric flask and diluted with water to a concentration of 0.06 mg / ml. A fresh solution was prepared weekly and stored in the refrigerator at 2-8°C. S-API: 0.4 mg / kg dose: Stock solution A was used as is. Solutions were prepared weekly as detailed above and stored in the refrigerator at 2-8°C.

[0126] Research Protocol: Male db / db mice (10–12 weeks old) from the Jackson Laboratory were fed a rodent diet throughout the study period. On day 1, ad libitum-fed blood glucose levels were recorded using a glucometer and body weight was measured using an animal weighing balance. Blood samples were collected for measurement of HbA1c levels. Based on blood glucose, body weight, and HbA1c levels, animals were grouped into different treatment groups as detailed above. · Animals from groups 2–4 were treated daily with the respective treatments for 28 days (dose volume: 1 ml / kg) by subcutaneous route (SC). Animals from group 5 were treated with semaglutide depot formulation by intramuscular injection only on day 1 (dose volume: 1 ml / kg). Ad libitum feeding blood glucose levels were recorded every 3 days throughout the study period, i.e. on days 1, 3, 6, 9, 12, 15, 18, 21, 24, and 27. Samples were collected 3 hours after treatment. On days 14 and 28, blood glucose levels were monitored in animals that were fasted overnight (approximately 14-16 hours). Semaglutide depot (Group 5): Plasma samples for measurement of test compound levels by bioanalysis were collected on days 1, 7, 14, 21, 28, 35, 42, 49 and 56. Semaglutide-API: Samples for measurement of semaglutide-API levels by bioanalysis were collected at 0 hours (pre-treatment) and 3 hours post-treatment on days 1, 7, 14, 21 and 28. On days 0 and 28, blood samples were collected for measurement of HbA1c levels. Body weight and food intake were recorded daily throughout the treatment period. At each time point, approximately 100 μl of blood was collected from the retro-orbital plexus of each animal into labeled tubes containing Li-heparin (10 IU / ml of blood). Tubes were mixed by manual inversion 4-5 times. Blood samples were kept on cold ice at all times and plasma was separated by centrifugation within 30 minutes of sample collection. Plasma was separated by centrifuging blood samples at 5,000 rpm for 5 minutes at 2-8°C. Plasma samples were stored at -80°C until bioanalysis.

[0127] result: Effects of test formulations on ad libitum-fed blood glucose levels in db / db mice: db / db mice had significantly higher blood glucose levels compared to normal mice. Semaglutide API at doses of 0.06 and 0.4 mg / kg / day induced a significant reduction in ad libitum blood glucose levels compared to vehicle controls. Semaglutide depot at 25 mg / kg IM once on day 1 only (dose 2 mg / kg API basis) induced a significant reduction in ad libitum blood glucose levels compared to vehicle controls.

[0128] The results are shown in FIG. 5 and summarized in Table 5.

[0129] [Table 14]

[0130] Effect of test formulations on fasting blood glucose levels in db / db mice: db / db mice had significantly higher fasting glucose levels compared to normal mice. Semaglutide API at doses of 0.06 and 0.4 mg / kg / day induced a significant reduction in fasting glucose levels on days 14 and 28 compared to vehicle control. Semaglutide depot (dose 2 mg / kg, API basis) at 25 mg / kg once IM on day 1 only induced a significant reduction in fasting glucose levels on days 14 and 28 compared to vehicle control. The results are shown in Figure 6 and summarized in Table 6.

[0131] [Table 15]

[0132] Effect of test formulations on HbA1c in db / db mice: db / db mice had significantly higher HbA1c levels compared to normal mice. Semaglutide API at doses of 0.06 and 0.4 mg / kg / day induced a significant reduction in HbA1c levels on day 28 compared to vehicle controls. Semaglutide depot at 25 mg / kg IM once on day 1 only (dose 2 mg / kg, API basis) induced a significant reduction in HbA1c levels on day 28 compared to vehicle controls. The results are shown in Figure 7 and summarized in Table 7.

[0133] [Table 16]

[0134] Effect of test formulation on body weight in db / db mice: db / db mice had significantly higher body weights compared to normal mice. Semaglutide API at doses of 0.06 and 0.4 mg / kg / day induced a significant reduction in body weight compared to vehicle control. Semaglutide depot at 25 mg / kg IM once on day 1 only (dose 2 mg / kg, API basis) induced a significant reduction in body weight compared to vehicle control. The results are shown in Figure 8 and summarized in Table 8.

[0135] [Table 17]

[0136] Effect of test formulation on food intake in db / db mice: db / db mice had significantly higher food intake compared to normal mice. Semaglutide API at doses of 0.06 and 0.4 mg / kg / day induced a significant reduction in daily food intake compared to vehicle control. Semaglutide depot at 25 mg / kg once IM on day 1 only (dose 2 mg / kg, API basis) induced a significant reduction in daily food intake compared to vehicle control. The results are shown in Figures 9A-9D and summarized in Table 9.

[0137] [Table 18]

[0138] Plasma concentrations of semaglutide formulations in db / db mice: Plasma concentrations on day 1 (0 hours) were below the limit of quantification, i.e., <10.2. Semaglutide API at a SC dose of 0.06 mg / kg / day administered daily showed increased plasma levels 3 hours after injection. Semaglutide API at a SC dose of 0.06 mg / kg / day administered daily showed plasma levels of approximately 100-500 ng / ml 24 hours after treatment. Semaglutide API at a SC dose of 0.4 mg / kg / day administered daily showed increased plasma levels 3 hours after injection. Semaglutide API at a SC dose of 0.4 mg / kg / day administered daily showed plasma levels of approximately 500-4,500 ng / ml 24 hours after treatment. This increase in plasma levels was dose-proportional at daily doses of 0.06-0.4 mg / kg.

[0139] Semaglutide depot (dose 2 mg / kg, API basis) at 25 mg / kg IM once on day 1 only showed reduced plasma levels from days 1 to 35. The results are shown in Figures 10A-10D and summarized in Tables 10A-10C.

[0140] [Table 19]

[0141] [Table 20]

[0142] [Table 21] BLQ<10.2

[0143] Thus, the depot composition of the invention maintains therapeutic plasma concentrations of semaglutide for at least 35 days following a single administration.

[0144] Example 5: Pharmacokinetic study of semaglutide formulations in minipigs The aim of this study was to test the pharmacokinetic effects of different formulations of semaglutide in male Göttingen minipigs.

[0145] [Table 22]

[0146] Research Protocol: Blood collection and sample storage Blood samples were collected via the jugular vein from minipigs in group 1 (IM) at 0, 6, 12, 24, 48, 72, 84, 144, 156, 216, 228, 288, 300, 360, 480, 600, 720 hours (day 30), 40, 50, and 60 days after dosing (total of 20 time points / minipigs) and from minipigs in group 2 (SC) at 0, 6, 12, 24, 48, 72 hours (pre-dose), 84, 144 hours (pre-dose), 156, 216 hours (pre-dose), 228, 288 hours (pre-dose), 300, 360 hours (pre-dose), 480 hours (pre-dose), 600 hours (pre-dose), and 720 hours (day 30) after dosing (total of 17 time points / minipigs). At each time point, approximately 2.0 ml of blood was collected and transferred into pre-labeled K2EDTA-coated vacutainers and mixed gently by inverting the tubes to facilitate mixing of the anticoagulant with the blood. Blood samples were kept on gel packs until centrifugation. Collected blood samples were centrifuged at 4,000 rpm for 10 minutes at 4°C. Plasma was separated after centrifugation. All plasma samples were transferred into pre-labeled (animal ID number, time point, study number, and group) tubes and stored at -70±10°C until analysis. Blood glucose levels were determined for all blood collection time points.

[0147] bioanalysis Bioanalysis was performed using a purpose-built liquid chromatography mass spectrometry (LC-MS / MS) method for quantification of semaglutide in plasma samples. The linear range was 1-204 ng / ml. Semaglutide was extracted from minipig plasma samples using solid-phase extraction techniques and quantified using LC-MS / MS with electrospray ionization (ESI) and multiple reaction monitoring (MRM) in positive ionization mode.

[0148] Pharmacokinetic analysis Plasma pharmacokinetic parameters of semaglutide were calculated using standard non-compartmental analysis using the linear trapezoidal method with linear interpolation (Phoenix® software, version 8.3, Pharsight Corporation, Mountain View, California 94040 / USA).

[0149] result: No clinical signs and mortality / morbidity were observed in the treatment groups. Plasma pharmacokinetic parameters of semaglutide in male Göttingen minipigs were assessed for groups treated with a single IM (10 mg / animal) semaglutide depot dose and repeated SC (0.008 mg / kg every 3 days until day 28) semaglutide API doses. Values ​​are expressed as mean ± SD and n=3 minipigs / time point / group. The mean ± SD plasma concentration-time profiles of semaglutide are shown in Figure 11 and summarized in Table 11. The corresponding pharmacokinetic parameters are summarized in Table 12.

[0150] [Table 23]

[0151] [Table 24] *Expressed as median (min-max) ** %AUC exp Values ​​> 25% and R sq Value < 0.8

[0152] In Group 1 (semaglutide depot: 10 mg / animal, single IM injection), the median time to peak plasma concentration (T max ) was 24 hours, and the terminal half-life (T 1 / 2 ) with a peak plasma concentration (C max ) was obtained. Mean exposure, AUC last and AUC inf were 54,400±19,200 and 55,500±19,500ng, respectively. * hr / mL.

[0153] In group 2 (semaglutide API: 0.008 mg / kg, repeated SC injections), the time to reach the mean peak plasma concentration (T max ) over 24 hours (12–48) with a peak plasma concentration (C max ) was obtained. The average exposure (AUC last ) is 17,600±5,440ng * hr / mL.

[0154] This study shows that a single intramuscular administration of a depot semaglutide formulation according to certain embodiments of the invention results in effective plasma concentrations over one month following administration.

[0155] Example 6: Comparison of semaglutide formulations in animal models of diabetes The aim of this study was to examine the pharmacokinetic and pharmacodynamic effects of different formulations of semaglutide in an animal model of diabetes (db / db mice).

[0156] treatment group Animals: Male db / db mice, 10-12 weeks old 1. Naive control, n=5 2. Vehicle administered daily SC (db / db mice) (Dose-(vehicle) 1ml / kg / day for 42 days), n=10 3. Semaglutide API administered daily SC (dose-0.02 mg / kg / day for 42 days), n=10 4. Semaglutide depot administered once IM (dose-2mg / kg, API basis), n=10 5. Semaglutide depot administered once SC (dose-2 mg / kg, API basis), n=10

[0157] Study Protocol Body weight and food consumption were monitored daily. Ad libitum-fed blood glucose levels were measured every 3 days. Fasting blood glucose levels were measured on days 14, 28, and 42. Glycated hemoglobin (HbA1c) levels were measured on days 0, 28, and 42. For PK analysis, samples were collected on days 1 (3 hours after treatment), 7, 14, 21, 28, and 35 and stored at -80°C until analysis.

[0158] result: The effect of semaglutide formulations on free-fed blood glucose levels is shown in Figure 12. db / db mice had significantly higher fed blood glucose levels compared to normal mice. Semaglutide API at a dose of 0.02 mg / kg / day induced a significant reduction in free-fed blood glucose levels compared to vehicle control. A single API-based semaglutide depot dose of 2 mg / kg IM on day 1 only induced a significant reduction in free-fed blood glucose levels at certain time points compared to vehicle control. A single API-based semaglutide depot dose of 2 mg / kg SC on day 1 only induced a significant reduction in free-fed blood glucose levels compared to vehicle control.

[0159] Figure 13 shows the effect of semaglutide formulations on fasting blood glucose levels. db / db mice had significantly higher fasting blood glucose levels compared to normal mice. Semaglutide API at a dose of 0.02 mg / kg / day induced a significant reduction in fasting blood glucose levels on days 14, 28, and 42 compared to vehicle control. Semaglutide API-based depot dose of 2 mg / kg once SC on day 1 only induced a significant reduction in fasting blood glucose levels on days 14 and 28 compared to vehicle control.

[0160] Figure 14 shows the effect of semaglutide formulations on HbA1c. db / db mice had significantly higher HbA1c compared to normal mice. Semaglutide API at a dose of 0.02 mg / kg / day induced a significant reduction in HbA1c levels on days 28 and 42 compared to vehicle control. A single API-based semaglutide depot dose of 2 mg / kg IM on day 1 only induced a significant reduction in HbA1c levels on day 42 compared to vehicle control. A single API-based semaglutide depot dose of 2 mg / kg SC on day 1 only induced a significant reduction in HbA1c levels on days 28 and 42 compared to vehicle control.

[0161] Figures 15 and 16 show the effect of semaglutide formulations on body weight and food intake, respectively. db / db mice had significantly higher body weights compared to normal mice. Semaglutide API at a dose of 0.02 mg / kg / day induced a trend toward reduced body weight compared to vehicle control, while both depot formulations administered IM and SC on day 1 only induced a significant reduction in body weight compared to vehicle control (Figure 15). All semaglutide formulations induced a significant reduction in daily food intake during the first week of treatment compared to vehicle control (Figure 16).

[0162] Figure 17A shows semaglutide plasma levels after daily administration of semaglutide API at 0.02 mg / kg / day and Figure 17B shows semaglutide plasma levels after a single SC administration of a semaglutide depot formulation. Daily SC administration of an immediate release formulation induced fluctuations with increased plasma levels 3 hours after injection, while SC administration of a depot formulation according to certain embodiments of the invention provided a decrease in plasma levels from day 1 to day 35, with fewer animals showing detectable plasma levels even 28 days after injection.

[0163] Although the present invention has been specifically described, those skilled in the art will appreciate that many variations and modifications may be made. Accordingly, the present invention is not to be construed as being limited to the specifically described embodiments, and the scope and spirit of the present invention will be more readily understood by reference to the following claims.

Claims

**Claim 1** A long-acting parenteral pharmaceutical composition comprising dry microparticles, wherein the long-acting parenteral pharmaceutical composition is a long-acting depot composition suitable for administration to a medically acceptable site in a subject in need thereof at a frequency of once every 4 weeks to once every 6 months, and the dry microparticles are formed by drying water-in-oil-in-water (w / o / w) double emulsion droplets containing an internal aqueous phase comprising a therapeutically effective amount of semaglutide or a pharmaceutically acceptable salt thereof, and a water-immiscible polymer phase comprising a biodegradable carrier selected from the group consisting of polylactide, polyglycolide, polycaprolactone, and combinations thereof, and an external aqueous phase, wherein the dry microparticles are characterized by a median particle size in the range of about 5 to about 20 μm, semaglutide is continuously released from the composition, and the composition releases less than 20% of the semaglutide or a pharmaceutically acceptable salt thereof in phosphate buffer at pH 7.4 over 24 hours and more than 80% of the semaglutide or a pharmaceutically acceptable salt thereof over 28 days. A long-acting parenteral pharmaceutical composition. **Claim 2** The long-acting parenteral pharmaceutical composition according to claim 1, wherein the dry microparticles are characterized by a median particle size in the range of about 7 to about 17 μm, preferably, the dry microparticles are characterized by a median particle size in the range of about 10 to about 15 μm. **Claim 3** The long-acting parenteral pharmaceutical composition according to claim 1 or 2, having physical properties such that less than 80% of the semaglutide or a pharmaceutically acceptable salt thereof is released in phosphate buffer at pH 7.4 over 14 days. **Claim 4** Having physical properties such that the average human steady-state plasma concentration (C ｓｓ，ａｖｇ ) of semaglutide is at least about 1 nmol / mL for about 4 weeks to about 6 months after a single administration, preferably, the average human steady-state plasma concentration (C ｓｓ，ａｖｇ ) of semaglutide is about 1 nmol / mL to about 5 nmol / mL for about 4 weeks to about 6 months after a single administration, the long-acting parenteral pharmaceutical composition according to claim 1 or 2. **Claim 5** The long-acting parenteral pharmaceutical composition according to claim 1 or 2, wherein the biodegradable carrier is a polymer selected from the group consisting of poly(D,L-lactide-co-glycolide) (PLGA), poly(D,L-lactide) (PLA), polyglycolide (PGA), polycaprolactone (PCL), and combinations thereof, preferably, the biodegradable carrier is poly(D,L-lactide-co-glycolide) (PLGA), more preferably, the PLGA has a molecular weight of less than 20 kDa, and most preferably, the PLGA has a molecular weight in the range of about 2 to about 18 kDa. **Claim 6** The ratio of semaglutide or a pharmaceutically acceptable salt thereof to the biodegradable carrier is in the range of about 1:2 to about 1:30 (w / w), preferably, the ratio of semaglutide or a pharmaceutically acceptable salt thereof to the biodegradable carrier is in the range of about 1:5 to about 1:20 (w / w), more preferably, the ratio of semaglutide or a pharmaceutically acceptable salt thereof to the biodegradable carrier is in the range of about 1:5 to about 1:15 (w / w), the long-acting parenteral pharmaceutical composition according to claim 1 or 2.

7. The long-acting parenteral pharmaceutical composition according to claim 1 or 2, having a drug loading capacity in the range of about 5% to about 15%.

8. The water-immiscible polymer phase further comprises a first surfactant comprising a fatty acid or a derivative thereof, preferably, the first surfactant is lecithin, hydrogenated lecithin, stearic acid, or a mixture or combination thereof, more preferably, the first surfactant comprises hydrogenated lecithin having a phosphatidylcholine content of more than 90% by weight, the long-acting parenteral pharmaceutical composition according to claim 1 or 2.

9. The long-acting parenteral pharmaceutical composition according to claim 1 or 2, comprising semaglutide or a pharmaceutically acceptable salt thereof as the sole active ingredient.

10. Each of the internal and external aqueous phases independently further comprises a second surfactant, preferably, the second surfactant is selected from the group consisting of polyvinyl alcohol (PVA), polysorbate, polyethylene oxide - polypropylene oxide block copolymer, polyethylene glycol, and cellulose ester, or each of the internal and external aqueous phases independently further comprises a tonicity modifier, preferably, the tonicity modifier is an ionic tonicity modifier comprising sodium chloride, or a non-ionic tonicity modifier comprising a sugar or a sugar alcohol, the long-acting parenteral pharmaceutical composition according to claim 1 or 2.

11. The composition releases the semaglutide active ingredient over a period of about 4 weeks to about 3 months, preferably, the composition releases the semaglutide active ingredient over a period of about 4 weeks to about 2 months, more preferably, the composition releases the semaglutide active ingredient over a period of about 4 weeks to about 6 weeks, the long-acting parenteral pharmaceutical composition according to claim 1 or 2.

12. A water-in-oil-in-water (w / o / w) double emulsion process, comprising: (i) dispersing an aqueous suspension or solution of semaglutide or a pharmaceutically acceptable salt thereof in a solution of the biodegradable carrier in a water-immiscible volatile organic solvent, thereby obtaining a water-in-oil emulsion; and (ii) dispersing the water-in-oil emulsion in a continuous external aqueous phase to form microparticles containing water-in-oil-in-water (w / o / w) double emulsion droplets. A long-acting parenteral pharmaceutical composition according to claim 1 or 2, prepared by the process.

13. The process further comprises (iii) collecting the thus formed microparticles by filtration or centrifugation, preferably, the process further comprises (iv) washing the collected microparticles, more preferably, the washing of the collected microparticles is carried out using an aqueous solution containing divalent cations, or the process further comprises drying the collected or washed microparticles. A long-acting parenteral pharmaceutical composition according to claim 12.

14. (1) Treatment of type 2 diabetes, preferably, the treatment of type 2 diabetes comprises reducing fasting blood glucose levels in a subject by at least about 5% over a period of about 4 weeks to about 6 months after a single administration, or the treatment of type 2 diabetes comprises reducing postprandial blood glucose levels in a subject by at least about 5% over a period of about 4 weeks to about 6 months after a single administration, or the treatment of type 2 diabetes comprises reducing hemoglobin A1c (HbA1c) levels in a subject by at least about 0.5% over a period of about 4 weeks to about 6 months after a single administration; (2) treatment of obesity; or (3) treatment of Parkinson's disease. A long-acting parenteral pharmaceutical composition according to claim 1 or 2, for use in the treatment.

15. A long-acting parenteral pharmaceutical composition comprising dry microparticles for use in a method of achieving an average human steady-state plasma concentration (C ｓｓ，ａｖｇ ) of at least about 1 nmol / mL of semaglutide for about 4 weeks to about 6 months after a single administration to a subject in need thereof, wherein the dry microparticles are formed by drying water-in-oil-in-water (w / o / w) double emulsion droplets containing an internal aqueous phase comprising a therapeutically effective amount of semaglutide or a pharmaceutically acceptable salt thereof, the dry microparticles, a water-immiscible polymer phase comprising a biodegradable carrier selected from the group consisting of polylactide, polyglycolide, polycaprolactone, and combinations thereof, and an external aqueous phase, preferably, the average human steady-state plasma concentration (C ｓｓ，ａｖｇ ) of the semaglutide is about 1 nmol / mL to about 5 nmol / mL for about 4 weeks to about 6 months after a single administration.

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