Sustained-release pharmaceutical composition comprising GLP-1 receptor agonist

Abstract

The present invention relates to a sustained-release pharmaceutical composition using a GLP-1 receptor agonist. Specifically, microparticles comprising semaglutide are prepared using a 5-hydroxydopamine-modified self-crosslinkable hyaluronic acid derivative (SAMH) as a carrier, and the semaglutide is gradually released over a long period of time upon injection into the body, thereby providing excellent effects on blood glucose control and weight management. The sustained-release pharmaceutical composition according to the present invention maintains a low viscosity before administration to the human body, thus being easily administered by injection, and is self-crosslinked and hydrogelated after administration to the human body, and thus can maintain the pharmacological effect of the semaglutide for a long period of time with only a single administration.

Description

Extended-release pharmaceutical composition containing a GLP-1 receptor agonist

[0001] The present invention relates to the pharmaceutical use of hyaluronic acid derivatives and hydrogels using the same. More specifically, the present invention relates to a sustained-release pharmaceutical composition comprising a GLP-1 receptor agonist.

[0002]

Technology underlying the invention

[0003] With the rapid growth of industries such as the medical, bio, and cosmetics industries, interest in functional biomaterials is increasing. In particular, to address the toxicity and side effects that can be caused by chemically synthesized polymers, the development of biocompatible materials using more stable natural polymers is emerging as a critical challenge. Against this backdrop, active research is being conducted on the natural polymer hyaluronic acid.

[0004] Hyaluronic acid is a hydrophilic, bio-derived polymer found abundantly in various layers of the skin. It possesses a complex range of functions, including hydration, supporting the formation of the extracellular matrix, acting as a filler, and participating in tissue regeneration mechanisms. For example, it is known to provide physical cushioning and friction reduction within joints, while in skin it maintains flexibility and protects against microbial invasion.

[0005] Hyaluronic acid is readily degraded in vivo by hyaluronidase, an enzyme that degrades hyaluronan. It can be combined with various drugs and utilized as a drug delivery system material. In particular, since receiving approval from the U.S. Food and Drug Administration (FDA), it has been widely used as a medical biomaterial, a scaffold for tissue engineering, and a polymer for drug delivery.

[0006] Conventional hyaluronic acid-related technologies have reported methods for synthesizing insoluble cross-linked hyaluronic acid derivatives using compounds with two functional groups, such as bis-epoxides, bis-halides, and formaldehyde. For example, U.S. Patent No. 4,582,865 discloses a hyaluronic acid cross-linking method using divinylsulfone, and U.S. Patent No. 4,713,448 proposes a cross-linking reaction using formaldehyde. Furthermore, U.S. Patent No. 5,356,883 discloses the synthesis of a hyaluronic acid derivative gel modified with O-acylurea or N-acylurea using carbodiimides. However, hyaluronic acid cross-links manufactured in the prior art have low resistance to hyaluronic acid degrading enzymes and a high possibility of residual unreacted chemicals, which may cause biotoxicity issues. In addition, it was difficult to control the degree of cross-linking or physical properties suitable for the intended application, which limited its use as a variety of medical materials.

[0007] The present inventors conducted research to overcome the limitations of existing hyaluronic acid-based technologies and develop functional hyaluronic acid derivatives with superior biocompatibility. Accordingly, we developed a hydrogel platform technology based on hyaluronic acid modified with pyrogallol groups and filed for Korean Patent Application No. 10-2019-7025688.

[0008] The present inventors have completed the present invention by manufacturing microparticles containing a drug requiring sustained release using a hyaluronic acid derivative according to Korean Patent Application No. 10-2019-7025688 and developing a sustained release drug delivery system through this.

[0009] Thus, the purpose of the present invention is to provide a hyaluronic acid derivative prepared by modifying hyaluronic acid with 5-hydroxyldopamine and its pharmaceutical use.

[0010] Another object of the present invention is to provide a hyaluronic acid derivative or a method for delivering a biomolecule or drug into a living body using the same.

[0011] Another object of the present invention relates to a modified hyaluronic acid derivative or its use for delivering a biomolecule or drug into a living body.

[0012] Another object of the present invention is to provide a drug delivery carrier (or vehicle) or drug delivery system (DDS) using the hyaluronic acid derivative for delivering a biomolecule or drug into a living body.

[0013] Another object of the present invention is to provide a drug delivery carrier or drug delivery system using the hyaluronic acid derivative for delivering a biomolecule or drug requiring sustained release into the body.

[0014] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

[0015] The term “biomolecule” used in the present invention comprehensively refers to biomolecules mentioned herein, including proteins, peptides, nucleic acids (DNA / RNA), antibodies, antibody fragments, siRNA, stem cells, etc.

[0016] In the present invention, “singular”, “a or an” may be interpreted to mean plural.

[0017] In the present invention, “sustained release” means a characteristic of being released slowly while maintaining an effective concentration for a specific period of time when administered into the body.

[0018] In the present invention, “carrier (or vehicle)” means a matrix or support for delivering a desired biomolecule or drug into the body by binding, adsorbing, mixing, or other methods.

[0019] In the present invention, the term “self-crosslinking or self-crosslinking type” means hyaluronic acid other than hyaluronic acid to which a crosslinking agent is bound, and which has the property of crosslinking and forming a hydrogel after injection into the body.

[0020] In the present invention, "injectable water" refers to pure water that can be directly injected for a specific purpose. It is purified water, primarily used to dissolve or dilute drugs for injection. This is necessary to maintain the effectiveness of the drug and ensure safe delivery.

[0021] In the present invention, “normal saline” means a solution manufactured to maintain an environment similar to that of human cells, which is a 0.9% sodium chloride (NaCl) solution.

[0022] The western-type pharmaceutical composition of the present invention may contain biomolecules and / or drugs in a range of 0.01 mg / kg to 50 mg / kg. However, the dosage may be adjusted to exceed or fall below this range depending on the condition of a specific patient.

[0023] The sustained-release pharmaceutical composition manufactured with microparticles of the present invention is dissolved in water for injection or saline solution and injected prior to human administration, and may be administered via various routes of administration depending on the patient's condition, therapeutic purpose, and drug properties. For example, it may be administered intramuscularly (IM), subcutaneously (SC), or topically, and, if necessary, transdermally, mucosally, nasally, or by inhalation.

[0024]

Means of solving the problem

[0025] In order to achieve the above-described purpose of the present invention, a hyaluronic acid derivative modified with 5-hydroxyldopamine was prepared according to the method described in the applicant's prior patent document, Korean Patent Application No. 10-2019-7025688, and used in the present invention.

[0026] The above hyaluronic acid derivative has the property of self-crosslinking depending on the oxidation reaction and pH conditions after injection into the body, and the inventors of the present invention named it SAMH, an abbreviation for “Self-Assembled Modified Hyaluronic Acids,” meaning a hyaluronic acid derivative modified by self-crosslinking.

[0027] The SAMH manufactured according to the above-mentioned prior patent document of the present applicant has a structure represented by the following chemical formula 1, wherein the molecular weight of the hyaluronic acid derivative may be 10,000 Da to 5,000,000 Da, and the 5-hydroxyldopamine substitution rate of the hyaluronic acid derivative may be 0.1% to 50%.

[0028]

[0029] In the above chemical formula 1, R1 is a hydroxyl group or 5-hydroxyldopamine, , and n is an integer from 1 to 1,000.

[0030] It has been confirmed that after injection into the body, SAMH can form a hydrogel through cross-linking as represented by the following chemical formula 2 or chemical formula 3.

[0031]

[0032] In the above chemical formula 2, HA' represents hyaluronic acid in which the carboxyl group is substituted with an amide group.

[0033]

[0034] In the above chemical formula 3, HA' represents hyaluronic acid in which the carboxyl group is replaced with an amide group.

[0035] In addition, the present invention provides a carrier containing a hyaluronic acid derivative of the above chemical formula 1, a biomolecule or a drug. In one embodiment of the present invention related thereto, the drug delivery carrier includes, as a biomolecule or drug, a small molecule chemical compound, an antibody, an antibody fragment, a protein, a peptide, a polypeptide, DNA and / or RNA, siRNA, a gene, and stem cells including stem cells, mesenchymal stem cells, or induced pluripotent stem cells (iPSCs), but is not limited thereto.

[0036] In this regard, in another embodiment of the present invention, the carrier comprising SAMH provides sustained release of a biomaterial or drug in vivo and in vitro.

[0037] In addition, the present invention provides a carrier for drug delivery, comprising a hyaluronic acid derivative (SAMH) of the above chemical formula 1; and a biomaterial or drug requiring sustained release in the body.

[0038] In addition, another embodiment of the present invention provides a sustained-release pharmaceutical composition comprising a hyaluronic acid derivative (SAMH) of the above chemical formula 1; and a biomaterial or drug requiring sustained release in the body.

[0039] In another embodiment of the present invention, the carrier or sustained-release pharmaceutical composition is provided as microparticles having a size of 1,000 μm or less. In the specification and claims of the present invention, the microparticles thus prepared are referred to as SAMH-MP.

[0040] In another embodiment of the present invention, the carrier or pharmaceutical composition can be dissolved in water for injection or physiological saline solution at 0.1% (w / v) to 15% (w / v) and then administered to the body.

[0041] In addition, the present invention provides a method for preparing a carrier for drug delivery or a sustained-release pharmaceutical composition, comprising the steps of preparing a hyaluronic acid derivative by reacting hyaluronic acid and 5-hydroxydopamine; and the step of preparing the hyaluronic acid derivative and a biomaterial and / or drug into microparticles.

[0042] In particular, the present invention relates to a sustained-release pharmaceutical composition comprising a GLP-1 receptor agonist (e.g., semaglutide). SAMH-based hydrogels or microparticles are cross-linked in the body, allowing for gradual drug release, thereby offering the advantage of maintaining a constant concentration over a long period of time with a single administration.

Effect of the invention

[0043] The sustained-release drug delivery carrier or sustained-release pharmaceutical composition according to the present invention is based on hyaluronic acid, which has excellent biocompatibility, and exhibits very low viscosity when dissolved in water for injection or saline solution before human administration, making it easy to administer by injection. After being injected into the body, it self-crosslinks to form a hydrogel, thereby providing a sustained-release effect over a long period of time. This can improve the convenience of taking and treatment compliance of a GLP-1 receptor agonist effective in the treatment of diabetes and obesity.

[0044] Figure 1 schematically illustrates the synthesis of a self-crosslinked modified hyaluronic acid derivative (SAMH) prepared by reacting with 5-hydroxyldopamine.

[0045] Figure 2 shows an electron microscope image of SAMH / semaglutide microparticles manufactured according to the present invention.

[0046] Figure 3 shows the blood concentration of semaglutide over time as measured by an ELISA kit. In each figure, SAMH-MP-SEMA stands for SAMH / semaglutide microparticles.

[0047] Figure 4 shows the results of blood sugar measurement.

[0048] Figure 5 shows the results of weight change observation.

[0049]

Specific details for carrying out the invention

[0050] Hereinafter, the present invention will be described in detail according to manufacturing examples and examples. Manufacturing example Manufacturing Example 1: Manufacturing of self-crosslinked modified hyaluronic acid derivatives

[0051] According to the method described in the detailed description of Korean Patent Application No. 10-2019-7025688 of the present applicant, as shown in Fig. 1, a hyaluronic acid derivative modified with a gallic acid group of the present invention was prepared. Specifically, hyaluronic acid (molecular weight 200K, Lifecore Biomedical, IL, USA) was first completely dissolved in triple-distilled water (TDW). Next, NHS (N-hydroxysuccinimide, Sigma, St. Louis, MO, USA) and EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (Thermo Scientific, Rockford, IL, USA)) were added, followed by the addition of 5'-hydroxydopamine (5-hydroxydopamine, Sigma) and the reaction for 24 hours. Next, the remaining EDC, NHS, and 5'-hydroxydopamine in the reaction solution were removed, and then freeze-dried to prepare a self-crosslinked modified hyaluronic acid derivative (SAMH) in the form of a white powder. Manufacturing Example 2: Manufacturing of SAMH / semaglutide microparticles (SAMH-MP)

[0052] Microparticles were prepared using SAMH obtained in Manufacturing Example 1 and semaglutide, a GLP-1 receptor agonist. To this end, the lyophilized SAMH obtained in Manufacturing Example 1 was dissolved in distilled water to a final concentration of 0.1-10% (w / v). Semaglutide (MCE, HY-114118), a GLP-1 receptor agonist, was added to the SAMH solution at a concentration of 0.1-10 mg / mL and mixed homogeneously. The solvent was evaporated using a spray lyophilizer or a spray dryer (Buchi Labortechnik AG, Flawil, Switzerland) according to the manufacturer's instructions, and microparticles composed of SAMH / semaglutide with an average diameter of 1-5 μm were prepared. SAMH and semaglutide microparticles were confirmed using an electron microscope (ref. ). The obtained SAMH / semaglutide microparticles were dissolved in physiological saline or water for injection to have a concentration in the range of 10-20 mg / mL and used in the following tests. Example Example 1: Induction of a diabetes model using SD rats

[0053] Twenty 6-week-old SD rats were purchased from Orient Bio (South Korea). They were divided into 10 cages, with two rats per experimental group. They were acclimated for 1 week under a 12-h light / dark cycle, at 20–25°C, with free access to solid food and water. All procedures were performed in accordance with the guidelines for the care and use of laboratory animals, and all protocols were approved by the applicant's Institutional Animal Care and Use Committee.

[0054] SD rats (7 weeks old, averaging 200-230 g) were fasted for 6 h and then injected with nicotinamide (NA), Sigma, N0636) and streptozotocin (STZ, Sigma, S0130). 110 mg / kg of NA was injected intraperitoneally, followed by 65 mg / kg of STZ. Water containing 10% sucrose (Sigma, S0389) was provided for 24 h after administration. After the STZ and NA treatments, blood samples were collected from the tail vein of the rats and blood glucose levels were measured using a portable Bayer Contour Next glucose monitor (Bayer). Rats were selected as diabetic rats if their blood glucose levels were measured in the range of 130-200 mg / dL for 3 or more consecutive times without fasting, and were used in the following experiments. For the experiment, rats were divided into five groups (2-5 rats per group). All samples were dissolved in phosphate buffered saline (PBS) and administered subcutaneously (sc) to rats: Category Experimental animals and sample Dosage Normal group Normal rat PBS 1 mL, single administration Negative control group 1 Diabetic rat PBS 0.5 mL, once daily Negative control group 2 Diabetic rat, SAMH microparticles 0.9 mg 1 mL, single administration Positive control group Diabetic rat, semaglutide 20 μg 0.5 mL, once daily Drug administration group Diabetic rat, SAMH / semaglutide microparticles 0.9 mg 1 mL, single administration Example 2: Confirmation of drug release and blood sugar changes

[0055] For all experimental groups, body weight and blood glucose levels were measured and recorded before and after drug administration. After drug administration, blood samples were collected from the tail vein, centrifuged (2,000 × g) for 15 minutes, and plasma was separated. The plasma was then stored at -80°C. Plasma was collected daily for the first 7 days and twice a week thereafter. Semaglutide concentrations in the collected plasma were analyzed using an ELISA kit (KRIBIOLISA, KBI5030). Measurement of semaglutide blood concentrations

[0056] Among the 20 rats used in the experiment, 18 were induced to have diabetes with NA / STZ, and showed an average of 17% body weight gain and high blood glucose levels of 155-173 mg / dL. Tail vein blood sampling was performed according to the experimental schedule after drug administration. The drug administration group that received a single dose of SAMH / semaglutide microparticles showed plasma semaglutide concentrations similar to those of the positive control group that received 20 μg of semaglutide once daily, and it was confirmed that semaglutide was released for up to 23 days (reference). Blood sugar changes

[0057] After the experiment, blood sugar levels were compared to those before drug administration. The positive control group, which received 20 μg of semaglutide daily, showed a 28% decrease compared to before drug administration. In contrast, the group administered a single dose of SAMH / semaglutide microparticles showed a decrease of approximately 30% (see reference). Weight changes

[0058] After monitoring and comparing body weight changes for 28 days, the positive control group treated with semaglutide injection showed a 45% increase, while the drug-administered group receiving SAMH / semaglutide injection showed a 39% increase (see reference).

[0059] In summary, the results of the above manufacturing examples and examples show that the SAMH / semaglutide microparticles of the present invention significantly exhibit blood sugar lowering and weight control effects through long-term sustained release.

[0060] The foregoing description of the present invention is provided for illustrative purposes only. Those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

A sustained-release pharmaceutical composition for treating diabetes and obesity, comprising a carrier composed of microparticles of a self-crosslinking hyaluronic acid derivative (SAMH) having a structure of the following chemical formula 1 and a pharmacologically effective amount of semaglutide: (In the above chemical formula 1, R1 is a hydroxyl group or 5-hydroxyldopamine, , and n is an integer from 1 to 1,000.) . In claim 1, the hyaluronic acid derivative is prepared by reacting hyaluronic acid with 5-hydroxydopamine, and is characterized in that it self-crosslinks by oxidation and pH change after being injected into the body, and is a sustained-release pharmaceutical composition. A sustained-release pharmaceutical composition according to claim 1 or 2, characterized in that the fine particles have an average diameter of 500 μm or less. A sustained-release pharmaceutical composition according to claim 1 or 2, wherein the microparticles are dissolved or suspended in water for injection or normal saline and administered in an injection form. A sustained-release pharmaceutical composition according to any one of claims 1 to 2, characterized in that the semaglutide is sustained-released in the body for 3 weeks or longer. A sustained-release pharmaceutical composition, characterized in that the diabetes in claim 1 or 2 is type 2 diabetes. A sustained-release pharmaceutical composition according to claim 1 or 2, characterized in that the sustained-release pharmaceutical composition is effective for both weight control and blood sugar control. A method for producing a sustained-release pharmaceutical composition for treating diabetes and obesity, comprising: a step of producing a self-crosslinking hyaluronic acid derivative (SAMH) by reacting hyaluronic acid and 5-hydroxydopamine; and a step of producing microparticles by mixing a GLP-1 receptor agonist into the hyaluronic acid derivative (SAMH). A method for producing a sustained-release pharmaceutical composition, characterized in that in claim 8, the microparticles are produced through a spray drying or spray freeze drying process. A method for producing a sustained-release pharmaceutical composition, characterized in that the concentration of semaglutide in claim 8 or 9 is in the range of 0.01 to 100 mg / mL.

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