Pharmaceutical composition for preventing or treating necrotic enteritis containing TGF-β1 and FGF2 genes or proteins as active ingredients

Abstract

The present invention relates to: a pharmaceutical composition for preventing or treating necrotic enteritis, the pharmaceutical composition containing TGF-β1 and FGF2 genes or proteins as active ingredients; and a pharmaceutical preparation and a health functional food that contain the pharmaceutical composition. According to the present invention, TGF-β1 and FGF2, which are known to increase the expression of the CFTR gene, can be used as novel agents for inducing intestinal maturation in premature infants, and various digestive complications such as necrotic enteritis and intestinal absorption disorders, which are intractable diseases, can be treated through the same.

Description

A pharmaceutical composition for the prevention or treatment of necrotizing enterocolitis comprising TGF-β1 and FGF2 genes or proteins as active ingredients

[0001] One example of the present invention relates to a pharmaceutical composition for the prevention or treatment of necrotizing enterocolitis comprising TGF-β1 and FGF2 genes or proteins as active ingredients, a pharmaceutical preparation comprising said pharmaceutical composition, and a health functional food.

[0002]

[0003] Necrotizing enterocolitis (NEC) is a representative intractable disease that primarily affects premature infants and is characterized by necrosis of the mucous membranes of the small or large intestine. It occurs in approximately 10% of extremely premature infants and is classified as a severe, intractable disease with a mortality rate reaching 25% even with treatment. However, there is currently no known cause of the disease, nor are there effective preventive or therapeutic methods. While prematurity is currently considered the cause of the disease, there has been little research conducted on suitable disease models for the development of treatments.

[0004] CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) is a cystic fibrosis membrane protein that plays a role in regulating the transport of chloride ions and substances. When CFTR functions normally in intestinal cells, fluid balance is maintained and the inflammatory response in the intestines can be suppressed; however, if the CFTR gene does not function properly, intestinal inflammation can easily occur, and the maturation process of the intestines may be inhibited, making the body vulnerable to NEC.

[0005] TGF-β1 (Transforming Growth Factor-Beta 1) is an anti-inflammatory cytokine that suppresses intestinal inflammation and promotes tissue repair. In the intestines of premature infants, there may be a deficiency of TGF-β1, which suppresses inflammatory responses, leading to a higher risk of developing NEC. Therefore, external supplementation or activation of TGF-β1 can play an important role in the prevention and treatment of NEC.

[0006] FGF2 (Fibroblast Growth Factor 2) is a growth factor that promotes cell proliferation and tissue regeneration, aiding in the maturation of intestinal tissue. Activation of FGF2 in the intestines of premature infants promotes the maturation of intestinal cells, which can prevent tissue damage caused by NEC and aid in intestinal regeneration.

[0007] The present invention relates to an inducer for promoting intestinal maturation in premature infants, and can be usefully applied in medical fields primarily related to neonatology, pediatrics, and the treatment of premature infants. In particular, it can be used as a preventive and therapeutic agent for various gastrointestinal complications, such as necrotizing enterocolitis and malabsorption caused by insufficient intestinal development in premature infants.

[0008] Necrotizing enterocolitis, which occurs due to insufficient intestinal development in premature infants, significantly reduces the survival rate of premature infants. Although it is a serious disease that can lead to intestinal dysfunction and growth retardation due to malabsorption and various gastrointestinal complications, there is currently no effective treatment available.

[0009] Therefore, the inventors intended to develop a therapeutic agent that induces the maturation of immature intestines, and through repeated research, arrived at the present invention.

[0010]

[0011] As a result of the inventors' diligent research,

[0012] A pharmaceutical composition for the prevention or treatment of necrotizing enterocolitis comprising TGF-β1 and FGF2 genes or proteins as active ingredients, a pharmaceutical preparation comprising said pharmaceutical composition, and a health functional food were designed.

[0013]

[0014] Accordingly, the present invention

[0015] The purpose is to provide a pharmaceutical composition for the prevention or treatment of necrotizing enterocolitis, comprising as active ingredients the TGF-β1 (Transforming Growth Factor-Beta 1) gene or protein; and the FGF2 (Fibroblast Growth Factor 2) gene or protein.

[0016]

[0017] In addition, the present invention

[0018] The purpose is to provide a pharmaceutical preparation for the prevention or treatment of necrotizing enteritis comprising the above pharmaceutical composition.

[0019]

[0020] In addition, the present invention

[0021] The purpose is to provide a health functional food for the prevention or improvement of necrotizing enteritis comprising the above pharmaceutical composition.

[0022]

[0023] In addition, the present invention

[0024] The purpose is to provide the above pharmaceutical composition for the prevention or treatment of necrotizing enteritis.

[0025]

[0026] In addition, the present invention

[0027] The purpose is to provide a method for preventing or treating necrotizing enteritis, comprising the step of administering the above pharmaceutical composition to an individual in need thereof.

[0028]

[0029] In order to achieve the above objective,

[0030] The present invention

[0031] A pharmaceutical composition for the prevention or treatment of necrotizing enterocolitis is provided, comprising as active ingredients TGF-β1 (Transforming Growth Factor-Beta 1) gene or protein; and FGF2 (Fibroblast Growth Factor 2) gene or protein.

[0032] In one embodiment of the present invention, the TGF-β1 and FGF2 genes or proteins may increase the expression of the CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) gene, but are not limited thereto.

[0033] In another embodiment of the present invention, the necrotizing enterocolitis may be neonatal necrotizing enterocolitis, but is not limited thereto.

[0034] In another embodiment of the present invention, the pharmaceutical composition may increase the expression level of a gene related to necrotizing enterocolitis, but is not limited thereto.

[0035] In another embodiment of the present invention, the necrotizing enteritis-related gene may be one or more selected from the group consisting of a mature intestinal cell-related gene, a digestive enzyme-related gene, a fatty acid transport-related gene, an amino acid transport-related gene, an inorganic solute-related gene, and an intercellular junction-related gene, but is not limited thereto.

[0036] In another embodiment of the present invention, the mature intestinal cell-related gene may be one or more selected from the group consisting of TM4SF20, THSD4, SLC46A1, LCT and ABTB2, but is not limited thereto.

[0037] In another embodiment of the present invention, the digestive enzyme-related gene may be one or more selected from the group consisting of TMPRSS15 and ANPEP, but is not limited thereto.

[0038] In another embodiment of the present invention, the fatty acid transport-related gene may be one or more selected from the group consisting of APOB, ACSL5, SLC27A4, and FABP, but is not limited thereto.

[0039] In another embodiment of the present invention, the amino acid transport-related gene may be one or more selected from the group consisting of SLC6A19, SLC6A6, SLC25A12, and SLC1A1, but is not limited thereto.

[0040] In another embodiment of the present invention, the inorganic solute-related gene may be one or more selected from the group consisting of CFTR, SLC26A3 and SLC12A2, but is not limited thereto.

[0041] In another embodiment of the present invention, the intercellular junction-related gene may be one or more selected from the group consisting of CLDN1 and ZO-1, but is not limited thereto.

[0042] In another embodiment of the present invention, the pharmaceutical composition may have effects such as promoting the maturation of intestinal cells, restoring nutrient digestion ability, promoting fatty acid absorption and metabolism, restoring amino acid absorption ability, maintaining electrolyte and fluid balance, strengthening barrier function, and reducing intestinal permeability, but is not limited thereto.

[0043]

[0044] In addition, the present invention

[0045] A pharmaceutical preparation for the prevention or treatment of necrotizing enteritis comprising the above pharmaceutical composition is provided.

[0046] In one embodiment of the present invention, the pharmaceutical formulation may further comprise a pharmaceutically acceptable carrier, excipient, or diluent, but is not limited thereto.

[0047] In another embodiment of the present invention, the pharmaceutical formulation may be an injectable formulation, an infusion formulation, a spray formulation, a liquid formulation, or a patch formulation, but is not limited thereto.

[0048] In another embodiment of the present invention, the necrotizing enterocolitis may be neonatal necrotizing enterocolitis, but is not limited thereto.

[0049] In another embodiment of the present invention, the pharmaceutical formulation may have effects such as promoting the maturation of intestinal cells, restoring nutrient digestion ability, promoting fatty acid absorption and metabolism, restoring amino acid absorption ability, maintaining electrolyte and fluid balance, strengthening barrier function, and reducing intestinal permeability, but is not limited thereto.

[0050]

[0051] In addition, the present invention

[0052] A health functional food for the prevention or improvement of necrotizing enteritis comprising the above pharmaceutical composition is provided.

[0053] In one embodiment of the present invention, the necrotizing enterocolitis may be neonatal necrotizing enterocolitis, but is not limited thereto.

[0054] In another embodiment of the present invention, the health functional food may have effects such as promoting the maturation of intestinal cells, restoring nutrient digestion ability, promoting fatty acid absorption and metabolism, restoring amino acid absorption ability, maintaining electrolyte and fluid balance, strengthening intestinal barrier function, and reducing intestinal permeability, but is not limited thereto.

[0055]

[0056] In addition, the present invention

[0057] The above pharmaceutical composition is used for the prevention or treatment of necrotizing enteritis.

[0058]

[0059] In addition, the present invention

[0060] A method for preventing or treating necrotizing enteritis is provided, comprising the step of administering the above pharmaceutical composition to an individual in need thereof.

[0061]

[0062] By the pharmaceutical composition for the prevention or treatment of necrotizing enterocolitis comprising the TGF-β1 and FGF2 genes or proteins of the present invention as active ingredients, the pharmaceutical preparation comprising the said pharmaceutical composition, and the health functional food, TGF-β1 and FGF2, which are known to increase the expression of the CFTR gene, can be utilized as new agents to induce intestinal maturation in premature infants, and thereby, various digestive system complications such as necrotizing enterocolitis and intestinal malabsorption, which are intractable diseases, can be treated.

[0063]

[0064] The effects of the present invention are not limited to those mentioned above, and should be understood to include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.

[0065]

[0066] Figures 1A and 1B relate to the correlation between the therapeutic effect of a necrotizing enterocolitis treatment agent and the CFTR gene. Specifically, Figure 1A is a graph of mRNA expression levels to confirm the therapeutic effect of extracellular vesicles derived from human umbilical cord-derived mesenchymal stem cells on necrotizing enterocolitis. Figure 1B is a graph of mRNA expression levels to confirm whether the therapeutic effect of extracellular vesicles derived from human umbilical cord-derived mesenchymal stem cells is due to the CFTR gene.

[0067] Figures 2A to 2F are graphs measuring the expression levels of genes related to necrotizing enterocolitis upon Forskolin treatment. Specifically, Figure 2A is a graph measuring the expression levels of genes related to inorganic solutes. Figure 2B is a graph measuring the expression levels of genes related to mature intestinal cells. Figure 2C is a graph measuring the expression levels of genes related to digestive enzymes. Figure 2D is a graph measuring the expression levels of genes related to fatty acid transport. Figure 2E is a graph measuring the expression levels of genes related to amino acid transport. Figure 2F is a graph measuring the expression levels of genes related to intercellular junctions.

[0068] Figures 3A to 3F are graphs measuring the expression levels of genes related to necrotizing enterocolitis upon treatment with TGF-β1 alone, FGF2 alone, and simultaneous treatment with TGF-β1 and FGF2. Specifically, Figure 3A is a graph measuring the expression levels of genes related to mature intestinal cells. Figure 3B is a graph measuring the expression levels of genes related to digestive enzymes. Figure 3C is a graph measuring the expression levels of genes related to fatty acid transport. Figure 3D is a graph measuring the expression levels of genes related to amino acid transport. Figure 3E is a graph measuring the expression levels of genes related to inorganic solutes. Figure 3F is a graph measuring the expression levels of genes related to intercellular junctions.

[0069] Figures 4A to 4F illustrate the effects of inhibiting intestinal leakage, increasing expression of intestinal cell functional markers, improving fat absorption capacity, and CFTR dependence in immature intestinal organoids treated simultaneously with TGF-β1 and FGF2. Specifically, Figure 4A is a graph showing the FITC-dextran permeability of the untreated group, the MSC-EV treated group, and the TGF-β1+FGF2 treated group, and Figure 4B is a graph comparing the FITC-dextran permeability of the TGF-β1+FGF2 treated group and the CFTR inhibitor combination group in GA27 and GA24 organoids. Figure 4C shows the ZO-1 and FABP expression images of the NT group, the EV39 group, and the TGF-β1+FGF2 treated group, and Figure 4D shows the ZO-1 and FABP expression images of the TGF-β1+FGF2 treated group and the CFTR inhibitor combination group. Figure 4E shows BODIPY-C16 uptake images of the NT group, EV39 group, and TGF-β1+FGF2 treatment group, and Figure 4F shows BODIPY-C16 uptake images of the TGF-β1+FGF2 treatment group and the CFTR inhibitor combination group in GA27 and GA24 organoids.

[0070]

[0071] The present invention will be described in detail below.

[0072] The present invention

[0073] A pharmaceutical composition for the prevention or treatment of necrotizing enterocolitis is provided, comprising as active ingredients TGF-β1 (Transforming Growth Factor-Beta 1) gene or protein; and FGF2 (Fibroblast Growth Factor 2) gene or protein.

[0074] As used herein, the term "TGF-β1 (Transforming Growth Factor-Beta 1)" refers to an anti-inflammatory cytokine that suppresses intestinal inflammation and promotes tissue repair. In the intestines of premature infants, there may be a deficiency of TGF-β1, which suppresses inflammatory responses, which increases the risk of developing NEC. Therefore, external supplementation or activation of TGF-β1 can play an important role in the prevention and treatment of NEC.

[0075] As used in this specification, the term "FGF2 (Fibroblast Growth Factor 2)" refers to a growth factor that promotes cell proliferation and tissue regeneration and helps in the maturation of intestinal tissue. Activation of FGF2 in the intestines of premature infants can promote the maturation of intestinal cells, thereby preventing tissue damage caused by NEC and helping intestinal regeneration.

[0076] As used herein, the term "Necrotizing enterocolitis (NEC)" refers to a disease that primarily occurs in premature infants and is a representative intractable disease characterized by necrosis of the mucous membranes of the small or large intestine. It occurs in approximately 10% of extremely premature infants and is classified as an intractable, severe disease with a mortality rate reaching 25% even with treatment. However, there is currently no known cause of the disease, nor effective preventive or therapeutic methods. Although the cause of the disease is currently attributed to prematurity, there has been little research conducted on suitable disease models for the development of therapeutic agents.

[0077] As used herein, the term "prevention" refers to any act of suppressing symptoms caused by necrotizing enteritis in an individual or delaying the onset of the disease by administering a pharmaceutical composition according to the present invention.

[0078] As used herein, the term "treatment" refers to any act in which symptoms caused by necrotizing enterocolitis in an individual are improved or beneficially altered by the administration of a pharmaceutical composition according to the present invention. The intended therapeutic effects include preventing the onset or recurrence of the disease, alleviating symptoms, reducing all direct or indirect pathological consequences associated with the disease, preventing metastasis, reducing the rate of disease progression, alleviating or temporarily resolving the disease state, and improving the disease state or prognosis. For the purposes of the present invention, the above treatment may be interpreted to include all acts of improving said symptoms by administering the pharmaceutical composition of the present invention to a patient with the onset of the disease, but is not specifically limited thereto.

[0079] As used herein, the term "administration" means introducing a pharmaceutical composition of the present invention to a subject by any appropriate method, by providing a composition of the present invention to the subject. The route of administration may be administered via various oral or parenteral routes as long as it can reach the target tissue.

[0080] The pharmaceutical composition of the present invention may be appropriately administered to an individual according to the conventional methods, routes of administration, and dosages used in the art, depending on the purpose or need. Examples of routes of administration may include oral, parenteral, subcutaneous, intraperitoneal, intrapulmonary, and nasal administration, and parenteral administration includes intramuscular, intravenous, intra-arterial, intraperitoneal, or subcutaneous administration.

[0081] In addition, an appropriate dosage and number of administrations may be selected according to methods known in the art, and the amount and number of administrations of the pharmaceutical composition of the present invention actually administered may be appropriately determined by various factors such as the type of symptom to be treated, the route of administration, gender, health status, diet, age and weight of the individual, and the severity of the disease.

[0082] As used herein, the term "individual" refers to a subject requiring treatment for a disease, and more specifically, to all animals including humans who are likely to develop or have already developed the disease. It may refer to mammals such as humans or non-human primates, mice, dogs, cats, horses, and cattle, but is not limited thereto.

[0083] Additionally, the above pharmaceutical composition may be provided as a pharmaceutical composition or pharmaceutical preparation comprising an active ingredient alone or comprising one or more pharmaceutically acceptable carriers, excipients, or diluents.

[0084] Specifically, the term “pharmaceuticalally acceptable carrier” as used herein refers to a material commonly used in formulations and may be a colloidal suspension, powder, saline solution, lipid, liposome, microsphere, or nano-spherical particle. It may also include, but is not limited to, saline solution, sterile water, Ringer’s solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposomes, etc., and may further include other conventional additives such as antioxidants, buffers, etc., as needed. These may form a complex with or be associated with a carrier and may be transported in vivo using a carrier system known in the art, such as lipids, liposomes, microparticles, gold, nanoparticles, polymers, condensation agents, polysaccharides, polyamino acids, dendrimers, saponins, adsorption-enhancing substances, or fatty acids.

[0085] Additionally, the term “pharmaceuticalally acceptable excipient” as used herein means any substance that is used as a carrier or medium for delivering a pharmaceutical composition according to the present invention, or is added to a formulation to improve handling or storage, or to facilitate the manufacture of a formulation in dosage units into articles such as capsules or tablets suitable for oral administration. Excipients may include, but are not limited to, diluents, disintegrants, binders, adhesives, wetting agents, lubricants, lubricants, fragrances, surfactants, and inclusion compounds.

[0086] When the above pharmaceutical composition is formulated, it may be prepared using excipients or diluents such as commonly used lubricants, sweeteners, flavorings, emulsifiers, suspending agents, preservatives, fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Regarding suitable pharmaceutically acceptable carriers and formulations, each component may be preferably formulated using the methods disclosed in Remington’s literature.

[0087] Solid dosage forms for oral administration may include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms may be prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc., with the above composition. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.

[0088] Liquid formulations for oral administration include suspensions, liquid formulations, emulsions, syrups, etc., and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as humectants, sweeteners, flavorings, and preservatives, may be included.

[0089] Preparations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. As non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As bases for suppositories, witepsol, macrogol, tween 61, cacao oil, laurin oil, glycerogelatin, etc. may be used, and when manufactured in the form of ophthalmic preparations, known excipients or diluents may be used.

[0090] In addition, the above pharmaceutical composition may be administered alone or in combination with other therapeutic agents. That is, the above pharmaceutical composition may be administered in conjunction with a known composition or other agent having a preventive or therapeutic effect on the said disease, and may be administered simultaneously, separately, or sequentially, and may be administered as a single or multiple doses. It is important to administer an amount that obtains maximum effect with a minimum amount without side effects by taking all of the above factors into consideration, and this can be easily determined by a person skilled in the art.

[0091] The above pharmaceutical composition may be administered orally or parenterally, and when administered parenterally, the method of administration may be selected as external application to the skin or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, intra-arterial injection, intramedullary injection, intracardiac injection, intrathecal injection, transdermal injection, nasal injection, enteral injection, local injection, sublingual injection, rectal injection, or thoracic injection.

[0092] The above pharmaceutical composition is administered in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level may be determined based on factors including the type and severity of the patient's disease, the patient's age, gender, condition, body weight, absorption of the active ingredient into the body, the drug's activity, inactivation rate and excretion rate, sensitivity to the drug, time of administration, route of administration and elimination rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. Additionally, the dosage may vary depending on the patient's condition and body weight, the degree of disease, the drug form, route of administration, and time, but can be appropriately selected by a person skilled in the art.

[0093] Specifically, the above pharmaceutical composition may be administered at a dose of 0.001 to 1000 mg / kg / day, more specifically at a dose of 0.1 to 100 mg / kg / day. The administration may be administered once a day or divided into several doses. For example, it may be administered every other day or once a week. However, since the dosage may be increased or decreased depending on the route of administration, severity of obesity, gender, body weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

[0094] In one embodiment of the present invention, the TGF-β1 and FGF2 genes or proteins may increase the expression of the CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) gene, but are not limited thereto.

[0095] As used in this specification, the term "CFTR (Cystic Fibrosis Transmembrane Conductance Regulator)" refers to a cystic fibrosis transmembrane protein that plays a role in regulating the transport of chloride ions and substances. When CFTR functions normally in intestinal cells, fluid balance is maintained and the inflammatory response in the intestine can be suppressed; however, if the CFTR gene does not function properly, intestinal inflammation can easily occur, and the maturation process of the intestine may be inhibited, making it vulnerable to NEC.

[0096] In another embodiment of the present invention, the necrotizing enterocolitis may be neonatal necrotizing enterocolitis, but is not limited thereto.

[0097] In another embodiment of the present invention, the pharmaceutical composition may increase the expression level of a gene related to necrotizing enterocolitis, but is not limited thereto.

[0098] In another embodiment of the present invention, the necrotizing enteritis-related gene may be one or more selected from the group consisting of a mature intestinal cell-related gene, a digestive enzyme-related gene, a fatty acid transport-related gene, an amino acid transport-related gene, an inorganic solute-related gene, and an intercellular junction-related gene, but is not limited thereto.

[0099] In another embodiment of the present invention, the mature intestinal cell-related gene may be one or more selected from the group consisting of TM4SF20, THSD4, SLC46A1, LCT and ABTB2, but is not limited thereto.

[0100] In another embodiment of the present invention, the digestive enzyme-related gene may be one or more selected from the group consisting of TMPRSS15 and ANPEP, but is not limited thereto.

[0101] In another embodiment of the present invention, the fatty acid transport-related gene may be one or more selected from the group consisting of APOB, ACSL5, SLC27A4, and FABP, but is not limited thereto.

[0102] In another embodiment of the present invention, the amino acid transport-related gene may be one or more selected from the group consisting of SLC6A19, SLC6A6, SLC25A12, and SLC1A1, but is not limited thereto.

[0103] In another embodiment of the present invention, the inorganic solute-related gene may be one or more selected from the group consisting of CFTR, SLC26A3 and SLC12A2, but is not limited thereto.

[0104] In another embodiment of the present invention, the intercellular junction-related gene may be one or more selected from the group consisting of CLDN1 and ZO-1, but is not limited thereto.

[0105] In another embodiment of the present invention, the pharmaceutical composition may have effects such as promoting the maturation of intestinal cells, restoring nutrient digestion ability, promoting fatty acid absorption and metabolism, restoring amino acid absorption ability, maintaining electrolyte and fluid balance, strengthening barrier function, and reducing intestinal permeability, but is not limited thereto.

[0106]

[0107] In addition, the present invention

[0108] A pharmaceutical preparation for the prevention or treatment of necrotizing enteritis comprising the above pharmaceutical composition is provided.

[0109] The above terms "necrotizing enterocolitis," "prevention," and "treatment," etc., may be within the scope described above.

[0110] In one embodiment of the present invention, the pharmaceutical formulation may further comprise a pharmaceutically acceptable carrier, excipient, or diluent, but is not limited thereto.

[0111] In another embodiment of the present invention, the pharmaceutical formulation may be an injectable formulation, an infusion formulation, a spray formulation, a liquid formulation, or a patch formulation, but is not limited thereto.

[0112] As used herein, the term "injectable" refers to a solution, suspension, emulsion, or solid sterile preparation that is administered directly into internal tissues or organs, such as subcutaneously, intramuscularly, or into blood vessels.

[0113] As used in this specification, the term "injectable" refers to a drug that is inserted into the body through the urethra, anus, vagina, etc., and dissolves due to body temperature or secretions to produce a medicinal effect.

[0114] As used in this specification, the term "spray" refers to a medicine that is sprayed out like a mist using a device.

[0115] As used in this specification, the term "liquid formulation" refers to a pharmaceutical form that provides a drug in liquid form, and is a formulation containing an active ingredient in liquid form.

[0116] As used in this specification, the term "patch" refers to a preparation designed to be attached to the skin to continuously exert a therapeutic effect. Patches have the advantage of having a lower risk of side effects such as gastrointestinal or liver damage associated with oral medications, and can be administered to patients for whom oral administration is difficult. Furthermore, since they are absorbed directly into the bloodstream through the skin without undergoing hepatic metabolism, they can exhibit the same therapeutic effect without side effects with a lower dose compared to oral medications.

[0117] In another embodiment of the present invention, the necrotizing enterocolitis may be neonatal necrotizing enterocolitis, but is not limited thereto.

[0118] In another embodiment of the present invention, the pharmaceutical formulation may have effects such as promoting the maturation of intestinal cells, restoring nutrient digestion ability, promoting fatty acid absorption and metabolism, restoring amino acid absorption ability, maintaining electrolyte and fluid balance, strengthening barrier function, and reducing intestinal permeability, but is not limited thereto.

[0119]

[0120] In addition, the present invention

[0121] A health functional food for the prevention or improvement of necrotizing enteritis comprising the above pharmaceutical composition is provided.

[0122] The above terms "necrotizing enterocolitis" and "prevention," etc., may be within the scope described above.

[0123] As used in this specification, the term "improvement" may refer to any action that at least reduces parameters related to the condition being treated, such as the severity of symptoms. In this case, the health functional food may be used for the prevention or improvement of the disease, either simultaneously with or separately from a therapeutic agent, either before or after the onset of the disease.

[0124] The health functional food defined in the present invention may be a health functional food that has sufficiently established functionality and safety for the human body, newly defined through the Health Functional Foods Act revised in 2008, and is listed in the regulations on the recognition of functional ingredients for health functional foods stipulated in KFDA Notice No. 2008-72.

[0125] In the above-mentioned health functional food, the active ingredient may be added directly to the food or used together with other foods or food ingredients, and may be used appropriately according to conventional methods. The amount of the active ingredient may be appropriately determined according to its purpose of use (for prevention or improvement). Generally, the active ingredient constituting the composition according to the present invention may be included in an amount of 0.01 to 15 weight%, preferably 0.2 to 10 weight% of the total food weight, and when manufactured as a beverage, it may be included in a ratio of 0.1 to 30 g, preferably 0.2 to 5 g, based on 100 mL, and the entire beverage may be composed of natural ingredients. However, in the case of long-term consumption for the purpose of health control and hygiene, the above amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount greater than the above range.

[0126] The health functional food according to the present invention may be formulated into a formulation of a conventional health functional food known in the art. The health functional food may further include one or more of a carrier, an excipient, a diluent, and an additive, and may be formulated into one selected from the group consisting of tablets, pills, powders, granules, powders, capsules, infusions, liquids, suspensions, emulsions, syrups, extracts, gums, teas, jellies, or beverages.

[0127] Foods to which compounds according to one aspect can be added include various types of food, powders, granules, tablets, capsules, syrups, beverages, gum, tea, vitamin complexes, health functional foods, etc.

[0128] As the above-mentioned food-grade acceptable carrier or additive, any carrier or additive known in the relevant art to be available for use in the preparation of the formulation to be prepared may be used. It may also include foods used as animal feed.

[0129] Specific examples of the above carrier, excipient, diluent, and additive may be at least one selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium phosphate, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0130] In addition to containing the above-mentioned active ingredient, the above-mentioned health functional food may contain other ingredients as essential components without special restrictions. For example, it may contain various flavorings or natural carbohydrates as additional ingredients, similar to ordinary beverages.

[0131] Examples of the natural carbohydrates described above may be monosaccharides, e.g., glucose, fructose, etc.; disaccharides, e.g., maltose, sucrose, etc.; polysaccharides, e.g., dextrin, cyclodextrin, etc., and conventional sugars, and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As flavoring agents other than those described above, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) may be advantageously used. The proportion of the natural carbohydrates may be appropriately determined by the choice of a person skilled in the art.

[0132] In addition to the above, a health functional food according to one aspect may contain various nutritional supplements, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. Furthermore, it may contain fruit pulp for the manufacture of natural fruit juices, fruit juice beverages, and vegetable beverages. In addition, the above health functional food composition may further include food additives, and unless otherwise stipulated, suitability as a "food additive" may be determined according to the specifications and standards for the relevant item in accordance with the general provisions and general test methods of the Food Additives Codex approved by the Ministry of Food and Drug Safety. These ingredients may be used independently or in combination, and the proportion of such additives may also be appropriately selected by a person skilled in the art.

[0133] In addition, the health functional food of the present invention may include additional ingredients that are commonly used in food compositions to improve odor, taste, visual appearance, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), and copper (Cu). In addition, it may include amino acids such as lysine, tryptophan, cysteine, and valine. In addition, food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dihydroacetate, etc.), disinfectants (bleaching powder and high-grade bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), etc.), coloring agents (tar dyes, etc.), colorants (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, potassium hydrogen tartrate, etc.), reinforcing agents, emulsifiers, thickeners (sizing agents), coating agents, gum bases, antifoaming agents, solvents, and improvers may be added. The above additives may be selected according to the type of food and used in appropriate amounts.

[0134] When the health functional food of the present invention is used as a food additive, it may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods.

[0135] In the health functional food of the present invention, the content of the pharmaceutical composition of the present invention is not particularly limited and may be varied depending on the condition of the subject to administration, the specific type of disease, the degree of progression, etc. If necessary, it may also be included in the total content of the food.

[0136] In one aspect, the above-mentioned health functional food may further include a health functional food for the prevention or improvement of the relevant disease.

[0137] The above-mentioned health functional food may be provided in combination with a conventionally known health functional food for the prevention or improvement of the relevant disease or a newly developed health functional food for the prevention or improvement of the relevant disease.

[0138]

[0139] If the above-mentioned health functional food further includes a health functional food for the prevention or improvement of the relevant disease, it is important that an amount is mixed such that the maximum effect can be obtained with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.

[0140] In addition, in one aspect, the health functional food may be consumed alone or in combination with a health functional food for the prevention or improvement of the relevant disease. The health functional food may be consumed in conjunction with a known composition or other health functional food having an effect of preventing or improving the relevant disease, and may be consumed simultaneously, separately, or sequentially, and may be consumed as a single or multiple times. It is important to consume an amount that obtains maximum effect with a minimum amount without side effects by taking all of the above factors into consideration, and this can be easily determined by a person skilled in the art.

[0141] In one embodiment of the present invention, the necrotizing enterocolitis may be neonatal necrotizing enterocolitis, but is not limited thereto.

[0142] In another embodiment of the present invention, the health functional food may have effects such as promoting the maturation of intestinal cells, restoring nutrient digestion ability, promoting fatty acid absorption and metabolism, restoring amino acid absorption ability, maintaining electrolyte and fluid balance, strengthening intestinal barrier function, and reducing intestinal permeability, but is not limited thereto.

[0143]

[0144] In addition, the present invention

[0145] The above pharmaceutical composition is used for the prevention or treatment of necrotizing enteritis.

[0146]

[0147] In addition, the present invention

[0148] A method for preventing or treating necrotizing enteritis is provided, comprising the step of administering the above pharmaceutical composition to an individual in need thereof.

[0149]

[0150] To aid in understanding the present invention, it will be explained in more detail below through examples. However, the following examples are merely illustrative for explaining the content of the present invention, and the scope of the present invention is not limited to the following examples. The examples of the present invention are provided to more completely explain the present invention to those with average knowledge in the technical field to which the invention belongs.

[0151]

[0152] [Example]

[0153] Example 1. Analysis of the correlation between the therapeutic effect of a necrotizing enterocolitis treatment and the CFTR gene

[0154] The expression levels of the relevant genes were measured by qRT-PCR in an immature intestinal organoid model (GA27) isolated from the intestinal tissue of a premature infant. The expression level of CFTR in the non-treated control group was set to 1, and the relative expression levels of the experimental group were measured. In the inventors' previous study, human umbilical cord-derived mesenchymal stem cell-derived extracellular vesicles (MSC-EVs), which demonstrated therapeutic effects in an animal model of necrotizing enterocolitis in premature infants, were used as the therapeutic control group.

[0155] Specifically, high-efficacy and low-efficacy MSC-EVs were treated to intestinal organoids of premature infants and cultured for 7 days. Afterward, RNA was extracted from the organoids, and the expression pattern of CFTR, one of the intestinal epithelial cell markers, was observed.

[0156] As a result, as shown in Figure 1A, there was no significant difference in the CFTR gene expression level of the group treated with low-efficacy MSC-EV (Negative control treatment) compared to the group not treated with treatment (Non-treated control), but it was confirmed that the CFTR gene expression level of the group treated with high-efficacy MSC-EV (Positive control treatment) was four times that of the group not treated with treatment (Non-treated control).

[0157] These results suggest that mesenchymal stem cell-derived extracellular vesicles significantly increase CFTR gene expression in immature intestinal organoids.

[0158] In addition, CFTR inhibitor 172 was additionally administered to confirm whether the therapeutic effect of mesenchymal stem cell-derived extracellular vesicles was due to the CFTR gene.

[0159] As a result, as shown in Figure 1B, the CFTR gene expression level in the group treated with high-efficacy MSC-EV (Positive control treatment) was five times that of the non-treated control group, but it was confirmed that the CFTR gene expression level in the group with added CFTR inhibitor (Positive control treatment + CFTR inhibitor) was significantly reduced.

[0160] These results suggest that the therapeutic effect of mesenchymal stem cell-derived extracellular vesicles is due to the CFTR gene.

[0161]

[0162] Example 2. Measurement of expression levels of necrotizing enterocolitis-related genes upon Forskolin treatment

[0163] To measure the expression levels of necrotizing enterocolitis-related genes upon Forskolin treatment, an immature intestinal organoid model (GA27) was treated with Forskolin, which induces CFTR activation, and the mRNA expression levels of necrotizing enterocolitis-related genes were measured by qRT-PCR.

[0164] Specifically, premature infant intestinal organoids were treated with Forskolin (10 μM) and cultured for 7 days. Afterward, RNA was extracted from the organoids, and the gene expression patterns of CFTR, an intestinal epithelial cell marker, as well as mature intestinal cell markers and functional intestinal epithelial cell markers (digestion, fatty acid transport, amino acid transport, intercellular junctions) that are abundant in adult intestinal epithelial cells were observed.

[0165] As a result, as shown in FIGS. 2A to 2F, compared to the Forskolin untreated group (NT), the Forskolin-treated group showed inorganic solute-related genes such as the CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) gene and the SLC26A3 (Solute Carrier Family 26 Member 3) gene regulated by CFTR (see FIG. 2A), mature intestinal cell-related genes such as the TM4SF20 (Transmembrane 4 L Six Family Member 20) gene, THSD4 (Thrombospondin Type 1 Domain Containing 4) gene, SLC46A1 (Solute Carrier Family 46 Member 1) gene, LCT (Lactase) gene, and ABTB2 (Ankyrin Repeat And BTB Domain Containing 2) gene (see FIG. 2B), and the TMPRSS15 (Transmembrane Protease, Serine 15) gene and digestive enzyme-related genes such as the ANPEP (Alanyl Aminopeptidase) gene (Digestive enzyme, see Fig. 2C), fatty acid transport-related genes such as the APOB (Apolipoprotein B) gene, ACSL5 (Acyl-CoA Synthetase Long Chain Family Member 5) gene and SLC27A4 (Solute Carrier Family 27 Member 4) gene (Fatty acid transport, see Fig. 2D), SLC6A19 (Solute Carrier Family 6 Member 19) gene, SLC6A6 (Solute Carrier Family 6 Member 6) gene,It was confirmed that the expression levels of amino acid transport-related genes, such as the SLC25A12 (Solute Carrier Family 25 Member 12) and SLC1A1 (Solute Carrier Family 1 Member 1) genes (see Fig. 2E), and cell junction-related genes, such as the CLDN1 (Claudin 1) and ZO-1 (Zonula Occludens 1) genes (see Fig. 2F), all significantly increased.

[0166] These results suggest that treatment with Forskolin increases the expression levels of necrotizing enterocolitis genes, including CFTR and SLC26A3, thereby demonstrating a therapeutic effect on necrotizing enterocolitis.

[0167]

[0168] Example 3. Measurement of expression levels of necrotizing enterocolitis-related genes upon simultaneous treatment with TGF-β1 and FGF2

[0169] To measure the expression levels of genes related to necrotizing enterocolitis when treated with TGF-β1 (Transforming Growth Factor-Beta 1) alone, FGF2 (Fibroblast Growth Factor 2) alone, and simultaneously treated with TGF-β1 and FGF2, mRNA expression levels of genes related to necrotizing enterocolitis were measured by qRT-PCR after treating an immature intestinal organoid model (GA27) with TGF-β1 alone, FGF2 alone, and simultaneously treated with TGF-β1 and FGF2. In the inventors' previous study, extracellular vesicles (EV39) derived from human umbilical cord mesenchymal stem cells, which demonstrated therapeutic effects in an animal model of necrotizing enterocolitis in premature infants, were used as a therapeutic control.

[0170] Specifically, intestinal organoids from premature infants were treated with EV39 alone and with TGF-β1 (0.1 ng / ml) and FGF2 (10 ng / ml) alone and in combination, and cultured for 7 days. Subsequently, RNA was extracted from the organoids, and the gene expression patterns of mature intestinal cell markers, which are frequently found in adult intestinal epithelial cells, and intestinal epithelial cell markers by function (digestion, fatty acid transport, amino acid transport, intercellular junctions) were observed.

[0171] As a result, as shown in FIGS. 3A to 3F, compared to the untreated group (NT), the TGF-β1-only treated group, and the FGF2-only treated group, in the group simultaneously treated with TGF-β1 and FGF2, genes related to mature intestinal cells such as TM4SF20 (Transmembrane 4 L Six Family Member 20), THSD4 (Thrombospondin Type 1 Domain Containing 4), SLC46A1 (Solute Carrier Family 46 Member 1), LCT (Lactase), and ABTB2 (Ankyrin Repeat And BTB Domain Containing 2) genes (Adult enterocyte gene, see FIG. 3A), genes related to digestive enzymes such as TMPRSS15 (Transmembrane Protease, Serine 15) and ANPEP (Alanyl Aminopeptidase) genes (Digestive enzyme, see FIG. 3B), and APOB (Apolipoprotein B) genes, Fatty acid transport-related genes such as the ACSL5 (Acyl-CoA Synthetase Long Chain Family Member 5) gene and the SLC27A4 (Solute Carrier Family 27 Member 4) gene (see Fig. 3C), amino acid transport-related genes such as the SLC6A19 (Solute Carrier Family 6 Member 19) gene, the SLC6A6 (Solute Carrier Family 6 Member 6) gene, the SLC25A12 (Solute Carrier Family 25 Member 12) gene and the SLC1A1 (Solute Carrier Family 1 Member 1) gene (see Fig. 3D), and the CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) gene,It was confirmed that the expression levels of inorganic solute-related genes, such as the SLC26A3 (Solute Carrier Family 26 Member 3) and SLC12A2 (Solute Carrier Family 12 Member 2) genes (see Fig. 3E), and cell junction-related genes, such as the CLDN1 (Claudin 1) and ZO-1 (Zonula Occludens 1) genes (see Fig. 3F), all significantly increased.

[0172] These results suggest that simultaneous treatment with TGF-β1 and FGF2 increases the expression levels of necrotizing enterocolitis genes, including CFTR and SLC26A3, thereby demonstrating a therapeutic effect on necrotizing enterocolitis.

[0173]

[0174] Example 4. Measurement of intestinal leakage inhibition and fat absorption capacity upon simultaneous treatment with TGF-β1 and FGF2

[0175] In two immature intestinal organoid models (GA27, GA24), inhibition of intestinal leakage related to necrotizing enterocolitis and fat absorption ability were measured upon simultaneous treatment with TGF-β1 and FGF2. In the inventors' previous study, extracellular vesicles (EV39) derived from human umbilical cord mesenchymal stem cells, which demonstrated therapeutic effects in an animal model of necrotizing enterocolitis in premature infants, were used as a therapeutic control. Additionally, to determine whether the therapeutic effect of the combined treatment of TGF-β1 and FGF2 was due to the CFTR gene, CFTR inhibitor 172 was additionally administered.

[0176]

[0177] Specifically, to evaluate the ability to inhibit intestinal leakage, intestinal organoids of premature infants cultured in a single layer using a transwell were treated with EV39 alone and with TGF-β1 (0.1 ng / ml) and FGF2 (10 ng / ml) in combination and cultured for 7 days. Afterward, fluorescein isothiocyanate (FITC)-dextran (4 kDa) was applied to the apical side, and the absorbance of the basolateral culture medium was measured after 30 minutes.

[0178] As a result, as shown in Figures 4A and 4B, the fluorescence level of FITC-dextran decreased in the group treated with TGF-β1 and FGF2 simultaneously compared to the untreated group (NT), similar to the EV39 treatment group, and it was confirmed that the reduction effect disappeared in the group with added CFTR inhibitor (TGF-β1 + FGF2 + CFTR inhibitor).

[0179] These results suggest that simultaneous treatment with TGF-β1 and FGF2 significantly reduces intestinal permeability in intestinal organoids of premature infants, thereby restoring barrier function, and that the therapeutic effect is mediated by the CFTR pathway.

[0180]

[0181] In addition, to identify functional enterocyte markers, intestinal organoid models cultured in a single layer were treated with EV39 alone and with TGF-β1 (0.1 ng / ml) and FGF2 (10 ng / ml) in combination and cultured for 7 days. Subsequently, the expression of FABP, a gene related to fatty acid transport, and ZO-1, a gene related to intercellular junctions, was measured using immunofluorescence staining.

[0182] As a result, as shown in Figures 4C and 4D, compared to the untreated group (NT), the expression levels of ZO-1 and FABP increased in the group treated with TGF-β1 and FGF2, similar to the EV39 treatment group, and it was confirmed that the expression levels of ZO-1 and FABP decreased in the group with added CFTR inhibitor (TGF-β1 + FGF2 + CFTR inhibitor).

[0183] These results suggest that combined treatment with TGF-β1 and FGF2 increases the expression of tight junction protein (ZO-1) and fatty acid binding protein (FABP), thereby improving barrier structural stability and lipid metabolism function, and that the mechanism of action is the restoration of intestinal cell function through the CFTR pathway.

[0184]

[0185] In addition, to evaluate lipid absorption capacity, a monolayer intestinal organoid model of a premature infant was cultured for 7 days with EV39 alone and with TGF-β1 (0.1 ng / ml) and FGF2 (10 ng / ml). Afterward, BODIPY-C16 was administered to induce intracellular lipid absorption, and intracellular fluorescence levels were measured.

[0186] As a result, as shown in Figures 4E and 4F, the absorption of BODIPY-C16 increased in the group treated with TGF-β1 and FGF2 simultaneously compared to the untreated group (NT), similar to the EV39 treatment group, and it was confirmed that this effect decreased in the group with added CFTR inhibitor (TGF-β1 + FGF2 + CFTR inhibitor).

[0187] These results suggest that simultaneous treatment with TGF-β1 and FGF2 significantly enhances lipid absorption capacity in intestinal organoids of premature infants, thereby improving intestinal nutrient absorption function, and that the mechanism involves the restoration of metabolic function of intestinal epithelial cells through the CFTR pathway.

[0188]

[0189] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

1. A pharmaceutical composition for the prevention or treatment of necrotizing enterocolitis, comprising as active ingredients TGF-β1 (Transforming Growth Factor-Beta 1) gene or protein; and FGF2 (Fibroblast Growth Factor 2) gene or protein.

2. In Claim 1, A pharmaceutical composition characterized in that the above TGF-β1 and FGF2 genes or proteins increase the expression of the CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) gene.

3. In Claim 1, A pharmaceutical composition characterized in that the above-mentioned necrotizing enteritis is neonatal necrotizing enteritis.

4. In Claim 1, The above pharmaceutical composition is characterized by increasing the expression level of a gene related to necrotizing enterocolitis.

5. In Claim 4, A pharmaceutical composition characterized in that the above-mentioned necrotizing enteritis-related gene is one or more selected from the group consisting of mature intestinal cell-related genes, digestive enzyme-related genes, fatty acid transport-related genes, amino acid transport-related genes, inorganic solute-related genes, and intercellular junction-related genes.

6. In Claim 5, A pharmaceutical composition characterized in that the above-mentioned mature intestinal cell-related gene is one or more selected from the group consisting of TM4SF20, THSD4, SLC46A1, LCT, and ABTB2.

7. In Claim 5, A pharmaceutical composition characterized in that the above digestive enzyme-related gene is one or more selected from the group consisting of TMPRSS15 and ANPEP.

8. In Claim 5, A pharmaceutical composition characterized in that the above fatty acid transport-related gene is one or more selected from the group consisting of APOB, ACSL5, SLC27A4, and FABP.

9. In Claim 5, A pharmaceutical composition characterized in that the above amino acid transport-related gene is one or more selected from the group consisting of SLC6A19, SLC6A6, SLC25A12, and SLC1A1.

10. In Claim 5, A pharmaceutical composition characterized in that the above-mentioned inorganic solute-related gene is one or more selected from the group consisting of CFTR, SLC26A3, and SLC12A2.

11. In Claim 5, A pharmaceutical composition characterized in that the above-mentioned intercellular junction-related gene is one or more selected from the group consisting of CLDN1 and ZO-1.

12. In Claim 5, The above pharmaceutical composition is characterized by having effects such as promoting the maturation of intestinal cells, restoring nutrient digestion ability, promoting fatty acid absorption and metabolism, restoring amino acid absorption ability, maintaining electrolyte and fluid balance, strengthening intestinal barrier function, and reducing intestinal permeability.

13. A pharmaceutical preparation for the prevention or treatment of necrotizing enteritis comprising the pharmaceutical composition of Claim 1.

14. In Claim 13, The above pharmaceutical formulation is characterized by further comprising a pharmaceutically acceptable carrier, excipient, or diluent.

15. In Claim 13, The above pharmaceutical preparation is characterized by being injectable, injectable, sprayable, liquid, or patch-type.

16. In Claim 13, A pharmaceutical preparation characterized in that the above-mentioned necrotizing enteritis is neonatal necrotizing enteritis.

17. In Claim 13, The above pharmaceutical preparation is characterized by having effects such as promoting the maturation of intestinal cells, restoring nutrient digestion ability, promoting fatty acid absorption and metabolism, restoring amino acid absorption ability, maintaining electrolyte and fluid balance, strengthening intestinal barrier function, and reducing intestinal permeability.

18. A health functional food for the prevention or improvement of necrotizing enteritis, comprising the pharmaceutical composition of Claim 1.

19. In Claim 18, A health functional food characterized in that the above-mentioned necrotizing enteritis is neonatal necrotizing enteritis.

20. In Claim 18, The above-mentioned health functional food is characterized by having effects such as promoting the maturation of intestinal cells, restoring nutrient digestion ability, promoting fatty acid absorption and metabolism, restoring amino acid absorption ability, maintaining electrolyte and fluid balance, strengthening intestinal barrier function, and reducing intestinal permeability.

Images