Sustained-release lipid pre-concentrate comprising sorbitan saturated fatty acid and method for preparing same

Abstract

The present invention relates to a lipid pre-concentrate and a method for preparing same, wherein the lipid pre-concentrate uses a biodegradable lipid having low injection force and low toxicity to delay the release rate of a drug, thereby exhibiting sustained release so that the administered drug can be continuously released for a long period of time.

Description

Sustained-release lipid precursor preparation containing sorbitan saturated fatty acid and method for preparing the same

[0001] The present invention relates to a lipid precursor formulation that exhibits sustained-release properties by delaying the release rate of a drug using sorbitan saturated fatty acid, a biodegradable lipid with low injectability and low toxicity, so that the administered drug can be continuously released for a long period of time, and a method for producing the same.

[0002]

[0003] Sustained-release concentrate is a formulation that continuously releases pharmacologically active substances with a single administration, preventing side effects that may occur with repeated administration and maintaining the effective concentration range of the pharmacologically active substance for a certain period of time or longer.

[0004] A representative sustained-release material currently in use due to its biodegradability is PLGA (poly(lactic-co-glycolic acid)), which has been approved by the Food and Drug Administration (FDA). U.S. Patent No. 5,480,656 discloses that PLGA, a biodegradable polymer, decomposes into lactic acid and glycolic acid over a certain period of time in the body, allowing for the sustained release of pharmacologically active substances. However, acidic substances, which are breakdown products of PLGA, can cause inflammatory responses and reduce cell proliferation rates (K. Athanasiou, GG Niederauer, and CM Agrawal, Biomaterials, 17, 93 (1996)). Furthermore, for sustained release, the drug must be encapsulated in PLGA solid particles measuring 10–100 micrometers and injected, but this can cause pain and inflammation during injection. Therefore, there is a need to develop a novel sustained-release formulation that can provide an effective concentration of pharmacologically active substances for a certain period of time while improving patient compliance.

[0005] As a formulation that can avoid the disadvantages of biodegradable polymer-based formulations, WO 2005 / 117830 discloses a liquid depot formulation comprising a low viscosity organic solvent comprising at least one neutral diacyl lipid (e.g., a diacylglycerol such as glyceryl dioleate) and / or at least one tocopherol; at least one phospholipid; and at least one biocompatible oxygen. However, formulations containing neutral diacyl lipids such as glyceryl dioleate have the problem of low biodegradability, are not bio-derived substances, have limited biocompatibility, and are highly likely to induce inflammation.

[0006] Meanwhile, Japanese Patent No. 5681626 discloses a sustained-release formulation comprising microspheres and a phospholipid component, wherein the microspheres are combined with a lipid component containing a sterol and a polylactide-co-glycolide (PLGA) polymer, and the release rate can be controlled. However, the lipid component is not sufficient to suppress the initial release, and its strong hydrophobicity has the disadvantage of deteriorating the properties of the microspheres. Furthermore, modification of the polylactide-co-glycolide (PLGA) polymer forming the microspheres causes difficulties in obtaining approval for drug use, limiting the practical application of the composition in a depot formulation.

[0007] Therefore, there is a need in the art to develop a sustained-release lipid formulation for injection that can prevent early drug release and has excellent biodegradability, biocompatibility, and injection power as a long-acting drug injection formulation.

[0008]

[0009] [Prior Art Literature]

[0010] 1. U.S. Patent No. 5,480,656

[0011] 2. International Patent Publication No. WO 2005 / 117830

[0012] 3. Japanese Patent No. 5681626

[0013]

[0014] The present invention aims to provide a lipid precursor formulation that exhibits sustained release properties by delaying the release rate of a drug while having a low injection force, so that the administered drug can be continuously released for a long period of time.

[0015] In addition, the present invention aims to provide a method for producing a lipid precursor preparation exhibiting the above-described effects.

[0016]

[0017] The present invention comprises phospholipids; sorbitan saturated fatty acids; and squalene,

[0018] A lipid precursor formulation that forms liquid crystals in an aqueous medium is provided.

[0019]

[0020] The present invention also provides a method for preparing the aforementioned lipid precursor formulation, comprising the step of preparing a lipid solution by mixing phospholipids; sorbitan saturated fatty acids; and squalene.

[0021]

[0022] The lipid precursor formulation of the present invention can form liquid crystals upon exposure to an aqueous medium, enabling sustained drug release, and can sustain drug release for approximately one week to one month. Furthermore, the liquid crystals formed by the lipid precursor formulation exhibit sustained drug release even at low viscosities, exhibit excellent viscoelasticity, and have low extrusion force during injection, providing excellent usability.

[0023]

[0024] Figure 1 is a photograph of the lipid precursor preparation manufactured in Manufacturing Example 1 injected into a pH 7.4 phosphate buffer solution.

[0025] Figure 2 is a photograph of the lipid precursor preparation manufactured in Manufacturing Example 2 injected into a pH 7.4 phosphate buffer solution.

[0026] Figure 3 is a photograph taken after a certain period of time after administering the lipid precursor preparation manufactured in Manufacturing Example 2 to SD RAT.

[0027] Figure 4 is a photograph of the phase (left) and gel phase (right) of the lipid precursor preparation manufactured in Manufacturing Example 3-1.

[0028] Figure 5 is a photograph of the properties (left) and gel properties (right) of the lipid precursor preparation manufactured in Manufacturing Example 3-13.

[0029] Figure 6 shows the results of PLM (polarized light microscopy) analysis of the lipid precursor preparation manufactured in Manufacturing Example 3-1.

[0030] Figure 7 shows the Cyro-TEM analysis results of the lipid precursor preparation manufactured in Manufacturing Example 3-1.

[0031] Figure 8 shows the results of animal PK evaluation of the lipid precursor preparation manufactured in Manufacturing Example 3-1 and the lipid precursor preparation manufactured in Comparative Example.

[0032] Figure 9 shows the results of bioimaging analysis of the lipid precursor preparation manufactured in Manufacturing Example 3-1.

[0033]

[0034] The present invention comprises phospholipids; sorbitan saturated fatty acids; and squalene,

[0035] It relates to a lipid precursor formulation that forms liquid crystals in an aqueous medium.

[0036]

[0037] Hereinafter, the present invention will be described in detail.

[0038] In the present invention, the pre-concentrate is a dosage form capable of releasing the drug over a long period of time, and corresponds to a sustained-release formulation. More specifically, it may be a sustained-release injectable formulation or oral formulation. A sustained-release formulation must gradually release the drug within it to the intended level over a certain period of time. In the present invention, the pre-concentrate is in the form of a pre-formulated lipid solution, and when exposed to an aqueous medium such as water or a biological fluid, it forms non-lamellar, specifically non-lamellar liquid crystals (hereinafter referred to as "liquid crystals"). In other words, the lipid precursor formulation of the present invention can form sustainable liquid crystals in the body. The liquid crystals may be used interchangeably with gels or depots.

[0039] In the present invention, the lipid precursor formulation comprises phospholipids; sorbitan saturated fatty acid; and squalene.

[0040] In the present invention, phospholipids can serve as a liquid crystal forming agent.

[0041] In one specific example, the phospholipid may be at least one selected from the group consisting of lecithin, soybean phospholipid, egg yolk phospholipid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidic acid, phosphatidylglycerine, sphingomyelin, phosphatidylinositol, hydrogenated phospholipids, and lysophospholipids.

[0042] In one specific example, the content of the phospholipid may be 30 to 60 wt%, or 30 to 45 wt%, based on the total weight (100 wt%) of the lipid precursor formulation. If the content of the phospholipid is less than 30 wt% or more than 60 wt%, the ability to form liquid crystals when the lipid precursor formulation is injected into a living body may be significantly reduced, and the ability of the drug to be released sustainably, as described later, may be reduced. As a result, there is a concern that the release control ability of the present invention may not be demonstrated for more than one week as desired.

[0043] In the present invention, sorbitan saturated fatty acid can play the role of a liquid crystal forming agent like phospholipid.

[0044] In one specific embodiment, the sorbitan saturated fatty acid may be at least one selected from the group consisting of sorbitan caprylate, sorbitan laurate, sorbitan monolaurate, sorbitan myristate, sorbitan palmitate, sorbitan monopalmitate, sorbitan stearate, sorbitan monostearate, sorbitan tristearate, sorbitan isostearate, and sorbitan monomyristate.

[0045] The content of the above sorbitan saturated fatty acid may be 5 to 45 wt%, 10 to 35 wt%, 15 to 30 wt%, or 25 to 30 wt% based on the total weight (100 wt%) of the lipid precursor formulation. Within the above content range, a release control ability of one week or longer can be secured.

[0046] In the present invention, squalene can serve as a liquid crystal strengthening agent. That is, squalene can help maintain the internal structure of the liquid crystal (stiffening) and can play a role in delaying the release rate of the drug.

[0047] In one specific example, the content of squalene may be 2.5 to 30 wt%, or 5 to 15 wt%, based on the total weight (100 wt%) of the lipid precursor formulation. Within the above content range, the internal structure of the liquid crystals can be strongly maintained.

[0048] The lipid precursor formulation of the present invention can function as a sustained-release formulation capable of long-term drug release and can be used in the form of a sustained-release injectable formulation. By utilizing the lipid precursor formulation as a sustained-release injectable formulation in the present invention, the drug contained in the formulation can be induced and controlled to be gradually released at the intended level.

[0049] The lipid precursor formulation of the present invention may additionally include a solvent. This solvent may serve to improve the solubility or injectability of the active ingredient. When a sustained-release lipid precursor formulation containing a solvent is injected into a living body, the solvent may be displaced or diluted by the biological fluid during the formation of liquid crystals and thus removed.

[0050] In one specific example, the type of solvent is not particularly limited, and any solvent that can be introduced into the human body in the form of an injection can be used. For example, the solvent may be one or more selected from the group consisting of water, ethanol, isopropanol, propylene glycol, N-methylpyrrolidone, glycerol, benzyl benzoate, acetic acid, and benzyl alcohol.

[0051] In one specific embodiment, the solvent content may be 5 to 25 wt%, or 10 to 20 wt%, based on the total weight of the lipid precursor formulation (100 wt%).

[0052] In one specific example, when water is used as a solvent, it may be used in the form of a mixture of water and a second solvent. The second solvent may be any of the solvents described above. In this case, the water content may be 5 wt% or less, or 3 wt% or less, and the second solvent content may be 5 to 20 wt%. Water within the above content range does not affect the formation of liquid crystals during the preparation and storage of the lipid precursor formulation.

[0053] The lipid precursor formulation of the present invention may additionally include a stabilizer. The stabilizer helps maintain the internal structure of the liquid crystals and may enable the production of strong liquid crystals.

[0054] In one specific example, the type of stabilizer is not particularly limited and may be one or more selected from the group consisting of tocopherol, tocopherol acetate, and cholesterol.

[0055] The lipid precursor formulation of the present invention may further comprise an antioxidant. The antioxidant may be used to improve the stability of the lipid precursor formulation and the drug.

[0056] In one specific example, the type of antioxidant is not particularly limited and may be at least one selected from the group consisting of sodium ascorbate (ViC_Na), ascorbic palmitate (ViC_palmitate), butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), methionine, monothioglycerol, sodium thiosulfate, and sodium metabisulfite.

[0057] The lipid precursor formulation of the present invention may additionally include a surfactant. The surfactant may serve to improve the dissolution of the main ingredient.

[0058] In one embodiment, the surfactant may be one or more selected from the group consisting of polysorbates (tween), cremophor, solutol, brize, Triton, glycolipid esters (e.g., sucrose oleate, sucrose palmitate and / or sucrose laurate) and surfactants having a polyoxyethylene head group.

[0059] The lipid precursor formulation of the present invention may additionally contain a drug.

[0060] In one specific embodiment, the drug may be selected from the group consisting of an anti-obesity agent, an analgesic agent, an anti-osteoarthritis agent, an anti-hair loss agent, or an anti-prostatic hypertrophy agent.

[0061] In one specific example, the obesity treatment agent may be semaglutide, tirzepatide, liraglutide, or the like.

[0062] In one specific example, the analgesic may be cerecoxib, meloxicam, or the like.

[0063] In one specific example, the osteoarthritis treatment agent may be betamethasone, dexamethasone, methylprednisolone, prednisolone, triamcinolone, or the like.

[0064] In one specific example, the hair loss and benign prostatic hyperplasia treatment may be Dutasteride and finasteride.

[0065] In the present invention, the name of the lipid precursor formulation may vary depending on the type of drug. For example, when semaglutide is used as the drug, the resulting lipid precursor formulation may be named "semaglutide lipid precursor formulation."

[0066] The lipid precursor formulation of the present invention can be used as a pharmaceutical composition depending on the effects, properties, uses, etc. of the drug. The pharmaceutical composition can delay the release of the drug to ensure sustained efficacy.

[0067] The lipid precursor formulation of the present invention may not contain diacylglycerol and / or sorbitan unsaturated fatty acid ester. The above diacylglycerol may be glyceryl dipalmitate, glyceryl phytanoate, glyceryl palmitoleate, glyceryl distearate, glyceryl dioleate, glyceryl dielaidiate, and glyceryl dilinoleate, and the above sorbitan unsaturated fatty acid ester may be sorbitan monooleate, sorbitan monolinoleate, sorbitan monopalmitoleate, sorbitan monomyristoleate, and sorbitan. It may be sorbitan sesquioleate, sorbitan sesquilinoleate, sorbitan sesquipalmitoleate, sorbitan sesquimyristoleate, sorbitan dioleate, sorbitan dilinoleate, sorbitan dipalmitoleate, and sorbitan dimyristoleate.

[0068] Alternatively, the lipid precursor formulation of the present invention may not contain an unsaturated fatty acid having 14 to 20 carbon atoms.

[0069] Specifically, the lipid precursor preparation of the present invention may be a pharmaceutical composition that does not contain one or more of diacylglycerol, sorbitan unsaturated fatty acid ester, and unsaturated fatty acid having 14 to 20 carbon atoms.

[0070] The lipid precursor formulation according to the present invention forms liquid crystals in an aqueous medium.

[0071] In one specific example, the aqueous medium may be water, a biological fluid, or a phosphate buffer solution (PBS).

[0072] The lipid precursor formulation according to the present invention can have an injection force of 1 to 30 N.

[0073] Injectability refers to the extrusion force at a comfortable injection speed for the patient. "Comfortable for the patient" is used to define an injection speed that does not cause injury or excessive pain to the patient during injection. As used herein, "comfort" encompasses not only the patient's comfort but also the comfort or ability of a physician or healthcare professional to inject the composition. In the present invention, injectability can be measured using the INTRONS 5569 device under the conditions of the experimental examples described below. Generally, a composition with a low injection force is less painful and easier to control when injected.

[0074] In the present invention, the injection force of the lipid precursor formulation may be 1 to 30 N or 1 to 13 N.

[0075]

[0076] The present invention also relates to a method for preparing the aforementioned lipid precursor formulation. The types and contents of the ingredients used in the method for preparing the lipid precursor formulation are as described for the lipid precursor formulation.

[0077] The method for preparing a lipid precursor formulation of the present invention may include a step of preparing a lipid solution by mixing phospholipids, sorbitan saturated fatty acids, and squalene (hereinafter, referred to as the lipid solution preparation step). In the above preparation method, the lipid solution may be used as a lipid precursor formulation.

[0078] In the lipid solution preparation step of the present invention, the lipid solution may additionally include a solvent. In this case, the lipid solution may be prepared by mixing and dissolving phospholipids, sorbitan saturated fatty acids, and squalene in the solvent.

[0079] In one embodiment, the lipid solution may additionally comprise a drug.

[0080] The above drug may be at least one selected from the group consisting of obesity treatment drugs, pain relievers, osteoarthritis treatment drugs, and hair loss and benign prostatic hyperplasia treatment drugs.

[0081] In one embodiment, the lipid solution may additionally comprise a stabilizer, an antioxidant, and / or a surfactant.

[0082] In the present invention, the lipid solution can be used by putting it into a container or syringe.

[0083]

[0084] In addition, the lipid precursor preparation of the present invention comprises a step of preparing a lipid solution by mixing phospholipids; sorbitan saturated fatty acid; and squalene (hereinafter, the lipid solution preparation step);

[0085] A step of preparing a drug solution containing a drug (hereinafter, drug solution preparation step); and

[0086] It can be prepared through a step of mixing a lipid solution and the drug solution (hereinafter, a mixing step). In the above manufacturing method, the mixed solution prepared through the mixing step can be used as a lipid precursor preparation.

[0087] In the lipid solution preparation step of the present invention, the lipid solution may additionally include a solvent. In this case, the lipid solution may be prepared by mixing and dissolving phospholipids, sorbitan saturated fatty acids, and squalene in the solvent.

[0088] In one embodiment, the lipid solution may additionally comprise a stabilizer, an antioxidant, and / or a surfactant.

[0089] In the present invention, the lipid solution can be used by putting it into a container or syringe.

[0090] In the drug solution manufacturing step of the present invention, the drug solution can be manufactured by mixing and dissolving the drug in a solvent.

[0091] In one specific example, the drug solution can be used by being injected into a container or syringe.

[0092] The mixing step of the present invention can be performed by a method of mixing a lipid solution introduced into a container or syringe and a drug solution introduced into the container or syringe.

[0093] In one specific embodiment, mixing may be performed by connecting two or more syringes and exchanging the solutions within the syringes. This exchange may be repeated two or more times, and specifically, may be performed 2 to 100 times, 10 to 80 times, 30 to 70 times, 40 to 60 times, or 50 times.

[0094]

[0095] The present invention also relates to a sustained-release injectable pharmaceutical composition comprising the aforementioned lipid precursor formulation.

[0096] In one embodiment, the sustained-release injectable pharmaceutical composition may be used for subcutaneous or intramuscular injection.

[0097] The present invention also relates to an oral pharmaceutical composition comprising the aforementioned sustained-release lipid precursor formulation.

[0098]

[0099] The present invention is susceptible to various modifications and various forms. The specific examples and descriptions set forth below are intended solely to aid understanding of the present invention and are not intended to limit the invention to any specific disclosed form. The scope of the present invention should be understood to encompass all modifications, equivalents, and alternatives falling within the spirit and technical scope of the present invention.

[0100]

[0101] Hereinafter, the present invention will be described in detail by way of examples. However, the following manufacturing examples and test examples specifically illustrate the present invention, and the content of the present invention is not limited by the following manufacturing examples and test examples.

[0102]

[0103] Example

[0104] Manufacturing Example 1. Manufacturing of a lipid precursor preparation using sorbitan saturated fatty acid.

[0105] A lipid precursor preparation was prepared with the ingredients and content (g) shown in Table 1 below.

[0106] Specifically, the ingredients in Table 1 below were added to a glass vial and mixed while stirring with a magnetic stirrer at room temperature to 60°C. A lipid precursor formulation was prepared by leaving it at room temperature for about 3 hours.

[0107]

[0108] Manufacturing Example 1-1 Manufacturing Example 1-2 Manufacturing Example 1-3 Manufacturing Example 1-4 Manufacturing Example 1-5 Manufacturing Example 1-6 Manufacturing Example 1-7 Manufacturing Example 1-8 Phosphatidyl choline 35 35 35 35 35 35 35 35 Sorbitan laurate (content, weight %) 0 (0 %) 5 (7.7%) 10 (14.3%) 15 (20.0%) 20 (25.0%) 30 (33.3%) 40 (40.0%) 30 Squalene 10 10 10 10 10 10 10 10 Ethanol 15 15 15 15 15 15 15 15

[0109]

[0110] Test Example 1. Confirmation of gel properties of lipid precursor formulation in water

[0111] The lipid precursor preparation prepared in Manufacturing Example 1 was injected into 5 mL of pH 7.4 phosphate buffer (PBS) using a pipette.

[0112]

[0113] Figure 1 shows a photograph of the lipid precursor preparation manufactured in Manufacturing Example 1 injected into a pH 7.4 phosphate buffer solution. As shown in Figure 1, it can be confirmed that as the content of sorbitan laurate, a sorbitan saturated fatty acid, increases, gel formation in water becomes stronger.

[0114] Additionally, it can be confirmed that no gel is formed when sorbitan laurate is not included or squalene is not included.

[0115] Through this, it can be confirmed that the lipid precursor preparation of the present invention normally exhibits a liquid state, but changes to a gel state when it comes into contact with an aqueous medium.

[0116]

[0117] Manufacturing Example 2. Manufacturing of a lipid precursor formulation using a liquid crystal forming agent

[0118] A lipid precursor preparation was prepared with the ingredients and content (g) shown in Table 2 below.

[0119] Specifically, the ingredients in Table 2 below were sequentially added to a glass vial, and then mixed while stirring with a magnetic stirrer at room temperature to 60°C. A lipid precursor formulation was prepared by leaving it at room temperature for approximately 3 hours.

[0120] In Table 2, oleic acid is an unsaturated fatty acid, sorbitan monooleate and sorbitan sesquioleate are sorbitan unsaturated fatty acids, stearic acid and myristic acid are saturated fatty acids, and glyceryl dioleate is a glyceryl unsaturated fatty acid.

[0121]

[0122] Manufacturing Example 1-6 Manufacturing Example 2-1 Manufacturing Example 2-2 Comparative Manufacturing Example 2-1 Comparative Manufacturing Example 2-2 Comparative Manufacturing Example 2-3 Comparative Manufacturing Example 2-4 Comparative Manufacturing Example 2-5 Comparative Manufacturing Example 2-6 Phosphatidyl Choline 35 35 35 35 35 35 35 35 35 35 Sorbitan Laurate 30 -------- Sorbitan Stearate - 30 ----- Sorbitan Caprylate - 30 ----- Oleic Acid - 30 ----- Sorbitan Monooleate - 30 ---- Sorbitan Sesquioleate - 30 ----- Stearic Acid - 30 ---- Myristic Acid - 30 ----- Glyceryl Dioleate--------30 Squalene101010101010101010 Ethanol151515151515151515

[0123]

[0124] Test Example 2. Confirmation of gel properties in water of lipid precursor formulation using liquid crystal forming agent

[0125] The lipid precursor preparation prepared in Manufacturing Example 2 was injected into 5 mL of pH 7.4 phosphate buffer using a pipette.

[0126]

[0127] Figure 2 shows a photograph of the lipid precursor preparation prepared in Manufacturing Example 2 injected into a pH 7.4 phosphate buffer solution.

[0128] As shown in Fig. 2, in the case of saturated fatty acids such as stearic acid (Comparative Manufacturing Example 2-4) or myristic acid (Comparative Manufacturing Example 2-5), a very weak gel is formed, but in the case of sorbitan laurate (Manufacturing Example 1-6), sorbitan stearate (Manufacturing Example 2-1), and sorbitan caprylate (Manufacturing Example 2-2), which are composed of sorbitan saturated fatty acids, it can be confirmed that strong liquid crystals are formed.

[0129] When sorbitan sesquioleate (Comparative Manufacturing Example 2-3), which is a sorbitan unsaturated fatty acid, was applied, it was confirmed that a relatively weak gel was formed and a phenomenon of spreading on the surface of the water appeared over time.

[0130]

[0131] Test Example 3. Test to confirm the injection power of lipid precursor preparation

[0132] The lipid precursor formulation was filled into a syringe to determine the effect of injection force.

[0133] Specifically, the injection force was evaluated by the load (N) measured when a 23-gauge syringe was mounted on INTRON's 5569 equipment and pressurized at a speed of 100 mm per minute.

[0134] The needle gauges primarily used in general liquid syringes are 23 gauge or larger, which are less painful and easier to use. Therefore, needles of 23 gauge or larger are used. In particular, when the load is 10 N or less, very small needles of 26 gauge or larger can be used, resulting in minimal pain for the patient and low injection force with minimal force.

[0135]

[0136] Table 3 below shows the results of measuring the injection force of the lipid precursor preparation manufactured in Manufacturing Example 2.

[0137]

[0138] (g) Manufacturing Example 1-6 Comparative Manufacturing Example 2-1 Comparative Manufacturing Example 2-2 Comparative Manufacturing Example 2-3 Comparative Manufacturing Example 2-4 Comparative Manufacturing Example 2-5 Comparative Manufacturing Example 2-6 Week Input (N) 9 N15 N32 N35 N9 N9 N41 N

[0139]

[0140] As shown in Table 3, it can be confirmed that formulations using sorbitan unsaturated fatty acid (Comparative Manufacturing Examples 2-2, 2-3), glyceryl unsaturated fatty acid (Comparative Manufacturing Example 2-6), or monounsaturated fatty acid (Comparative Manufacturing Example 2-1) require a strong injection force to be injected compared to formulations using sorbitan saturated fatty acid (Manufacturing Example 1-6) of the present invention. In the case of formulations using saturated fatty acid (Comparative Manufacturing Examples 2-4, 2-5), although they have a low injection force, they have low gel-forming ability in water, and therefore are not suitable as components of lipid precursor formulations.

[0141] That is, it can be confirmed that sorbitan saturated fatty acid has excellent gel-forming ability in water despite having low injection power.

[0142]

[0143] Test Example 4. Evaluation of local irritation after subcutaneous administration of lipid precursor formulation in SD RAT

[0144] The lipid precursor preparation prepared in Manufacturing Example 2 was filled into a syringe and administered subcutaneously to SD rats, etc. After a certain period of time, skin irritation at the administered local site was confirmed.

[0145]

[0146] Figure 3 shows a photograph taken after a certain period of time (7 days) after administering the lipid precursor preparation manufactured in Manufacturing Example 2 to SD RAT.

[0147] As shown in Figure 3, Comparative Manufacturing Example 2-6, which used GDO (glyceryl dioleate), a glyceryl polyunsaturated fatty acid, showed an inflammatory response due to pus and excessive blood vessel formation. In contrast, when the sorbitan saturated fatty acid according to the present invention was used, it was confirmed that almost no inflammatory response was observed.

[0148] In addition, when glyceryl unsaturated fatty acid is applied, there is a problem that the formulation remains after 35 days because it is not completely biodegradable, and when sorbitan unsaturated fatty acid is applied (Comparative Manufacturing Example 2-2), it is not biodegradable and a small amount of the formulation may remain. When sorbitan saturated fatty acid according to the present invention is applied, most of it is biodegradable, so stability can be secured.

[0149]

[0150] Manufacturing Examples 3 and 4. Manufacturing of lipid precursor formulations using semaglutide and tirzepatide drugs.

[0151] According to the ingredients and contents (g) of Tables 4 to 6 below, a lipid precursor preparation containing a drug was prepared.

[0152] Specifically, in Comparative Manufacturing Examples 3-1 and 3-2, Manufacturing Examples 3-1 to 3-12, Comparative Manufacturing Example 4-1, and Manufacturing Examples 4-1 to 4-4, the ingredients (content: g) in Table 4 below were sequentially added to a glass vial, and then mixed while stirring with a magnetic stirrer at room temperature to 60°C. A lipid precursor formulation was prepared by leaving it at room temperature for about 3 hours.

[0153] In Manufacturing Examples 3-13 and 3-14, only the remaining components except for semaglutide, which corresponds to the drug, were mixed and dissolved in the composition of Table 5 below to prepare a solution (preparation of a lipid solution), and then semaglutide was separately dissolved in water for injection to prepare a solution (preparation of a drug solution). After that, the lipid solution and the drug solution were each filled into a syringe, and the syringes were connected, and the drug in the syringes was mixed by alternately moving back and forth between the two syringes more than 50 times to prepare a lipid precursor formulation.

[0154]

[0155] Comparative Manufacturing Example 3-1 Comparative Manufacturing Example 3-2 Manufacturing Example 3-1 Manufacturing Example 3-2 Manufacturing Example 3-3 Manufacturing Example 3-4 Manufacturing Example 3-5 Manufacturing Example 3-6 Semaglutide 55555555 Water------2.5-Acetic acid 55555555 Phosphatidyl choline 35 35 35 40 50 60 35 35 Sorbitan laurate--2 5 20 20 15 22.5 25 Sorbitan monooleate 25 Glyceryl dioleate 25 Ascorbic acid palmitate-------- Tocopherol acetate------- 5 Cholesterol 55 55--5-Squalene 10 10 10 10 10 10 10 Ethanol 15 15 15 15 10 10 15 15 Tween 80-------- Gel formation result Gel formation Gel formation Gel Forming gel Forming gel Forming gel Forming gel Forming gel

[0156]

[0157] Manufacturing Example 3-7 Manufacturing Example 3-8 Manufacturing Example 3-9 Manufacturing Example 3-10 Manufacturing Example 3-11 Manufacturing Example 3-12 Manufacturing Example 3-13 Manufacturing Example 3-14 Semaglutide 55555522 Water--5---33 Acetic acid 55-555--Phosphatidyl choline 4035353525304040 Sorbitan laurate 3025252045152525 Sorbitan monooleate Glyceryl dioleate Ascorbic acid palmitate---5---- Tocopherol acetate-5-55533 Cholesterol 2.555555--Squalene 2.55101020301212 Ethanol 151515155101515 Tween 80--0.5-----Gel formation resultGel formationGel formationGel formationGel formationGel formationWeak gel formationWeak gel formationGel formation

[0158]

[0159] Manufacturing Example 4-1 Manufacturing Example 4-2 Manufacturing Example 4-3 Manufacturing Example 4-4 Comparative Manufacturing Example 4-1 Tirzepatide 11111 Phosphatidyl choline 3535404035 Sorbitan laurate 30252020 Ascorbic acid palmitate 5-tocopherol acetate 5-5-squalene 51515155 Cholesterol 55-5 Ethanol 1010101010 DMSO 99999 Gel formation result Gel formation Gel formation Gel formation Gel formation No gel formation

[0160]

[0161] The lipid precursor preparation was injected into 5 mL of pH 7.4 phosphate buffer using a pipette.

[0162] As a result of confirming gel formation, it can be confirmed that the lipid precursor preparation of Manufacturing Example 3-1 normally exhibits a liquid state, but changes to a gel state when it comes into contact with a pH 7.4 phosphate buffer (Fig. 4).

[0163] In addition, in the case of a method of alternately mixing and injecting a syringe containing a drug dissolved in water for injection and a syringe containing a lipid solution, as in Manufacturing Example 3-13, it can be confirmed that the two solutions are well mixed and exhibit a transparent liquid phase, which then changes to a gel phase in a pH 7.4 phosphate buffer solution (Fig. 5).

[0164]

[0165] Test Example 5. Confirmation of the liquid crystal structure of semaglutide lipid precursor preparation.

[0166] The lipid precursor preparation was applied to a slide glass, and a pH 7.4 phosphate buffer solution was sprayed over the lipid precursor preparation, and left for approximately 1 minute. After slightly removing moisture, a coverslip was placed to secure the preparation, and observation was performed under a polarizing microscope (PLM, NICON / LV100Pol, 5x magnification).

[0167]

[0168] Figure 6 is a photograph observed under a polarizing microscope after the semaglutide lipid precursor preparation prepared in Manufacturing Example 3-1 was sprayed with a pH 7.4 phosphate buffer solution. The photograph appears as a dark black screen, which corresponds to a typical characteristic of a cubic structure among non-lamellar structures.

[0169]

[0170] Additionally, the structure of the lipid precursor preparation was confirmed by Cyro-TEM.

[0171] Specifically, a gel was formed in a pH phosphate buffer solution, dispersed by sonication, and dispensed onto a mesh grid. The sample was then frozen in ethane at -170°C using liquid nitrogen. The grid was then fixed in a cassette in liquid nitrogen and observed in a Cyro-TEM (Thermo Scientific™, Glacios 2) at 200 kV.

[0172] Figure 7 shows the Cyro-TEM analysis results of the lipid precursor preparation of Manufacturing Example 3-1.

[0173] As shown in Fig. 7, a lattice structure is observed within the particle, which confirms that it has typical characteristics of forming a non-lamellar structure.

[0174]

[0175] Test Example 6. Confirmation of sustained release in a release test of semaglutide and tirzetapide lipid precursor formulations.

[0176] Long-term release tests (in vitro dissolution tests) of lipid precursor formulations were conducted by adding 5 mL of pH 7.4 phosphate buffer containing 0.05% Tween 80 to a 6 cc glass vial, and pipetting the lipid precursor formulations of Comparative Manufacturing Example 3-1, Comparative Manufacturing Example 3-2, and Manufacturing Example 3-1 to form gels. The tests were performed using a rotary rotator (PTR-35, lab23009) at a rotation speed of 30 rpm and a temperature of 37°C for 28 days. The collected samples were analyzed by HPLC under the following conditions.

[0177]

[0178] <HPLC 조건>

[0179] Column: Aegispak C18-L (4.6 x 250 mm, 3.0 μm) or equivalent column

[0180] Mobile phase: A solution of 700 mL of 10 mM phosphate buffer A and 300 mL of acetonitrile, adjusted to pH 2.0 with phosphoric acid.

[0181] A solution prepared by mixing 200 mL of B-10 mM phosphate buffer and 800 mL of acetonitrile and adjusting the pH to 2.0 with phosphoric acid.

[0182] Gradient condition mobile phase A(%): B(%)

[0183]

[0184] Time (min)MP A (%)MP B (%)0901019010105050110100200100219010309010

[0185] Flow rate: 1.2 mL / min Temperature: 50℃

[0186] Injection volume: 20 μL

[0187] Wavelength: 200 nm

[0188]

[0189] The release test results of the semaglutide lipid precursor formulation are shown in Table 8 below.

[0190]

[0191] Long-term release day 1 (%)Long-term release day 14 (%)Long-term release day 21 (%)Long-term release day 28 (%)Long-term release day 35 (%)Comparative manufacturing example 3-15.576.4100.0100.0100.0Comparative manufacturing example 3-24.326.327.641.571.5Manufacturing example 3-111.558.175.587.5100.0

[0192]

[0193] As shown in Table 8, in the case of Comparative Manufacturing Example 3-1, it can be confirmed that 100% release is quickly completed on the 21st day, and in the case of Comparative Manufacturing Example 3-2, it can be confirmed that the drug release is very slow with a release of less than 30% for up to 3 weeks and a low release of less than 50% even on the 28th day.

[0194] On the other hand, Manufacturing Example 3-1 took 28 days to reach a release rate of 85% or more, showing superior stable sustained-release properties for one month.

[0195]

[0196] Additionally, the release test results of the tirzetapide lipid precursor formulation are shown in Table 9 below.

[0197]

[0198] Long-term release day 1 (%) Long-term release day 14 (%) Long-term release day 21 (%) Long-term release day 28 (%) Long-term release day 35 (%) Manufacturing example 4-18.548.165.589.5100.0 Comparative manufacturing example 4-15.5100.0100.0100.0100.0

[0199]

[0200] As shown in Table 9, in the case of Comparative Manufacturing Example 4-1, it can be confirmed that the release rate quickly reaches 100% on the 14th day. On the other hand, it can be confirmed that Manufacturing Example 4-1 takes 28 days to reach a release rate of 85% or more, demonstrating superior stable sustained-release properties for one month.

[0201]

[0202] Test Example 7. Confirmation of sustained-release properties in animal PK tests of semaglutide lipid precursor formulations.

[0203] The sustained-release properties of lipid precursor formulations were tested by conducting animal PK experiments to determine the drug release behavior.

[0204] The lipid precursor preparation of Manufacturing Example 3-1 was used, and Novo Nordisk's Ozempic injection was used as a control drug. The drug was injected subcutaneously into the back of four male SD rats, each 7-8 weeks old and weighing an average of 300 g. The control drug was administered at 2.5 mpk and the test drug was administered at 10 mpk. After administration, blood samples were collected at 0, 1, 3, 6, 12, 24, 48, 96, 168, 336, 576, and 672 hours, and plasma separation was performed at 12,000 rpm for approximately 2 minutes at 4°C. The plasma concentration of SD rats was analyzed using LC-MS / MS, and pharmacokinetic analysis was performed by a non-compartmental method using Phoenix® WinNonlin® software (Ver. 8.3, Certara). The analysis conditions are as follows.

[0205]

[0206] <LC-MS / MS 조건>

[0207] Column: ACE C18 (2.1 x 50 mm, 3.0 μm), ACE

[0208] Mobile phase: A-10 mM ammonium acetate buffer

[0209] Acetonitrile containing B-0.1% formic acid

[0210] Gradient condition mobile phase A(%): B(%)

[0211] Temperature: 40℃

[0212]

[0213] Time(min)Flow(ml / min)AB00.455451.50.45952.50.45952.510.4554540.45545

[0214]

[0215] Figure 8 shows changes in the blood concentration of semaglutide.

[0216] As shown in Fig. 8, it can be confirmed that the lipid precursor preparation of Manufacturing Example 3-1 maintains the blood concentration of the drug for 28 days compared to the control drug.

[0217]

[0218] Test Example 8. Bioimaging Test of Semaglutide Lipid Progenitor Formulation in Animals

[0219] Bioimaging tests were conducted in mice by loading fluorescent materials into lipid precursor preparations.

[0220] In Manufacturing Example 3-1, a lipid precursor formulation was prepared using 30 ug of DilC Dye (Invitrogen) instead of the drug. After dissolving the lipid precursor formulation, approximately 200 ul of the prepared sample was administered subcutaneously to BALB / c nude mice (male, 5-6 weeks old). Images were taken using a bioimaging device (lumina X5, PerkinElmer) before administration and at 1, 24, 48, 72, 168, 504, and 672 hours after administration. During the photography, isoflurane was used for inhalation anesthesia, and the images were taken at the wavelength of the dye (ex 620 nm / em 670 nm).

[0221] Figure 9 shows the change in the luminescence level of the fluorescent substance of the formulation loaded with the fluorescent substance administered to the mouse.

[0222] Specifically, G1 is DilC Dye (Invitrogen) dissolved in a solvent and administered subcutaneously at a concentration of 0.15 ul in the corresponding amount (200 ul), and G2-1 and G2-2 are the same DilC Dye dissolved in a lipid precursor preparation at a concentration of 0.15 ul and administered subcutaneously.

[0223] As shown in the figure, in the case of G1, it was confirmed to have an overall low sensitivity because it rapidly spread throughout the body and gradually disappeared, whereas in the case of G2-1 and G2-2, it was confirmed that the fluorescent substance was emitting light only in the local area where the formulation loaded with the fluorescent substance was administered. In addition, it was confirmed that the fluorescent substance was emitted more strongly on the 7th day than in the beginning. This seems to be a result discovered as the fluorescent substance was emitted weakly in the beginning and gradually over time.

[0224] Therefore, compared to G1, the G2-1 and G2-2 formulations act to release the drug slowly while maintaining its presence at the administered site according to local administration, and this effect can be visually confirmed through bioimaging that it has the effect of providing sustained-release through the formulation instead of a general liquid. Based on these results, it can be confirmed that the drug loaded into the lipid precursor formulation of the present invention can be slowly released in the body.

[0225]

[0226] Manufacturing Example 5. Manufacturing of a lipid precursor preparation containing an analgesic and an osteoarthritis treatment agent.

[0227] Celecoxib and meloxicam were used as analgesics, and dexamethasone and triamcinolone were used as osteoarthritis treatment.

[0228] The ingredients (content: g) in Table 11 below were sequentially added to a glass vial, and mixed while stirring with a magnetic stirrer at room temperature to 60°C. A lipid precursor formulation was prepared by leaving it at room temperature for about 3 hours.

[0229]

[0230] Manufacturing Example 5-1 Comparative Manufacturing Example 5-1 Manufacturing Example 5-2 Comparative Manufacturing Example 5-2 Manufacturing Example 5-3 Comparative Manufacturing Example 5-3 Manufacturing Example 5-4 Comparative Manufacturing Example 5-4 Celecoxib 55-----Meloxicam--55---Dexamethasone----55-Triamcinolone------55 Phosphatidyl Choline 35 35 45 45 35 35 35 35 35 35 35 35 35 Sorbitan Laurate 20-15-15-15-Ascorbic Acid Palmitate--------Tocopherol Acetate--------Squalene 15 15 15 15 15 15 15 15 Cholesterol 5 5 5 5 5 5 5 5 Ethanol 10 10 10 10 10 10 10 DMSO 10 5 5 5 5 15 15 15 15 Gel Formation Result Gel Formation Gel Not Formed. Gel Formation Gel Not Formed No gel formationNo gel formationNo gel formationNo gel formation

[0231]

[0232] As a result of the gel forming ability evaluation, it was confirmed that the lipid precursor preparation containing sorbitan laurate, a saturated fatty acid of sorbitan, normally exhibits a liquid state, but changes to a gel state when it comes into contact with a pH 7.4 phosphate buffer.

[0233]

[0234] Manufacturing Example 6. Manufacturing of a lipid precursor preparation containing a hair loss and benign prostatic hyperplasia treatment agent.

[0235] Dutasteride and finasteride were used as treatments for benign prostatic hyperplasia.

[0236] The ingredients in Table 12 (content: g) were sequentially added to a glass vial, and mixed while stirring with a magnetic stirrer at room temperature to 60°C. A lipid precursor formulation was prepared by leaving it at room temperature for approximately 3 hours.

[0237]

[0238] Manufacturing Example 6-1 Manufacturing Example 6-2 Manufacturing Example 6-3 Manufacturing Example 6-4 Manufacturing Example 6-5 Comparative Manufacturing Example 6-1 Comparative Manufacturing Example 6-2 Dutasteride 555--5 Finasteride---55-5 Phosphatidyl Choline 30353530353030 Sorbitan Laurate 2015152015--Ascorbic Acid Palmitate--10-10 Squalene 20201020102020 Cholesterol 5555555 Ethanol 10101010101010 DMSO 10101010101010 Gel Formation Result Gel Formation Gel Formation Gel Formation Gel Formation No gel formation. No gel formation.

[0239]

[0240] As a result of the gel forming ability evaluation, it was confirmed that the lipid precursor preparation containing sorbitan laurate, a saturated fatty acid of sorbitan, normally exhibits a liquid phase, but changes to a gel phase when it comes into contact with a pH 7.4 phosphate buffer.

[0241]

[0242] Test Example 9. Confirmation of sustained release in a release test of a lipid precursor formulation containing a hair loss and benign prostatic hyperplasia treatment agent.

[0243] Long-term release testing (in vitro dissolution testing) was performed by adding 5 mL of pH 7.4 phosphate buffer containing 0.05% Tween 80 to a 6 cc glass vial, and pipetting the lipid precursor formulation to form a gel. The test was performed using a rotary rotator (PTR-35, lab23009) at a rotation speed of 30 rpm and a temperature of 37°C for 28 days. The collected samples were analyzed by HPLC under the following conditions.

[0244]

[0245] <HPLC 조건>

[0246] Column: phenomenox C18 (4.6 x 250 mm, 5.0 μm) or equivalent column

[0247] Mobile phase: A solution of 100 mL of water and 900 mL of methanol

[0248] Flow rate: 1.0 mL / min

[0249] Temperature: Room temperature

[0250] Injection volume: 20 μL

[0251] Wavelength: 235 nm

[0252]

[0253] Table 13 shows the results of the long-term release test.

[0254]

[0255] Long-term release day 1 (%)Long-term release day 14 (%)Long-term release day 21 (%)Long-term release day 28 (%)Long-term release day 35 (%)Manufacturing example 6-12.754.374.690.7100.0Comparative manufacturing example 6-110.6100.0100.0100.0100.0Manufacturing example 6-43.845.980.389.5100.0Comparative manufacturing example 6-212.2100.0100.0100.0100.0

[0256]

[0257] As shown in Table 13, in the case of Comparative Manufacturing Examples 6-1 and 6-2, it can be confirmed that 100% release is quickly completed on the 14th day. On the other hand, in the case of Manufacturing Examples 6-1 and 6-4, it can be confirmed that release of less than 55% is shown for up to 2 weeks, and drug release of more than 85% is shown on the 28th day.

[0258]

[0259] Although the present invention has been described in detail only with respect to the described embodiments, it will be apparent to those skilled in the art that various modifications and variations are possible within the technical scope of the present invention, and it is natural that such modifications and variations fall within the scope of the appended claims.

[0260]

[0261] The lipid precursor formulation of the present invention can form liquid crystals upon exposure to an aqueous medium, enabling sustained drug release, and can sustain drug release for approximately one week to one month. Furthermore, the liquid crystals formed by the lipid precursor formulation exhibit sustained drug release even at low viscosities, exhibit excellent viscoelasticity, and have low extrusion force during injection, providing excellent usability.

Claims

1. Contains phospholipids; sorbitan saturated fatty acids; and squalene. A lipid precursor preparation that forms liquid crystals in an aqueous medium.

2. In paragraph 1, A lipid precursor formulation having a phospholipid content of 30 to 60 wt% based on the total weight of the lipid precursor formulation.

3. In paragraph 1, A lipid precursor preparation wherein the sorbitan saturated fatty acid is at least one selected from the group consisting of sorbitan caprylate, sorbitan laurate, sorbitan monolaurate, sorbitan myristate, sorbitan palmitate, sorbitan monopalmitate, sorbitan stearate, sorbitan monostearate, sorbitan tristearate, sorbitan isostearate, and sorbitan monomyristate.

4. In paragraph 1, A lipid precursor preparation having a content of sorbitan saturated fatty acid of 5 to 45 wt% based on the total weight of the lipid precursor preparation.

5. In paragraph 1, A lipid precursor preparation having a squalene content of 2.5 to 30 wt% based on the total weight of the lipid precursor preparation.

6. In paragraph 1, Including additional solvent, A lipid precursor formulation wherein the solvent is at least one selected from the group consisting of water, ethanol, isopropanol, propylene glycol, N-methylpyrrolidone, glycerol, benzyl benzoate, acetic acid, and benzyl alcohol.

7. In paragraph 1, A lipid precursor formulation additionally comprising a drug.

8. In paragraph 7, The drug is a lipid precursor selected from the group consisting of semaglutide, liraglutide, tirzepatide, cerecoxib, meloxicam, betamethasone, dexamethasone, methylprednisolone, prednisolone, triamcinolone, dutasteride, and finasteride.

9. A method for preparing a lipid precursor preparation according to claim 1, comprising the step of preparing a lipid solution by mixing phospholipids, sorbitan saturated fatty acids, and squalene.

10. In paragraph 9, A method for preparing a lipid precursor formulation, wherein the lipid solution additionally comprises a solvent.

11. In paragraph 9, A method for preparing a lipid precursor formulation, wherein the lipid solution additionally contains a drug.

12. In paragraph 9, A step of preparing a drug solution containing a drug; and A method for preparing a lipid precursor formulation, further comprising the step of mixing a lipid solution and the drug solution.

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