Drug sustained release microsphere composition as well as preparation method and application thereof

Abstract

The invention provides a drug sustained release microsphere composition as well as a preparation method and application thereof. The drug sustained release microsphere composition comprises an active component, a first lactide-glycolide and / or a second lactide-glycolide copolymer and other optional pharmaceutically acceptable auxiliary materials. The drug sustained-release microsphere composition prepared by the invention is round in form, high in drug loading capacity, capable of realizing long-term stable release and high in application value.

Description

[0001] This application claims priority to the Chinese patent application filed with the China Patent Office on November 9, 2023, with application number 202311494268.0 and application name “Baricitinib sustained-release microsphere composition, preparation method and use thereof”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present invention relates to the field of pharmaceutical preparations, and in particular to a pharmaceutical sustained-release microsphere composition, a preparation method thereof, and an application thereof. Background Art

[0003] Janus kinase (JAK) is a non-receptor tyrosine protein kinase whose main substrate is signal transducer and activator of transcription (STAT). JAK-STAT is involved in the post-receptor signal transduction of many cytokines. A variety of cytokines, including interleukins, interferons, and erythropoietin, activate the JAK-STAT pathway after binding to the corresponding receptors. The above cytokines and signaling pathways always play an important role in the body's innate and adaptive immune processes. However, when the signal level is unbalanced, it often causes excessive differentiation, proliferation, and even apoptosis resistance of specific immune cells, releasing excessive levels of cytokines and leading to inflammatory responses in different parts of the body, which in turn manifests as a variety of autoimmune-mediated inflammatory diseases in clinical practice, including blood tumors, rheumatic immune diseases, skin and gastrointestinal diseases.

[0004] Baricitinib, as a selective JAK1 and JAK2 inhibitor, has a strong inhibitory effect on JAK1 and JAK2. The currently marketed dosage form is a tablet, mainly used for adults with moderate to severe active rheumatoid arthritis and adult-onset severe alopecia areata. Rheumatoid arthritis (RA) is a common chronic, systemic autoimmune disease characterized by chronic synovitis and cartilage destruction. Patients often suffer from persistent joint pain, swelling, and stiffness. Severe cases can cause cardiovascular, pulmonary, and skeletal complications. RA symptoms are recurrent and difficult to cure, and the treatment cycle is long. Patients generally require a 12-week treatment cycle of baricitinib, taking 2 mg orally daily. Effective relief of symptoms depends to a certain extent on the patient's adherence to medication. Alopecia areata is the most common type of hair loss after androgenic alopecia. It has a significant impact on appearance, social interaction, and mental stress, and is often accompanied by symptoms such as reduced quality of life and sleep, generalized anxiety disorder, and severe depression. After taking 2 mg of baricitinib orally daily for up to one year, 22.6% of patients achieved significant scalp hair regrowth, with 67.5% achieving 90% or greater hair coverage.

[0005] For both of these conditions, which require long-term medication, patient adherence is a significant challenge to disease progression. Forgetting or missing doses can significantly negatively impact disease control. Therefore, a sustained-release formulation has been developed. Compared to daily oral tablets, this formulation can significantly reduce dosing frequency, minimize fluctuations in blood drug concentrations, mitigate toxic side effects, and improve patient compliance.

[0006] Poly(lactide-co-glycolide) (PLGA) is a high-molecular-weight copolymer formed by the polymerization of lactide (LA) and glycolide (GA) in the presence of a catalyst. It degrades into lactic acid, water, and carbon dioxide in the body. PLGA microspheres, with their excellent biocompatibility and biodegradability, have demonstrated significant value in the field of long-acting sustained-release medications.

[0007] In a previous study (Anwer MK, Ali EA, Iqbal M, Ahmed MM, Aldawsari MF, Saqr AA, Ansari MN, Aboudzadeh MA. Development of Sustained Release Baricitinib Loaded Lipid-Polymer Hybrid Nanoparticles with Improved Oral Bioavailability. Molecules. 2021 Dec 28; 27(1):168.), PLGA was used as a carrier material to prepare a baricitinib oral suspension nanoparticle. However, the nanoparticles released 89% of the drug within 12 hours, which obviously cannot improve the dilemma of long-term medication adherence during baricitinib treatment.

[0008] Existing patent document CN115969796A mentions a method for preparing baricitinib microspheres, but baricitinib has low solubility in benzyl alcohol and ethyl acetate, and a large amount of drug powder will be deposited in the lower layer of the oil phase, making this method not feasible.

[0009] Therefore, there is an urgent need to develop a baricitinib sustained-release microsphere preparation composition and its preparation process to achieve long-term stable release of the drug, reduce the frequency of administration, and reduce the treatment risks caused by large fluctuations in blood drug concentrations. Summary of the Invention

[0010] The present invention aims to, at least to some extent, address the technical problems existing in the prior art. To this end, the present invention provides a baricitinib sustained-release microsphere formulation composition, its preparation method, and its use. In the present invention, a "drug sustained-release microsphere composition" may also be referred to as a "baricitinib sustained-release microsphere composition" or a "microsphere formulation."

[0011] On the one hand, the present invention provides a drug sustained-release microsphere composition, comprising: an active ingredient, wherein the active ingredient includes baricitinib and / or a pharmaceutically acceptable salt thereof; a lactide-glycolide copolymer, wherein the lactide-glycolide copolymer includes a first lactide-glycolide and / or a second lactide-glycolide copolymer; and optionally, other pharmaceutically acceptable excipients.

[0012] According to an embodiment of the present invention, the other pharmaceutically acceptable excipients include a lyoprotectant.

[0013] According to an embodiment of the present invention, the lyoprotectant is selected from one or more of mannitol, sucrose and lactose. In some specific embodiments, the lyoprotectant is selected from mannitol.

[0014] According to an embodiment of the present invention, based on the total weight of the active ingredient and the lactide-glycolide copolymer, the content of the active ingredient is 5-25wt%, preferably 7-20wt%, and more preferably 10-15wt%. In some embodiments, the content of the active ingredient is 5wt%, 6wt%, 7wt%, 8wt%, 9wt%, 10wt%, 11wt%, 12wt%, 13wt%, 14wt%, 15wt%, 16wt%, 17wt%, 18wt%, 19wt%, 20wt%, 21wt%, 22wt%, 23wt%, 24wt%, 25wt%, or a range value between any two of these values ​​as endpoints.

[0015] According to an embodiment of the present invention, based on the total weight of the active ingredient and the lactide-glycolide copolymer, the content of the lactide-glycolide copolymer is 75-95wt%, preferably 80-93wt%, and more preferably 85-95wt%. In some embodiments, the total content of the lactide-glycolide copolymer is 75wt%, 76wt%, 77wt%, 78wt%, 79wt%, 80wt%, 81wt%, 82wt%, 83wt%, 84wt%, 85wt%, 86wt%, 87wt%, 88wt%, 89wt%, 90wt%, 91wt%, 92wt%, 93wt%, 94wt%, 95wt% or a range between any two of these values ​​as endpoints. In some specific embodiments, the total content of the lactide-glycolide copolymer is 84wt%. In some specific embodiments, the total content of the lactide-glycolide copolymer is 85wt%. In some specific embodiments, the total content of the lactide-glycolide copolymer is 88 wt %. In some specific embodiments, the total content of the lactide-glycolide copolymer is 94 wt %.

[0016] According to an embodiment of the present invention, the intrinsic viscosity of the lactide-glycolide copolymer is 0.08 to 0.90 dl / g, preferably 0.16 to 0.60 dl / g. In some embodiments, the viscosity of the lactide-co-glycolide copolymer is 0.08 dl / g, 0.09 dl / g, 0.10 dl / g, 0.16 dl / g, 0.19 dl / g, 0.20 dl / g, 0.24 dl / g, 0.30 dl / g, 0.35 dl / g, 0.40 dl / g, 0.46 dl / g, 0.50 dl / g, 0.55 dl / g, 0.57 dl / g, 0.60 dl / g, 0.68 dl / g, 0.70 dl / g, 0.75 dl / g, 0.80 dl / g, 0.84 dl / g, 0.90 dl / g, or a range between any two of these values ​​as endpoints. In some specific embodiments, the viscosity of the lactide-co-glycolide copolymer is 0.20 dl / g.

[0017] According to an embodiment of the present invention, the molar ratio of lactide to glycolide in the lactide-glycolide copolymer is 90:10 to 25:75, preferably 75:25 to 50:50. In some embodiments, the molar ratio of lactide to glycolide in the lactide-glycolide copolymer is 90:10, 80:20, 75:25, 70:30, 65:35, 60:40, 55:45, 50:50, 45:55, 40:60, 35:65, 30:70, 25:75, or a range between any two of these values ​​as endpoints, for example, 75:25 to 50:50. In some embodiments, the molar ratio of lactide to glycolide in the lactide-glycolide copolymer is 75:25. In some embodiments, the molar ratio of lactide to glycolide in the lactide-glycolide copolymer is 50:50.

[0018] According to an embodiment of the present invention, the lactide-glycolide copolymer is selected from one or both of a first lactide-glycolide copolymer and a second lactide-glycolide copolymer, wherein, when the lactide-glycolide copolymer is selected from both of the first lactide-glycolide copolymer and the second lactide-glycolide copolymer, the viscosity of the first lactide-glycolide copolymer is less than the viscosity of the second lactide-glycolide copolymer. In some embodiments, the lactide-glycolide copolymer is the first lactide-glycolide copolymer. In some embodiments, the lactide-glycolide copolymer is the second lactide-glycolide copolymer. In some embodiments, the lactide-glycolide copolymer is a mixture of the first lactide-glycolide copolymer and the second lactide-glycolide copolymer.

[0019] According to an embodiment of the present invention, the viscosity of the first lactide-glycolide copolymer is 0.08 to 0.30 dl / g, and the viscosity of the second lactide-glycolide copolymer is 0.32 to 0.90 dl / g. In some specific embodiments, the viscosity of the first lactide-glycolide copolymer is 0.20 dl / g, and the viscosity of the second lactide-glycolide copolymer is 0.57 dl / g.

[0020] According to an embodiment of the present invention, when the lactide-glycolide copolymer is selected from two of a first lactide-glycolide copolymer and a second lactide-glycolide copolymer, the mass ratio of the first lactide-glycolide copolymer to the second lactide-glycolide copolymer is 1:9 to 9:1. In some embodiments, the mass ratio of the first lactide-glycolide copolymer to the second lactide-glycolide copolymer is 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or a range between any two of these values ​​as endpoints, for example, 1:1 to 2:1. In some specific embodiments, the mass ratio of the first lactide-glycolide copolymer to the second lactide-glycolide copolymer is 1:1.

[0021] In another aspect, the present invention provides a method for preparing a baricitinib sustained-release microsphere composition, the method comprising:

[0022] (1) dissolving a lactide-glycolide copolymer in a first organic solvent to prepare a carrier solution;

[0023] (2) dissolving baricitinib or a pharmaceutically acceptable salt thereof in a second organic solvent to prepare a drug solution;

[0024] (3) dissolving the hydrophilic polymer in deionized water to prepare an external aqueous phase solution;

[0025] (4) adding the carrier solution obtained in step (1) to the drug solution obtained in step (2), mixing, and obtaining an oil phase solution;

[0026] (5) adding the oil phase obtained in step (4) dropwise to the external aqueous phase solution obtained in step (3) under shear conditions to prepare an O / W emulsion;

[0027] (6) stirring to evaporate the first organic solvent and the second organic solvent, collecting by filtration, and washing with deionized water to obtain a microsphere suspension.

[0028] According to an embodiment of the present invention, the method further comprises:

[0029] (7) adding a lyophilization protectant to the microsphere suspension obtained in step (6), and lyophilizing to obtain a baricitinib sustained-release microsphere composition.

[0030] Furthermore, the lactide-glycolide copolymer in step (1) is selected from one or both of a first lactide-glycolide copolymer and a second lactide-glycolide copolymer.

[0031] Furthermore, in step (1), the concentration of the lactide-co-glycolide copolymer in the carrier solution is 120 to 500 mg / mL. In some specific embodiments, the concentration of the lactide-co-glycolide copolymer in the carrier solution is 120 mg / mL, 150 mg / mL, 200 mg / mL, 250 mg / mL, 300 mg / mL, 350 mg / mL, 400 mg / mL, 450 mg / mL, or 500 mg / mL.

[0032] Furthermore, in step (5), the oil phase solution obtained in step (4) is added dropwise to the external aqueous phase solution in step (3) under shear conditions.

[0033] Furthermore, in step (5), the shearing conditions include a rotation speed of 3000 to 6000 rpm, and a dropping speed of 0.1 to 2.0 mL / min.

[0034] Furthermore, the hydrophilic polymer includes at least one of methylcellulose, polyvinyl pyrrolidone, carboxymethyl cellulose, lecithin, gelatin, polyvinyl alcohol, polyoxyethylene-polyoxypropylene block copolymer, polyoxyethylene sorbitan fatty acid ester and polyoxyethylene. In some specific embodiments, the hydrophilic polymer is polyvinyl alcohol.

[0035] Furthermore, the step (5) comprises: injecting the oil phase solution into the external aqueous phase solution at a uniform speed of 0.1 to 2.0 mL / min through a syringe pump at 3000 to 6000 rpm. In some embodiments, the shear condition is to inject the oil phase solution into the external aqueous phase solution through a syringe pump at a uniform speed of 0.1 mL / min, 0.2 mL / min, 0.3 mL / min, 0.4 mL / min, 0.5 mL / min, 0.6 mL / min, 0.7 mL / min, 0.8 mL / min, 0.9 mL / min, 1.0 mL / min, 1.1 mL / min, 1.2 mL / min, 1.3 mL / min, 1.4 mL / min, 1.5 mL / min, 1.6 mL / min, 1.7 mL / min, 1.8 mL / min, 1.9 mL / min, or 2.0 mL / min at 3000 rpm, 3500 rpm, 4000 rpm, 4500 rpm, 5000 rpm, 5500 rpm, or 6000 rpm.

[0036] Furthermore, in step (6), the first organic solvent and the second organic solvent are evaporated by stirring.

[0037] Furthermore, the freeze-drying process in step (7) specifically includes:

[0038] S1: pre-freezing the microsphere suspension after adding the lyoprotectant;

[0039] S2: further sublime the product obtained in S1;

[0040] S3: Dry the product obtained in S2 to obtain a lyophilized preparation of baricitinib sustained-release microspheres.

[0041] Furthermore, the pre-freezing process parameters are: setting temperature -30 to -50°C, holding time 4 to 6 hours; preferably, the pre-freezing process parameters are: setting temperature -40°C, holding time 5 hours.

[0042] Furthermore, the drying process includes sublimation drying and desorption drying. The sublimation drying process includes: controlling the vacuum degree at 0.1-0.3 mbar, heating to -10--20°C at 0.4-0.6°C / min, and maintaining at -10--20°C for 10-12 hours; the desorption drying process includes: controlling the vacuum degree at 0.1-0.3 mbar, heating to 10-30°C at 0.4-0.6°C / min, and maintaining at 10-30°C for 9-11 hours; preferably, the sublimation drying process includes: controlling the vacuum degree at 0.2 mbar, heating to -15°C at 0.5°C / min, and maintaining at -15°C for 11 hours. The desorption drying process is: controlling the vacuum degree at 0.2 mbar, heating to 20°C at 0.5°C / min, and maintaining at 20°C for 10 hours.

[0043] According to an embodiment of the present invention, the concentration of the lactide-glycolide copolymer in the carrier solution is 120-500 mg / mL.

[0044] According to an embodiment of the present invention, the first organic solvent is selected from one or more of dichloromethane, chloroform, ethyl acetate, and methanol; and the second organic solvent is selected from one or two of dimethyl sulfoxide, dimethylformamide, and N-methylpyrrolidone. In some specific embodiments, the first organic solvent is selected from dichloromethane, and the second organic solvent is selected from dimethyl sulfoxide.

[0045] According to an embodiment of the present invention, the volume ratio of the second organic solvent to the first organic solvent is 1:7 to 1:2. In some embodiments, the volume ratio of the second organic solvent to the first organic solvent is 1:7, 1:6, 1:5, 1:4, 1:3, 1:2 or a range value between any two of these values ​​as endpoint values, for example, 1:6 to 1:2. In some specific embodiments, the volume ratio of the second organic solvent to the first organic solvent is 1:5. In some embodiments, the volume ratio of the first organic solvent to the second organic solvent is 3:10. In some specific embodiments, the volume ratio of the first organic solvent to the second organic solvent is 2:5.

[0046] According to an embodiment of the present invention, the concentration of the hydrophilic polymer in the external aqueous phase solution is 0.5% to 2%. In some embodiments, the concentration of the hydrophilic polymer in the external aqueous phase solution is 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, or a range between any two of these values ​​as endpoints. In some specific embodiments, the concentration of the hydrophilic polymer in the external aqueous phase solution is 0.5%.

[0047] According to an embodiment of the present invention, the concentration of the hydrophilic polymer in the external aqueous phase solution is 0.5% to 2%. In some embodiments, the concentration of the hydrophilic polymer in the external aqueous phase solution is 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, or a range between any two of these values ​​as endpoints. In some specific embodiments, the concentration of the hydrophilic polymer in the external aqueous phase solution is 0.5%.

[0048] According to an embodiment of the present invention, the volume ratio of the oil phase solution to the external aqueous phase solution is 1.0:200 to 2.0:200. In some embodiments, the volume ratio of the oil phase solution to the external aqueous phase solution is 1.0:200, 1.1:200, 1.2:200, 1.3:200, 1.4:200, 1.5:200, 1.6:200, 1.7:200, 1.8:200, 1.9:200, 2.0:200, or a range value between any two of these values ​​as endpoints, for example, 1.1:200 to 1.8:200. In some specific embodiments, the volume ratio of the oil phase solution to the external aqueous phase solution is 1.2:200. In some specific embodiments, the volume ratio of the oil phase solution to the external aqueous phase solution is 1.3:200. In some specific embodiments, the volume ratio of the oil phase solution to the external aqueous phase solution is 1.4:200. In some specific embodiments, the volume ratio of the oil phase solution to the external aqueous phase solution is 1.8:200.

[0049] The present invention provides a drug sustained-release microsphere composition, a preparation method thereof, an application thereof, and a method for preventing or treating alopecia areata or rheumatoid arthritis, which will be described in detail below.

[0050] In one aspect, the present invention provides the use of the aforementioned sustained-release microsphere composition in the preparation of a medicament. According to an embodiment of the present invention, the medicament is used to prevent or treat alopecia areata, rheumatoid arthritis or atopic dermatitis.

[0051] In another aspect, the present invention provides a method for preventing or treating alopecia areata or rheumatoid arthritis. According to an embodiment of the present invention, the method comprises: administering the aforementioned microsphere preparation to a subject.

[0052] According to an embodiment of the present application, the drug is administered by injection.

[0053] Beneficial effects of the present invention:

[0054] The advantages of the baricitinib sustained-release microsphere composition prepared by the present invention will be partially listed in the following description or understood through the practice of the present invention.

[0055] First, the pharmaceutical composition obtained by the present invention can release the drug stably for a long time, can better exert the sustained-release effect, reduce the frequency of administration, and improve the patient's medication compliance.

[0056] Second, the microsphere preparation method of the present invention is simple and suitable for industrialization. The prepared drug microspheres have a round shape and a high drug loading capacity, achieving long-term stable release, reducing the treatment risk caused by fluctuations in blood drug concentration, and reducing toxic side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] Figure 1This is a microscope photograph of baricitinib microspheres in Example 2;

[0058] Figure 2 This is a microscope photograph of baricitinib microspheres in Example 5;

[0059] Figure 3 This is a microscope photograph of baricitinib microspheres in Example 8;

[0060] Figure 4 This is the in vitro cumulative release curve of the microspheres prepared in Example 1;

[0061] Figure 5 This is the in vitro cumulative release curve of the microspheres prepared in Example 2;

[0062] Figure 6 This is the in vitro cumulative release curve of the microspheres prepared in Example 4;

[0063] Figure 7 This is the in vitro cumulative release curve of the microspheres prepared in Example 6;

[0064] Figure 8 This is the in vitro cumulative release curve of the microspheres prepared in Example 8;

[0065] Figure 9 This is the in vitro cumulative release curve of the microspheres prepared in Example 10;

[0066] Figure 10 This is the in vitro cumulative release curve of the microspheres prepared in Example 11;

[0067] Figure 11 This is the in vitro cumulative release curve of the microspheres prepared in Example 13.

[0068] Terminology

[0069] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood to indicate or imply relative importance or implicitly specify the number of the technical features indicated. Therefore, features defined as "first" or "second" may explicitly or implicitly include one or more of such features. Furthermore, in the description of the present invention, unless otherwise specified, "plurality" means two or more.

[0070] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0071] In this document, the terms “include” or “comprising” are open expressions, that is, including the contents specified in the present invention, but not excluding other contents.

[0072] PVA stands for polyvinyl alcohol; PLGA stands for poly(lactic-co-glycolic acid) or poly(lactide-co-glycolide); and DMSO stands for dimethyl sulfoxide. DETAILED DESCRIPTION

[0073] The present invention is further described in detail below by way of specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the present invention to the following examples. Any technology implemented based on the present invention in accordance with common technical knowledge and customary means in the art falls within the scope of the present invention.

[0074] For ease of description, the lactide-glycolide molar ratio in PLGA and the intrinsic viscosity of PLGA are indicated in parentheses after PLGA. For example, "PLGA (50 / 50, 0.2 dl / g)" indicates a lactide-glycolide copolymer having a lactide-glycolide molar ratio of 50:50 and an intrinsic viscosity of 0.2 dl / g.

[0075] Table 1 Experimental parameters of freeze-drying process

[0076]

[0077] The "drug loading" used in this invention refers to the actual drug loading, calculated according to the following equation: Drug loading = drug amount in microspheres / (drug amount in microspheres + PLGA amount in microspheres) × 100%. Encapsulation efficiency = drug amount encapsulated in the system / total encapsulated and unencapsulated drug amount in the system × 100%. Release was determined using a shaker method and high-performance liquid chromatography (HPLC) using a C18 column, a mobile phase of pH 6.5 phosphate buffer-acetonitrile (70:30), and a detection wavelength of 220 nm.

[0078] Table 2 Prescription composition of Examples 1 to 13

[0079]

[0080]

[0081] Example 1

[0082] Weigh 400 mg of PLGA (50 / 50, 0.20 dl / g) and dissolve it in 1 mL of dichloromethane to obtain a carrier solution. Weigh 50 mg of baricitinib and dissolve it in 0.2 mL of dimethyl sulfoxide to obtain a drug solution. Dissolve 1 g of polyvinyl alcohol in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). Mix the carrier solution and the drug solution to obtain an oil phase. Inject this oil phase into 200 mL of external aqueous phase solution at a uniform speed of 1 mL / min through a syringe pump at 5000 rpm and homogenize and emulsify for 5 minutes. Then stir at a low speed of 300 rpm for 3 to 5 hours to remove dichloromethane and dimethyl sulfoxide. Collect the microspheres by filtration, wash with deionized water, add mannitol, and freeze-dry to obtain baricitinib sustained-release microsphere powder. The obtained microspheres are round in shape, and no drug crystals are precipitated. The drug loading is 9.51% and the encapsulation efficiency is 87.6%. Its in vitro cumulative release curve is as shown in Figure 2. Figure 4 As shown in the figure, the drug-loaded microspheres had a low burst release and showed a uniform slow release rate within 8 days.

[0083] Example 2

[0084] Weigh 400 mg of PLGA (50 / 50, 0.20 dl / g) and dissolve it in 1 mL of dichloromethane to obtain a carrier solution; weigh 50 mg of baricitinib and dissolve it in 0.3 mL of dimethyl sulfoxide to obtain a drug solution. Dissolve 1 g of polyvinyl alcohol in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). Mix the carrier solution and the drug solution to obtain an oil phase. Inject this oil phase into 200 mL of external aqueous phase solution at a uniform speed of 1 mL / min through an injection pump at 5000 rpm and homogenize and emulsify for 5 minutes. Then stir at a low speed of 300 rpm for 3 to 5 hours to remove dichloromethane and dimethyl sulfoxide. Collect the microspheres by filtration, wash with deionized water, add mannitol, and freeze-dry to obtain baricitinib sustained-release microsphere powder. The obtained microspheres are round in shape, with no drug crystals precipitated, a drug loading of 10.02%, and an encapsulation efficiency of 93.8%. The microscopic photograph is shown below. Figure 1 Its in vitro cumulative release curve is shown in Figure 5 As shown in the figure, the drug-loaded microspheres had a low burst release and showed a uniform slow release rate within 6 days.

[0085] Example 3

[0086] 400 mg of PLGA (50 / 50, 0.20 dl / g) was dissolved in 1 mL of dichloromethane to obtain a carrier solution. 50 mg of baricitinib was dissolved in 0.4 mL of dimethyl sulfoxide to obtain a drug solution. 1 g of polyvinyl alcohol was dissolved in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). The carrier and drug solutions were mixed to obtain an oil phase. This oil phase was homogenized at 5000 rpm using a syringe pump at a rate of 1 mL / min into 200 mL of the external aqueous phase solution for 5 minutes. The mixture was then stirred at a low speed of 300 rpm for 3-5 hours to remove the dichloromethane and dimethyl sulfoxide. The microspheres were collected by filtration, washed with deionized water, added with mannitol, and lyophilized to obtain baricitinib sustained-release microsphere powder. The resulting microspheres were rounded with no drug crystals, had a drug loading of 8.80%, and an encapsulation efficiency of 81.1%.

[0087] The results of Examples 1-3 show that changes in DMSO volume have little effect on the in vitro release behavior of microspheres. In general, DMSO can effectively encapsulate the drug. However, in order to ensure that baricitinib can be completely dissolved, the volume of DMSO needs to be reasonably adjusted according to the dosage.

[0088] Example 4

[0089] Weigh 400 mg of PLGA (50 / 50, 0.20 dl / g) and dissolve it in 1 mL of dichloromethane to obtain a carrier solution; weigh 25 mg of baricitinib and dissolve it in 0.3 mL of dimethyl sulfoxide to obtain a drug solution. Dissolve 1 g of polyvinyl alcohol in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). Mix the carrier solution and the drug solution to obtain an oil phase. Inject this oil phase into 200 mL of external aqueous phase solution at a uniform speed of 1 mL / min through a syringe pump at 5000 rpm and homogenize and emulsify for 5 minutes. Then stir at a low speed of 300 rpm for 3 to 5 hours to remove dichloromethane and dimethyl sulfoxide. Collect the microspheres by filtration, wash with deionized water, add mannitol, and freeze-dry to obtain baricitinib sustained-release microsphere powder. The obtained microspheres are round in shape, and no drug crystals are precipitated. The drug loading is 4.56% and the encapsulation efficiency is 79.8%. Its in vitro cumulative release curve is as shown in Figure 2. Figure 6 As shown in the figure, the drug-loaded microspheres had a low burst release and showed a uniform slow release rate within 6 days.

[0090] Example 5

[0091] Weigh 400 mg of PLGA (50 / 50, 0.20 dl / g) and dissolve it in 1 mL of dichloromethane to obtain a carrier solution. Weigh 60 mg of baricitinib and dissolve it in 0.3 mL of dimethyl sulfoxide to obtain a drug solution. Dissolve 1 g of polyvinyl alcohol in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). Mix the carrier solution and the drug solution to obtain an oil phase. Inject this oil phase into 200 mL of external aqueous phase solution at a uniform speed of 1 mL / min through an injection pump at 5000 rpm and homogenize and emulsify for 5 minutes. Then stir at a low speed of 300 rpm for 3 to 5 hours to remove dichloromethane and dimethyl sulfoxide. Collect the microspheres by filtration, wash with deionized water, add mannitol, and freeze-dry to obtain baricitinib sustained-release microsphere powder. The obtained microspheres are round in shape, and no drug crystals are precipitated. The drug loading is 9.87% and the encapsulation efficiency is 76.4%. Its microscopic photograph is as shown below. Figure 2 shown.

[0092] Example 6

[0093] 400 mg of PLGA (50 / 50, 0.20 dl / g) was weighed and dissolved in 1 mL of dichloromethane to obtain a carrier solution; 75 mg of baricitinib was weighed and dissolved in 0.3 mL of dimethyl sulfoxide to obtain a drug solution. 1 g of polyvinyl alcohol was dissolved in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). The carrier solution and the drug solution were mixed to obtain an oil phase. This oil phase was homogenized at 5000 rpm by injecting a syringe pump at a speed of 1 mL / min into 200 mL of the external aqueous phase solution and homogenized for 5 minutes. Subsequently, the mixture was stirred at a low speed of 300 rpm for 3 to 5 hours to remove dichloromethane and dimethyl sulfoxide. The microspheres were collected by filtration, washed with deionized water, added with mannitol, and freeze-dried to obtain baricitinib sustained-release microsphere powder. The obtained microspheres were round in shape, with no drug crystals precipitated, the drug loading was 13.87%, and the encapsulation efficiency was 89.0%. Its in vitro cumulative release curve is shown in Figure 2. Figure 7 As shown in the figure, the drug-loaded microspheres had a low burst release and showed a uniform slow release rate within 6 days.

[0094] Example 7

[0095] 400 mg of PLGA (50 / 50, 0.20 dl / g) was dissolved in 1 mL of dichloromethane to obtain the carrier solution. 100 mg of baricitinib was dissolved in 0.4 mL of dimethyl sulfoxide to obtain the drug solution (the DMSO volume needed to be increased to 0.4 mL for a 100 mg dose to ensure complete drug dissolution). 1 g of polyvinyl alcohol was dissolved in 200 mL of purified water to obtain the external aqueous phase solution (0.5% PVA). The carrier and drug solutions were mixed to obtain the oil phase. This oil phase was homogenized at 5000 rpm using a syringe pump at a constant rate of 1 mL / min into 200 mL of the external aqueous phase solution and homogenized for 5 minutes. The mixture was then stirred at a low speed of 300 rpm for 3-5 hours to remove the dichloromethane and dimethyl sulfoxide. The microspheres were collected by filtration, washed with deionized water, added with mannitol, and lyophilized to obtain baricitinib sustained-release microsphere powder. The obtained microspheres were round in shape without drug crystal precipitation, with a drug loading of 15.87% and an encapsulation efficiency of 80.4%.

[0096] The results of Examples 4-7 showed that as the dosage of baricitinib in the drug solution increased, the drug loading of the microspheres gradually increased, and the high drug loading microspheres did not show an obvious burst release effect. The in vitro release behaviors of microspheres with different drug loadings were basically the same.

[0097] Example 8

[0098] 400 mg of PLGA (50 / 50, 0.20 dl / g) was weighed and dissolved in 1.5 mL of dichloromethane to obtain a carrier solution; 50 mg of baricitinib was weighed and dissolved in 0.3 mL of dimethyl sulfoxide to obtain a drug solution. 1 g of polyvinyl alcohol was dissolved in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). The carrier solution and the drug solution were mixed to obtain an oil phase. This oil phase was homogenized at 5000 rpm by injecting a syringe pump at a uniform speed of 1 mL / min into 200 mL of the external aqueous phase solution and homogenized and emulsified for 5 minutes. Subsequently, the mixture was stirred at a low speed of 300 rpm for 3 to 5 hours to remove dichloromethane and dimethyl sulfoxide. The microspheres were collected by filtration, washed with deionized water, mannitol was added, and freeze-dried to obtain baricitinib sustained-release microsphere powder. The obtained microspheres were round in shape, with no drug crystals precipitated, the drug loading was 6.76%, and the encapsulation efficiency was 62.0%. The microscopic photograph is shown below. Figure 3 Its in vitro cumulative release curve is shown in Figure 8 As shown in the figure, the drug-loaded microspheres had a low burst release and showed a uniform slow release rate within 8 days.

[0099] The results showed that increasing the volume of dichloromethane in the carrier solution resulted in a decrease in the viscosity of the oil phase and a significant decrease in the microsphere encapsulation efficiency.

[0100] Example 9

[0101] 300 mg of PLGA (50 / 50, 0.20 dl / g) was dissolved in 1 mL of dichloromethane to obtain a carrier solution. 50 mg of baricitinib was dissolved in 0.3 mL of dimethyl sulfoxide to obtain a drug solution. 1 g of polyvinyl alcohol was dissolved in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). The carrier and drug solutions were mixed to obtain an oil phase. This oil phase was homogenized at 5000 rpm using a syringe pump at a constant rate of 1 mL / min into 200 mL of the external aqueous phase solution for 5 minutes. The mixture was then stirred at a low speed of 300 rpm for 3-5 hours to remove the dichloromethane and dimethyl sulfoxide. The microspheres were collected by filtration, washed with deionized water, added with mannitol, and lyophilized to obtain a baricitinib sustained-release microsphere powder. The resulting microspheres were rounded with no drug crystals, had a drug loading of 10.45%, and an encapsulation efficiency of 74.6%.

[0102] Example 10

[0103] Weigh 400 mg of PLGA (75 / 25, 0.19 dl / g) and dissolve it in 1 mL of dichloromethane to obtain a carrier solution; weigh 50 mg of baricitinib and dissolve it in 0.3 mL of dimethyl sulfoxide to obtain a drug solution. Dissolve 1 g of polyvinyl alcohol in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). Mix the carrier solution and the drug solution to obtain an oil phase. Inject this oil phase into 200 mL of external aqueous phase solution at a uniform speed of 1 mL / min through a syringe pump at 5000 rpm and homogenize and emulsify for 5 minutes. Then stir at a low speed of 300 rpm for 3 to 5 hours to remove dichloromethane and dimethyl sulfoxide. Collect the microspheres by filtration, wash with deionized water, add mannitol, and freeze-dry to obtain baricitinib sustained-release microsphere powder. The obtained microspheres are round in shape, and no drug crystals are precipitated. The drug loading is 8.13% and the encapsulation efficiency is 75.2%. Its in vitro cumulative release curve is as shown in Figure 2. Figure 9 As shown in the figure, the drug-loaded microspheres had a low burst release and showed a uniform slow release rate within 8 days.

[0104] The results showed that as the lactide ratio in PLGA increased, the hydrophobicity of the polymer increased, the degradation and release rate of the microspheres slowed down, and the drug release period was prolonged.

[0105] Example 11

[0106] 150 mg of PLGA (50 / 50, 0.20 dl / g) and 150 mg of PLGA (50 / 50, 0.57 dl / g) were dissolved in 1 mL of dichloromethane to obtain a carrier solution. 50 mg of baricitinib was dissolved in 0.3 mL of dimethyl sulfoxide to obtain a drug solution. 1 g of polyvinyl alcohol was dissolved in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). The carrier solution and drug solution were mixed to obtain an oil phase. This oil phase was homogenized at 5000 rpm using a syringe pump at a constant rate of 1 mL / min into 200 mL of the external aqueous phase solution and homogenized for 5 minutes. The mixture was then stirred at a low speed of 300 rpm for 3-5 hours to remove the dichloromethane and dimethyl sulfoxide. The microspheres were collected by filtration, washed with deionized water, added with mannitol, and lyophilized to obtain baricitinib sustained-release microsphere powder. The microspheres were round and free of drug crystals. The drug loading was 10.46% and the encapsulation efficiency was 74.7%. Figure 10 As shown in the figure, the drug-loaded microspheres had a low burst release and showed a uniform slow release within 14 days without a significant release stagnation stage.

[0107] The results showed that a mixture of lactide-glycolide copolymer selected from PLGA (50 / 50, 0.20 dl / g) and PLGA (50 / 50, 0.57 dl / g) could significantly prolong the drug release period and improve the sustained-release effect of microspheres.

[0108] Example 12

[0109] 150 mg of PLGA (50 / 50, 0.20 dl / g) and 150 mg of PLGA (50 / 50, 0.57 dl / g) were dissolved in 1 mL of dichloromethane to prepare the carrier solution. 100 mg of baricitinib was dissolved in 0.4 mL of dimethyl sulfoxide to prepare the drug solution (the DMSO volume should be increased to 0.4 mL for a 100 mg dose to ensure complete drug dissolution). 1 g of polyvinyl alcohol was dissolved in 200 mL of purified water to prepare the external aqueous phase solution (0.5% PVA). The carrier solution and drug solution were mixed to prepare the oil phase. This oil phase was homogenized at 5000 rpm using a syringe pump at a constant rate of 1 mL / min into 200 mL of the external aqueous phase solution and homogenized for 5 minutes. The mixture was then stirred at a low speed of 300 rpm for 3-5 hours to remove the dichloromethane and dimethyl sulfoxide. The microspheres were collected by filtration, washed with deionized water, added with mannitol, and lyophilized to obtain baricitinib sustained-release microsphere powder. The resulting microspheres were round in shape with no drug crystals precipitated, with a drug loading of 17.99% and an encapsulation efficiency of 72.2%.

[0110] Example 13

[0111] 400 mg of PLGA (50 / 50, 0.20 dl / g) was dissolved in 1 mL of a mixture of dichloromethane and ethyl acetate (2:1, v / v) to obtain a carrier solution. 50 mg of baricitinib was dissolved in 0.3 mL of dimethyl sulfoxide to obtain a drug solution. 1 g of polyvinyl alcohol was dissolved in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). The carrier and drug solutions were mixed to obtain an oil phase. This oil phase was homogenized at 4000 rpm using a syringe pump at a rate of 2 mL / min into 200 mL of the external aqueous phase solution and homogenized for 5 minutes. The mixture was then stirred at a low speed of 300 rpm for 3-5 hours to remove the dichloromethane, ethyl acetate, and dimethyl sulfoxide. The microspheres were collected by filtration, washed with deionized water, added with mannitol, and lyophilized to obtain baricitinib sustained-release microsphere powder. The microspheres were round and free of drug crystals. The drug loading of the obtained microspheres was 7.56% and the encapsulation efficiency was 68.8%. Figure 11 As shown in the figure, the drug-loaded microspheres had a low burst release and showed a uniform slow release rate within 7 days.

[0112] The results showed that the use of a mixture of dichloromethane and ethyl acetate as the organic solvent in the carrier solution would reduce the encapsulation efficiency of the microspheres to a certain extent.

[0113] Comparative study on the effect of poloxamer 407 on encapsulation efficiency and drug loading

[0114] Patent document CN115969796A mentions a method for preparing baricitinib microspheres. The present invention repeated this experiment and found that baricitinib has low solubility in benzyl alcohol and ethyl acetate, and a large amount of drug powder will be deposited in the lower layer of the oil phase, making this method unfeasible. The patent also mentions the addition of poloxamer 407. Given that the patented method is not feasible, the present invention added poloxamer 407 to the self-developed formulation process to investigate whether the presence of poloxamer 407 has a significant beneficial effect on the encapsulation efficiency and drug loading of baricitinib microspheres.

[0115] Table 2 Poloxamer 407 comparative prescription table

[0116]

[0117] Comparative Example 1

[0118] 12.5 mg of poloxamer 407 and 400 mg of PLGA (50 / 50, 0.20 dl / g) were dissolved in 1 mL of dichloromethane to prepare a carrier solution. 50 mg of baricitinib was dissolved in 0.3 mL of dimethyl sulfoxide to prepare a drug solution. 1 g of polyvinyl alcohol was dissolved in 200 mL of purified water to prepare an external aqueous phase solution (0.5% PVA). The carrier solution and drug solution were mixed to prepare an oil phase. This oil phase was homogenized at 5000 rpm using a syringe pump at a constant rate of 1 mL / min into 200 mL of the external aqueous phase solution and homogenized for 5 minutes. The mixture was then stirred at a low speed of 300 rpm for 3-5 hours to remove the dichloromethane and dimethyl sulfoxide. The microspheres were collected by filtration, washed with deionized water, added with mannitol, and lyophilized to obtain baricitinib sustained-release microsphere powder. The resulting microspheres had a drug loading of 8.42% and an encapsulation efficiency of 77.2%.

[0119] Comparative Example 2

[0120] 20 mg of poloxamer 407 and 400 mg of PLGA (50 / 50, 0.20 dl / g) were dissolved in 1 mL of dichloromethane to obtain a carrier solution. 50 mg of baricitinib was dissolved in 0.3 mL of dimethyl sulfoxide to obtain a drug solution. 1 g of polyvinyl alcohol was dissolved in 200 mL of purified water to obtain an external aqueous phase solution (0.5% PVA). The carrier solution and drug solution were mixed to obtain an oil phase. This oil phase was homogenized at 5000 rpm using a syringe pump at a constant rate of 1 mL / min into 200 mL of the external aqueous phase solution and homogenized for 5 minutes. The mixture was then stirred at a low speed of 300 rpm for 3-5 hours to remove the dichloromethane and dimethyl sulfoxide. The microspheres were collected by filtration, washed with deionized water, added with mannitol, and lyophilized to obtain baricitinib sustained-release microsphere powder. The resulting microspheres had a drug loading of 8.34% and an encapsulation efficiency of 78.2%.

[0121] The comparison results showed that the addition of poloxamer 407 to the formulation had no significant beneficial effect on the drug encapsulation properties of baricitinib microspheres.

Claims

1. A drug sustained-release microsphere composition, characterized in that: include: a) an active ingredient, the active ingredient comprising baricitinib and / or a pharmaceutically acceptable salt thereof; b) a lactide-glycolide copolymer, wherein the lactide-glycolide copolymer comprises a first lactide-glycolide and / or a second lactide-glycolide copolymer; c) optionally other pharmaceutically acceptable excipients.

2. The drug sustained-release microsphere composition according to claim 1, characterized in that: The other pharmaceutically acceptable excipients include lyoprotectants; The lyophilization protective agent is selected from one or more of mannitol, sucrose and lactose.

3. The drug sustained-release microsphere composition according to claim 1, characterized in that: The content of the active ingredient is 5 to 25 wt % based on the total weight of the active ingredient and the lactide-glycolide copolymer; Based on the total weight of the active ingredient and the lactide-glycolide copolymer, the content of the lactide-glycolide copolymer is 75-95wt%.

4. The drug sustained-release microsphere composition according to claim 1, characterized in that: The viscosity of the lactide-glycolide copolymer is 0.08 to 0.90 dl / g; preferably, the viscosity of the lactide-glycolide copolymer is 0.16 to 0.60 dl / g; more preferably, the viscosity of the lactide-glycolide copolymer is 0.20 dl / g; The molar ratio of lactide to glycolide in the lactide-glycolide copolymer is 90:10 to 25:75; preferably, the molar ratio of lactide to glycolide in the lactide-glycolide copolymer is 75:25 to 50:

50.

5. The drug sustained-release microsphere composition according to claims 1-4, characterized in that: The viscosity of the first lactide-glycolide copolymer is 0.08 to 0.30 dl / g, and the viscosity of the second lactide-glycolide copolymer is 0.32 to 0.90 dl / g; Preferably, when the lactide-glycolide copolymer is selected from two of a first lactide-glycolide copolymer and a second lactide-glycolide copolymer, the viscosity of the first lactide-glycolide copolymer is less than the viscosity of the second lactide-glycolide copolymer; Preferably, when the lactide-glycolide copolymer is selected from two of a first lactide-glycolide copolymer and a second lactide-glycolide copolymer, the mass ratio of the first lactide-glycolide copolymer to the second lactide-glycolide copolymer is 1:9 to 9:1; preferably, the mass ratio of the first lactide-glycolide copolymer to the second lactide-glycolide copolymer is 1:

1.

6. A method for preparing the drug sustained-release microsphere composition according to any one of claims 1 to 5, characterized in that: The method comprises the following steps: (1) dissolving a lactide-glycolide copolymer in a first organic solvent to prepare a carrier solution; (2) dissolving baricitinib or a pharmaceutically acceptable salt thereof in a second organic solvent to prepare a drug solution; (3) dissolving a hydrophilic polymer in deionized water to prepare an external aqueous phase solution; (4) adding the carrier solution obtained in step (1) to the drug solution obtained in step (2), mixing, and obtaining an oil phase solution; (5) adding the oil phase solution obtained in step (4) dropwise to the external water phase solution obtained in step (3) under shear conditions to prepare an O / W emulsion; (6) stirring to evaporate the first organic solvent and the second organic solvent, filtering to collect, and washing to obtain a microsphere suspension; (7) adding a lyophilization protectant to the microsphere suspension obtained in step (6), and lyophilizing to obtain a baricitinib sustained-release microsphere composition; Wherein, the lactide-glycolide copolymer is selected from one or both of a first lactide-glycolide copolymer and a second lactide-glycolide copolymer.

7. The preparation method according to claim 6, characterized in that: In the step (1), the concentration of the lactide-glycolide copolymer in the carrier solution is 120 to 500 mg / mL; In the step (3), the hydrophilic polymer comprises at least one of methyl cellulose, polyvinyl pyrrolidone, carboxymethyl cellulose, lecithin, gelatin, polyvinyl alcohol, polyoxyethylene-polyoxypropylene block copolymer, polyoxyethylene sorbitan fatty acid ester and polyoxyethylene; preferably, the hydrophilic polymer is selected from polyvinyl alcohol; further, the concentration of the hydrophilic polymer in the external aqueous phase solution is 0.5% to 2%; In the step (5), the shearing conditions include a rotation speed of 3000 to 6000 rpm, and a dripping speed of 0.1 to 2.0 mL / min; further, the volume ratio of the oil phase solution to the external aqueous phase solution is 1.0:200 to 2.0:200; In the step (6), the first organic solvent is selected from one or more of dichloromethane, chloroform, ethyl acetate and methanol; the second organic solvent is selected from one or more of dimethyl sulfoxide, dimethylformamide and N-methylpyrrolidone; further, the volume ratio of the second organic solvent to the first organic solvent is 1:7 to 1:

2.

8. The preparation method according to claim 6, characterized in that: The freeze-drying process in step (7) specifically includes: S1: pre-freezing the microsphere suspension after adding the lyophilization protective agent; S2: further subliming the product obtained in S1; S3: drying the product obtained in S2 to obtain a baricitinib sustained-release microsphere lyophilized preparation; The pre-freezing process parameters in S1 are: setting temperature -30 to -50°C, holding time 4 to 6 hours; preferably, the pre-freezing process parameters are: setting temperature -40°C, holding time 5 hours; The drying process in S3 includes sublimation drying and analytical drying, wherein the sublimation drying process includes: controlling the vacuum degree at 0.1-0.3 mbar, heating to -10--20°C at 0.4-0.6°C / min, and maintaining at -10--20°C for 10-12 hours; the analytical drying process includes: controlling the vacuum degree at 0.1-0.3 mbar, heating to 10-30°C at 0.4-0.6°C / min, and maintaining at 10-30°C for 9-11 hours.

9. Use of the drug sustained-release microsphere composition according to any one of claims 1 to 5 or the drug sustained-release microsphere composition obtained by the preparation method according to any one of claims 6 to 8 in the preparation of drugs, characterized in that: The medicine is used for preventing or treating alopecia areata, rheumatoid arthritis or atopic dermatitis.

10. The use according to claim 9, characterized in that: The drug is administered by injection.

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