Sustained-release drug-loaded particles

Abstract

Sustained release drug-loaded microparticles, pharmaceutical compositions comprising them, methods of making them, and methods of treatment using them are described.

Description

Cross-reference to related applications

[0001] This application claims the priority benefit of U.S. Provisional Patent Application No. 63 / 528,015, filed July 20, 2023, the entire contents of which are incorporated herein by reference. Technical Field

[0002] The sustained-release drug-loaded microparticles, pharmaceutical compositions containing them, methods for preparing them, and treatment methods using them are described. Background Technology

[0003] Formulating drugs in injectable compositions that provide sustained-release properties can be challenging. Polymer microparticle-based formulations, such as parenteral formulations based on poly(D,L-lactide-co-glycolic acid) (PLGA) microparticles, have been described for various drugs, but only 19 PLGA microsphere-based drug products are commercially available. See, for example, Li et al., Bridging the gap between fundamental research and product development of long-acting injectable PLGA microspheres, Exp. Op. Drug. Deliv. (2022).

[0004] Triptorelin is a synthetic decapeptide agonist analog of gonadotropin-releasing hormone (GnRH) and reversibly inhibits luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Triptorelin can be used to treat hormone-responsive cancers, such as breast or prostate cancer; manage endometriosis, female infertility, and uterine fibroids; and treat precocious puberty.

[0005] Injectable formulations of triptorelin are approved for the treatment of prostate cancer (TRELSTAR®, Verity Pharmaceuticals) and central precocious puberty (TRIPTODUR®, Arbor Pharmaceuticals, LLC). U.S. Patent 10,166,181 describes an injectable pharmaceutical composition comprising a mixture of triptorelin PLGA microparticles, wherein each type of triptorelin microparticle contains triptorelin dihydroxynaphthyl acetate in a different lactic and glycolic acid copolymer (PLGA), such as a PLGA containing about 85% lactide and 15% glycolide or a PLGA containing about 75% lactide and 25% glycolide, wherein the mixture of microparticles allegedly provides prolonged triptorelin release after injection. U.S. Patent No. 7,252,842 discloses microparticles containing a hydrophilic active agent, including triptorelin-loaded microparticles comprising a PLGA containing about 85% lactide and 15% glycolide. However, the microparticles prepared according to Example 28 of U.S. Patent No. 7,252,842 have low core loading (approximately 2%), poor encapsulation efficiency (approximately 17%), and an average particle size of < 20 µm. Therefore, these microparticles may not be suitable for developing depot formulations for one, three, or six months, as large quantities of microparticles are required to achieve the target dose.

[0006] There is still a need for sustained-release injectable drug compositions, such as those that provide drug release over an extended period of time (e.g., 1 week, 1 month, 6 weeks, 3 months, or 6 months) after injection. Summary of the Invention

[0007] In one aspect, a drug-carrying microparticle is provided, the drug-carrying microparticle comprising: (a) Medicine; (b) A polymer component comprising one or more polymers selected from ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymers with a lactide:co-lactide ratio of about 50:50, ester-terminated PLGA polymers with a lactide:co-lactide ratio of about 75:25, and ester-terminated polylactide (PLA) polymers. (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release the drug in the body for at least one week after being administered via subcutaneous or intramuscular injection.

[0008] In some embodiments, the microparticles release the drug in the body for at least one month after administration via subcutaneous injection. In some embodiments, the microparticles release the drug in the body for at least three months after administration via subcutaneous injection. In some embodiments, the microparticles release the drug in the body for at least six months after administration via subcutaneous or intramuscular injection.

[0009] In one aspect, a drug-carrying microparticle is provided, the drug-carrying microparticle comprising: (a) Medicine; (b) Lactide: One or more ester-terminated PLGA polymers with a lactide ratio of about 50:50; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release the drug in the body for at least one week after being administered via subcutaneous or intramuscular injection.

[0010] In one aspect, a drug-carrying microparticle is provided, the drug-carrying microparticle comprising: (a) Medicine; (b) Lactide: One or more ester-terminated PLGA polymers with a lactide ratio of about 50:50; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release the drug in the body for at least one month after being administered via subcutaneous or intramuscular injection.

[0011] In one aspect, a drug-carrying microparticle is provided, the drug-carrying microparticle comprising: (a) Medicine; (b) Lactide: One or more ester-terminated PLGA polymers with a lactide ratio of about 50:50; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release the drug in the body for at least 6 weeks after administration via subcutaneous or intramuscular injection.

[0012] One or more ester-terminated PLGA polymers may be selected from one or more of RG502, RG503, and RG504 (as a single polymer or a mixture thereof). One or more ester-terminated PLGA polymers may comprise one ester-terminated PLGA polymer. The ester-terminated PLGA polymer may be RG503, or the ester-terminated PLGA polymer may be RG504. One or more ester-terminated PLGA polymers may comprise two different ester-terminated PLGA polymers. The two different ester-terminated PLGA polymers may be present in a weight ratio of lower molecular weight PLGA to higher molecular weight PLGA of about 99:1 to about 51:49, such as ratios of about 99:1, about 90:10, about 80:20, about 70:30, about 60:40, about 55:45, or about 51:49. The two different ester-terminated PLGA polymers may be RG504 and RG503. The two different ester-terminated PLGA polymers can be RG503 and RG502. The weight ratio of triptorelin to one or more ester-terminated PLGA polymers can be from about 4 / 100 to about 10 / 100. The weight ratio of FSH to the ester-terminated PLGA polymer can be from about 5 / 1000 to about 5 / 100 or higher.

[0013] In another aspect, a drug-loaded microparticle is provided, the drug-loaded microparticle comprising: (a) Medicine; (b) Lactide: an ester-terminated PLGA polymer with a lactide ratio of approximately 75:25; (c) Surfactants; and (d) One or both of basic salts and polyols. The microparticles release the drug in the body for at least 3 months after being administered via subcutaneous or intramuscular injection.

[0014] The ester-terminated PLGA polymer can be RG753S. The weight ratio of triptorelin to the ester-terminated PGLA polymer can be from about 5 / 100 to about 10 / 100. The weight ratio of FSH to the ester-terminated PLGA polymer can be from about 5 / 1000 to about 5 / 100 or higher.

[0015] In another aspect, a drug-loaded microparticle is provided, the drug-loaded microparticle comprising: (a) Medicine; (b) Two different ester-terminated PLA polymers; (c) Surfactants; and (d) One or both of basic salts and polyols. The microparticles release the drug in the body for at least 6 months after administration via subcutaneous or intramuscular injection.

[0016] Two different ester-terminated PLA polymers may exist in a weight ratio of lower molecular weight PLA to higher molecular weight PLA of approximately 90:10 to approximately 60:40, such as ratios of approximately 90:10, approximately 85:15, approximately 80:20, approximately 75:25, approximately 70:30, approximately 65:35, or approximately 60:40. The two different ester-terminated polylactide PLA polymers may be R203S and R205S. The weight ratio of triptorelin to PLA polymer may be approximately 5 / 100 to approximately 15 / 100. The weight ratio of FSH to PLA polymer may be approximately 5 / 1000 to approximately 5 / 100 or higher.

[0017] In some embodiments, the drug is selected from one or more small molecule drugs, peptide drugs, and protein drugs. In some embodiments, the drug is selected from one or more of the following: triptorelin, follicle-stimulating hormone (FSH), luteinizing hormone (LH), human chorionic gonadotropin (hCG), atosiban, insulin, abiraterone, barusiban, degarelix, desmopressin, ganirelix (e.g., ganirelix acetate), gemcitabine, gonadorelin (e.g., gonadorelin acetate), menotropin, or human menopausal gonadotropin (hMG), merotocin, progesterone, quinolones, somatostatin, growth hormone, and tolterodine. In some embodiments, the drug is triptorelin, optionally wherein triptorelin is triptorelin acetate. In some embodiments, the drug is FSH, optionally wherein FSH is recombinant FSH, optionally wherein FSH is selected from follicle-stimulating hormone α, follicle-stimulating hormone β, and follicle-stimulating hormone δ. In other specific embodiments, FSH is follicle-stimulating hormone (FSH) delta. In some embodiments, the medicament is two or more medicaments. In some embodiments, the medicament comprises FSH (e.g., follicle-stimulating hormone delta) and LH and / or hCG.

[0018] In any drug-loaded microparticles described herein, the surfactant may comprise poloxamer (e.g., poloxamer 188) or polyvinyl alcohol. In any drug-loaded microparticles described herein, the basic salt may comprise sodium chloride. In any drug-loaded microparticles described herein, the polyol may comprise sucrose or polyethylene glycol.

[0019] The average particle size of the drug-loaded microparticles described herein can be from about 20 µm to about 180 µm, including from about 30 µm to about 150 µm, such as from about 30 µm to about 120 µm, including from about 30 µm to about 80 µm.

[0020] The drug-loaded microparticles described herein may further contain residual amounts of the components used to prepare the microparticles. Therefore, the drug-loaded microparticles may further contain approximately 0.01% w / w to approximately 1.0% w / w of an acid such as acetic acid or citric acid. The drug-loaded microparticles may have approximately 0.1% w / w to approximately 5% w / w of a polyol, such as approximately 0.20% w / w to approximately 5% w / w of sucrose or such as 0.20% w / w to approximately 5% w / w of polyethylene glycol. The drug-loaded microparticles may further contain approximately 0.01% w / w to approximately 1.0% w / w of a buffer salt such as citric acid. The drug-loaded microparticles may further contain approximately 50 ppm to approximately 20,000 ppm of a residual organic solvent, such as approximately 50 ppm to approximately 20,000 ppm of ethyl acetate. The drug-loaded microparticles may have approximately 0% w / w to approximately 4% w / w of a surfactant.

[0021] This document also provides pharmaceutical compositions comprising any drug-loaded microparticles disclosed herein. Pharmaceutical compositions may be formulated for subcutaneous injection. Pharmaceutical compositions may be formulated for intramuscular injection. Pharmaceutical compositions may comprise drug-loaded microparticles and one or more pharmaceutically acceptable carriers, diluents, and / or excipients. Pharmaceutical compositions may comprise drug-loaded microparticles and a pharmaceutically acceptable diluent selected from carboxymethyl cellulose, dextran, and hyaluronic acid and its salts. Pharmaceutical compositions may comprise drug-loaded microparticles and a pharmaceutically acceptable diluent comprising hyaluronic acid and / or its salts, optionally wherein the hyaluronic acid and / or its salts have an average molecular weight of about 1 MDa to about 5 MDa, further optionally wherein the hyaluronic acid and / or its salts have an average molecular weight of about 3.2 MDa, and further optionally wherein the composition comprises about 0.01% (w / v) to about 5% (w / v) of hyaluronic acid and / or its salts based on the volume of the pharmaceutical composition, including about 1% (w / v) to about 3% (w / v), or about 1.5 mg of hyaluronic acid and / or its salts per mL of the pharmaceutical composition.

[0022] The pharmaceutical compositions described herein can be formulated to contain a certain amount of drug-loaded microparticles, thereby providing a therapeutically effective amount of the drug in a single dose. The pharmaceutical compositions can be formulated to provide a therapeutically mean plasma concentration of the drug for at least one week, at least one month, at least six weeks, at least three months, or at least six months following a single dose administered subcutaneously or intramuscularly.

[0023] Pharmaceutical compositions containing triptorelin microparticles, formulated to release triptorelin in the body for approximately one week after subcutaneous or intramuscular injection, can be formulated to contain an amount of triptorelin microparticles, thereby providing approximately 1 mg of triptorelin in a single dose. Pharmaceutical compositions containing triptorelin microparticles, formulated to release triptorelin in the body for approximately one month after subcutaneous or intramuscular injection, can be formulated to contain an amount of triptorelin microparticles, thereby providing 3.75 mg of triptorelin in a single dose. Pharmaceutical compositions containing triptorelin microparticles, formulated to release triptorelin in the body for approximately three months after subcutaneous or intramuscular injection, can be formulated to contain an amount of triptorelin microparticles, thereby providing 11.25 mg of triptorelin in a single dose. Pharmaceutical compositions containing triptorelin-loaded microparticles, formulated to release triptorelin in vivo for approximately 6 months after subcutaneous or intramuscular injection, can be formulated to contain a specific amount of triptorelin-loaded microparticles to provide 22.5 mg of triptorelin in a single dose. Such pharmaceutical compositions can provide a mean plasma concentration of at least 0.1 ng / mL of triptorelin over a period of at least 1 week, at least 1 month, at least 6 weeks, at least 3 months, or at least 6 months following a single dose administered via subcutaneous or intramuscular injection.

[0024] Pharmaceutical compositions containing FSH-loaded microparticles, formulated to release FSH in vivo for approximately one week after subcutaneous or intramuscular injection, can be formulated to contain an amount of FSH-loaded microparticles to provide approximately 85 µg or more of FSH (e.g., follicle-stimulating hormone δ) in a single dose. Pharmaceutical compositions containing FSH-loaded microparticles, formulated to release FSH in vivo for approximately one month after subcutaneous or intramuscular injection, can be formulated to contain an amount of FSH-loaded microparticles to provide approximately 0.35 mg or more of FSH (e.g., follicle-stimulating hormone δ) in a single dose. Pharmaceutical compositions containing FSH-loaded microparticles, formulated to release FSH in vivo for approximately six weeks after subcutaneous or intramuscular injection, can be formulated to contain an amount of FSH-loaded microparticles to provide approximately 0.5 mg or more of FSH (e.g., follicle-stimulating hormone δ) in a single dose. Pharmaceutical compositions containing FSH-loaded microparticles, formulated to release FSH in vivo for approximately 3 months after subcutaneous or intramuscular injection, may be formulated to contain an amount of FSH-loaded microparticles to provide approximately 1 mg or more of FSH (e.g., follicle-stimulating hormone delta) in a single dose. Pharmaceutical compositions containing FSH-loaded microparticles, formulated to release FSH in vivo for approximately 6 months after subcutaneous or intramuscular injection, may be formulated to contain an amount of FSH-loaded microparticles to provide approximately 2 mg or more of FSH (e.g., follicle-stimulating hormone delta) in a single dose. Such pharmaceutical compositions may provide a mean plasma concentration of at least 14 mIU / ml of FSH over a period of at least 1 week, at least 1 month, at least 6 weeks, at least 3 months, or at least 6 months following administration of a single dose via subcutaneous or intramuscular injection.

[0025] This article also provides treatment methods that include administering the pharmaceutical compositions disclosed herein to a subject in need via subcutaneous or intramuscular injection. The method may include injecting a single dose of the drug to provide a therapeutically effective amount. The method may provide a therapeutically mean plasma concentration of the drug for at least one week, at least one month, at least six weeks, at least three months, or at least six months following the administration of the single dose via subcutaneous or intramuscular injection.

[0026] For triptorelin embodiments, a weekly approach may include an injection delivering a single dose of about 1 mg triptorelin, a monthly approach may include an injection delivering a single dose of 3.75 mg triptorelin, a 3-month approach may include an injection delivering a single dose of 11.25 mg triptorelin, and a 6-month approach may include an injection delivering a single dose of 22.5 mg triptorelin. Such approaches may deliver a mean plasma concentration of at least 0.1 ng / mL of triptorelin over a period of at least 1 week, at least 1 month, at least 6 weeks, at least 3 months, or at least 6 months following a single subcutaneous or intramuscular injection. Subjects may require treatment for one or more of hormone-responsive cancers, endometriosis, female infertility, uterine fibroids, and central precocious puberty. Subjects may require treatment for hormone-responsive breast cancer or hormone-responsive prostate cancer.

[0027] For FSH embodiments, a weekly approach may include a single injection of about 85 µg or more of FSH (e.g., follicle-stimulating hormone delta), a monthly approach may include a single injection of about 0.35 mg or more of FSH (e.g., follicle-stimulating hormone delta), a 6-week approach may include a single injection of about 1 mg or more of FSH (e.g., follicle-stimulating hormone delta), a 3-month approach may include a single injection of about 1 mg or more of FSH (e.g., follicle-stimulating hormone delta), and a 6-month approach may include a single injection of about 2 mg or more of FSH (e.g., follicle-stimulating hormone delta). Such approaches may provide a mean plasma concentration of at least 14 mIU / ml FSH for at least 1 week, at least 1 month, at least 6 weeks, at least 3 months, or at least 6 months following administration of a single dose via subcutaneous or intramuscular injection. Subjects may require treatment for female or male infertility.

[0028] Also provided are the triptorelin-loaded microparticles described herein and compositions comprising them for the treatment of one or more of hormone-responsive cancers, endometriosis, female infertility, uterine fibroids, and central precocious puberty. Also provided are the uses of the triptorelin-loaded microparticles described herein and compositions comprising them in the preparation of a medicament for the treatment of one or more of hormone-responsive cancers, endometriosis, female infertility, uterine fibroids, and central precocious puberty.

[0029] Also provided are the FSH-loaded microparticles described herein and compositions comprising them for the treatment of female or male infertility. Also provided are the uses of the FSH-loaded microparticles described herein and compositions comprising them in the preparation of medicaments for the treatment of female or male infertility.

[0030] This document also provides kits that contain (a) a container containing the drug-loaded microparticles disclosed herein and (b) instructions for preparing pharmaceutical compositions comprising microparticles and pharmaceutically acceptable carriers, such as those suitable for intramuscular or subcutaneous injection.

[0031] Methods for preparing drug-loaded microparticles are also provided, including: (a) Adding (i) a drug solution containing the drug in an aqueous solvent to (ii) a polymer solution containing one or more polymers in an ethyl acetate solution, the polymers being selected from ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymers with a lactide:glycol ratio of about 50:50, ester-terminated PLGA polymers with a lactide:glycol ratio of about 75:25, and ester-terminated polylactide (PLA) polymers, and mixing to obtain a dispersion of the drug solution in the polymer solution; and (b) Add (iii) a phase inversion solution comprising a surfactant and optionally one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension of drug-loaded microparticles comprising the drug and one or more polymers selected from ester-terminated PLGA polymers with a lactide:glycol ratio of about 50:50, ester-terminated PLGA polymers with a lactide:glycol ratio of about 75:25, and ester-terminated PLA polymers.

[0032] This article also provides methods for preparing the triptorelin-loaded microparticles disclosed herein, including: (a) Adding (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution of a mixture of one or more ester-terminated PLGA polymers containing lactide: glycolide in an ethyl acetate solution of about 50:50, and mixing to obtain a dispersion of the triptorelin solution in the polymer solution; and (b) Add (iii) a phase inversion solution containing a surfactant and optionally one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension of triptorelin-loaded microparticles containing triptorelin and one or more PLGA polymers.

[0033] This article also provides methods for preparing the triptorelin-loaded microparticles disclosed herein, including: (a) Adding (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution of an ester-terminated PLGA polymer containing lactide:glycol in an ethyl acetate solution of about 75:25, and mixing to obtain a dispersion of the triptorelin solution in the polymer solution; and (b) Add (iii) a phase inversion solution containing a surfactant and one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension of triptorelin-loaded microparticles containing triptorelin and PLGA polymer.

[0034] This article also provides methods for preparing the triptorelin-loaded microparticles disclosed herein, including: (a) Adding (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution containing a mixture of two different ester-terminated PLA polymers in an ethyl acetate solution, and mixing to obtain a dispersion of the triptorelin solution in the polymer solution; and (b) Add (iii) a phase inversion solution containing a surfactant and one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension of triptorelin-loaded microparticles containing triptorelin and PLA polymer.

[0035] Triptorelin can be produced as triptorelin acetate by any method. A triptorelin solution may have a pH of approximately 4.

[0036] This paper also provides methods for preparing the FSH-loaded microparticles disclosed herein, including: (a) Adding (i) an FSH solution containing FSH in an aqueous solvent (such as an aqueous buffer solution) to (ii) a polymer solution of a mixture of one or more ester-terminated PLGA polymers containing lactide:glycol in an ethyl acetate solution at a ratio of about 50:50, and mixing to obtain a dispersion of the FSH solution in the polymer solution; and (b) Add (iii) a phase inversion solution containing a surfactant and optionally one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension containing FSH-loaded microparticles of FSH and one or more PLGA polymers.

[0037] This paper also provides methods for preparing the FSH-loaded microparticles disclosed herein, including: (a) Adding (i) an FSH solution containing FSH in an aqueous solvent (such as an aqueous buffer solution) to (ii) a polymer solution of an ester-terminated PLGA polymer containing lactide:glycol in an ethyl acetate solution at a ratio of about 75:25, and mixing to obtain a dispersion of triptorelin solution in the polymer solution; and (b) Add (iii) a phase inversion solution containing a surfactant and one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension containing FSH-loaded microparticles of FSH and PLGA polymer.

[0038] This paper also provides methods for preparing the FSH-loaded microparticles disclosed herein, including: (a) Adding (i) an FSH solution containing FSH in an aqueous solvent (such as an aqueous buffer solution) to (ii) a polymer solution containing a mixture of two different ester-terminated PLA polymers in an ethyl acetate solution, and mixing to obtain a dispersion of the FSH solution in the polymer solution; and (b) Add (iii) a phase inversion solution containing a surfactant and one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension of FSH-loaded microparticles containing FSH and PLA polymers.

[0039] According to any method, the aqueous solvent can be an aqueous acidic buffer, such as an aqueous acetate buffer or an aqueous citrate buffer. The phase inversion solution may contain a buffering substance, such as a buffer salt. The phase inversion solution may contain poloxamer (e.g., poloxamer 188) and / or polyvinyl alcohol as a surfactant. The phase inversion solution may contain sodium chloride as a basic salt. The phase inversion solution may contain sucrose and / or polyethylene glycol as a polyol.

[0040] According to any method, the method may further include, after step (b), adding a wash phase solution, which optionally contains an aqueous buffer and a surfactant. The wash phase solution may contain a citrate buffer and a surfactant such as poloxamer (e.g., poloxamer 188) or polyvinyl alcohol.

[0041] According to any method, the method may further include one or more of the following steps after step (b): removing the solvent (such as by vacuum), separating the particles from the suspension, drying the particles (optionally by freeze-drying), and sterilizing the particles (optionally by ionizing radiation, such as by gamma radiation). Attached Figure Description

[0042] Figure 1 The preparation of the drug-loaded microparticles described herein is illustrated.

[0043] Figure 2 The mean plasma concentration-time curves after subcutaneous administration of formulation 1 (triptorelin formulation for 1 month) to rats as described in Example 1 are shown.

[0044] Figure 3 The mean plasma concentration-time curves after subcutaneous administration of formulation 2 (a 1-month triptorelin formulation) to rats as described in Example 1 are shown.

[0045] Figure 4 The mean plasma concentration-time curves after subcutaneous administration of formulation 3 (a 1-month triptorelin formulation) to rats as described in Example 1 are shown.

[0046] Figure 5 The mean plasma concentration-time curves after subcutaneous administration of formulation 9 (a 1-month triptorelin formulation) to rats as described in Example 2 are shown.

[0047] Figure 6 The mean plasma concentration-time curves after subcutaneous administration of formulation 12 (a 1-month triptorelin formulation) to rats as described in Example 2 are shown.

[0048] Figure 7 The mean plasma concentration-time curves after subcutaneous administration of formulation 16 (a 1-month triptorelin formulation) to rats as described in Example 2 are shown.

[0049] Figure 8 The mean plasma concentration-time curves after subcutaneous administration of formulation 19 (a 1-month triptorelin formulation) to rats as described in Example 2 are shown.

[0050] Figure 9 The mean plasma concentration-time curves after subcutaneous administration of formulation 28 (a 3-month triptorelin formulation) to rats as described in Example 3 are shown.

[0051] Figure 10 The mean plasma concentration-time curves after subcutaneous administration of formulation 29 (a 3-month triptorelin formulation) to rats as described in Example 3 are shown.

[0052] Figure 11 The mean plasma concentration-time curves after subcutaneous administration of formulation 37 (a 3-month triptorelin formulation) to rats as described in Example 4 are shown.

[0053] Figure 12 The mean plasma concentration-time curves after subcutaneous administration of formulation 43 (6-month triptorelin formulation) to rats as described in Example 5 are shown.

[0054] Figure 13 The mean plasma concentration-time curves after subcutaneous administration of formulation 45 (a 3-month rFSH formulation) to rats as described in Example 6 are shown.

[0055] Figure 14 The mean plasma concentration-time curves after subcutaneous administration of formulation 51 (a 3-month rFSH formulation) to rats as described in Example 6 are shown.

[0056] Figure 15 The mean plasma concentration-time curves of FSH after subcutaneous administration to rats in hyaluronic acid or dextran carriers as described herein for 3 months are shown.

[0057] Figure 16 An example illustrates the IL-4 response after subcutaneous administration of FSH microparticles formulated in hyaluronic acid or dextran carriers to rats for 3 months, as described herein. Detailed Implementation

[0058] The sustained-release drug-loaded microparticles, pharmaceutical compositions containing them, methods for preparing them, and treatment methods using them are described. definition

[0059] Unless otherwise defined, the technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art.

[0060] As used in this article, the singular forms “a” and “the” can refer to both singular and plural unless explicitly stated otherwise.

[0061] As used herein, when defining numbers or ranges, the term “about” means, as will be understood by one of ordinary skill in the art, depending on the context of the number or range used, that the number or range is not limited to the exact number or range stated, but covers values ​​near the listed number or range. Unless otherwise apparent from the context or convention in the art, “about” means plus or minus 10% (± 10%) of a particular term.

[0062] As used herein, the term “administer (administration or administering)” means, for example, being provided, given, administered, and / or prescribed by a healthcare professional or their authorized agent or under their guidance; and, for example, being administered, taken, or consumed by a healthcare professional or subject or patient.

[0063] As used herein, the terms “subject” and “patient” refer to any mammal, including but not limited to humans, pets and laboratory animals (e.g., dogs, cats, rodents, rabbits, guinea pigs, primates, etc.), as well as farm animals and livestock (e.g., horses, camels, donkeys, cattle, sheep, pigs, goats, etc.).

[0064] As used herein, the phrase "therapeuticly effective amount" refers to a dose that is provided or has been determined to provide a specific pharmacological effect (such as pain relief) when administered to a subject requiring such treatment. However, even if a person skilled in the art considers such a dose to be therapeutically effective, a "therapeuticly effective amount" is not always effective in treating a given subject's condition. Exemplary doses and therapeutically effective amounts are provided below with reference to adult subjects. Such amounts can be adjusted by a person skilled in the art according to standard practices required to treat a particular subject and / or condition. Sustained-release drug-loaded microparticles

[0065] This article describes sustained-release drug-loaded microparticles and pharmaceutical compositions comprising them, which can release a therapeutically effective amount of drug over an extended period of time after administration, such as at least one week, at least one month, at least six weeks, at least three months, or at least six months after injection (e.g., subcutaneous or intramuscular). These microparticles are prepared by the method described herein, which involves mixing a solution of a polymer in an organic solvent with a solution of a drug in an aqueous solvent to obtain a dispersion of the drug in the polymer solution, and adding an aqueous solution of a surfactant (an aqueous surfactant solution) to the dispersion of the drug in the polymer solution. The aqueous surfactant solution acts as an extraction medium for the organic solvent and as a continuous phase for the microparticle suspension that immediately forms due to the precipitation of the polymer onto the drug. While not wishing to be bound by theory, it is believed that the addition of the aqueous surfactant solution induces a phase transition from the organic phase to the aqueous phase and immediately forms the microparticle suspension. Therefore, this method does not result in the formation of a double emulsion (W / O / W) because the solvent must be removed from the double emulsion before microparticle formation. Surprisingly, it has been found that by using the polymers, solvents and other excipients described herein, microparticles that release a therapeutically effective amount of drug over extended time periods after administration (such as at least one week, at least one month, at least six weeks, at least three months or at least six months after injection) can be obtained.

[0066] There are no particular limitations on the types of drugs that can be formulated according to this disclosure, and they include small molecule drugs, peptide drugs, and protein drugs. In specific embodiments, the drugs formulated as described herein are selected from one or more peptide drugs or protein drugs. In other specific embodiments, the drugs formulated as described herein are selected from one or more of the following: triptorelin, follicle-stimulating hormone (FSH), luteinizing hormone (LH), human chorionic gonadotropin (hCG), atosiban, insulin, abiraterone, balusiban, degarelix, desmopressin, ganirelix (e.g., ganirelix acetate), gemcitabine, gonadotropin (e.g., gonadotropin acetate), urinary gonadotropin or human menopausal gonadotropin (hMG), meropenem, progesterone, quinolones, somatostatin, growth hormone, and tolterodine. In other specific embodiments, the drug formulated as described herein is triptorelin. In other specific embodiments, the drug formulated as described herein is FSH. In other specific embodiments, the drug formulated as described herein is LH. In other specific embodiments, the drug formulated as described herein is hCG. In other specific embodiments, the drug formulated as described herein is atosiban. In other specific embodiments, the drug formulated as described herein is insulin. In other specific embodiments, the drug formulated as described herein is abiraterone. In other specific embodiments, the drug formulated as described herein is balusiban. In other specific embodiments, the drug formulated as described herein is degarelix. In other specific embodiments, the drug formulated as described herein is desmopressin. In other specific embodiments, the drug formulated as described herein is ganirelix (e.g., ganirelix acetate). In other specific embodiments, the drug formulated as described herein is gemcitabine. In other specific embodiments, the drug formulated as described herein is gonadotropin or human menopausal gonadotropin (hMG). In other specific embodiments, the drug formulated as described herein is meropenem. In other specific embodiments, the drug formulated as described herein is progesterone. In other specific embodiments, the drug formulated as described herein is quigolide. In other specific embodiments, the drug formulated as described herein is somatostatin. In other specific embodiments, the drug formulated as described herein is growth hormone. In other specific embodiments, the drug formulated as described herein is tolterodine.

[0067] In any embodiment where the drug is triptorelin, triptorelin may be triptorelin acetate.

[0068] In any embodiment where the medicament is FSH, the FSH may be human FSH, including isolated human FSH or recombinant human FSH. In any embodiment where the medicament is FSH, the FSH may be recombinant human FSH selected from follicle-stimulating hormone (FSH), follicle-stimulating hormone (FSH) α, follicle-stimulating hormone (FSH) β, and follicle-stimulating hormone (FSH) δ. In a specific embodiment, the FSH is FSH δ. FSH δ is a recombinant FSH of human cell line origin approved as REKOVELLE® (FSH δ) and can be produced by the method disclosed in WO 2009 / 127826.

[0069] In any embodiment, the drug may comprise two or more drugs. In a specific embodiment, the drug comprises FSH (e.g., follicle-stimulating hormone delta) and LH and / or hCG.

[0070] As discussed below, the methods described herein can be used to formulate different drugs. Parameters typically tailored for a given drug typically include any solvents used (including solvents for drug solutions), the concentration of the drug used to prepare the drug solution (if applicable), the polymer used for the polymer component, the solvent used for the polymer component, and the excipients used (e.g., surfactants, pH adjusters (if any), stabilizers (if any), etc.). For example, all solvents used should be compatible with the drug and, for example, should not denature the drug, which may be a problem for protein drugs. Alternatively or alternatively, process parameters can be tailored for a given drug, such as the time and stirring rate for dispersion and phase inversion steps, the composition of the solution used for phase inversion and washing steps, the drying steps (e.g., by lyophilization) and the nature of the excipients used for drying, and the nature of the sterilization steps. For example, the washing solution can be tailored to reduce the drug's solubility, thereby improving encapsulation efficiency, which can be achieved, for example, by adjusting the pH and / or using excipients that reduce the drug's solubility in solution. On the other hand, as discussed below, polymers can be selected to achieve target sustained-release properties, such as 1 week, 1 month, 6 weeks, 3 months, 6 months, or longer. The following discussion highlights other parameters that can be selected and controlled to achieve various purposes.

[0071] The methods disclosed herein are described with reference to drugs that are very different from a physicochemical point of view: triptorelin (1.3 kDa decapeptide) and FSH (35 kDa glycoprotein dimer). Those skilled in the art will understand that the methods described herein can be applied to other drugs, including other protein and peptide drugs, including LH, hCG, atosiban, insulin, degarelix, desmopressin, ganirelin (e.g., ganirelin acetate), gonadorelin (e.g., gonadorelin acetate), hMG, meropenem, progesterone, somatostatin, and growth hormone, as well as non-protein drugs such as abiraterone, balusiban, gemcitabine, quigolide, and tolterodine. Triptorelin

[0072] In some embodiments, the drug is triptorelin or a pharmaceutically acceptable salt thereof, such as a water-soluble salt, such as acetate. Triptorelin is a decapeptide having the following formula: pGlu-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2. Its systemic (IUPAC) name is L-pyroglutamyl-L-histyl-L-tryptophanyl-L-seryl-L-tyrosyl-D-tryptophanyl-L-leucyl-L-arginyl-L-prolyl-glycamide or [D-Trp6]GnRH. Triptorelin has the molecular formula C 64 H 82 N 18 O 13 And a molecular weight of 1311.5 g / mol. It is registered with CAS Registry No. 57773-63-4. As used herein, “triptorelin” refers to triptorelin decapeptide and its pharmaceutically acceptable salts, including its water-soluble salts, such as triptorelin acetate. As used herein, “triptorelin decapeptide” refers to triptorelin decapeptide.

[0073] Triptorelin is an agonist analog of gonadotropin-releasing hormone (GnRH) and reversibly inhibits luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Triptorelin can be used to treat hormone-responsive cancers, such as breast or prostate cancer; manage endometriosis, female infertility, and uterine fibroids; and treat precocious puberty. Injectable formulations of triptorelin are approved for the treatment of prostate cancer (TRELSTAR®, Verity Pharmaceuticals) and central precocious puberty (TRIPTODUR®, Arbor Pharmaceuticals, LLC). Solvents used in triptorelin solutions

[0074] Solvents used for triptorelin solutions include water and can be acidic aqueous solutions such as acetic acid, succinic acid, lactic acid, citric acid, or phosphoric acid solutions, optionally including salts of the corresponding acid anions to increase or provide buffering capacity, such as acetate, succinate, lactate, citrate, or phosphate. Therefore, solvents used for triptorelin solutions can include acids such as acetic acid, succinic acid, lactic acid, citric acid, or phosphoric acid (and optionally buffer salts), which can be present in any amount to provide a triptorelin solution with a desired pH, such as a target pH of about 4, as discussed in more detail below.

[0075] The pH of the triptorelin solution can be from about 3.5 to about 5.0, or from about 4.0 to about 4.8, including about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4.0, about 4.1, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, or about 4.8. For example, the triptorelin solution may contain about 0.1% v / v to about 30% v / v of acid. The triptorelin solution may have a pH from about 3.9 to about 4.9, including pH from 3.9 to 4.9, including pH from 3.9 to 4.8, including pH from about 4, including pH from 4. It has been found that the pH of the triptorelin solution affects the core loading, encapsulation efficiency, and triptorelin release of the triptorelin-loaded microparticles.

[0076] In a specific embodiment, the solvent used for the triptorelin solution is an aqueous acidic solution that provides a triptorelin solution with a pH of 3.9 to 4.9 (including pH 3.9 to 4.8, including pH about 4, including pH 4). For example, the solvent may be an aqueous acetic acid solution containing about 2% to about 20% v / v, including about 9% v / v, including about 5.3% v / v, including about 4.4% v / v, including about 3.5% v / v of acetic acid, or an equivalent amount of a different acid. Alternatively, the solvent may be an aqueous acetate buffer solution with an acetate concentration of about 10 mM to about 200 mM (e.g., to achieve the target pH as needed), or an equivalent amount of another aqueous acidic buffer solution.

[0077] In the examples where triptorelin is triptorelin acetate, the total molar concentration of acetate is the sum of acetate contributed by triptorelin acetate and acetate present in the aqueous acidic acetate buffer. In the examples where triptorelin is triptorelin acetate and the solvent used for the triptorelin solution is an aqueous acidic acetate buffer, the total molar concentration of acetate has been determined to have a significant effect on the in vitro release of triptorelin from the triptorelin-loaded microparticles, with higher molar concentrations of acetate ions associated with slower in vitro triptorelin release. A similar effect is expected for in vivo release. Furthermore, similar effects are expected with other buffers.

[0078] The solvent volume used in the triptorelin solution can affect the encapsulation efficiency of triptorelin-loaded microparticles and the in vitro release of triptorelin. Specifically, reducing the solvent volume has been determined to lead to reduced encapsulation efficiency and slower in vitro triptorelin release. Similar effects are expected for in vivo release. In some experiments, it has also been found that avoiding the use of sodium ions in the triptorelin solution is advantageous and results in, for example, increased encapsulation efficiency, increased core loading, and slower in vitro release of triptorelin-loaded microparticles. Therefore, in some embodiments, the solvent used in the triptorelin solution does not contain sodium ions.

[0079] The concentration of triptorelin (e.g., triptorelin acetate) in the triptorelin solution can range from about 1% to about 60% w / v, including any values ​​in between, such as about 10% to about 40% w / v, about 20% to about 60% w / v, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, and about 55% w / v. The concentration of triptorelin in the triptorelin solution affects the properties of the microparticles and may therefore affect the release of triptorelin from the triptorelin-loaded microparticles. In some embodiments, when preparing microparticles that provide triptorelin release over a period of 1 month, the concentration of triptorelin (e.g., triptorelin acetate) in the triptorelin solution can be from about 10% to about 30% w / v. In some embodiments, when preparing microparticles that provide triptorelin release over a period of 1 month, the concentration of triptorelin (e.g., triptorelin acetate) in the triptorelin solution may be from about 50 mM to about 250 mM. In some embodiments, when preparing microparticles that provide triptorelin release over a period of 3 months, the concentration of triptorelin (e.g., triptorelin acetate) in the triptorelin solution may be from about 15% to about 30% w / v. In some embodiments, when preparing microparticles that provide triptorelin release over a period of 3 months, the concentration of triptorelin (e.g., triptorelin acetate) in the triptorelin solution may be from about 100 mM to about 250 mM. In some embodiments, when preparing microparticles that provide triptorelin release over a period of 6 months, the concentration of triptorelin (e.g., triptorelin acetate) in the triptorelin solution may be from about 15% to about 40% w / v. In some embodiments, when preparing microparticles that provide triptorelin release over a period of 6 months, the concentration of triptorelin (e.g., triptorelin acetate) in the triptorelin solution may be from about 100 mM to about 300 mM.

[0080] Triptorelin solutions may include viscosity enhancers, such as one or more polyols, such as glucose, fructose, lactose, sucrose, mannitol, dextran, or polyethylene glycol (PEG). It has been found that adding a viscosity enhancer such as mannitol to a triptorelin solution affects the in vitro release of triptorelin from loaded triptorelin microparticles. Increasing the viscosity of the triptorelin solution was expected to result in a slower in vitro release of triptorelin; however, the opposite effect was observed. Specifically, the addition of mannitol (e.g., 20% w / v) was found to lead to a burst release of triptorelin, followed by a faster in vitro release. A similar high burst effect is expected in vivo. Without being bound by theory, it is assumed that mannitol is encapsulated within the microparticles, thus providing a highly porous structure that leads to a faster in vitro release of triptorelin. Alternatively or concurrently, the encapsulated mannitol may generate an osmotic pressure gradient that promotes triptorelin release. FSH

[0081] As described above, in some embodiments, the drug formulated as described herein is FSH. FSH is a 35.5 kDa glycoprotein heterodimer composed of two polypeptide units, α and β. In some embodiments, FSH is human FSH. In some embodiments, FSH is human FSH isolated from a human donor. In some embodiments, FSH is recombinant FSH. In some embodiments, recombinant FSH has been generated / expressed in mammalian cell lines. In some embodiments, recombinant FSH has been generated / expressed in Chinese hamster ovary cells. In some embodiments, recombinant FSH has been generated / expressed in human cell lines. Therefore, the FSH formulated as described herein can be human FSH, including isolated human FSH (e.g., human urine-derived FSH) or recombinant human FSH. In any embodiment where the drug is FSH, FSH can be recombinant human FSH selected from follicle-stimulating hormone α, follicle-stimulating hormone β, and follicle-stimulating hormone δ. In a specific embodiment, FSH is follicle-stimulating hormone δ. Follicle-stimulating hormone (FSH) δ is a recombinant FSH of human cell line origin approved as REKOVELLE® (FSH δ) and can be produced by the method disclosed in WO 2009 / 127826.

[0082] Surprisingly, it was found that FSH can be formulated as described herein (e.g., encapsulated in microparticles as described herein) to obtain FSH-loaded microparticles and pharmaceutical compositions containing them, which achieve sustained release of the biologically active form of FSH. This is particularly surprising for FSH, as it is known to be a “brittle” molecule, for example, as a non-covalent dimer. It was also surprising that the FSH formulated as described herein exhibited low antigenicity. This contrasts with unencapsulated FSH, which has been reported to induce antibodies in mice. Solvents used in FSH solutions

[0083] Suitable solvents for FSH include solvents in which FSH is stable and solvents that are physically and / or chemically stable for FSH. Solvents for FSH solutions include water and can be aqueous buffer solutions, including aqueous buffer solutions with a slightly acidic pH, such as buffer solutions containing phosphoric acid, acetic acid, succinic acid, citric acid, or lactic acid, optionally including salts of the corresponding acid anions to increase or provide buffering capacity, such as phosphates, citrates, acetates, succinates, or lactates. Therefore, solvents for FSH solutions can include acids, such as phosphoric acid or citric acid or acetic acid or succinic acid or lactic acid (and optionally buffer salts), which can be present in any amount to provide an FSH solution with a desired pH, such as a target pH of about 6.75, as discussed in more detail below.

[0084] The pH of the FSH solution can be from about 4.5 to about 9, including about 4.5, about 5.0, about 5.5, about 6.0, about 6.5, about 6.75, about 7.0, about 7.5, about 8.0, about 8.5, or about 9.0, or any value between these. FSH dissociates relatively rapidly below pH 4.5. In some embodiments, the pH of the FSH solution is between 6 and 8, for example, pH 6.5 to 7.0. In some embodiments, the FSH solution has a pH of about 6.75, including pH 6.75.

[0085] The FSH solution may optionally contain one or more additional components, such as one or more additional components that can stabilize FSH. In some embodiments, the FSH solution contains an antioxidant (such as methionine). Thus, in some embodiments, the FSH solution includes methionine, such as 0.5 mg / mL methionine. In some embodiments, the mass ratio of FSH to antioxidant (such as methionine) is about 1:1. Additionally or alternatively, in some embodiments, the FSH solution includes sodium sulfate, which can stabilize FSH.

[0086] FSH solutions may include viscosity enhancers, such as one or more polyols, such as glucose, fructose, lactose, sucrose, mannitol, dextran, and polyethylene glycol (PEG). As discussed above with reference to triptorelin microparticles, the addition of viscosity enhancers (such as mannitol) to drug solutions may affect the in vitro drug release from drug-loaded microparticles.

[0087] The FSH solution may contain one or more surfactants, such as polysorbate 20, polysorbate 80, and / or poloxamer (e.g., poloxamer 188). In some embodiments, the FSH solution contains polysorbate 20 and / or polysorbate 80 at a concentration of about 0.0005 mg / mL to about 0.1 mg / mL. In some embodiments, the FSH solution includes poloxamer (e.g., poloxamer 188) at a concentration of about 0.0005 mg / mL to about 0.1 mg / mL. In other embodiments, the FSH solution contains one or more other surfactants in functionally equivalent amounts.

[0088] In some embodiments, the FSH solution comprises water, phosphoric acid (e.g., Na2HPO4·2H2O), methionine, and polysorbate 20. In some embodiments, the FSH solution comprises water, phosphoric acid (e.g., Na2HPO4·2H2O), methionine, and poloxamer 188. Solvents used for other drugs

[0089] Solvents used to prepare drug solutions for other drugs typically include water and can be aqueous buffer solutions, including aqueous buffer solutions with a slightly acidic or acidic pH or a slightly alkaline or alkaline pH, depending on the solubility and stability properties of the drug.

[0090] Drug solutions can have any pH suitable for the drug, such as a pH that allows the drug to be appropriately stable and soluble. As mentioned above, the pH of the drug solution can affect core loading, encapsulation efficiency, and drug release from the particles.

[0091] The volume of solvent used in the drug solution can affect encapsulation efficiency and in vitro drug release. Specifically, reducing the solvent volume can lead to decreased encapsulation efficiency and slower in vitro drug release. A similar effect is expected for in vivo release.

[0092] The drug concentration in the drug solution can be any suitable amount to achieve the target particulate properties. For example, the drug concentration in the drug solution can be from about 0.01% to about 60% w / v, including any value in between, such as about 1% to about 60% w / v, about 10% to about 40% w / v, about 20% to about 60% w / v, about 0.01%, about 0.05%, about 0.1%, about 0.5%, about 1%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, and about 55% w / v. In some embodiments, when preparing particles that provide drug release over a period of 1 week, the drug concentration in the drug solution can be from about 0.01% to about 55% w / v, or from about 0.05 mM to about 500 mM. In some embodiments, when preparing microparticles that provide drug release over a period of 1 month, the drug concentration in the drug solution may be from about 10% to about 30% w / v, or from about 50 mM to about 250 mM. In some embodiments, when preparing microparticles that provide drug release over a period of 3 months, the drug concentration in the drug solution may be from about 15% to about 30% w / v, or from about 100 mM to about 250 mM. In some embodiments, when preparing microparticles that provide drug release over a period of 6 months, the drug concentration in the drug solution may be from about 15% to about 40% w / v, or from about 100 mM to about 300 mM.

[0093] Drug solutions may include viscosity enhancers, such as one or more polyols, such as glucose, fructose, lactose, sucrose, mannitol, dextran, or polyethylene glycol (PEG). As discussed above with reference to triptorelin and / or FSH, the addition of viscosity enhancers (such as mannitol) to drug solutions may affect the in vitro drug release from drug-loaded microparticles. polymer

[0094] Polymers suitable for forming the microparticles described herein include biodegradable polymers, such as biodegradable lactide / glycolic acid polymers, including polylactide (PLA), poly(D,L-lactide-co-glycolic acid) (PLGA) polymers, and mixtures thereof. Such polymers include RESORB® polymers from Evonik Industries, EXPANSORB® polymers from Sequins Pharmaceuticals, PURASORB® polymers from Corbion, and VIATEL polymers from Ashland. TM Polymer. Optionally, the polymer used to prepare the microparticles as described herein is a “purified” polymer, wherein “purified” means a reduced level of residual monomer content (e.g., less than about 0.11% glycolide and less than about 0.04% lactide, including less than 0.11% glycolide and less than 0.04% lactide).

[0095] Exemplary polymers include, but are not limited to, RESOMER® 502H, RESOMER® 502, and RESOMER. ® Select5050 DLG 2E-P, RESOMER® 503H, RESOMER® 503, RESOMER ® Select 5050 DLG 3.8EP, RESOMER ® Select 4852 DLG 3.2EP, RESOMER ® Select 4852 DLG 4.4EP, RESOMER ® Select 5248 DLG 3.2EP, RESOMER ® Select 5248 DLG 4.4EP, RESOMER® 504H, RESOMER®504, RESOMER ® Select 5050 DLG 5E-P, RESOMER® RG752H, RESOMER® RG752S, RESOMER® RG753H, RESOMER® RG753S, RESOMER ® Select 7525 DLG 3.8 EP, RESOMER ® Select 7723DLG 3E-P, RESERVED ® Select 7723 DLG 4.5EP, RESOMER ®Select 7327 DLG 3E-P、RESOMER ® Select 7327 DLG 4.5E-P、RESOMER® RG756S、RESOMER® R202H、RESOMER® R202S、RESOMER® R203H、RESOMER® R203S、RESOMER® R205S、EXPANSORB® DLG 85-7A、EXPANSORB®DLG 85-7E、EXPANSORB® DL 100-5A、EXPANSORB® DL 100-7A、 VIATEL TM DLG 5002A、VIATEL TM DLG 5002E、VIATEL TM DLG 5003A、VIATEL TM DLG 5003E、VIATEL TM DLG 5005A、VIATEL TM DLG 5005 E、 VIATEL TM DLG 7502A、VIATEL TM DLG 7502E、VIATEL TM DLG 7503A、VIATEL TM DLG 7503E、VIATEL TM DLG 7505A、VIATEL TM DLG 7505E、VIATEL TM Ultrapure DLG5002A、VIATEL TM Ultrapure DLG 5002 E、VIATEL TM Ultrapure DLG 5003A、VIATEL TM Ultrapure DLG 5003E、VIATEL TM Ultrapure DLG 5005A、VIATEL TM Ultrapure DLG 5005E、VIATEL TM Ultrapure DLG 7502A、VIATEL TM Ultrapure DLG 7502E、VIATEL TM UltrapureDLG 7503A、VIATEL TMUltrapure DLG 7503E, VIATEL TM Ultrapure DLG 7505A, VIATEL TM Ultrapure DLG 7505E, VIATEL TM DL 02A, VIATEL TM DL 02E, VIATEL TM DL 03A, VIATEL TM DL03E, VIATEL TM DL 05A, VIATEL TM DLG 05 E, VIATEL TM Ultrapure DL 02A, VIATEL TM Ultrapure DL 02E, VIATEL TM Ultrapure DL 03A, VIATEL TM Ultrapure DL 03E, VIATEL TM Ultrapure DL 05A and VIATEL TM Ultrapure DLG 05 E. The properties of exemplary polymers are shown in the table below. Table A The intrinsic viscosity was determined at 25°C with a polymer concentration of 0.5% in CHCl3.

[0096] The polymer can be an ester-terminated PLA or PLGA polymer, a carboxyl-terminated PLA or PLGA polymer, or a polyethylene glycol (PEG)-terminated PLA or PLGA polymer. The polymer can have a lactide / glycol ratio of about 100:0 to about 50:50, including about 100:0, about 90:10, about 85:15, about 75:25, about 65:35, about 55:45, or about 50:50.

[0097] One or more PLGA polymers with a lactide:glycol ratio of about 50:50, such as ester-terminated PLGA polymers, can be used. In some embodiments, a single PLGA polymer, such as a single ester-terminated PLGA polymer, such as RG502, RG503, or RG504, is used. In some embodiments, a mixture of two or more different PLGA polymers is used, including a mixture of two different ester-terminated PLGA polymers. Ester-terminated PLGA polymers (such as RG502) are particularly suitable for preparing microparticles that provide drug release over a period of one week. Ester-terminated PLGA polymers (such as RG503) are particularly suitable for preparing microparticles that provide drug release over a period of one month. In some embodiments, the ester-terminated PLGA polymer used to prepare microparticles that provide drug release over a period of one week has a lower molecular weight (such as RG502 and RG503) than the ester-terminated PLGA polymer used to prepare microparticles that provide drug release over a period of one month. Alternatively, a mixture of two or more ester-terminated PLGA polymers (such as RG503 and RG504, RG502 and RG503, and RG502 and RG504) can be used to prepare microparticles that provide drug release (e.g., triptorelin or FSH release) over a period of one month, as illustrated in the examples below. Similar polymers can be used to prepare microparticles that provide drug release (e.g., triptorelin or FSH release) over a period of six weeks. For any mixture of two ester-terminated PLGA polymers (e.g., RG503 and RG504, RG502 and RG503, or a mixture of RG502 and RG504), the ratio of the lower molecular weight polymer to the higher molecular weight polymer can be from about 10:90 to about 90:10, or any value between therewith, including about 80:20, about 75:25, about 70:30, about 60:40, about 50:50, about 40:60, or about 20:80. In some embodiments, a mixture of ester-terminated PLGA polymer RG503 and ester-terminated PLGA polymer RG504 is used at a ratio of about 10:90 to about 90:10. In some embodiments, a mixture of ester-terminated PLGA polymer RG503 and ester-terminated PLGA polymer RG504 is used at a ratio of about 85:15 to about 30:60 or any value between therewith (including about 80:20, about 60:40, about 50:50, or about 40:60). In some embodiments, a mixture of ester-terminated PLGA polymer RG502 and ester-terminated PLGA polymer RG503 is used at a ratio of about 10:90 to about 90:10.In some embodiments, a mixture of ester-terminated PLGA polymer RG502 and ester-terminated PLGA polymer RG503 is used at a ratio of about 85:15 to about 30:60 or any value between therewith (including about 80:20, about 60:40, about 50:50, or about 40:60). In specific embodiments, a mixture of ester-terminated PLGA polymer RG503 and ester-terminated PLGA polymer RG504 is used at a ratio of about 99:1 to about 51:49 or any value between therewith (including about 99:1, about 90:10, about 80:20, about 70:30, about 60:40, about 55:45, and about 51:49). It has been found that the residual monomer content of the polymer can affect the formation of drug-related impurities. Therefore, in some embodiments, polymers with low residual monomer content are used.

[0098] In some embodiments, such as for microparticles providing triptorelin release over a period of one month, the weight ratio of triptorelin to the ester-terminated PLGA polymer is about 4 / 100 to about 10 / 100 or higher. In some embodiments, such as for microparticles providing FSH release over a period of one week or one month, the weight ratio of FSH to the ester-terminated PLGA polymer is about 5 / 1000 to about 5 / 100 or higher.

[0099] The polymer can be an ester-terminated PLGA polymer with a lactide / glycol ratio of about 75:25. In some embodiments, the polymer is RG753S. In other embodiments, the polymer is RG753H. In other embodiments, the polymer is RG752S. Ester-terminated PLGA polymers with a lactide / glycol ratio of about 75:25 (such as RG753S) can be used to prepare microparticles that provide drug release (e.g., triptorelin or FSH release) over a 3-month period, as illustrated in the following examples. In other embodiments, the polymer used for microparticles that provide drug release (e.g., triptorelin or FSH release) over a 3-month period is a mixture of RG753S and RG752S. In some embodiments, such as for microparticles providing triptorelin release over a 3-month period, the ratio of triptorelin to the ester-terminated PGLA polymer is about 5 / 100 to about 10 / 100 or higher. In some embodiments, such as for microparticles that provide FSH release over a period of 3 months, the ratio of FSH to ester-terminated PGLA polymer is about 5 / 1000 to about 5 / 100 or higher.

[0100] A mixture of two or more PLA or PLGA polymers may be used, including a mixture of two PLA or PLGA polymers. In some embodiments, a mixture of two or more PLA polymers is used, including a mixture of two ester-terminated PLA polymers. The ratio of the lower molecular weight polymer to the higher molecular weight polymer may be from about 10:90 to about 90:10, or any value between thereand, including about 80:20, about 75:25, about 70:30, about 65:35, about 60:40, about 55:45, or about 50:50. In some embodiments, a mixture of ester-terminated PLA polymer R203S and ester-terminated PLA polymer R205S is used at a ratio of about 10:90 to about 90:10. In some embodiments, a mixture of ester-terminated PLA polymer R203S and ester-terminated PLA polymer R205S is used at a ratio of about 85:15 to about 70:30 or any value between therewith (including about 85:15, about 80:20, about 75:25, or about 70:30). A mixture of two or more ester-terminated PLA polymers (such as R203S and R205S) can be used to prepare microparticles that provide drug release (e.g., triptorelin or FSH release) over a period of 6 months, as illustrated in the following examples. In some embodiments, to prepare microparticles that provide drug release over a 6-month period, a mixture of two or more ester-terminated PLA polymers (such as R203S and R205S) may be used at a ratio of about 85:15 to about 70:30 or any value between therewith (including about 85:15, about 80:20, about 75:25, or about 70:30). In some embodiments, such as for microparticles that provide triptorelin release over a 6-month period, the ratio of triptorelin to the ester-terminated PLA polymer is about 5 / 100 to about 15 / 100 or higher. In some embodiments, such as for microparticles that provide FSH release over a 6-month period, the ratio of FSH to the ester-terminated PLA polymer is about 5 / 1000 to about 10 / 100 or higher.

[0101] The in vivo degradation rate of the microparticles described herein is influenced by the ratio of lactide to glycolide in the polymer and the molecular weight of the polymer. Although polymers with higher molecular weights generally exhibit lower degradation rates, it has been surprisingly found that polymers with lower molecular weights can be used to prepare microparticles as described herein that release a therapeutically effective amount of drug over extended time periods after administration, such as at least 1 week, at least 1 month, at least 6 weeks, at least 3 months, or at least 6 months after injection. Solvents for polymers

[0102] The solvent used for the polymer solution is a halogen-free organic solvent or a solvent mixture that is partially miscible with water. The organic solvent or solvent mixture may have a solubility between 1.5% w / w and 40% w / w in water or in an aqueous surfactant phase with or without a buffer at 20°C. A solvent with a water solubility greater than 40% w / w can be mixed with another solvent with a lower water solubility, such that the overall water solubility of the solvent mixture is less than 40% w / w. The solvent used for the polymer solution may be acetone, ethyl formate (EtFo), ethyl acetate (EtAc), methyl acetate (MetAc), or methyl ethyl ketone (MEK). In some embodiments, the solvent used for the polymer solution is ethyl acetate or acetone. In some embodiments, the solvent used for the polymer solution is ethyl acetate. It has been found that the solvent used for the polymer solution can affect particle properties, including particle size, core loading, drug release, etc.

[0103] For embodiments where the drug is a non-covalent polymer, such as a non-covalent dimer like FSH, the solvent used for the polymer solution is advantageously a solvent that does not denature or dissociate the non-covalent polymer. Examples of potentially denaturing solvents that may not be suitable include acetonitrile, dichloromethane (DCM), dimethyl sulfoxide (DMSO), and N-methyl-2-pyrrolidone (NMP). Butanone and ethyl formate may also be harmful to brittle proteins such as FSH. In some embodiments, such as when the drug is a non-covalent polymer (such as a non-covalent dimer, such as FSH), the solvent used for the polymer solution is ethyl acetate or acetone.

[0104] For a one-week embodiment, a typical polymer solution may contain about 10% w / v to about 30% w / v of dissolved polymer, such as about 20% w / v of polymer. For other embodiments (e.g., one-month, three-month, and six-month embodiments), a typical polymer solution may contain about 20% w / v to about 60% w / v of dissolved polymer, such as about 30% w / v to about 50% w / v of polymer, or about 35% w / v to about 45% w / v of polymer.

[0105] In some embodiments, such as for preparing microparticles that provide drug release over a period of one week, the polymer solution contains about 20% w / v to about 50% w / v of dissolved polymer, including about 20% w / v, about 25% w / v, about 30% w / v, about 35% w / v, about 40% w / v, about 45% w / v, or about 50% w / v, and any amount therein. In some embodiments, such as for preparing microparticles that provide drug release over a period of one month, the polymer solution contains about 20% w / v to about 45% w / v of dissolved polymer, including about 20% w / v, about 25% w / v, about 30% w / v, about 35% w / v, about 40% w / v, and any amount therein. In some embodiments, for preparing microparticles that provide drug release over a period of one month, the polymer solution contains about 30% w / v of dissolved polymer. In some embodiments, for preparing microparticles that provide drug release over a 3-month period, the polymer solution contains about 30% w / v to about 50% w / v of dissolved polymer, including about 30% w / v, about 35% w / v, about 40% w / v, or about 45% w / v, or any amount therein. In some embodiments, for preparing microparticles that provide drug release over a 3-month period, the polymer solution contains about 40% w / v of dissolved polymer. In some embodiments, for preparing microparticles that provide drug release over a 6-month period, the polymer solution contains about 30% w / v to about 50% w / v of dissolved polymer, including about 30% w / v, about 35% w / v, about 40% w / v, or about 45% w / v, or any amount therein. In some embodiments, for preparing microparticles that provide drug release over a 6-month period, the polymer solution contains about 40% w / v of dissolved polymer. Aqueous surfactant solution

[0106] Aqueous surfactant solutions for phase inversion may contain cationic, anionic, or nonionic surfactants. The surfactant may be a poloxamer surfactant (e.g., poloxamer 188). Alternatively or alternatively, the surfactant may be a polyvinyl alcohol (e.g., MOWIOL®) surfactant. Alternatively or alternatively, the surfactant may be a polyalkoxylated symmetrical block polymer of POLOXAMNINE® ethylenediamine surfactant, polyethylene glycol alkyl ether, polysorbate (e.g., TWEEN® or SPAN®), sucrose ester (e.g., SISTERNA® or Ryoto glycoester), fatty alcohol polyglycoside, 3-[(3-cholanopropyl)dimethylammonium]-1-propanesulfonate (CHAPS), 3-([3-cholanopropyl]dimethylammonium)-2-hydroxy-1-propanesulfonate (CHAPSO), decyl-PD-glucopyranoside, decyl-PD-maltopyranoside, dodecyl-PD-maltopyranoside, sodium oleate, polyvinyl alcohol, polyoxylated fatty acid ether (e.g., BRIJ®), polyethylene glycol tert-octylphenyl ether (e.g., Triton® X-100), and mixtures of any two or more thereof.

[0107] As described above, the surfactant may be or may contain poloxamer, which is a block copolymer of polyoxyethylene (also known as poly(ethylene oxide) or poly(ethylene glycol) block) and polyoxypropylene (also known as poly(propylene oxide) or poly(propylene glycol) block). Poloxamer typically has a central polyoxypropylene block flanked by polyoxyethylene blocks. According to the conventional nomenclature of poloxamer, the first two numbers multiplied by 100 correspond to the approximate molecular weight of the polyoxypropylene block, while the last number multiplied by 10 corresponds to the relative content of polyoxyethylene. Therefore, poloxamer 188 is a polyoxyethylene-polyoxypropylene-polyoxyethylene triblock polymer with an approximate polyoxypropylene molecular weight of 1800 g / mol and a polyoxyethylene content of 80%. In some embodiments, the surfactant is or may contain poloxamer 188. In some embodiments, the aqueous surfactant solution for phase inversion contains about 1% w / v to about 10% w / v of poloxamer, such as poloxamer 188, or functionally equivalent amounts of different poloxamers, or, for example, a mixture of poloxamer 188 and different poloxamers. In some embodiments, for preparing microparticles that provide drug release over a period of 3 months, the aqueous surfactant solution for phase inversion contains about 3% w / v of poloxamer, such as poloxamer 188, or functionally equivalent amounts of different poloxamers, or, for example, a mixture of poloxamer 188 and different poloxamers. It has been found that the amount of surfactant used can affect microparticle properties, including size, core loading, drug release, etc.

[0108] As described above, the surfactant may be or include polyvinyl alcohol (PVA). Suitable examples of PVA include those with a molecular weight of about 130,000, such as MOWIOL® 18-88. In some embodiments, the surfactant solution for phase inversion contains about 0.5% w / v to about 5% w / v of PVA, such as MOWIOL® 18-88. In some embodiments, for the purpose of preparing microparticles that provide triptorelin release over a period of one month, the surfactant solution for phase inversion contains about 1% w / v or about 2% w / v of PVA, such as MOWIOL® 18-88.

[0109] Aqueous surfactant solutions for phase inversion may contain viscosity modifiers, such as one or more polyols, such as glucose, fructose, lactose, sucrose, mannitol, dextran, or polyethylene glycol. Aqueous surfactants may contain polyethylene glycols, such as those with a molecular weight of about 6000 g / mol or less, including about 6000 g / mol, about 5000 g / mol, about 4000 g / mol, about 3000 g / mol, about 2000 g / mol, and about 1000 g / mol. Therefore, aqueous surfactant solutions may contain viscosity modifiers of about 0% w / v to about 50% w / v, including about 1% w / v to about 50% w / v, such as about 10% w / v to about 40% w / v, or about 20% w / v to about 30% w / v. In some embodiments, the surfactant solution for phase inversion contains about 10% w / v, about 15% w / v, about 20% w / v, about 25% w / v, about 30% w / v, about 35% w / v, about 40% w / v, about 45% w / v, or about 50% w / v of sucrose, or any amount thereof, or an equivalent amount of another viscosity modifier. In some embodiments, the surfactant solution for phase inversion contains about 25% w / v of sucrose, including 25% w / v of sucrose, or an equivalent amount of another viscosity modifier. In some embodiments, the surfactant solution for phase inversion contains about 5% w / v, about 10% w / v, about 15% w / v, about 20% w / v, about 25% w / v, or about 30% w / v of PEG (such as PEG3000, PEG3350, PEG4000, or PEG6000), or any amount thereof, or an equivalent amount of another viscosity modifier. In some embodiments, the surfactant solution for phase inversion contains about 25% w / v PEG (such as PEG3000, PEG3350, PEG4000, or PEG6000), including 25% w / v PEG (such as PEG3000, PEG3350, PEG4000, or PEG6000) or any amount thereof, or an equivalent amount of another viscosity modifier. In some embodiments, the aqueous surfactant solution does not contain a viscosity modifier.

[0110] Additionally or alternatively, the aqueous surfactant solution may contain a salt, such as an alkaline salt (e.g., an alkali metal salt such as sodium chloride). Thus, the aqueous surfactant solution may contain about 0% w / v to about 20% w / v, including about 1% w / v to about 20% w / v, or about 5% w / v to about 15% w / v of an alkaline salt (e.g., sodium chloride). In some embodiments, the surfactant solution for phase inversion contains about 2% w / v, about 5% w / v, about 10% w / v, or about 15% w / v of sodium chloride, or any amount thereof, or an equivalent amount of another salt. In some embodiments, the surfactant solution for phase inversion contains about 10% w / v of sodium chloride, including 10% w / v of sodium chloride, or an equivalent amount of another salt. In some embodiments, the surfactant solution for phase inversion contains about 5% w / v of sodium chloride, including 5% w / v of sodium chloride, or an equivalent amount of another salt. In some embodiments, the surfactant solution for phase inversion contains about 15% w / v of sodium chloride, including 15% w / v of sodium chloride, or an equivalent amount of another salt. In some embodiments, the surfactant solution for phase inversion contains about 2% w / v sodium chloride, including 2% w / v sodium chloride, or an equivalent amount of another salt. It has been found that adding an appropriate amount (including, for example, about 5% w / v) of sodium chloride to the aqueous surfactant solution for phase inversion increases the average particle size without affecting particle morphology. It has also been found that adding an appropriate amount (including, for example, about 5% w / v) of sodium chloride results in a slowing of the in vitro triptorelin release from the triptorelin particles. In some embodiments, the aqueous surfactant solution does not contain salt. In some embodiments, the aqueous surfactant solution contains about 2% w / v sodium chloride and optionally 25% w / v sucrose.

[0111] Aqueous surfactant solutions may contain a co-solvent miscible with water and a solvent for the polymer solution, but not the polymer solvent. When using a co-solvent, the aqueous surfactant can extract more polymer solvent from the polymer solution compared to an equal volume of aqueous surfactant solution. This reduces the volume fraction of aqueous surfactant required to form a particulate suspension, thereby reducing the amount of surfactant to be removed from the particulate suspension. The amount of co-solvent used can be selected based on the polymer and the polymer solvent. Typically, about 1 w / w% to 40 w / w% of co-solvent can be added. Suitable co-solvents include, but are not limited to, alcohols, such as ethanol.

[0112] Alternatively or alternatively, the aqueous surfactant solution may contain an aqueous buffer solution, such as a citrate buffer, with a specific pH (e.g., pH 6.5). The buffer concentration can alter the solubility of the polymer solvent in the surfactant solution used for phase inversion. Therefore, it may affect the drug release performance of the drug-loaded particles. Furthermore, the buffer concentration may affect the solubility of unencapsulated drug molecules (e.g., triptorelin or FSH) that may be present in the surfactant solution after phase inversion. Alternatively or alternatively, the drug solubility can be a function of pH, which can be used to change the drug's solubility in the surfactant solution by altering the pH. As a specific example, in the case of encapsulated FSH, the pH of the surfactant solution used for phase inversion can be adjusted to pH 4.5 to reduce the solubility of FSH in the surfactant solution.

[0113] It has been determined that the surfactant phase composition can affect the properties of microparticles in influencing drug release. For example, in the production of triptorelin microparticles, it has been found that adding a viscosity modifier such as sucrose at 50% w / v to the aqueous surfactant solution used for phase inversion reduces the average particle size of triptorelin-loaded microparticles. When no viscosity modifier is added, a lower viscosity is obtained, and spongy microparticles and microcapsules are formed. In the production of FSH microparticles, it has been found that adding a viscosity modifier such as polyethylene glycol at 25% w / v to the aqueous surfactant solution used for phase inversion reduces the average particle size of FSH-loaded microparticles. When using lower concentrations of polyethylene glycol, a lower viscosity is obtained, and spongy microparticles and / or microcapsules are formed. Furthermore, the solubility of FSH in aqueous solutions has been investigated, and it has been found that polyethylene glycol significantly reduces the solubility of FSH, which can reduce or eliminate leakage of FSH from the microparticles during solvent removal by solvent evaporation. Other components

[0114] One or more of the drug solutions, polymer solutions, and aqueous surfactant solutions may contain one or more additional components suitable for the pharmaceutical composition, such as fillers, buffers, chelating agents, preservatives, antioxidants, cosolvents, and cryoprotectants. Such additional components may be used in amounts that will not unduly negatively affect the target properties of the particles, such as drug loading and release properties. Manufacturing method

[0115] The drug-loaded microparticles described herein can be prepared by any suitable method. The drug-loaded microparticles described herein can be prepared by combining the drug solution described herein with the polymer solution described herein, and then adding the aqueous surfactant solution described herein to initiate a phase inversion. For example... Figure 1As shown, drug solutions can be prepared, for example, by dissolving the drug in an aqueous buffer solution as described herein. Polymer solutions can be prepared, for example, by dissolving the polymer in a halogen-free organic solvent or a mixture of solvents partially miscible with water as described herein. Figure 1 As shown, a drug solution is added to a polymer solution in a container and mixed to obtain a dispersion of the drug solution in the polymer solution (e.g., an emulsion). Then, a phase inversion solution containing a surfactant is added to the container and mixed to induce a phase inversion, resulting in the formation of a suspension of polymer particles containing the drug in an aqueous surfactant solution. Although Figure 1 This refers to triptorelin solution, but other drug-loaded microparticles (including FSH-loaded microparticles) can be prepared using the same method.

[0116] This article provides a method for preparing drug-loaded microparticles, the method comprising: (a) Adding (i) a drug solution containing the drug in an aqueous solvent to (ii) a polymer solution containing one or more polymers in an ethyl acetate solution, the polymers being selected from ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymers with a lactide:glycol ratio of about 50:50, ester-terminated PLGA polymers with a lactide:glycol ratio of about 75:25, and ester-terminated polylactide (PLA) polymers, and mixing to obtain a dispersion of the drug solution in the polymer solution; and (b) Add (iii) a phase inversion solution containing a surfactant and optionally a basic salt and two of a polyol to the dispersion and mix to obtain a suspension of drug-loaded microparticles containing the drug and one or more polymers, for example, the one or more polymers being selected from ester-terminated PLGA polymers with a lactide: glycolide ratio of about 50:50, ester-terminated PLGA polymers with a lactide: glycolide ratio of about 75:25, and ester-terminated PLA polymers.

[0117] In one aspect, a method is provided for preparing triptorelin-loaded microparticles that provide triptorelin release over a period of one week. For example, the method for preparing triptorelin-loaded microparticles may include: (a) Adding (i) a triptorelin solution containing triptorelin in an aqueous buffer solution to (ii) a polymer solution containing a mixture of one or more ester-terminated PLGA polymers in an ethyl acetate solution, and mixing to obtain a dispersion of the triptorelin solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing one or both of a surfactant and optional viscosity modifier (such as polyethylene glycol) and buffer salt (such as sodium citrate) to the dispersion and mix to obtain a suspension of triptorelin-loaded microparticles containing triptorelin and one or more PLGA polymers.

[0118] On the other hand, a method is provided for preparing triptorelin-loaded microparticles that provide triptorelin release over a period of one week. For example, the method for preparing triptorelin-loaded microparticles may include: (a) Adding (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution containing ester-terminated PLGA polymer in an ethyl acetate solution, and mixing to obtain a dispersion of the triptorelin solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing a surfactant to the dispersion and mix to obtain a suspension of triptorelin-loaded microparticles containing triptorelin and PLGA polymer.

[0119] On the other hand, a method is provided for preparing triptorelin-loaded microparticles that provide triptorelin release over a period of one month. For example, the method for preparing triptorelin-loaded microparticles may include: (a) Adding (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution containing a mixture of one or more ester-terminated PLGA polymers in an ethyl acetate solution, and mixing to obtain a dispersion of the triptorelin solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing one or both of a surfactant and optionally a viscosity modifier (such as sucrose) and an alkaline salt (such as sodium chloride) to the dispersion and mix to obtain a suspension of triptorelin-loaded microparticles containing triptorelin and one or more PLGA polymers.

[0120] On the other hand, a method is provided for preparing triptorelin-loaded microparticles that provide triptorelin release over a period of one month. For example, the method for preparing triptorelin-loaded microparticles may include: (a) Adding (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution containing a mixture of two different ester-terminated PLGA polymers in an ethyl acetate solution, and mixing to obtain a dispersion of the triptorelin solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing a surfactant to the dispersion and mix to obtain a suspension of triptorelin-loaded microparticles containing triptorelin and PLGA polymer.

[0121] On the other hand, a method is provided for preparing triptorelin-loaded microparticles that provide triptorelin release over a period of one month. For example, the method for preparing triptorelin-loaded microparticles may include: (a) Adding (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution containing ester-terminated PLGA polymer in an ethyl acetate solution, and mixing to obtain a dispersion of the triptorelin solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing one or both of a surfactant and a viscosity modifier (such as sucrose and polyethylene glycol) and an alkaline salt (such as sodium chloride) to the dispersion and mix to obtain a suspension of triptorelin-loaded microparticles containing triptorelin and PLGA polymer.

[0122] On the other hand, a method is provided for preparing triptorelin-loaded microparticles that provide triptorelin release over a period of 3 months. For example, the method for preparing triptorelin-loaded microparticles may include: (a) Add (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution of an ester-terminated PLGA polymer containing lactide:glycol in an ethyl acetate solution at a ratio of about 75:25, and mix to obtain a dispersion of the triptorelin solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing one or both of a surfactant and a viscosity modifier (such as sucrose) and an alkaline salt (such as sodium chloride) to the dispersion and mix to obtain a suspension of triptorelin-loaded microparticles containing triptorelin and PLGA polymer.

[0123] On the other hand, a method is provided for preparing triptorelin-loaded microparticles that provide triptorelin release over a period of 6 months. For example, the method for preparing triptorelin-loaded microparticles may include: (a) Adding (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution containing a mixture of two different ester-terminated PLA polymers in an ethyl acetate solution, and mixing to obtain a dispersion of the triptorelin solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing one or both of a surfactant and a viscosity modifier (such as sucrose) and an alkaline salt (such as sodium chloride) to the dispersion and mix to obtain a suspension of triptorelin-loaded microparticles containing triptorelin and PLA polymer.

[0124] On the other hand, a method is provided for preparing FSH-loaded microparticles that provide FSH release over a period of one week. For example, the method for preparing FSH-loaded microparticles may include: (a) Adding (i) an FSH solution containing FSH in an aqueous buffer solution to (ii) a polymer solution containing a mixture of one or more ester-terminated PLGA polymers in an ethyl acetate solution, and mixing to obtain a dispersion of the FSH solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing one or both of a surfactant and optional viscosity modifier (such as polyethylene glycol) and buffer salt (such as sodium citrate) to the dispersion and mix to obtain a suspension containing FSH-loaded microparticles of FSH and one or more PLGA polymers.

[0125] On the other hand, a method is provided for preparing FSH-loaded microparticles that provide FSH release over a period of one week. For example, the method for preparing FSH-loaded microparticles may include: (a) Adding (i) the FSH solution containing FSH in an aqueous acidic solvent to (ii) the polymer solution containing the ester-terminated PLGA polymer in an ethyl acetate solution, and mixing to obtain a dispersion of the FSH solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing a surfactant to the dispersion and mix to obtain a suspension containing FSH-loaded microparticles of FSH and PLGA polymer.

[0126] On the other hand, a method is provided for preparing FSH-loaded microparticles that provide FSH release over a period of one month. For example, the method for preparing FSH-loaded microparticles may include: (a) Adding (i) an FSH solution containing FSH in an aqueous buffer solution to (ii) a polymer solution containing a mixture of one or more ester-terminated PLGA polymers in an ethyl acetate solution, and mixing to obtain a dispersion of the FSH solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing one or both of a surfactant and optional viscosity modifier (such as polyethylene glycol) and buffer salt (such as sodium citrate) to the dispersion and mix to obtain a suspension containing FSH-loaded microparticles of FSH and one or more PLGA polymers.

[0127] On the other hand, a method is provided for preparing FSH-loaded microparticles that provide FSH release over a period of one month. For example, the method for preparing FSH-loaded microparticles may include: (a) Adding (i) the FSH solution containing FSH in an aqueous buffer solution to (ii) a polymer solution containing a mixture of two different ester-terminated PLGA polymers in an ethyl acetate solution, and mixing to obtain a dispersion of the FSH solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing a surfactant to the dispersion and mix to obtain a suspension containing FSH-loaded microparticles of FSH and PLGA polymer.

[0128] On the other hand, a method is provided for preparing FSH-loaded microparticles that provide FSH release over a period of one month. For example, the method for preparing FSH-loaded microparticles may include: (a) Adding (i) the FSH solution containing FSH in an aqueous acidic solvent to (ii) the polymer solution containing the ester-terminated PLGA polymer in an ethyl acetate solution, and mixing to obtain a dispersion of the FSH solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing a surfactant to the dispersion and mix to obtain a suspension containing FSH-loaded microparticles of FSH and PLGA polymer.

[0129] On the other hand, a method is provided for preparing FSH-loaded microparticles that provide FSH release over a period of 3 months. For example, the method for preparing FSH-loaded microparticles may include: (a) Add (i) the FSH solution containing FSH in an aqueous buffer solution to (ii) a polymer solution of an ester-terminated PLGA polymer containing lactide:glycol in an ethyl acetate solution at a ratio of approximately 75:25, and mix to obtain a dispersion of the FSH solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing one or both of a surfactant and a viscosity modifier (such as polyethylene glycol) and a buffer salt (such as sodium citrate) to the dispersion and mix to obtain a suspension containing FSH-loaded microparticles of FSH and PLGA polymers.

[0130] On the other hand, a method is provided for preparing FSH-loaded microparticles that provide FSH release over a period of 6 months. For example, the method for preparing FSH-loaded microparticles may include: (a) Adding (i) the FSH solution containing FSH in an aqueous buffer solution to (ii) a polymer solution containing a mixture of two different ester-terminated PLA polymers in an ethyl acetate solution, and mixing to obtain a dispersion of the FSH solution in the polymer solution (dispersion step); and (b) Add (iii) a phase inversion solution containing one or both of a surfactant and a viscosity modifier (such as polyethylene glycol) and a buffer salt (such as sodium citrate) to the dispersion and mix to obtain a suspension of FSH-loaded microparticles containing FSH and PLA polymers.

[0131] For all the methods presented herein, the respective volume fractions of the polymer solution and the drug solution are selected such that a particulate suspension is formed immediately upon combination with an aqueous surfactant solution.

[0132] Several process parameters were found to affect the properties of the particles (such as particle size and release properties), including the temperature of dispersion step (a), which is typically carried out at ambient temperature, such as about 20°C–25°C, the mixing time and mixing rate of dispersion step (a), and the phase inversion solution transfer rate in step (b). The specific effects of these parameters may vary from drug to drug, but the principles discussed in this paper are considered applicable to different drugs.

[0133] As described above, there are two mixing times: the mixing time for step (a), which provides a dispersion of the drug solution in the polymer solution; and the mixing time for step (b), which obtains a suspension of drug-loaded microparticles containing the drug and at least one polymer. The mixing time for step (a) can be selected and adjusted as needed to disperse the drug in the polymer solution and to partially evaporate the solvent from the polymer solution. The mixing time for step (b) can be selected and adjusted as needed to obtain a suspension of drug-loaded microparticles and is generally shorter than the mixing time for step (a).

[0134] Step (a) (e.g., preparing a dispersion of the drug solution in a polymer solution) can be performed under a nitrogen flow (or under any other inert gas flow). Increasing the nitrogen flow rate has been determined to result in the formation of larger particles with smaller open structures, higher core loading, and slower in vitro release rates. Similar effects are expected on in vivo release.

[0135] The mixing speed in steps (a) and (b) can be selected independently, but can conveniently be the same speed, and can be any suitable speed, including about 500 rpm to about 8000 rpm, about 800 rpm to about 2400 rpm, or about 4000 rpm to about 8000 rpm, and can vary with production scale.

[0136] Generally, increasing the stirring speed in at least one of steps (a) and (b) can lead to the formation of smaller particles, which in turn can lead to accelerated drug release in vitro and in vivo. It has also been determined that increasing the stirring speed can lead to increased core loading and improved encapsulation efficiency. Adjusting the stirring speed and mixing time has also been determined to lead to increased average particle size and thus slower drug release in vitro and in vivo.

[0137] The phase inversion solution transfer rate can be any suitable rate to achieve the desired phase transfer and particle formation, and can vary with production scale. Increasing the phase inversion solution transfer rate has been determined to result in the formation of larger particles with higher core loading and slower in vitro release rates. Similar effects are expected on in vivo release rates.

[0138] To remove excess excipients after particle formation, a washing phase may be added to the particle suspension after step (b) (e.g., after obtaining a suspension of drug-loaded particles). The washing phase is typically an aqueous solution and may contain an aqueous buffer (such as an aqueous acidic buffer (e.g., a citrate buffer at pH 4.5 to pH 6.5) or an aqueous buffer with a pH greater than 7.0 (e.g., a TRIS buffer at pH 7.4)), a surfactant (such as a polyethylene glycol-polypropylene glycol surfactant (e.g., poloxamer, such as POLOXAMER® 188) or polyvinyl alcohol), and a salt (e.g., sodium chloride). In some embodiments, the washing phase includes a viscosity modifier, such as PEG as described above. Therefore, in some embodiments, an aqueous washing phase is added after step (b). Without being bound by theory, the washing phase is believed to act as a colloidal stabilizer for the particles and also as an additional extraction medium for any residual organic polymer solvents still present in the particles.

[0139] The organic solvent component of the polymer solvent can be removed, for example, by vacuum and / or increased temperature. Typically, the evaporation of the organic solvent is increased by decreasing the vacuum pressure and increasing the temperature. In some cases, the formation of peptide-related impurities is temperature-dependent. Solvent evaporation under vacuum conditions increases the transition of the organic solvent (e.g., ethyl acetate) from the particles to the continuous phase of the suspension, resulting in complete hardening of the particles. In some embodiments, the particle suspension is agitated by vibration mixing during solvent evaporation under vacuum conditions. In some embodiments, after step (b), the organic solvent (e.g., ethyl acetate) is removed by vacuum.

[0140] Particulates can be removed from the suspension by any suitable means, such as centrifugation and / or tangential / cross-flow filtration. For example, in some embodiments, after step (b), the particulates are separated from the suspension, such as by centrifugation or filtration. In some embodiments, the particulates are separated from the suspension after the solvent is removed by vacuum. In some embodiments, the particulates are concentrated in a processing chamber using tangential / cross-flow filtration, followed by washing the particulates with water to remove excess surfactant from the suspension.

[0141] The particles can be further processed, such as freeze-drying, spray drying, or drying in a fluidized bed. In some embodiments, the particles are typically freeze-dried together with protective excipients, such as one or more of ascorbic acid, amino acids, mannitol, and tocopheryl acetate.

[0142] Particulate matter can be sterilized by any suitable means, such as by ionizing radiation, such as gamma radiation. In some embodiments, particulate matter can be sterilized by gamma radiation doses of about 15 kGy to 45 kGy, including 20 kGy ± 10% (e.g., 18 kGy - 22 kGy), 25 kGy ± 10% (e.g., 22.5 kGy - 27.5 kGy), 27.75 kGy ± 10% (e.g., 25 kGy - 30.5 kGy), 35 kGy ± 10% (e.g., 31.5 kGy - 38.5 kGy), 40 kGy ± 10% (e.g., 36 kGy - 44 kGy), and 45 kGy ± 10% (e.g., 41.5 kGy - 49.5 kGy). In some embodiments, particulate matter can be sterilized by an electron beam (β radiation). Surprisingly, it was found that drug-loaded microparticles, as described herein, that have undergone gamma irradiation are stable, for example, against drug degradation. This is particularly surprising for FSH-loaded microparticles, as FSH is generally unstable and readily degraded. Therefore, the gamma-irradiated FSH-loaded microparticles described herein advantageously exhibit higher FSH stability and represent advantageous embodiments. However, in some embodiments, the microparticles can be aseptically manufactured, i.e., without final sterilization by ionizing radiation.

[0143] The encapsulation efficiency of the method described herein is typically at least 80% by weight, including 90% to 95% by weight, based on the weight of triptorelin. Particle characteristics

[0144] In one aspect, a drug-carrying microparticle is provided, the drug-carrying microparticle comprising: (a) Medicine; (b) A polymer component comprising one or more polymers selected from ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymers with a lactide:co-lactide ratio of about 50:50, ester-terminated PLGA polymers with a lactide:co-lactide ratio of about 75:25, and ester-terminated polylactide (PLA) polymers. (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release the drug in the body for at least one week after administration via subcutaneous or intramuscular injection. In some embodiments, the microparticles release the drug in the body for at least one month after administration via subcutaneous or intramuscular injection. In some embodiments, the microparticles release the drug in the body for at least three months after administration via subcutaneous or intramuscular injection. In some embodiments, the microparticles release the drug in the body for at least six months after administration via subcutaneous or intramuscular injection.

[0145] In some embodiments, the drug is selected from one or more small molecule drugs, peptide drugs, and protein drugs. In some embodiments, the drug is selected from one or more of the following: triptorelin, follicle-stimulating hormone (FSH), luteinizing hormone (LH), human chorionic gonadotropin (hCG), atosiban, insulin, abiraterone, balusiban, degarelix, desmopressin, ganirelix (e.g., ganirelix acetate), gemcitabine, gonadorelin (e.g., gonadorelin acetate), urinary gonadotropin or human menopausal gonadotropin (hMG), meropenem, progesterone, quinolones, somatostatin, growth hormone, and tolterodine. In some embodiments, the drug is triptorelin. In some embodiments, the drug is follicle-stimulating hormone (FSH). In some embodiments, the drug comprises two or more drugs. In some embodiments, the drug comprises FSH and LH and / or hCG.

[0146] In one aspect, triptorelin-loaded microparticles are provided, which comprise: (a) Triptorelin; (b) Lactide: One or more ester-terminated PLGA polymers with a lactide ratio of about 50:50; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release triptorelin FSH in the body for at least one week after administration via subcutaneous or intramuscular injection.

[0147] In one aspect, triptorelin-loaded microparticles are provided, which comprise: (a) Triptorelin; (b) One or more polymers selected from ester-terminated PLGA polymers with a lactide:glycol ratio of about 50:50, ester-terminated PLGA polymers with a lactide:glycol ratio of about 75:25, and two different ester-terminated PLA polymers. (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0148] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) Lactide: One or more ester-terminated PLGA polymers with a lactide ratio of about 50:50; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0149] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) Lactide: ester-terminated PLGA polymers with a lactide ratio of approximately 50:50, such as RG503; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0150] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) Lactide: Ester-terminated PLGA polymers with a lactide ratio of approximately 50:50, such as RG503; and (c) Polosham, such as Polosham 188, The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0151] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) Lactide: Ester-terminated PLGA polymers with a lactide ratio of approximately 50:50, such as RG503; and (c) Polosham, such as Polosham 188; and (d) One or both of sodium chloride and polyethylene glycol. The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0152] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) Lactide: Ester-terminated PLGA polymers with a lactide ratio of approximately 50:50, such as RG503; and (c) Polyvinyl alcohol, The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0153] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) Lactide: Ester-terminated PLGA polymers with a lactide ratio of approximately 50:50, such as RG503; and (c) Polyvinyl alcohol; and (d) One or both of sodium chloride and polyethylene glycol. The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0154] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) Lactide: A mixture of two different ester-terminated PLGA polymers, such as RG504 and RG503, with a lactide ratio of about 50:50. (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0155] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) A mixture of two different ester-terminated PLGA polymers in a ratio of approximately 50:50, wherein the two different ester-terminated polylactide PLGA polymers are RG504 and RG503; and (c) Polosham, such as Polosham 188; and (d) One or both of sodium chloride and polyethylene glycol; The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0156] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) A mixture of two different ester-terminated PLGA polymers in a ratio of approximately 50:50, wherein the two different ester-terminated polylactide PLGA polymers are RG504 and RG503; and (c) Polyvinyl alcohol; and (d) One or both of sodium chloride and polyethylene glycol; The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0157] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) Lactide: A mixture of three different ester-terminated PLGA polymers, such as RG504, RG503 and RG502, in a ratio of about 50:50. (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0158] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) A mixture of three different ester-terminated PLGA polymers in a ratio of approximately 50:50, wherein the three different ester-terminated polylactide PLGA polymers are RG504, RG503, and RG502; and (c) Polosham, such as Polosham 188; The microparticles release triptorelin in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0159] In one aspect, triptorelin-loaded microparticles are provided, which comprise: (a) Triptorelin; (b) Lactide: an ester-terminated PLGA polymer with a lactide ratio of approximately 75:25; (c) Surfactants; and (d) One or both of basic salts and polyols. The microparticles release triptorelin in the body for at least 3 months after administration via subcutaneous or intramuscular injection.

[0160] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin acetate; (b) Lactide: ester-terminated PLGA polymers with a lactide ratio of about 75:25, such as RG753S; (c) Polosham, such as Polosham 188; and (d) One or both of sodium chloride and sucrose. The microparticles release triptorelin in the body for at least 3 months after administration via subcutaneous or intramuscular injection.

[0161] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin; (b) Two different ester-terminated PLA polymers; (c) Surfactants; and (d) One or both of basic salts and polyols. The microparticles release triptorelin in the body for at least 6 months after administration via subcutaneous or intramuscular injection.

[0162] On the other hand, triptorelin-loaded microparticles are provided, which contain: (a) Triptorelin acetate; (b) Two different ester-terminated polylactide (PLA) polymers, wherein the two different ester-terminated polylactide PLA polymers are R203S and R205S. (c) Polosham, such as Polosham 188; and (d) One or both of sodium chloride and sucrose. The microparticles release triptorelin in the body for at least 6 months after administration via subcutaneous or intramuscular injection.

[0163] In one aspect, FSH-carrying microparticles are provided, which contain: (a) FSH; (b) One or more polymers selected from ester-terminated PLGA polymers with a lactide:glycol ratio of about 50:50, ester-terminated PLGA polymers with a lactide:glycol ratio of about 75:25, and two different ester-terminated PLA polymers. (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release FSH in the body for at least one week after administration via subcutaneous or intramuscular injection.

[0164] In one aspect, FSH-carrying microparticles are provided, which contain: (a) FSH; (b) One or more polymers selected from ester-terminated PLGA polymers with a lactide:glycol ratio of about 50:50, ester-terminated PLGA polymers with a lactide:glycol ratio of about 75:25, and two different ester-terminated PLA polymers. (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release FSH in the body for at least one month after administration via subcutaneous or intramuscular injection, or optionally, the microparticles release FSH in the body for at least six weeks after administration via subcutaneous or intramuscular injection.

[0165] On the other hand, FSH-carrying microparticles are provided, which contain: (a) FSH; (b) Lactide: One or more ester-terminated PLGA polymers with a lactide ratio of about 50:50; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release FSH in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0166] On the other hand, FSH-carrying microparticles are provided, which contain: (a) FSH; (b) Lactide: ester-terminated PLGA polymers with a lactide ratio of approximately 50:50, such as RG503; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release FSH in the body for at least one month after administration via subcutaneous or intramuscular injection.

[0167] On the other hand, FSH-carrying microparticles are provided, which contain: (a) FSH; (b) Lactide: One or more ester-terminated PLGA polymers with a lactide ratio of about 50:50; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release FSH in the body for at least 6 weeks after administration via subcutaneous or intramuscular injection.

[0168] On the other hand, FSH-carrying microparticles are provided, which contain: (a) FSH; (b) Lactide: ester-terminated PLGA polymers with a lactide ratio of approximately 50:50, such as RG502; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release FSH in the body for at least 6 weeks after administration via subcutaneous or intramuscular injection.

[0169] In one aspect, FSH-carrying microparticles are provided, which contain: (a) FSH; (b) Lactide: an ester-terminated PLGA polymer with a lactide ratio of approximately 75:25; (c) Surfactants; and (d) One or both of basic salts and polyols. The microparticles release FSH in the body for at least 3 months after administration via subcutaneous or intramuscular injection.

[0170] On the other hand, FSH-carrying microparticles are provided, which contain: (a) FSH; (b) Lactide: ester-terminated PLGA polymers with a lactide ratio of about 75:25, such as RG753S; (c) Polosham, such as Polosham 188; and (d) One or both of sodium chloride and polyethylene glycol. The microparticles release FSH in the body for at least 3 months after administration via subcutaneous or intramuscular injection.

[0171] On the other hand, FSH-carrying microparticles are provided, which contain: (a) FSH; (b) Two different ester-terminated PLA polymers; (c) Surfactants; and (d) One or both of basic salts and polyols. The microparticles release FSH in the body for at least 6 months after administration via subcutaneous or intramuscular injection.

[0172] On the other hand, FSH-carrying microparticles are provided, which contain: (a) FSH; (b) Two different ester-terminated PLA polymers, wherein the two different ester-terminated polylactide PLA polymers are R203S and R205S. (c) Polosham, such as Polosham 188; and (d) One or both of sodium chloride and polyethylene glycol. The microparticles release FSH in the body for at least 6 months after administration via subcutaneous or intramuscular injection.

[0173] Residual contentThe microparticles described herein may have residual amounts of one or more components used to prepare the microparticles, which are present in one or more of a polymer solution, a drug solution, a phase inversion solution, and a washing phase, such as the solvent of the polymer solution; the acid and / or buffer salt of the aqueous acid solution used in the triptorelin solution; the solvent, surfactant, basic salt, viscosity modifier, etc. of the phase inversion solution; and the surfactant or buffer salt of the washing phase. For example, the microparticles described herein may have residual amounts of one or more of the following: (i) organic solvents in polymer solutions, from about 50 ppm to about 20,000 ppm; (ii) acids and / or buffer salts in aqueous acid solutions used in pharmaceutical solutions, from about 0.01% w / w to about 1.0% w / w; (iii) viscosity modifiers, from about 0.1% w / w to about 5% w / w; (iv) surfactants, from about 0% w / w to about 4% w / w; (v) basic salts, from about 0% to about 0.5% w / w; (vi) buffer salts in phase inversion solutions / surfactant solutions or washing phases, from about 0.01% w / w to about 1.0% w / w. For example, microparticles prepared using the components described in the examples may have residual amounts of one or more of ethyl acetate, acetate / acetate, sucrose, NaCl, citric acid / citric acid salt, poloxamer (e.g., poloxamer 188), or polyvinyl alcohol and polyethylene glycol. For example, microparticles prepared using the components described in the examples may have a residual acetic acid content of about 0.01% w / w to about 1.0% w / w; a residual sucrose content of about 0.20% w / w to about 5.0% w / w; a residual citric acid content of about 0.01% w / w to about 1.0% w / w; a residual ethyl acetate content of about 50 ppm to about 20,000 ppm; a residual surfactant content of about 0% w / w to about 4% w / w; a residual polyethylene glycol content of about 0.20% w / w to about 5% w / w; and / or a residual NaCl content of about ≤ 0.5% w / w.

[0174] Particle size: As described above, the particle size can be adjusted by regulating various process parameters. For example, the particles described herein may have an average particle size of about 20 µm to about 180 µm, including about 30 µm to about 150 µm, such as about 30 µm to about 120 µm, including about 30 µm to about 80 µm, including average particle sizes of 20 µm to 180 µm, 30 µm to 150 µm, 30 µm to 120 µm, and 30 µm to 80 µm.

[0175] Target doseThe microparticles described herein can be formulated into compositions as described herein to deliver a target dose of the formulated drug over a period of 6 months, 3 months, 1 month, or 1 week. For triptorelin, the target dose for a typical human patient over 6 months may be 22.5 mg; the target dose for a typical human patient over 3 months may be 11.25 mg; the target dose for a typical human patient over 1 month may be 3.75 mg; and the target dose for a typical human patient over 1 week may be approximately 1 mg. For FSH, such as follicle-stimulating hormone delta, the target dose of the formulated drug over a period of 6 months, 3 months, 6 weeks, 1 month, or 1 week may be a dose delivered daily at approximately 12 µg. For FSH, such as follicle-stimulating hormone delta (FSH), the target dose for a typical human patient (e.g., a typical male patient) over 6 months may be about 2 mg or more; the target dose for a typical human patient over 3 months may be about 1 mg or more; the target dose for a typical human patient over 6 weeks may be about 0.5 mg or more; the target dose for a typical human patient over 1 month may be about 0.35 mg or more; and the target dose for a typical human patient over 1 week may be about 85 µg or more. For example, taking into account the potential burst release, the target dose for a 1-month product may be about 1 mg to about 5 mg, and the target dose for a 6-month product may be about 2 mg to about 10 mg. The target dose for products of other durations (e.g., 6 weeks) can be adjusted similarly.

[0176] Released in the body As described above, the microparticles described herein are sustained-release drug-loaded microparticles. For example, as described above, depending on the polymer used, the microparticles described herein can provide in vivo release of the drug over a prolonged period (such as 1 week, 1 month, 3 months, or 6 months). Microparticles can be prepared to provide therapeutically effective plasma levels of the drug over a prolonged period (such as 1 week, 1 month, 3 months, or 6 months). For triptorelin, microparticles can be prepared to provide a mean plasma concentration of at least 0.1 ng / mL of triptorelin in typical human patients over a prolonged period (e.g., 1 month, 3 months, or 6 months). For FSH, microparticles can be prepared to provide a mean plasma concentration of at least 14 mIU / mL of FSH (e.g., follicle-stimulating hormone δ) in typical human patients (e.g., typical male patients) over a prolonged period (e.g., 1 week, 1 month, 3 months, or 6 months). Pharmaceutical Composition

[0177] The microparticles described herein can be formulated into pharmaceutical compositions comprising one or more pharmaceutically acceptable carriers, diluents, and / or excipients, including fillers, buffers, chelating agents, antioxidants, preservatives, cosolvents, etc.

[0178] Examples of suitable excipients include polyethylene glycol (PEG), polyols (such as trehalose or mannitol), and preservatives (such as phenol, benzyl alcohol, m-cresol, methylparaben, propylparaben, benzalkonium chloride, and benzyl chloride).

[0179] The microparticles described in this article can be freeze-dried and then reconstituted before use.

[0180] In specific embodiments, the microparticles are formulated for subcutaneous or intramuscular injection. In some embodiments, the drug-loaded microparticles described herein are formulated in a carrier comprising carboxymethyl cellulose. In some embodiments, the drug-loaded microparticles described herein are formulated in a carrier comprising dextran. In some embodiments, the drug-loaded microparticles described herein are formulated in a carrier comprising hyaluronic acid or a salt thereof (e.g., sodium hyaluronate). As discussed below and illustrated in the examples, it has been surprisingly found that formulating drug-loaded microparticles as described herein with hyaluronic acid or a salt thereof provides several advantages.

[0181] As used herein, hyaluronic acid refers to a polymer of D-glucuronic acid and N-acetyl-D-glucosamine disaccharide linked by alternating β-(1→4) and β-(1→3) glycosidic bonds. As used herein, salts of hyaluronic acid (also known as hyaluronic salts or hyaluronates) include metal salts of hyaluronic acid, such as sodium and potassium salts. In the following discussion, hyaluronic acid and its salts are collectively referred to as "HA".

[0182] The HA used as a carrier / diluent in the pharmaceutical compositions described herein may have an average molecular weight greater than about 1 MDa or greater than about 3 MDa, such as an average molecular weight of about 1 MDa to about 5 MDa or about 2 MDa to about 4 MDa, including about 1.5 MDa, about 2.0 MDa, about 2.5 MDa, about 3.0 MDa, about 3.5 MDa, about 4 MDa, or about 5 MDa. In a specific embodiment, the HA has a molecular weight of about 3 MDa. In other specific embodiments, the HA has a molecular weight of 3.2 ± 0.6 g / mol or greater.

[0183] When HA is used as a carrier / diluent in the pharmaceutical compositions described herein, it can be used in any suitable amount to obtain a composition with the desired properties. For example, based on the volume of the pharmaceutical composition, HA can be used in amounts from about 0.01% (w / v) to about 5% (w / v), including from about 1% (w / v) to about 3% (w / v). In some embodiments, HA is used in an amount of about 1.5 mg / mL of the pharmaceutical composition. In other specific embodiments, HA with a molecular weight of 3.2 ± 0.6 g / mol is used in an amount of about 1.5 mg / mL of the pharmaceutical composition.

[0184] As described above, formulating the drug-loaded microparticles described herein with HA offers several advantages. For example, it has been found that formulating the drug-loaded microparticles described herein in a carrier containing HA (e.g., containing an HA diluent) provides favorable shear-thinning / viscoelastic behavior compared to formulations prepared with dextran (e.g.), resulting in higher microparticle suspension stability and better injectability. Specifically, it has been found that a carrier containing HA (e.g., containing an HA diluent) provides high viscosity at low shear rates, which delays microparticle sedimentation, while also exhibiting low viscosity at high shear rates, i.e., allowing easy injection with syringes and needles when injected. Additionally, formulating the drug-loaded microparticles described herein with HA increases the zeta potential. Therefore, formulating the drug-loaded microparticles described herein in a composition containing HA can provide compositions with good physicochemical stability and good injectability.

[0185] Furthermore, it was surprisingly found that formulation of drug-loaded microparticles as described herein with HA reduced the immunogenicity of the composition, as reflected in one or both of the following: (i) a reduced local immune response to the composition at the injection site (e.g., reduced local inflammation) and (ii) a reduced systemic immune response, which may be reflected in an increased IL-4 (anti-inflammatory) response and / or a reduced level of neutralizing antidrug antibodies, for example, compared to compositions not formulated with HA (such as those formulated with dextran), as shown in Example 7. Without being bound by theory, it is believed that formulations containing HA reduce the adsorption of plasma proteins to microparticles. Plasma proteins are known to act as complement with potentially pro-inflammatory effects; therefore, this reduction in plasma protein adsorption to microparticles may play a role in reducing the immunogenicity of microparticle formulations formulated with HA. Therefore, formulation of drug-loaded microparticles as described herein in a composition containing HA can provide a composition with reduced (e.g., low) immunogenicity.

[0186] Furthermore, for some drugs, it has been surprisingly found that formulations of the drug-loaded microparticles described herein using HA may affect pharmacokinetic characteristics. For example, as described in Example 7 below, formulations of the FSH-loaded microparticles described herein using HA increased the plasma concentration-time profile of FSH after injection compared to formulations using dextran. On the other hand, formulations of the triptorelin-loaded microparticles described herein using HA did not show any effect on the plasma concentration-time profile of triptorelin after injection compared to formulations using dextran.

[0187] In the following discussion, it should be understood that any pharmaceutical composition described herein may contain the carrier as described above, even if not specifically mentioned.

[0188] In some aspects, a composition comprising drug-loaded microparticles is provided, the composition being formulated to contain a certain amount of drug-loaded microparticles to provide, for example, a target dose of medicine effective for 1 week, 1 month, 6 weeks, 3 months, or 6 months, such as an amount providing a therapeutically effective plasma level within a target time, via a single dose administered subcutaneously or intramuscularly. A pharmaceutical composition comprising drug-loaded microparticles is also provided, which additionally or alternatively provides a therapeutically effective mean plasma concentration of medicine in typical human patients for at least 1 week after administration of a single dose via subcutaneous or intramuscular injection. A pharmaceutical composition comprising drug-loaded microparticles is also provided, which additionally or alternatively provides a therapeutically effective mean plasma concentration of medicine in typical human patients for at least 1 month after administration of a single dose via subcutaneous or intramuscular injection. A pharmaceutical composition comprising drug-loaded microparticles is also provided, which additionally or alternatively provides a therapeutically effective mean plasma concentration of medicine in typical human patients for at least 6 weeks after administration of a single dose via subcutaneous or intramuscular injection. A pharmaceutical composition comprising drug-loaded microparticles is also provided, which, upon administration of a single dose via subcutaneous or intramuscular injection, additionally or alternatively provides mean plasma concentrations of the drug in typical human patients for at least 3 months. A pharmaceutical composition comprising drug-loaded microparticles is also provided, which, upon administration of a single dose via subcutaneous or intramuscular injection, additionally or alternatively provides mean plasma concentrations of the drug in typical human patients for at least 6 months.

[0189] Therefore, in one aspect, a pharmaceutical composition comprising triptorelin-loaded microparticles is provided, the pharmaceutical composition being formulated to contain a certain amount of triptorelin-loaded microparticles, thereby providing, for example, about 1 mg of triptorelin in a single dose via subcutaneous or intramuscular injection. A pharmaceutical composition comprising triptorelin-loaded microparticles is also provided, which, upon administration of a single dose via subcutaneous or intramuscular injection, additionally or alternatively provides a mean plasma concentration of at least 0.1 ng / mL of triptorelin in typical human patients over a period of at least 1 week.

[0190] On the other hand, a pharmaceutical composition comprising triptorelin microparticles is provided, the pharmaceutical composition being formulated to contain a certain amount of triptorelin microparticles, thereby providing 3.75 mg of triptorelin in a single dose, for example, by subcutaneous or intramuscular injection. A pharmaceutical composition comprising triptorelin microparticles is also provided, which, following administration of a single dose by subcutaneous or intramuscular injection, additionally or alternatively provides a mean plasma concentration of at least 0.1 ng / mL of triptorelin in typical human patients over a period of at least one month.

[0191] On the other hand, a pharmaceutical composition comprising triptorelin microparticles is provided, the pharmaceutical composition being formulated to contain a certain amount of triptorelin microparticles, thereby providing 11.25 mg of triptorelin in a single dose, for example, by subcutaneous or intramuscular injection. A pharmaceutical composition comprising triptorelin microparticles is also provided, which, following administration of a single dose by subcutaneous or intramuscular injection, additionally or alternatively provides a mean plasma concentration of at least 0.1 ng / mL of triptorelin in typical human patients over a period of at least 3 months.

[0192] On the other hand, a pharmaceutical composition comprising triptorelin microparticles is provided, the pharmaceutical composition being formulated to contain a certain amount of triptorelin microparticles, thereby providing 22.5 mg of triptorelin in a single dose, for example, by subcutaneous or intramuscular injection. A pharmaceutical composition comprising triptorelin microparticles is also provided, which, following administration of a single dose by subcutaneous or intramuscular injection, additionally or alternatively provides a mean plasma concentration of at least 0.1 ng / mL of triptorelin in typical human patients over a period of at least 6 months.

[0193] On the other hand, a pharmaceutical composition comprising FSH-loaded microparticles is provided, the pharmaceutical composition being formulated to contain a certain amount of FSH-loaded microparticles, thereby providing approximately 12 µg of FSH per day in a single dose, for example, by subcutaneous or intramuscular injection.

[0194] On the other hand, a pharmaceutical composition comprising FSH-loaded microparticles is provided, the pharmaceutical composition being formulated to contain a certain amount of FSH-loaded microparticles, thereby providing, for example, about 85 µg or more of FSH (e.g., follicle-stimulating hormone δ) in a single dose via subcutaneous or intramuscular injection. A pharmaceutical composition comprising FSH-loaded microparticles is also provided, which, upon administration of a single dose via subcutaneous or intramuscular injection, additionally or alternatively provides a mean plasma concentration of at least 14 mlU / mL of FSH in typical human patients (e.g., typical male patients) over a period of at least 1 week.

[0195] On the other hand, a pharmaceutical composition comprising FSH-loaded microparticles is provided, the pharmaceutical composition being formulated to contain a certain amount of FSH-loaded microparticles, thereby providing, for example, about 0.35 mg or more of FSH (e.g., follicle-stimulating hormone δ) in a single dose via subcutaneous or intramuscular injection. A pharmaceutical composition comprising FSH-loaded microparticles is also provided, which, upon administration of a single dose via subcutaneous or intramuscular injection, additionally or alternatively provides a mean plasma concentration of at least 14 mlU / mL of FSH in typical human patients (e.g., typical male patients) over a period of at least 1 month.

[0196] On the other hand, a pharmaceutical composition comprising FSH-loaded microparticles is provided, the pharmaceutical composition being formulated to contain a certain amount of FSH-loaded microparticles, thereby providing, for example, about 0.5 mg or more of FSH (e.g., follicle-stimulating hormone δ) in a single dose via subcutaneous or intramuscular injection. A pharmaceutical composition comprising FSH-loaded microparticles is also provided, which, upon administration of a single dose via subcutaneous or intramuscular injection, additionally or alternatively provides a mean plasma concentration of at least 14 mlU / mL of FSH in typical human patients (e.g., typical male patients) over a period of at least 6 weeks.

[0197] On the other hand, a pharmaceutical composition comprising FSH-loaded microparticles is provided, the pharmaceutical composition being formulated to contain a certain amount of FSH-loaded microparticles, thereby providing, for example, about 1 mg or more of FSH (e.g., follicle-stimulating hormone δ) in a single dose via subcutaneous or intramuscular injection. A pharmaceutical composition comprising FSH-loaded microparticles is also provided, which, upon administration of a single dose via subcutaneous or intramuscular injection, additionally or alternatively provides a mean plasma concentration of at least 14 mlU / mL of FSH in typical human patients (e.g., typical male patients) over a period of at least 3 months.

[0198] On the other hand, a pharmaceutical composition comprising FSH-loaded microparticles is provided, the pharmaceutical composition being formulated to contain a certain amount of FSH-loaded microparticles, thereby providing, for example, about 2 mg or more of FSH (e.g., follicle-stimulating hormone δ) in a single dose via subcutaneous or intramuscular injection. A pharmaceutical composition comprising FSH-loaded microparticles is also provided, which, upon administration of a single dose via subcutaneous or intramuscular injection, additionally or alternatively provides a mean plasma concentration of at least 14 mlU / mL of FSH in typical human patients (e.g., typical male patients) over a period of at least 6 months.

[0199] As described above, the FSH dose can be adjusted (e.g., increased) to address any burst release of FSH, thereby providing a sustained release of FSH over a predetermined period of time (e.g., 1 week, 1 month, 6 weeks, 3 months, 6 months, etc.).

[0200] A kit is also provided comprising (a) a container containing drug-loaded microparticles in lyophilized form as described herein, in an amount providing a therapeutically effective amount of the drug, and (b) instructions for preparing a pharmaceutical composition comprising microparticles and a pharmaceutically acceptable carrier, such as one suitable for administration by subcutaneous or intramuscular injection.

[0201] A kit is also provided comprising (a) a container containing lyophilized triptorelin microparticles as described herein, in amounts providing 3.75 mg, 11.25 mg, or 22.5 mg triptorelin, and (b) instructions for preparing a pharmaceutical composition comprising microparticles and a pharmaceutically acceptable carrier, such as one suitable for administration by subcutaneous or intramuscular injection.

[0202] A kit is also provided comprising (a) a container containing lyophilized FSH-loaded microparticles (e.g., follicle-stimulating hormone delta microparticles) as described herein, provided in amounts such as 85 µg FSH, 0.35 mg FSH, 0.5 mg FSH, 1 mg FSH, or 2 mg FSH or more, and (b) instructions for preparing a pharmaceutical composition comprising microparticles and a pharmaceutically acceptable carrier, such as suitable for administration by subcutaneous or intramuscular injection. Therapeutic methods

[0203] This document also provides treatment methods using the pharmaceutical compositions described herein. These methods may include administering the pharmaceutical composition described herein to a subject in need of it, for example by injecting (administering) a therapeutically effective amount of the composition described herein to a subject in need of it.

[0204] The amount of the drug-loaded microparticle composition applied can be any amount of drug that effectively delivers the target dose, such as the amount of drug that delivers the target dose effectively within 1 week, 1 month, 6 weeks, 3 months or 6 months, such as the amount of drug plasma level that delivers the therapeutically effective dose within the target time (such as 1 week, 1 month, 3 months or 6 months), such as those discussed above for each of triptorelin and FSH.

[0205] For the triptorelin composition, the subject may have and / or require treatment for one or more conditions selected from hormone-responsive cancers such as breast or prostate cancer, endometriosis, female infertility, uterine fibroids, or precocious puberty. In a specific embodiment, the subject treated with the triptorelin-loaded microparticles disclosed herein had hormone-sensitive prostate cancer or hormone-sensitive breast cancer.

[0206] The amount of the composition comprising triptorelin microparticles as disclosed herein may be any amount that effectively provides a target dose of triptorelin, such as an amount that provides a target dose of triptorelin effective over 1 week, 1 month, 6 weeks, 3 months, or 6 months, or an amount that provides a therapeutically effective plasma level of triptorelin over a target time (such as a mean plasma concentration of at least 0.1 ng / mL of triptorelin over a longer period (e.g., 1 week, 1 month, 6 weeks, 3 months, or 6 months). The typical volume of the composition for subcutaneous injection is about 0.5 to 2 mL. The typical volume of the composition for intramuscular injection is about 0.5 to 3 mL, and in some examples, it is 0.5 to 5 mL.

[0207] Therefore, this article provides treatment methods comprising administering a pharmaceutical composition comprising the triptorelin-loaded microparticles disclosed herein to a human subject in need of such treatment by subcutaneous or intramuscular injection of a single dose of approximately 1 mg triptorelin. In some embodiments, the method provides a mean plasma concentration of at least 0.1 ng / mL of triptorelin over a period of at least 1 week following a single subcutaneous or intramuscular injection.

[0208] This article also provides treatment methods that include administering a single dose of 3.75 mg triptorelin to a human subject in need of this treatment via subcutaneous or intramuscular injection of a pharmaceutical composition comprising the triptorelin-loaded microparticles disclosed herein. In some embodiments, the method provides a mean plasma concentration of at least 0.1 ng / mL of triptorelin over a period of at least one month following a single subcutaneous or intramuscular injection.

[0209] This article also provides treatment methods comprising administering a single dose of 11.25 mg triptorelin to a human subject in need of this treatment via subcutaneous or intramuscular injection of a pharmaceutical composition comprising the triptorelin-loaded microparticles disclosed herein. In some embodiments, the method provides a mean plasma concentration of at least 0.1 ng / mL of triptorelin over a period of at least 3 months following a single subcutaneous or intramuscular injection.

[0210] This article also provides treatment methods comprising administering a single dose of 22.5 mg triptorelin to a human subject in need of this treatment via subcutaneous or intramuscular injection of a pharmaceutical composition comprising the triptorelin-loaded microparticles disclosed herein. In some embodiments, the method provides a mean plasma concentration of at least 0.1 ng / mL of triptorelin over a period of at least 6 months following a single subcutaneous or intramuscular injection.

[0211] For FSH-loaded microparticle compositions, subjects may have and / or require treatment for female infertility or male infertility.

[0212] The amount of FSH composition administered can be any amount that effectively delivers the target dose of FSH, such as the amount that delivers the target dose of FSH effectively over 1 week, 1 month, 6 weeks, 3 months, or 6 months, or the amount that delivers the therapeutically effective FSH plasma level (such as a mean plasma concentration of at least 14 mlU / mL of FSH over a longer period, such as 1 week, 1 month, 6 weeks, 3 months, or 6 months) over the target time period. The typical volume of the composition for subcutaneous injection is about 0.1 to 2 mL. The typical volume of the composition for intramuscular injection is about 0.1 to 3 mL, and in some examples, it is 0.1 to 3 mL.

[0213] Therefore, this article provides treatment methods comprising administering a single dose of approximately 85 µg or more of FSH (e.g., follicle-stimulating hormone δ) to a human subject in need of this treatment via subcutaneous or intramuscular injection of a pharmaceutical composition comprising the FSH-loaded microparticles disclosed herein. In some embodiments, the method provides a mean plasma concentration of at least 14 mlU / mL of FSH over a period of at least 1 week following a single subcutaneous or intramuscular injection.

[0214] This article also provides treatment methods that include administering a single dose of about 0.35 mg or more of FSH (e.g., follicle-stimulating hormone delta) to a human subject in need of this treatment via subcutaneous or intramuscular injection of a pharmaceutical composition comprising the FSH-loaded microparticles disclosed herein. In some embodiments, the method provides a mean plasma concentration of at least 14 mlU / mL of FSH over a period of at least 1 month following a single subcutaneous or intramuscular injection.

[0215] This article also provides treatment methods that include administering a single dose of about 0.5 mg or more of FSH (e.g., follicle-stimulating hormone delta) to a human subject in need of this treatment via subcutaneous or intramuscular injection of a pharmaceutical composition comprising the FSH-loaded microparticles disclosed herein. In some embodiments, the method provides a mean plasma concentration of at least 14 mlU / mL of FSH over a period of at least 6 weeks following a single subcutaneous or intramuscular injection.

[0216] This article also provides treatment methods that include administering a single dose of about 1 mg or more of FSH (e.g., follicle-stimulating hormone δ) to a human subject in need of this treatment via subcutaneous or intramuscular injection of a pharmaceutical composition comprising the FSH-loaded microparticles disclosed herein. In some embodiments, the method provides a mean plasma concentration of at least 14 mlU / mL of FSH over a period of at least 3 months following a single subcutaneous or intramuscular injection.

[0217] This article also provides treatment methods that include administering a single dose of about 2 mg or more of FSH (e.g., follicle-stimulating hormone delta) to a human subject in need of this treatment via subcutaneous or intramuscular injection of a pharmaceutical composition comprising the FSH-loaded microparticles disclosed herein. In some embodiments, the method provides a mean plasma concentration of at least 14 mlU / mL of FSH over a period of at least 6 months following a single subcutaneous injection. Example

[0218] The following specific examples are included to illustrate the invention described herein. These examples are in no way intended to limit the scope of the invention. Example 1: Triptorelin microparticles for 1 month (laboratory scale)

[0219] A 1-month triptorelin microparticle formulation was prepared on a laboratory scale (3 g) using the following general method: Step 1: Prepare the feed solution for the particle formation step; Step 2: Particle formation; Step 3: Remove ethyl acetate from the polymer particles by evaporation; Step 4: Separate the particles from the suspension and wash them; and Step 5: Freeze-drying.

[0220] The prepared polymer solution and a known amount of drug solution were sequentially transferred to a dispersion vessel. The drug solution was then dispersed in the polymer solution at a controlled mixing rate. After a dispersion time, a surfactant solution containing poloxamer 188 was transferred to the dispersion vessel. This action caused a phase inversion, leading to the formation of triptorelin-loaded microparticles. As described above, the dispersion step temperature, dispersion step mixing rate, dispersion step mixing time, and phase inversion rate can affect particle size and triptorelin release rate, and these can be selected and adjusted to prepare the triptorelin-loaded microparticles described in this example. The drug-loaded microparticle suspension contained excess organic polymer solvent as well as unencapsulated drug and excipients, which were removed in subsequent solvent evaporation and washing steps. A washing phase solution containing 16% poloxamer 188 at pH 6.5 was added to stabilize the microparticles. Excess polymer solvent was removed by applying vacuum and increasing temperature during continuous stirring using magnetic stirring. Ethyl acetate was removed by evaporation, thereby increasing the transition of ethyl acetate from the microparticles to the aqueous surfactant solution (continuous phase), resulting in complete hardening of the polymer microparticles.

[0221] For this laboratory-scale method, the microparticles were separated from the continuous phase by centrifugation, in which the supernatant containing residual solvent and excipients was discarded, the microparticles were resuspended in ultrapure water and centrifuged again, the supernatant was discarded, and then the microparticles were resuspended in a surfactant solution (0.25% poloxamer 188) and centrifuged again. Aliquots of the microparticle suspension were transferred to lyophilized vials, and the vials containing the microparticle suspension were lyophilized under appropriate conditions to provide the triptorelin-loaded microparticle formulation. The triptorelin-loaded microparticle formulation was sterilized by gamma radiation at a dose of 25 kGy ± 10%.

[0222] The table below lists the components and properties of the particles used to prepare a 1-month formulation on a laboratory scale. Table 1. Triptorelin formulations after 1 month (laboratory scale; 3 g scale) F68 = Poloxamer 188 (F68) (KOLLIPHOR® P188), European Pharmacopoeia, derived from BASF / BTC (Europe, Gesmar, USA). nd = Undetermined Formulation 1 (Laboratory Scale) Formulation 2 (Laboratory Scale) Formulation 3 (Laboratory Scale) Formulation 4 (Laboratory Scale) Formulation 5 (Laboratory Scale) Formulation 6 (Laboratory Scale) Formulation 7 (Laboratory Scale) Formulation 8 (Laboratory Scale) In vitro release of triptorelin microparticles after 1 month (laboratory scale)

[0223] The in vitro release of the triptorelin-loaded microparticle formulations prepared as described above was evaluated as follows. Ten aliquots of 10 mg microparticles were weighed into polypropylene screw containers, and 1.5 mL of aqueous buffer was added to each aliquot as the release medium. The samples were placed on a top-mounted shaker and stirred at 37°C. At appropriate sampling times (0 h, 4 h, 24 h, 3 days, and 7 days), two samples were centrifuged to separate the microparticles from the release medium. The release medium was filtered through a syringe pre-filter, and the triptorelin content was determined by HPLC analysis. Formulations 1, 2, and 3 exhibited different drug release characteristics over seven days in vitro. A one-month pharmacokinetic study of triptorelin microparticles (laboratory scale).

[0224] For in vivo evaluation, the aforementioned 1-month triptorelin-loaded microparticles (formulations 1–8) were administered as a single subcutaneous (sc) injection to Sprague-Dawley (SD) rats to provide a dose of 1 mg triptorelin, with 8 rats in each treatment group. The study lasted for three (3) months. Blood samples were collected at the following time points for bioanalysis and pharmacokinetic assessment: days 1, 3, 5, 7, 14, 28, 42, 56, 70, and 84 post-administration. For the first four blood sampling time points, blood was collected from 4 animals in each group, allowing for staggered blood sampling. Body weight was measured on the day of administration and again on days 28, 56, and 84. Injection sites were carefully monitored.

[0225] The results for formulations 1 (polymer RG503), 2 (polymer RG504), and 3 (polymer RG503 / RG502) are shown in the figures below. Figures 2 to 4In all test groups, mean plasma triptorelin concentrations above 100 pg / mL (target minimum concentration) were achieved at 1 month (672 hours). Two months after triptorelin administration, mean plasma triptorelin concentrations were close to or below the 100 pg / mL threshold, with the mean concentration in PK test group 8 / formulation 8 significantly above the 100 pg / mL threshold (not shown). Three months after triptorelin administration, all mean plasma triptorelin concentrations were below the 100 pg / mL threshold. Stability study of triptorelin microparticles over one month (laboratory scale)

[0226] The storage stability of the selected 1-month triptorelin-loaded microparticles in this example was assessed by monitoring the evolution of major impurities over time using HPLC analysis. The primary focus of the stability study was the evolution of the following major individual impurities exhibiting the largest relative peak areas at the following relative retention times (RRTs), which are relative to the retention time of the triptorelin peak: at RRT 1.09 / 1.10, RRT 1.13 / 1.14, RRT 1.16 / 1.17, RRT 1.18 / 1.19, and RRT 1.22 / 1.23. The evolution of these individual impurities was examined over 6 months of storage (T0, T1, T3, T6 months) and exposure to different conditions (5°C ± 3°C, 25°C ± 2°C / 60% RH, 30°C ± 2°C / 65% RH). According to the ICH Q3B guidelines, for a daily dose of 3.75 mg of a drug, the identification threshold is 0.5%, and the acceptable threshold is 1.0%. Therefore, it is very important that a single impurity does not exceed 1.0%.

[0227] All formulations prepared with ester-terminated polymers (RG504, RG503, RG502, or mixtures thereof) exhibited good stability, with individual impurities remaining well below the acceptable threshold of 1.0% (e.g., formulation 2 (RG504) and formulation 3 (RG503 / RG502)). Furthermore, by comparing the results for individual impurities in different formulations, the following observations were obtained: ■ Individual impurities at RRT 1.09 / 1.10 and RRT 1.13 / 1.14 are most relevant because they increase over time (only slowly and still well below the 1.0% acceptable threshold). ■ For polymers with lower molecular weights, the individual impurities at RRT 1.09 / 1.10 are relatively large. ■ For higher polymer / drug ratios, individual impurities at RRT 1.09 / 1.10 are larger. ■ Gamma radiation appears to have no significant effect on individual impurities at RRT 1.09 / 1.10. ■ Gamma radiation has a strong effect on individual impurities at RRT 1.13 / 1.14; such impurities are not observed in unradiated materials; i.e., < 0.1%. Example 2: Triptorelin microparticles (medium size) for 1 month

[0228] Using the components and process parameters described in the table below, a 1-month triptorelin microparticle formulation was prepared on a medium scale (51 g scale) in a manner similar to that described in Example 1. Unlike Example 1, the following steps were used to separate the microparticles from the suspension: after the evaporation step, the microparticle suspension was concentrated in a processing chamber by tangential / cross-flow filtration. The microparticle suspension in the processing chamber was washed with water to remove excess surfactant and unencapsulated API. Lyophilization and sterilization were performed in a manner similar to that described in Example 1.

[0229] The table below lists the components and properties of the particles used to prepare a 1-month formulation on a medium scale. Table 2. Triptorelin microparticles after 1 month (medium size; 51 g size) nd = Undetermined. Poloxamer 188 (EMPROVE® Expert), European Pharmacopoeia, obtained from Merck. PEG 6000, polyethylene glycol 6000, European Pharmacopoeia, obtained from Carl Roth. MOWIOL® 18-88, polyvinyl alcohol, sourced from Aldrich / Merck. Formulation 9 (Medium-scale) Formulation 10 ( Medium size) Formulation 11 (Medium-scale) Formulation 12 (Medium-scale) Formulation 13 (Medium-scale) Formulation 14 (Medium-scale) preparation 15 (Medium size) Formulation 16 (Medium-scale) Formulation 17 (Medium-scale) Formulation 18 (Medium-scale) Formulation 19 (Medium-scale) Formulation 20 (Medium-scale) Formulation 21 (Medium-scale) Formulation 22 (Medium Scale) Formulation 23 (Medium-scale) Formulation 24 (Medium-scale) Formulation 25 (Medium-scale) Formulation 26 (Medium-scale) Formulation 27 (Medium-scale) In vitro release (medium scale) of triptorelin microparticles after 1 month

[0230] The in vitro release of triptorelin microparticle formulations prepared at a medium scale was evaluated as described in Example 1. Formulations 9 to 27 (before gamma irradiation) exhibited significant drug release in vitro over seven days. A 1-month pharmacokinetic study of triptorelin microparticles (medium-sized)

[0231] For in vivo evaluation, a 1-month-old triptorelin-loaded microparticle formulation (formulation 9-20) prepared at a medium scale as described above was administered as a single subcutaneous (sc) injection to Sprague-Dawley (SD) rats to provide a dose of 1 mg triptorelin, with 8 rats in each treatment group. The study lasted for three (3) months. Blood samples were collected at the following time points for bioanalysis and pharmacokinetic assessment: days 1, 3, 5, 7, 14, 28, 42, 56, 70, and 84 post-administration. For the first four blood sampling time points, blood was collected from 4 animals in each group, allowing for staggered blood sampling. Body weight was measured on the day of administration and again on days 28, 56, and 84. Injection sites were carefully monitored.

[0232] Figures 5 to 8 Results for formulations 9 (RG503 (purified), PEG6000, F68), 12 (RG503 (purified) / RG502, PEG6000, F68), 16 (RG503 (purified), PEG4000, polyvinyl alcohol), and 19 (RG503, PEG3350, F68) are presented separately. All test groups achieved mean triptorelin plasma concentrations above 100 pg / mL at 1 month (672 hours). Two months after triptorelin administration, mean triptorelin plasma concentrations in all test groups were below the 100 pg / mL threshold. Stability study of triptorelin microparticles over one month (medium-sized)

[0233] The storage stability of the selected 1-month triptorelin-loaded microparticles in this example was assessed by monitoring the evolution of major impurities over time using HPLC analysis. The primary focus of the stability study was the evolution of the following major individual impurities with the largest relative peak areas at the following relative retention times (RRTs), which are relative to the retention time of the triptorelin peak: at RRT 1.09 / 1.10, RRT 1.13 / 1.14, RRT 1.16 / 1.17, RRT 1.18 / 1.19, and RRT 1.22 / 1.23. The evolution of these individual impurities was examined over 12 months of storage (T0, T1, T3, T6, T12 months) and exposure to different conditions (5°C ± 3°C, 25°C ± 2°C / 60% RH, 30°C ± 2°C / 75% RH). According to the ICH Q3B guidelines, for a daily dose of 3.75 mg of a drug, the identification threshold is 0.5%, and the acceptable threshold is 1.0%. Therefore, it is very important that a single impurity does not exceed 1.0%.

[0234] In summary, we observed the following results: Nuclear loadAfter an initial decrease due to gamma radiation, the core loading exhibited good stability over 12 months, almost independent of storage conditions. The molecular weight of PEG had no effect on the core loading, showing the same curves under all storage conditions for 12 months (e.g., formulation 14 (PEG 6000) and formulation 16 (PEG 4000)). Total impurities Following a strong increase in gamma radiation, the total impurities in samples stored at 5°C were, in most cases, slightly lower than those in samples stored at 25°C / 60% RH or 30°C / 65% RH. Largest single impurity All formulations prepared using the ester-terminated polymer (RG503, or a mixture of RG503 and RG502) exhibited good stability, with individual impurities remaining below the acceptable threshold of 1.0%. Furthermore, by comparing the results of individual impurities in different formulations, the following observations were obtained: ■ Individual impurities at RRT 1.09 / 1.10 depend on storage conditions above 25°C. ■ When using polymer blends (e.g., formulation 12 (RG503 / RG502)), the individual impurities at RRT 1.09 / 1.10 are relatively large. ■ Gamma radiation appears to have no significant effect on individual impurities at RRT 1.09 / 1.10. ■ Gamma radiation has a strong effect on individual impurities at RRT 1.17 / 1.18; such impurities are not observed in unradiated materials; i.e., < 0.1%. ■ When using PVA (e.g., formulations 14 and 16), the individual impurities at RRT 1.17 / 1.18 are relatively large. Residual ethyl acetate content After an initial decrease due to gamma radiation, the ethyl acetate content became very uniform and showed a continuous decreasing trend over 12 months at 25°C and above. A slight increase in residual EtAc was observed during 12 months of storage at 5°C ± 3°C. Residual water content Almost identical values ​​were observed at T0 and T12; the conclusion is that no water entered the vial over time. In vitro release curve All in vitro release curves showed very similar shapes. Gamma radiation caused a significant slowdown in the IVR curves. The IVR curves of samples stored for 12 months at 25°C / 60% RH or 30°C / 75% RH accelerated slightly, but remained lower than the pre-radiation IVR curves. Example 3: Triptorelin microparticles (medium size) for 3 months

[0235] Triptorelin microparticles for 3 months were prepared on a medium scale (51 g) using the following general method: Step 1: Prepare the feed solution for the particle formation step; Step 2: Particle formation; Step 3: Remove ethyl acetate from the polymer particles by evaporation; Step 4: Separate the particles from the suspension and wash them; and Step 5: Freeze-drying.

[0236] The prepared polymer solution and a known amount of drug solution were sequentially transferred to a dispersion vessel. The drug solution was then dispersed in the polymer solution under a nitrogen flow. After a dispersion time, a surfactant solution containing poloxamer 188 and sucrose was transferred to the dispersion vessel. This action caused a phase inversion, leading to the formation of triptorelin-loaded microparticles. As described above, the dispersion step temperature, dispersion step mixing rate, dispersion step mixing time, nitrogen flow rate, and phase inversion rate can affect the particle size and triptorelin release rate, and these can be selected and adjusted to prepare the triptorelin-loaded microparticles described in this example. The drug-loaded microparticle suspension contained excess organic polymer solvent as well as unencapsulated drug and excipients, which were removed in subsequent solvent evaporation and washing steps. A washing phase solution containing 16% poloxamer 188 at pH 6.5 was added to stabilize the microparticles. Excess polymer solvent was removed by applying vacuum and increasing temperature during continuous stirring with vibration. Ethyl acetate was removed by evaporation, thereby increasing the transition of ethyl acetate from the microparticles to the aqueous surfactant solution (continuous phase), resulting in complete hardening of the polymer microparticles. Following the evaporation step, the particulate suspension is concentrated in a processing chamber via tangential / cross-flow filtration. The particulate suspension in the processing chamber is washed with water to remove excess surfactant and unencapsulated API. Aliquots of the particulate suspension are transferred to lyophilized vials, which are then lyophilized under appropriate conditions to provide the triptorelin-loaded particulate formulation. The triptorelin-loaded particulate formulation is sterilized by gamma radiation at a dose of 28 kGy ± 10%.

[0237] The components and particle properties used to prepare a medium-scale 3-month triptorelin formulation are shown in the table below. Table 3. Triptorelin microparticle formulations after 3 months (medium scale) F68 = Poloxamer 188 (F68) (KOLLIPHOR® P188), European Pharmacopoeia, derived from BASF / BTC (Europe, GESMA, USA). Formulation 29 was reconstituted in different diluents and then tested in vivo. The particles of formulation 30 are separated during the manufacturing process: formulation 31 is prepared by subjecting a portion of the particles of formulation 30 to an additional 140 µm sieving step. The particles of formulation 34 are separated during the manufacturing process: formulation 35 is prepared by subjecting a portion of the particles of formulation 34 to γ-radiation sterilization. Formulation 28 (Medium-scale) Formulation 29 (Medium-scale) Formulation 30 (medium scale) Formulation 31 (Medium-scale) Example 32 (Medium Scale) Formulation 33 (Medium-scale) Formulation 34 (Medium-scale) Formulation 35 (Medium-scale) Example 36 (Medium Size) In vitro release (medium scale) of triptorelin microparticles over 3 months.

[0238] As described in Example 1, the in vitro release of triptorelin microparticle formulations prepared at a moderate scale (before gamma irradiation) for 3 months was evaluated. All formulations prepared at the moderate scale (formulations 28-36) showed significant drug release in vitro over seven days. A 3-month pharmacokinetic study of triptorelin microparticles (medium-sized)

[0239] For in vivo evaluation, 3-month-old triptorelin-loaded microparticle formulations (formulations 28-36) prepared at a medium scale as described above were administered as a single subcutaneous (sc) injection to Sprague-Dawley (SD) rats to provide a dose of 3 mg triptorelin, with 8 rats in each treatment group. (Formulation 35 was sterilized by gamma radiation prior to injection). The study duration was 112 days (4 months). The table below summarizes the PK study groups in this study. Blood samples were collected for bioanalysis and PK assessment at the following time points: days 1, 3, 5, 7, 14, 28, 42, 56, 84, and 112 post-administration. For the first four blood sampling time points, blood was collected from 4 animals in each group, allowing for staggered blood sampling. Body weight was measured on the day of administration and again on days 28, 84, and 112. Injection sites were carefully monitored.

[0240] The results of formulation 28 are as follows Figure 9 As shown, all test groups achieved mean triptorelin plasma concentrations above 100 pg / mL for at least 3 months (2016 hours).

[0241] Three-month triptorelin microparticles of formulation 29 were formulated for injection in two different carriers / diluents: (i) dextran 70 / polysorbate 80 diluent or (ii) sodium hyaluronate / polysorbate 80 diluent. PK results for these different compositions containing formulation 29 microparticles are as follows: Figure 10 As shown. Long-term stability study of triptorelin microparticles over 3 months (medium-sized)

[0242] The long-term stability of triptorelin microparticles in formulations 28–36 was evaluated over 24 months under the following three storage conditions: 5°C ± 3°C, 25°C ± 2°C / 60% RH ± 5% RH; 30°C ± 2°C / 65% RH ± 5% RH. Samples were analyzed after gamma irradiation (T0) and at the following time points: 1 month (T1), 3 months (T3), 6 months (T6), 9 months (T9), 12 months (T12), 18 months (T18), and 24 months (T24). The following properties were evaluated: core loading, purity, in vitro release, particle size distribution, morphology, appearance, residual water, residual solvent, and molecular weight distribution. Overall, we observed the following results: Nuclear load After an initial decrease due to gamma radiation, the nuclear load fluctuates around the T0 value, showing a slight decreasing trend over time, which is less noticeable for samples stored at 5°C. However, these differences are only minor. Total impuritiesFollowing a strong increase in gamma radiation, the total impurities in samples stored at 5°C were, in most cases, slightly lower than those in samples stored at 25°C / 60% RH or 30°C / 65% RH. Largest single impurity The largest single impurity among all gamma-radiation batches was found at RRT 1.08 / 1.09. The peak signals individually increased with storage time (up to approximately 12 months) and then remained at a constant level or even decreased slightly. Furthermore, temperature effects (more significant increases at higher storage temperatures) and dose effects (more significant increases at higher radiation doses) were observed. Residual ethyl acetate content After an initial decrease due to gamma radiation, the ethyl acetate content in samples stored at 5°C generally remained constant. However, a more significant reduction in ethyl acetate was observed with increasing storage temperature and time. Residual water content Almost identical values ​​were observed at T0, T12, and T24; the conclusion is that no water entered the vial over time. molecular weight of polymer Based on only a few data points (T6 and T12 values), a slight decrease in the molecular weight of the polymer was observed over time. This decrease became more pronounced with increasing storage temperature and gamma radiation dose. In vitro release curve All in vitro release profiles exhibited very similar shapes. Starting from a very low initial burst, the range of fluctuations increased with storage time, but remained within a reasonable range. Typically, gamma radiation was observed to result in a slower release rate. Example 4: Triptorelin microparticles for 3 months (commercial scale; 510 g)

[0243] Triptorelin microparticles for 3 months were prepared on a commercial scale (510 g) using the following general method: Step 1: Prepare the feed solution for the particle formation step; Step 2: Particle formation; Step 3: Remove ethyl acetate from the polymer particles by evaporation; Step 4: Separate the particles from the suspension and wash them; and Step 5: Freeze-drying.

[0244] The prepared polymer solution and a known amount of drug solution were transferred to a dispersion vessel, and the drug solution was dispersed in the polymer solution under a nitrogen flow. After a dispersion time, a surfactant solution containing poloxamer 188 and sucrose was transferred to the dispersion vessel. This caused a phase inversion, which in turn led to the formation of triptorelin-loaded microparticles. As described above, the dispersion step temperature, dispersion step mixing rate, dispersion step mixing time, nitrogen flow rate, and phase inversion rate can affect the particle size and triptorelin release rate, and are selected and adjusted to prepare the triptorelin-loaded microparticles described in this example. The drug-loaded microparticle suspension contained excess organic polymer solvent as well as unencapsulated drug and excipients, which were removed in subsequent solvent evaporation and washing steps, respectively. A washing phase solution containing 16% poloxamer 188 at pH 6.5 was added to stabilize the microparticles. Excess polymer solvent was removed by applying vacuum and increasing temperature during continuous stirring with vibration. Ethyl acetate was removed by evaporation, thereby increasing the transition of ethyl acetate from the microparticles to the aqueous surfactant solution (continuous phase), resulting in complete hardening of the polymer microparticles. Following the evaporation step, the particulate suspension is concentrated in a processing chamber via tangential / cross-flow filtration. The particulate suspension in the processing chamber is washed with water to remove excess surfactant and unencapsulated drug. Aliquots of the particulate suspension are transferred to lyophilized vials, which are then lyophilized under appropriate conditions to provide triptorelin-loaded particulate formulations. The triptorelin-loaded particulate formulations are sterilized by gamma radiation at a dose of 25 kGy ± 10%.

[0245] The components and particle properties used to prepare a commercial-scale 3-month triptorelin formulation are shown in the table below. Table 4. 3-month microparticle formulations (commercial scale; 510 g scale) F68 = Poloxamer 188 (F68) (KOLLIPHOR® P188), European Pharmacopoeia, from BASF / BTC (Europe, Gesmar, USA), or Poloxamer 188, from ThermoFisher Scientific (formerly Alfa Aesar). Formulation 37 (Commercial Scale) Formulation 38 (Commercial Scale) Formulation 39 (Commercial Scale) Formulation 40 (commercial scale) Formulation 41 (Commercial Scale) Formulation 42 (Commercial Scale) In vitro release of triptorelin microparticles over 3 months (commercial scale)

[0246] As described in Example 1, the in vitro release of triptorelin-loaded microparticles from formulations 37 and 38 over 3 months (before and after gamma irradiation) was evaluated, and significant amounts of triptorelin were released over a seven-day period. A 3-month pharmacokinetic study of triptorelin microparticles (commercial scale).

[0247] The first commercial-scale study investigated the release profiles of a commercially prepared 3-month triptorelin microparticle formulation (Formulation 37) by evaluating plasma concentrations obtained after a single subcutaneous (sc) injection into spragdolli (SD) rats. The study was conducted in eight male spragdolli rats and lasted for six months.

[0248] The second commercial-scale study compared the release profiles of a commercially prepared 3-month triptorelin microparticle formulation (Formulation 38) with those of the approved 3-month release reference formulation (Decapeptyl SR, 11.25 mg), assessed by evaluating plasma concentrations obtained after a single subcutaneous (sc) injection into Spraguedoli (SD) rats. This study was conducted in 16 male Spraguedoli rats, divided into two treatment groups of eight rats each. The study lasted for six months.

[0249] In both studies, blood samples were collected at the following time points for bioanalysis and pharmacokinetic (PK) assessment: 1, 3, 5, 7, 14, 28, 42, 56, 70, 84, 112, 140, and 168 days post-drug administration. For the first four blood sampling time points, blood was collected from four animals in each group, allowing for staggered blood sampling.

[0250] In the first study, the test formulation (formulation 37) had a confirmed sustained release profile in which individual and mean triptorelin plasma concentrations were above the clinically relevant 100 pg / mL threshold for at least 4 months. Figure 11In the first study, the test formulation (formulation 38) exhibited a confirmed sustained release profile, with individual and mean triptorelin plasma concentrations above the clinically relevant 100 pg / mL threshold for at least 4 months, and the concentration-time profile subsequently decreased steadily to approximately 100 pg / mL or lower for up to 6 months (data not shown). The reference formulation (Decapeptyl SR) showed significant variation in its profile up to 3 months prior, but all individual measurements remained below the limit of detection after 4 months. PK study of triptorelin microparticles after 3 months of storage (commercial scale)

[0251] To investigate the effect of potential degradation during 12 and 22 months of storage at 25°C on the pharmacokinetics of a commercially prepared 3-month triptorelin microparticle formulation, plasma concentrations obtained after a single subcutaneous (sc) injection into Spraguedale (SD) rats were evaluated. Specifically, this study evaluated the release profiles of a commercially prepared 3-month triptorelin microparticle formulation (Formulation 38; PK group 4) after 12 months of storage at 25°C, and a medium-scale prepared 3-month triptorelin microparticle formulation (Formulation 28; PK group 5) after 22 months of storage at 25°C. Each treatment group consisted of 8 rats and included administration of 3.0 mg triptorelin. The study lasted for up to 6 months.

[0252] Blood samples were collected at the following time points for bioanalysis and pharmacokinetic (PK) assessment: 1, 3, 5, 7, 14, 28, 42, 56, 70, 84, 112, 140, and 168 days post-drug administration. For the first four blood sampling time points, blood was collected from four animals in each group, allowing for staggered blood sampling. The test group achieved mean triptorelin plasma concentrations above 100 pg / mL for at least 3 months (2016 hours). At 6 months, the mean concentration in the test group decreased to below 100 pg / mL. Example 5: Triptorelin microparticles for 6 months (laboratory scale)

[0253] The formulation containing triptorelin microparticles prepared in this example provided drug release above the minimum target level (e.g., 0.1 ng / mL) over 6 months after subcutaneous (SC) injection into rats. The components used to prepare the 6-month formulation containing triptorelin microparticles are shown below. Table 5. R203S (Evonik Industries): lactide / glycolic acid ratio of 100:0, with ester-terminal groups. R205S (Evonik Industries): lactide / glycolic acid ratio of 100:0, with ester-terminal groups. F68 = Poloxamer 188 (F68) (KOLLIPHOR® P188), European Pharmacopoeia, derived from BASF / BTC (Europe, Gesmar, USA).

[0254] The method for manufacturing these triptorelin formulations is similar to that described in Example 1. Particle formation and solvent removal are performed in a manner similar to that described in Example 1. In this example, the particles are separated from the continuous phase by centrifugation. The supernatant containing residual solvent and excipients is discarded. The particles are resuspended in ultrapure water and centrifuged again. The supernatant is discarded, and the particles are subsequently resuspended in a surfactant solution (0.25% poloxamer 188) and centrifuged again. Lyophilization and sterilization are performed in a manner similar to that described in Example 1.

[0255] The table below summarizes the physical properties of the triptorelin-loaded microparticle formulation prepared in this example.

[0256] Six-month-old triptorelin-loaded microparticles were administered to Spraguedoli (SD) rats via a single subcutaneous (sc) injection, and plasma levels of triptorelin were monitored. Figure 12 Mean pharmacokinetic curves and standard deviations are shown. All individual animals achieved plasma concentrations of triptorelin higher than 100 pg / mL for at least 6 months after administration. Figure 12 ). Example 6: Triptorelin microparticles (medium size) for 6 months

[0257] A formulation containing triptorelin-loaded microparticles, designed to provide drug release over 6 months, was prepared on a medium-scale basis. The components used to prepare this 6-month formulation are shown in the table below. Table 6. R203S (Evonik Industries): lactide / glycolic acid ratio of 100:0, with ester-terminal groups. R205S (Evonik Industries): lactide / glycolic acid ratio of 100:0, with ester-terminal groups. F68 = Poloxamer 188 (F68) (KOLLIPHOR® P188), European Pharmacopoeia, derived from BASF / BTC (Europe, Gesmar, USA).

[0258] The microparticle manufacturing method used to prepare these triptorelin formulations is similar to that described in Example 1. Microparticle formation and solvent removal are performed in a manner similar to that described in Example 3. Lyophilization and sterilization are carried out in a manner similar to that described in Example 3.

[0259] The table below summarizes the physical properties of the triptorelin-loaded microparticle formulation prepared in this example.

[0260] The in vitro release of the 6-month triptorelin-loaded microparticle formulation prepared on a medium scale was evaluated as described in Example 1, and significant drug release was observed in vitro over 7 days. The results indicate that this example, prepared on a medium scale using 51 g of polymer, reproduced the in vitro release profile of Example 5, prepared on a smaller scale. Example 6: 3-month FSH microparticles (laboratory scale; 3 g)

[0261] Using a method similar to that described in Example 1, follicle-stimulating hormone (FSH) delta was used as the drug (API) instead of triptorelin to prepare FSH-loaded microparticles designed to provide drug release over at least 3 months.

[0262] Follicle-stimulating hormone (FSH) microparticles were prepared from the FSH-δ drug substance and provided in the form of a buffer solution with a concentration of 0.6 mg / mL. The FSH-δ drug substance solution was concentrated by ultracentrifugation prior to microparticle preparation.

[0263] The formation of the microparticles and the removal of the solvent were carried out in a manner similar to that described in Example 1, but using the components and process parameters described in the table below. In this example, the microparticles were separated from the continuous phase by centrifugation, wherein the supernatant containing residual solvent and excipients was discarded. The microparticles were resuspended in ultrapure water and centrifuged again, the supernatant was discarded, and the microparticles were resuspended in a surfactant solution (0.25% poloxamer 188) and centrifuged again. Lyophilization and sterilization were performed in a manner similar to that described in Example 1.

[0264] The properties of the compositions used to prepare the formulations and the FSH microparticles are shown in the table below. Table 7. FSH microparticle formulations after 3 months (laboratory scale) F68 = Poloxamer 188 (F68) (KOLLIPHOR® P188), European Pharmacopoeia, sourced from BASF / BTC (Europe, GESMA, USA); PEG3000, European Pharmacopoeia, sourced from Merck; PEG6000, European Pharmacopoeia, sourced from Carl Roth. Formulation 45 (Laboratory Scale) Formulation 46 (Laboratory Size) Formulation 47 (Laboratory Scale) Formulation 48 (Laboratory Size) Formulation 49 (Laboratory Scale) Formulation 50 (laboratory scale) Formulation 51 (Laboratory Scale)

[0265] The in vitro release of follicle-stimulating hormone (FSH)-loaded delta-microparticle formulations over 3 months was evaluated as follows. For each sampling point, an aliquot of 30 mg microparticles was weighed into a polypropylene screw container, and 1.5 mL of aqueous buffer was added to each aliquot as the release medium. The samples were placed on a top-mounted shaker and stirred at 37°C. At appropriate sampling times (0 h, 4 h, 24 h, 3 days, 7 days, and 10 days), both samples were centrifuged to separate the microparticles from the release medium. The release medium was filtered through a syringe pre-filter, and the FSH content was determined by HPLC analysis. The in vitro release of FSH-loaded delta-microparticles from formulation 45 over 3 months showed a significant release of FSH over 7 days in vitro. Size exclusion chromatography of formulations 45 and 46 confirmed that the FSH released in the in vitro studies was structurally intact. It was found that the volume of the aqueous phase of the drug (e.g., drug volumetric loading) affected the core loading and the in vitro release profile. PK study of FSH particles over 3 months

[0266] The 3-month-old follicle-stimulating hormone (FSH)-loaded microparticles prepared as described above were administered to Sprague-Dawley (SD) rats (n=8 per group) via a single subcutaneous (sc) injection (0.218 mg / 0.6 mL), and plasma FSH-loaded FSH levels were monitored. Figure 13 and Figure 14 The mean FSH plasma concentrations of formulations 45 and 51 over 12 weeks are shown. Formulations 45, 46, and 51 exhibited prolonged exposure over 12 weeks, with release profiles comparable to those of triptorelin microparticles. Example 7: Diluent Study

[0267] Studies were conducted to evaluate formulations containing dextran or sodium hyaluronate diluents. Viscosity study Formulations having the following components were prepared to compare the viscosity properties of formulations having dextran diluent or sodium hyaluronate diluent. preparation

[0268] Viscosity tests were performed on a research rheometer (DHR2, TA Instruments) equipped with a 40 mm 1° cone measuring system, with a test gap set to 29 µm. A solvent trap cap was used to minimize drying of the sample at the exposed edges. After equilibration at 25°C for 30 seconds, the sample was exposed to a 30-second pre-shear at a rate of 0.1 s⁻¹. This immediately resulted in a shear rate scan, from 0.1 s⁻¹ to 10000 s⁻¹, logarithmically scaled, with 8 points per ten times the shear rate, applying shear for 30 seconds at each rate, and calculating the viscosity in the last 5 seconds of each step.

[0269] An important parameter derived from viscosity data is the degree of viscoelasticity, which is the ratio of viscosity at low shear (“zero-shear viscosity”) to viscosity at high shear. HA has been found to provide high viscosity at low shear rates, making it suitable as a non-flowing support for delaying the settling of drug-loaded particles. However, for injectable formulations, the diluent also advantageously exhibits low viscosity at high shear rates (i.e., when it is injected), making injection via syringes and needles relatively easy. The data below present viscoelasticity data calculated as the ratio of viscosity at a shear rate of 0.1 s⁻¹ (near zero shear) to viscosity at a shear rate of 1000 s⁻¹, illustrating the advantageous properties of the HA diluent. Injectability studies

[0270] To assess injectability, approximately 235 mg of lyophilized 3-month-old follicle-stimulating hormone-loaded delta-microparticles (with an average volume diameter of 49 µm) were weighed into vials. Ten vials were prepared, sealed with stoppers, and capped with aluminum caps. Then, 1.2 ml of diluent (containing the above-described components) was loaded into a 2 ml syringe, transferred to each vial vial adapter, and the microparticles were reconstituted by gently rotating and tapping the vials. After reconstitution, the resulting microparticle suspension was aspirated back into the syringe, and a 13 mm 27G ultrathin-walled needle was attached to the syringe. Testing was performed on a Stable Micro Systems TA.XTplusC texture analyzer operated at a fixed plunger speed of 2.5 mm / s. The formulation with the HA diluent exhibited better injectability, determined by the maximum force required to expel the syringe contents. plasma protein adsorption

[0271] Under reducing conditions, the adsorption of human plasma proteins to 3-month-old follicle-stimulating hormone (FSH)-loaded microparticles was assessed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) combined with Coomassie brilliant blue staining. For this study, duplicate samples were prepared by incubating 10 mg of lyophilized FSH-loaded microparticles with 0.7 mL of a solution containing (A) 10 mg / mL Tween 80, 10 mM sodium phosphate (pH 7.2) and (B) a dextran-based diluent (see above) or (C) 10 mg / mL Tween 80, 10 mM sodium phosphate, and 15 mg sodium hyaluronate (MW 3.2 M g / mol). 0.3 mL of human plasma was then added to each vial and incubated further at 37°C for 1 hour, followed by washing the microparticles three times with purified water (by centrifugation at 3000 rpm and resuspending three times in 1 mL of purified water). The resulting microparticles were incubated with SDS buffer under reducing conditions, and the extracted proteins were applied to molecularly labeled 4%–12% BisTris gels in lane 1.

[0272] The results (not shown) indicated that microparticles pre-incubated with a dextran-based diluent exhibited significantly greater adsorption of plasma proteins, while microparticles pre-incubated with a HA-based diluent showed the lowest adsorption. As mentioned above, this could potentially influence the immune response, since plasma proteins are known to function as complement with potentially pro-inflammatory effects. PK Research

[0273] The 3-month follicle-stimulating hormone (FSH)-loaded microparticles prepared as described above and formulated with dextran-based or HA-based diluents (as described above) were administered to Sprague-Dawley (SD) rats (n=8 per group) via a single subcutaneous (sc) injection (0.218 mg / 0.6 mL), and plasma FSH levels were monitored. Figure 15 The average FSH plasma concentration over 12 weeks is shown. As the figure illustrates, the diluent has a surprising effect on FSH plasma concentration, with the HA-based formulation maintaining higher plasma levels for a longer period. A similar study was conducted using triptorelin microparticles for 3 months, as described above. Figure 10 The mean plasma concentration of triptorelin over 12 weeks is shown. Unlike results obtained with follicle-stimulating hormone (FSH)-loaded microparticles, the plasma concentration-time curves for triptorelin-loaded microparticles are not affected by the carrier (e.g., results obtained with both formulations are substantially the same). immune response studies

[0274] The immune response of rats to subcutaneous injection of follicle-stimulating hormone (FSH)-loaded delta microparticles prepared as described above and diluted with either the dextran-based or HA-based diluents as described above was also evaluated. The HA formulation was found to possess anti-inflammatory properties and reduced local inflammatory responses of FSH-loaded delta microparticles to infiltrating neutrophils, monocytes, and macrophages by interacting with CD44 receptors on these cell types (data not shown). Although the levels of IFN-γ, TNF-α, and IL-6 were similar in both groups, the HA group showed a higher level of IL-4, indicating an anti-inflammatory response. Figure 16 In addition, although the total antibody signals were similar in both groups, the higher plasma concentration of FSH in the HA group indicated that the amount of neutralizing antibodies in the HA group was lower than that in the dextran group, further supporting the reduced immunogenicity of the formulation in the HA-based diluent.

Claims

1. A drug-loaded microparticle, the drug-loaded microparticle comprising: (a) Medicine; (b) A polymer component comprising one or more polymers selected from ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymers with a lactide:co-lactide ratio of about 50:50, ester-terminated PLGA polymers with a lactide:co-lactide ratio of about 75:25, and ester-terminated polylactide (PLA) polymers. (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release the drug in the body for at least one week after being administered via subcutaneous injection.

2. The drug-loaded microparticles of claim 1, wherein the drug is selected from one or more small molecule drugs, peptide drugs, and protein drugs.

3. The drug-loaded microparticles according to any one of claims 1-2, wherein the drug is selected from one or more of the following: triptorelin, follicle-stimulating hormone (FSH), luteinizing hormone (LH), human chorionic gonadotropin (hCG), atosiban, insulin, abiraterone, balusiban, degarelix, desmopressin, ganirelix (e.g., ganirelix acetate), gemcitabine, gonadorelin (e.g., gonadorelin acetate), urinary gonadotropin or human menopausal gonadotropin (hMG), meropenem, progesterone, quinolones, somatostatin, growth hormone, and tolterodine.

4. The drug-loaded microparticles of claim 1, wherein the drug comprises two or more drugs, optionally wherein the two or more drugs comprise FSH and LH and / or hCG.

5. The drug-loaded microparticles according to any one of claims 1-4, wherein the average particle size of the drug-loaded microparticles is from about 2 µm to about 180 µm.

6. The drug-loaded microparticles according to any one of claims 1-5, wherein the drug is triptorelin.

7. The drug-loaded microparticles of claim 6, wherein the triptorelin is triptorelin acetate.

8. The drug-loaded microparticle as described in any one of claims 6-7, wherein the drug-loaded microparticle comprises: (a) Triptorelin; (b) Lactide: One or more ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymers with a lactide ratio of about 50:50; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release triptorelin in the body for at least one month after being administered via subcutaneous injection.

9. The drug-loaded microparticles of claim 8, wherein the lactide:glycol ratio of one or more ester-terminated PLGA polymers comprising one or more selected from RG502, RG503 and RG504 in an amount of about 50:

50.

10. The drug-loaded microparticles of claim 8 or claim 9, wherein the lactide:glycol ratio of one or more ester-terminated PLGA polymers is an ester-terminated PLGA polymer with a lactide:glycol ratio of about 50:

50.

11. The drug-loaded microparticles of claim 9, wherein the ester-terminated PLGA polymer having a lactide:glycol ratio of about 50:50 is RG503.

12. The drug-loaded microparticles of claim 8 or claim 9, wherein the one or more ester-terminated PLGA polymers having a lactide:glycol ratio of about 50:50 comprise two different ester-terminated PLGA polymers having a lactide:glycol ratio of about 50:

50.

13. The drug-loaded microparticles of claim 12, wherein the two different ester-terminated PLGA polymers are present in a weight ratio of lower molecular weight PLGA to higher molecular weight PLGA of about 99:1 to about 51:49, such as about 99:1, about 90:10, about 80:20, about 70:30, about 60:40, about 55:45, or about 51:

49.

14. The drug-loaded microparticles of claim 12 or claim 13, wherein the two different ester-terminated PLGA polymers having a lactide:glycol ratio of about 50:50 are RG504 and RG503.

15. The drug-loaded microparticles according to any one of claims 5-14, wherein the weight ratio of triptorelin to one or more ester-terminated PGLA polymers is about 4 / 100 to about 10 / 100.

16. The drug-loaded microparticles as described in any one of claims 6-7, wherein the drug-loaded microparticles comprise: (a) Triptorelin; (b) Lactide: an ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymer with a lactide ratio of approximately 75:25; (c) Surfactants; and (d) One or both of basic salts and polyols. The microparticles release triptorelin in the body for at least 3 months after being administered via subcutaneous injection.

17. The drug-loaded microparticles of claim 16, wherein the ester-terminated PLGA polymer is RG753S.

18. The drug-loaded microparticles of any one of claims 16-17, wherein the weight ratio of triptorelin to the ester-terminated PGLA polymer is about 5 / 100 to about 10 / 100.

19. The drug-loaded microparticle as described in any one of claims 6-7, wherein the drug-loaded microparticle comprises: (a) Triptorelin; (b) Two different ester-terminated polylactide (PLA) polymers; (c) Surfactants; and (d) One or both of basic salts and polyols. The microparticles release triptorelin in the body for at least 6 months after being administered via subcutaneous injection.

20. The drug-loaded microparticles of claim 19, wherein the two different ester-terminated PLA polymers are present in a weight ratio of lower molecular weight PLA to higher molecular weight PLA of about 90:10 to about 60:40, such as about 90:10, about 85:15, about 80:20, about 75:25, about 70:30, about 65:35, or about 60:

40.

21. The drug-loaded microparticles of claim 19 or claim 20, wherein the two different ester-terminated polylactide PLA polymers are R203S and R205S.

22. The drug-loaded microparticles according to any one of claims 19-21, wherein the weight ratio of triptorelin to PLA polymer is about 5 / 100 to about 15 / 100.

23. The drug-loaded microparticles according to any one of claims 1-5, wherein the drug is FSH, optionally wherein the FSH is recombinant FSH, optionally wherein the FSH is selected from follicle-stimulating hormone α, follicle-stimulating hormone β and follicle-stimulating hormone δ, and further optionally wherein the FSH is follicle-stimulating hormone δ.

24. The drug-loaded microparticle of claim 23, wherein the drug-loaded microparticle comprises: (a) FSH; (b) Lactide: One or more ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymers with a lactide ratio of about 50:50; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release FSH in the body for at least one month after being administered via subcutaneous injection.

25. The drug-loaded microparticle of claim 23, wherein the drug-loaded microparticle comprises: (a) FSH; (b) Lactide: One or more ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymers with a lactide ratio of about 50:50; (c) Surfactants; and (d) One or both of the following: an optional basic salt and a polyol. The microparticles release FSH in the body for at least 6 weeks after being administered via subcutaneous injection.

26. The drug-loaded microparticles of claim 24 or claim 25, wherein the lactide:glycol ratio of one or more ester-terminated PLGA polymers comprising one or more selected from RG502, RG503 and RG504 in a ratio of about 50:

50.

27. The drug-loaded microparticle of claim 23, wherein the drug-loaded microparticle comprises: (a) FSH; (b) Lactide: an ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymer with a lactide ratio of approximately 75:25; (c) Surfactants; and (d) One or both of basic salts and polyols. The microparticles release FSH in the body for at least 3 months after being administered via subcutaneous injection.

28. The drug-loaded microparticles of claim 27, wherein the ester-terminated PLGA polymer is RG753S.

29. The drug-loaded microparticles according to any one of claims 27-28, wherein the weight ratio of FSH to the ester-terminated PGLA polymer is about 5 / 100 to about 10 / 100.

30. The drug-loaded microparticle of claim 23, wherein the drug-loaded microparticle comprises: (a) FSH; (b) Two different ester-terminated polylactide (PLA) polymers; (c) Surfactants; and (d) One or both of basic salts and polyols. The microparticles release FSH in the body for at least 6 months after being administered via subcutaneous injection.

31. The drug-loaded microparticles of claim 30, wherein the two different ester-terminated PLA polymers are present in a weight ratio of lower molecular weight PLA to higher molecular weight PLA of about 90:10 to about 60:40, such as about 90:10, about 85:15, about 80:20, about 75:25, about 70:30, about 65:35, or about 60:

40.

32. The drug-loaded microparticles according to any one of claims 30-31, wherein the two different ester-terminated polylactide PLA polymers are R203S and R205S.

33. The drug-loaded microparticles according to any one of claims 30-32, wherein the weight ratio of FSH to PLA polymer is about 5 / 100 to about 15 / 100.

34. The drug-loaded microparticles according to any one of claims 1-33, wherein the drug-loaded microparticles comprise poloxamer 188 or polyvinyl alcohol as a surfactant.

35. The drug-loaded microparticles according to any one of claims 1-34, wherein the drug-loaded microparticles comprise sodium chloride as an alkaline salt.

36. The drug-loaded microparticles according to any one of claims 1-35, wherein the drug-loaded microparticles comprise sucrose as a polyol.

37. The drug-loaded microparticle as claimed in any one of claims 1-36, wherein the drug-loaded microparticle comprises one or more of the following: The content of polyols is from about 0.1% w / w to about 5% w / w, such as the content of sucrose and / or polyethylene glycol from about 0.20% w / w to about 5% w / w. The residual content of acids such as acetic acid is from about 0.01% w / w to about 1.0% w / w. Residual content of buffer salts such as citric acid, ranging from approximately 0.01% w / w to approximately 1.0% w / w; The content of residual organic solvents, such as ethyl acetate, is from about 50 ppm to about 20,000 ppm; and Surfactant content of approximately 0% w / w to approximately 4% w / w, such as poloxamer 188 and / or polyethylene glycol content.

38. A pharmaceutical composition comprising drug-loaded microparticles as described in any one of claims 1-37.

39. The pharmaceutical composition of claim 38, wherein the pharmaceutical composition is formulated for subcutaneous injection.

40. The pharmaceutical composition of claim 39, wherein the pharmaceutical composition is formulated for intramuscular injection.

41. The pharmaceutical composition of any one of claims 38-40, wherein the pharmaceutical composition comprises the drug-loaded microparticles and one or more pharmaceutically acceptable carriers, diluents and / or excipients.

42. The pharmaceutical composition according to any one of claims 38-41, wherein the pharmaceutical composition comprises the drug-loaded microparticles and a pharmaceutically acceptable diluent selected from carboxymethyl cellulose, dextran, and hyaluronic acid and its salts.

43. The pharmaceutical composition of any one of claims 38-42, wherein the pharmaceutical composition comprises the drug-loaded microparticles and a pharmaceutically acceptable diluent comprising hyaluronic acid and / or a salt thereof, optionally wherein the hyaluronic acid and / or the salt thereof has an average molecular weight of about 1 MDa to about 5 MDa, further optionally wherein the hyaluronic acid and / or the salt thereof has an average molecular weight of about 3.2 MDa, and further optionally wherein the composition comprises about 0.01% (w / v) to about 5% (w / v) of the hyaluronic acid and / or the salt thereof based on the volume of the pharmaceutical composition, including about 1% (w / v) to about 3% (w / v), or about 1.5 mg of hyaluronic acid and / or the salt thereof per mL of the pharmaceutical composition.

44. A pharmaceutical composition comprising triptorelin-loaded microparticles as claimed in any one of claims 1-7 or 34-37, the pharmaceutical composition being formulated to contain an amount of such triptorelin-loaded microparticles to provide about 1 mg of triptorelin in a single dose, and / or to provide a mean plasma concentration of at least 0.1 ng / mL of triptorelin for at least one week following administration of a single dose by subcutaneous or intramuscular injection.

45. A pharmaceutical composition comprising triptorelin-loaded microparticles as claimed in any one of claims 1-15 or 34-37, the pharmaceutical composition being formulated to contain an amount of such triptorelin-loaded microparticles to provide 3.75 mg of triptorelin in a single dose, and / or to provide a mean plasma concentration of at least 0.1 ng / mL of triptorelin for at least one month following administration of a single dose via subcutaneous or intramuscular injection.

46. ​​A pharmaceutical composition comprising triptorelin-loaded microparticles as claimed in any one of claims 16-18 or 34-37, the pharmaceutical composition being formulated to contain an amount of such triptorelin-loaded microparticles to provide 11.25 mg of triptorelin in a single dose, and / or to provide a mean plasma concentration of at least 0.1 ng / mL of triptorelin for at least 3 months following administration of a single dose via subcutaneous or intramuscular injection.

47. A pharmaceutical composition comprising triptorelin-loaded microparticles as claimed in any one of claims 19-22 or 34-37, the pharmaceutical composition being formulated to contain an amount of such triptorelin-loaded microparticles to provide 22.5 mg of triptorelin in a single dose, and / or to provide a mean plasma concentration of at least 0.1 ng / mL of triptorelin for at least 6 months following administration of a single dose via subcutaneous or intramuscular injection.

48. The pharmaceutical composition of any one of claims 44-47, wherein the pharmaceutical composition comprises the drug-loaded microparticles and one or more pharmaceutically acceptable carriers, diluents and / or excipients.

49. The pharmaceutical composition of any one of claims 44-47, wherein the pharmaceutical composition comprises the drug-loaded microparticles and a pharmaceutically acceptable diluent selected from carboxymethyl cellulose, dextran, and hyaluronic acid and its salts.

50. The pharmaceutical composition of any one of claims 44-47, wherein the pharmaceutical composition comprises the drug-loaded microparticles and a pharmaceutically acceptable diluent comprising hyaluronic acid and / or its salt, optionally wherein the hyaluronic acid and / or its salt has an average molecular weight of about 1 MDa to about 5 MDa, further optionally wherein the hyaluronic acid and / or its salt has an average molecular weight of about 3.2 MDa, and further optionally wherein the composition comprises about 0.01% (w / v) to about 5% (w / v) of the hyaluronic acid and / or its salt based on the volume of the pharmaceutical composition, including about 1% (w / v) to about 3% (w / v), or about 1.5 mg of hyaluronic acid and / or its salt per mL of the pharmaceutical composition.

51. A pharmaceutical composition comprising FSH-loaded microparticles as claimed in any one of claims 23, 24, 26 or 34-37, the pharmaceutical composition being formulated to contain an amount of such FSH-loaded microparticles to provide about 0.35 mg or more of FSH (e.g., follicle-stimulating hormone delta) in a single dose, and / or to provide a mean plasma concentration of at least 14 mlU / mL of FSH for at least one month following administration of a single dose via subcutaneous or intramuscular injection.

52. A pharmaceutical composition comprising FSH-loaded microparticles as claimed in any one of claims 23-26 or 34-37, the pharmaceutical composition being formulated to contain an amount of such FSH-loaded microparticles to provide about 0.5 mg or more of FSH (e.g., follicle-stimulating hormone delta) in a single dose, and / or to provide a mean plasma concentration of at least 14 mlU / mL of FSH for at least 6 weeks following administration of a single dose via subcutaneous or intramuscular injection.

53. A pharmaceutical composition comprising FSH-loaded microparticles as claimed in any one of claims 27-29 or 34-37, the pharmaceutical composition being formulated to contain an amount of such FSH-loaded microparticles to provide about 1-5 mg of FSH (e.g., follicle-stimulating hormone delta) in a single dose, and / or to provide a mean plasma concentration of at least 14 mlU / mL of FSH for at least 3 months following administration of a single dose via subcutaneous or intramuscular injection.

54. A pharmaceutical composition comprising FSH-loaded microparticles as claimed in any one of claims 30-37, the pharmaceutical composition being formulated to contain an amount of such FSH-loaded microparticles to provide about 2-10 mg of FSH (e.g., follicle-stimulating hormone delta) in a single dose, and / or to provide a mean plasma concentration of at least 14 mlU / ml of FSH for at least 6 months following administration of a single dose via subcutaneous or intramuscular injection.

55. The pharmaceutical composition of any one of claims 51-54, wherein the pharmaceutical composition comprises the drug-loaded microparticles and one or more pharmaceutically acceptable carriers, diluents and / or excipients.

56. The pharmaceutical composition of any one of claims 51-54, wherein the pharmaceutical composition comprises the drug-loaded microparticles and a pharmaceutically acceptable diluent selected from carboxymethyl cellulose, dextran, and hyaluronic acid and its salts.

57. The pharmaceutical composition of any one of claims 51-54, wherein the pharmaceutical composition comprises the drug-loaded microparticles and a pharmaceutically acceptable diluent comprising hyaluronic acid and / or a salt thereof, optionally wherein the hyaluronic acid and / or the salt thereof has an average molecular weight of about 1 MDa to about 5 MDa, further optionally wherein the hyaluronic acid and / or the salt thereof has an average molecular weight of about 3.2 MDa, and further optionally wherein the composition comprises about 0.01% (w / v) to about 5% (w / v) of the hyaluronic acid and / or the salt thereof based on the volume of the pharmaceutical composition, including about 1% (w / v) to about 3% (w / v), or about 1.5 mg of hyaluronic acid and / or the salt thereof per mL of the pharmaceutical composition.

58. A treatment method comprising administering, by subcutaneous or intramuscular injection, the pharmaceutical composition as described in any one of claims 38-57 to a subject in need of the treatment.

59. A method of administering triptorelin to a subject in need of treatment, the method comprising administering, by subcutaneous or intramuscular injection, triptorelin-loaded microparticles as claimed in any one of claims 1-22 or 34-37, or a composition comprising triptorelin-loaded microparticles as claimed in any one of claims 38-50, optionally wherein the subject requires treatment for one or more of hormone-responsive cancer, endometriosis, female infertility, uterine fibroids, and central precocious puberty.

60. A method of administering FSH to a subject in need of it, the method comprising administering, by subcutaneous or intramuscular injection, an FSH-loaded microparticle as claimed in any one of claims 23-37 or a composition comprising FSH-loaded microparticles as claimed in any one of claims 38-43 or 51-57, optionally wherein the subject requires treatment for female infertility or male infertility.

61. A kit comprising (a) a container containing drug-loaded microparticles in lyophilized form as described in any one of claims 1-37 and (b) instructions for preparing a pharmaceutical composition comprising the microparticles and a pharmaceutically acceptable carrier.

62. A method for preparing drug-loaded microparticles, the method comprising: (a) Add (i) the drug solution containing the drug in an aqueous solvent to (ii) a polymer solution containing one or more polymers in an ethyl acetate solution, the one or more polymers being selected from ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymers with a lactide:co-lactide ratio of about 50:50, ester-terminated PLGA polymers with a lactide:co-lactide ratio of about 75:25, and ester-terminated polylactide (PLA) polymers, and mix to obtain a dispersion of the drug solution in the polymer solution; as well as (b) Add (iii) a phase inversion solution comprising a surfactant and optionally one or two of an alkaline salt and a polyol to the dispersion, and mix to obtain a suspension comprising the drug and drug-loaded microparticles of the one or more polymers selected from ester-terminated PLGA polymers with a lactide: glycolide ratio of about 50:50, ester-terminated PLGA polymers with a lactide: glycolide ratio of about 75:25, and ester-terminated PLA polymers.

63. The method of claim 62, wherein the drug is selected from one or more small molecule drugs, peptide drugs, and protein drugs.

64. The method of claim 62 or claim 63, wherein the drug is selected from one or more of the following: triptorelin, follicle-stimulating hormone (FSH), luteinizing hormone (LH), human chorionic gonadotropin (hCG), atosiban, insulin, abiraterone, balusiban, degarelix, desmopressin, ganirelix (e.g., ganirelix acetate), gemcitabine, gonadorelin (e.g., gonadorelin acetate), urinary gonadotropin or human menopausal gonadotropin (hMG), meropenem, progesterone, quinolones, somatostatin, growth hormone, and tolterodine.

65. The method of any one of claims 62-64, wherein the drug is triptorelin, optionally wherein the triptorelin is triptorelin acetate.

66. The method of any one of claims 62-64, wherein the drug is FSH, optionally wherein the FSH is recombinant FSH, optionally wherein the FSH is selected from follicle-stimulating hormone α, follicle-stimulating hormone β and follicle-stimulating hormone δ, and further optionally wherein the FSH is follicle-stimulating hormone δ.

67. The method of any one of claims 62-66, wherein the drug comprises two or more drugs, optionally wherein the drug comprises FSH and LH and / or hCG.

68. A method for preparing triptorelin-loaded microparticles, the method comprising: (a) Add (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution of a mixture of one or more ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymers containing lactide: glycolide in an ethyl acetate solution of about 50:50, and mix to obtain a dispersion of the triptorelin solution in the polymer solution; as well as (b) Add (iii) a phase inversion solution containing a surfactant and optionally one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension containing triptorelin and triptorelin-loaded microparticles of the one or more PLGA polymers.

69. A method for preparing triptorelin-loaded microparticles, the method comprising: (a) Add (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution of an ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymer containing a lactide:co-lactide ratio of about 75:25 in an ethyl acetate solution, and mix to obtain a dispersion of the triptorelin solution in the polymer solution; as well as (b) Add (iii) a phase inversion solution containing a surfactant and one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension containing triptorelin and triptorelin-loaded microparticles of the PLGA polymer.

70. A method for preparing triptorelin-loaded microparticles, the method comprising: (a) Add (i) a triptorelin solution containing triptorelin in an aqueous acidic solvent to (ii) a polymer solution containing a mixture of two different ester-terminated polylactide (PLA) polymers in an ethyl acetate solution, and mix to obtain a dispersion of triptorelin in the polymer solution; as well as (b) Add (iii) a phase inversion solution containing a surfactant and one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension containing triptorelin and triptorelin-loaded microparticles of these PLA polymers.

71. The method of any one of claims 68-70, wherein the triptorelin is triptorelin acetate.

72. The method of any one of claims 68-71, wherein the pH of the triptorelin solution is about 4.

73. The method of any one of claims 68-72, wherein the aqueous acidic solvent is an aqueous acetic acid solvent.

74. A method for preparing FSH-loaded microparticles, the method comprising: (a) Add (i) the FSH solution containing FSH in an aqueous solvent to (ii) a polymer solution of an ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymer containing a lactide:co-lactide ratio of about 50:50 in an ethyl acetate solution, and mix to obtain a dispersion of the FSH solution in the polymer solution. as well as (b) Add (iii) a phase inversion solution containing a surfactant and one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension containing FSH and FSH-loaded microparticles of the PLGA polymer.

75. A method for preparing FSH-loaded microparticles, the method comprising: (a) Add (i) the FSH solution containing FSH in an aqueous solvent to (ii) a polymer solution of an ester-terminated poly(D,L-lactide-co-lactide) (PLGA) polymer containing a lactide:co-lactide ratio of about 75:25 in an ethyl acetate solution and mix to obtain a dispersion of the FSH solution in the polymer solution. as well as (b) Add (iii) a phase inversion solution containing a surfactant and one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension containing FSH and FSH-loaded microparticles of the PLGA polymer.

76. A method for preparing FSH-loaded microparticles, the method comprising: (a) Add (i) an FSH solution containing FSH in an aqueous solvent to (ii) a polymer solution containing a mixture of two different ester-terminated polylactide (PLA) polymers in an ethyl acetate solution, and mix to obtain a dispersion of the FSH in the polymer solution; as well as (b) Add (iii) a phase inversion solution containing a surfactant and one or two of an alkaline salt and a polyol to the dispersion and mix to obtain a suspension containing FSH-loaded particles of the PLA polymers.

77. The method of any one of claims 74-76, wherein the phase inversion solution comprises poloxamer or polyvinyl alcohol as a surfactant.

78. The method of any one of claims 74-77, wherein the phase inversion solution comprises sodium chloride as an alkaline salt.

79. The method of any one of claims 74-78, wherein the phase inversion solution comprises sucrose or polyethylene glycol as a polyol.

80. The method of any one of claims 62-79, further comprising, after step (b), adding a wash phase solution optionally comprising an aqueous buffer and a surfactant.

81. The method of claim 80, wherein the washing phase solution comprises citrate buffer and poloxamer 188 and / or polyvinyl alcohol.

82. The method of any one of claims 62-81, further comprising, optionally, removing the solvent by vacuum after step (b).

83. The method of any one of claims 62-82, further comprising, after step (b), optionally separating the particles from the suspension by centrifugation or filtration.

84. The method of any one of claims 62-83, further comprising optionally drying the particles by freeze-drying.

85. The method of any one of claims 62-84, further comprising sterilizing the particles by ionizing radiation, optionally by gamma radiation.

86. The triptorelin-loaded microparticles of any one of claims 1-22 or 34-37, or the composition containing triptorelin-loaded microparticles of any one of claims 38-50, for the treatment of one or more of hormone-responsive cancers, endometriosis, female infertility, uterine fibroids, and central precocious puberty.

87. The FSH-loaded microparticles of any one of claims 23-37 or the composition containing FSH-loaded microparticles of any one of claims 38-43 or 51-57, for the treatment of female infertility or male infertility.

88. Use of triptorelin-loaded microparticles as claimed in any one of claims 1-22 or 34-37, or of a composition comprising triptorelin-loaded microparticles as claimed in any one of claims 38-50, in the preparation of a medicament for treating one or more of hormone-responsive cancers, endometriosis, female infertility, uterine fibroids, and central precocious puberty.

89. Use of the FSH-loaded microparticles of any one of claims 23-37 or the composition containing FSH-loaded microparticles of any one of claims 38-43 or 51-57 in the preparation of a medicament for treating female infertility or male infertility.

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