Pharmaceutical composition for preventing or treating cancer comprising inflammatory macrophages

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating cancer, the pharmaceutical composition comprising inflammatory macrophages. Inflammatory macrophages according to an aspect have cancer prevention or treatment effects, such as significantly reducing the size of a tumor while increasing the proportion of inflammatory macrophages, such as M1-type macrophages, in the tumor. In addition, when the pharmaceutical composition is administered in combination with an anticancer drug or further includes the anticancer drug, remarkable cancer prevention or treatment effects are exhibited, even if a relatively small amount of the anticancer drug is included, compared to when each substance is used alone, and side effects of the anticancer drug are not exhibited. Therefore, the pharmaceutical composition according to an aspect can be used in various industrial fields as a monotherapy or combination therapy against cancer.

Description

Pharmaceutical composition for the prevention or treatment of cancer containing inflammatory macrophages

[0001] The present invention relates to a pharmaceutical composition for the prevention or treatment of cancer comprising inflammatory macrophages.

[0002]

[0003] Ovarian cancer is a malignant tumor that occurs in the ovaries, which play a crucial role in female reproduction and hormone secretion, and it most commonly develops between the ages of 50 and 70. It is known to have a high mortality rate because approximately 60% of cases are diagnosed when the disease has already progressed. Currently, surgical resection and first-line chemotherapy are the treatment methods; however, recurrence within a few years of initial treatment is common due to resistance to chemotherapy. Therefore, research is needed on new therapies that can have a tumor-specific and direct impact, rather than relying on existing treatment techniques.

[0004] Cancer immunotherapy is a treatment method that treats cancer by improving the immune system to fight the disease. Macrophages, a type of immune cell, can undergo polarity shifts from M1 (pro-inflammatory) to M2 (anti-inflammatory and reparative) depending on the microenvironment, and are known to play dual roles in inflammation and cancer, respectively. In particular, within tumor tissue, an increase in macrophages called tumor-associated macrophages (TAMs) influences tumor progression and metastasis.

[0005] TAMs play a role in promoting tumor growth, invasion, immune evasion, and angiogenesis by interacting with tumor cells within tumor tissue. These are primarily classified as M2 macrophages, which facilitate tumor growth and metastasis by creating a favorable tumor environment through interactions with tumor cells. In contrast, M1 macrophages directly mediate cytotoxicity that kills tumor cells and inhibit tumor growth by activating the immune system's response. Recent studies suggest that macrophage immunotherapy is considered a novel approach for treating deadly cancers, including ovarian cancer, but further research into the underlying mechanisms is required.

[0006] Therefore, this study investigated the effects on tumor growth and metastasis by injecting M1-induced bone marrow-derived cells (BMDMs), which serve as a contrasting role to ovarian cancer xenograft models characterized by increased M2 or M2-like macrophages, thereby altering the role of macrophages within the tumor. Based on these findings, we confirmed that the combined administration of M1 macrophages and anticancer drugs exhibits superior tumor-suppressing effects, thereby proposing a novel strategy for the treatment of ovarian cancer.

[0007]

[0008] One aspect provides a pharmaceutical composition for the prevention or treatment of cancer comprising inflammatory macrophages.

[0009] Another aspect is to provide a method for the prevention or treatment of cancer comprising the step of administering inflammatory macrophages to an individual in need.

[0010] Another aspect is providing health functional foods for the prevention or improvement of cancer that contain inflammatory macrophages.

[0011] Another aspect is to provide the use of the aforementioned inflammatory macrophages for the manufacture of drugs for the prevention or treatment of cancer.

[0012]

[0013] One aspect provides a pharmaceutical composition for the prevention or treatment of cancer comprising inflammatory macrophages.

[0014] The term "inflammatory macrophage" refers to a type of macrophage that plays an important role in the immune response, primarily performing the function of fighting against pathogens and promoting inflammatory responses. These cells become activating in response to the presence of pathogens or tissue damage, and enhance the inflammatory response by secreting inflammatory cytokines TNF-α, IL-6, or IL-1β.

[0015] In one embodiment, the inflammatory macrophage may be one or more selected from the group consisting of M1 type macrophages and M1-like macrophages.

[0016] The above "classically activated macrophage (M1)" refers to a type of tumor-associated macrophage (TAM) that is activated by inflammatory cytokines such as LPS, IFN-γ, IL-1β, TNF-α, or TLR engagement. M1 macrophages secrete pro-inflammatory cytokines or chemokines such as IL-6, IL-12, and TNF-α, and function as antigen-presenting cells (APCs) to activate T cells, or to induce pathogen death or cancer cell death by activating tumor-infiltrating lymphocytes (TILs) or NK cells, or to directly kill cancer cells through phagocytosis.

[0017] The aforementioned "M1-like macrophages" possess characteristics similar to the M1-type macrophages, but differ depending on the environment or activation conditions. M1-like macrophages promote inflammatory responses and play a crucial role in immune defense against pathogens and tumor cells by producing cytokines such as TNF-α, IL-6, and IL-12. Like M1-type macrophages, M1-like macrophages are activated by stimuli such as IFN-γ or LPS; however, unlike M1-type macrophages, the role they perform varies depending on the actual tumor microenvironment.

[0018] In one embodiment, the pharmaceutical composition for the prevention or treatment of cancer may further include one or more other active ingredients as active ingredients, in addition to the inflammatory macrophages, that exhibit the same or similar function or assist the same.

[0019] In one embodiment, the pharmaceutical composition may further include a suitable carrier, excipient, or diluent commonly used in the manufacture of pharmaceutical compositions.

[0020] The above pharmaceutical composition may have any one dosage form selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, liquids, emulsions, syrups, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories, and may be various dosage forms for oral or parenteral administration. When formulating, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms are prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., with one or more compounds. In addition, lubricants such as magnesium stearate and talc are also used in addition to simple excipients. Liquid preparations for oral administration include suspensions, oral liquids, emulsions, and syrups; in addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients, such as humectants, sweeteners, flavorings, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleates may be used as non-aqueous solvents and suspension solvents. Witepsol, Macrogol, Tween 61, cocoa paste, laurin paste, and glycerogelatin may be used as bases for suppositories.

[0021] The above pharmaceutical composition is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" means an amount sufficient to prevent or treat cancer with a reasonable benefit / risk ratio applicable to medical use, and the effective dose level may be determined based on factors including individual type and severity, age, gender, drug activity, weight, 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. In this case, the content of the active ingredient included in the above pharmaceutical composition may be 0.0001 weight % to 10 weight %, specifically 0.001 weight % to 1 weight %, based on the total weight of the composition.

[0022] In one embodiment, the inflammatory macrophages are 1 x 10 7 Up to 3 x 10 9 cells / kg may be administered, specifically, the inflammatory macrophages are 1 x 10 7 Up to 3 x 10 9 cells / kg, 1.5 x 10 7 Up to 3 x 10 9 cells / kg, 2 x 10 7 Up to 3 x 10 9 cells / kg, 2.5 x 10 7 Up to 3 x 10 9 cells / kg, 1 x 10 8 Up to 3 x 10 9 cells / kg, 1.5 x 10 8 Up to 3 x 10 9 cells / kg, 2 x 10 8 Up to 3 x 10 9 cells / kg, 2.5 x 10 8 Up to 3 x 10 9 cells / kg, 1 x 10 8 Up to 3 x 10 8 cells / kg, 1.5 x 10 8 Up to 3 x 10 8cells / kg, 2 x 10 8 Up to 3 x 10 8 cells / kg, 2.5 x 10 8 Up to 3 x 10 8 cells / kg, 1 x 10 8 Up to 2.5 x 10 8 cells / kg, 1.5 x 10 8 Up to 2.5 x 10 8 cells / kg, 2 x 10 8 Up to 2.5 x 10 8 cells / kg, 1 x 10 9 Up to 3 x 10 9 cells / kg, 1.5 x 10 9 Up to 3 x 10 9 cells / kg, 2 x 10 9 Up to 3 x 10 9 cells / kg or 2.5 x 10⁶ 9 Up to 3 x 10 9 cells / kg may be administered. More specifically, the inflammatory macrophages are 1 x 10 8 Up to 3 x 10 8 cells / kg, 1.5 x 10 8 Up to 3 x 10 8 cells / kg, 2 x 10 8 Up to 3 x 10 8 cells / kg, 2.5 x 10 8 Up to 3 x 10 8 cells / kg, 1 x 10 8 Up to 2.5 x 10 8 cells / kg, 1.5 x 10 8 Up to 2.5 x 10 8 cells / kg or 2 x 10 8 Up to 2.5 x 10 8 cells / kg, can be administered.

[0023] In one embodiment, after injecting the inflammatory macrophages (M1-BMDMs) into an animal model with ovarian cancer and checking the tumor size on the 21st day, the tumor size of the control group not treated with M1-BMDMs was 1848.41 mm 3 On the other hand, M1-BMDMs 1 x 10 6 The tumor size of the experimental group treated with cells / mouse was 1477.67 mm 3 and 2.5 x 10 M1-BMDMs 6 The tumor size of the experimental group treated with cells / mouse was 1057.46 mm 3 It was confirmed that the tumor size decreased significantly as time increased in proportion to the concentration of injected inflammatory macrophages (see Example 4).

[0024] In one embodiment, the pharmaceutical composition may increase the proportion of inflammatory macrophages within a tumor. This may be due to a natural increase in the proportion of M1-type macrophages within the tumor by injecting inflammatory macrophages, or it may be the result of M2-type macrophages being repolarized into M1-type macrophages upon injecting inflammatory macrophages.

[0025] In one embodiment, when the inflammatory macrophages (M1-BMDMs) were injected into an animal model with ovarian cancer, it was confirmed that the proportion of M1-type macrophages in the tumor increased over time. In particular, it was confirmed that at 12 hours after the injection of the inflammatory macrophages, the proportion of M1-type macrophages in the tumor increased to 25.36%, and the proportion of M2-type macrophages decreased to approximately 0% (see Example 5).

[0026] In one embodiment, the pharmaceutical composition may further include a pharmaceutical composition for the prevention or treatment of cancer other than the inflammatory macrophage, i.e., an anticancer agent.

[0027] The above pharmaceutical composition may be provided mixed with other anticancer agents, and the anticancer agent may be a conventionally known anticancer agent or a newly developed anticancer agent.

[0028] If the above pharmaceutical composition further includes other anticancer agents, it is important that the amounts mixed allow for maximum effect to be obtained with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.

[0029] When the above pharmaceutical composition further includes other anticancer agents, a synergistic effect may occur in which the preventive or therapeutic effects of cancer, such as the tumor formation inhibitory effect or the tumor size inhibitory effect, become more pronounced than when the pharmaceutical composition containing the above inflammatory macrophage is included only as an active ingredient.

[0030] The term “anticancer agent” above refers to a drug that inhibits the growth or proliferation of cancer cells, and is classified into cytotoxic anticancer agents, targeted anticancer agents, immunotherapy agents, etc., and an anticancer agent can be selected depending on the type or stage of cancer, the patient's condition, etc., and two or more drugs may be used simultaneously to enhance the effect.

[0031] In one aspect, the above anticancer agent may be a cytotoxic anticancer agent. The “cytotoxic anticancer agent” refers to an anticancer agent that inhibits cell proliferation and induces apoptosis by inhibiting the replication or synthesis of DNA and RNA or protein synthesis required at each stage of the cell division cycle of cancer cells. Not only cancer cells but also normal cells are affected by the cytotoxic anticancer agent, resulting in side effects such as anemia, immunosuppression, hair loss, or diarrhea. The above cytotoxic anticancer agent includes alkylating agents, metabolic antagonists, DNA rotase inhibitors, or microtubule inhibitors, and specifically, it may be a microtubule inhibitor.

[0032] The term “microtubule-targeting agents” refers to drugs that inhibit the proliferation of cancer cells by interfering with the function of microtubules, which are crucial in the process of cell division. These drugs bind to the microtubules of cancer cells, inhibiting cell division and ultimately inducing the death of the cancer cells.

[0033] In one embodiment, the microtubule inhibitor may be one or more selected from the group consisting of cabazitaxel, paclitaxel, docetaxel, vinblastine, vincristine, and vinorelbine, specifically one or more selected from the group consisting of cabazitaxel, paclitaxel, and docetaxel, and more specifically paclitaxel.

[0034] The aforementioned “paclitaxel” acts to stop the cell division process by inhibiting the breakdown of microtubules, and is an anticancer drug primarily used in the treatment of solid tumors such as breast cancer, lung cancer, or ovarian cancer.

[0035] In one embodiment, the pharmaceutical composition may be administered alone or in combination with other anticancer agents. That is, the pharmaceutical composition may be administered in conjunction with other known anticancer agents having a preventive or therapeutic effect against cancer, 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, taking all of the above factors into consideration, and this can be easily determined by a person skilled in the art.

[0036] The term “concurrent administration” above refers to the administration of two or more drugs simultaneously or during the same treatment period.

[0037] When the above pharmaceutical composition is administered in combination with other anticancer agents, a synergistic effect may occur in which the preventive or therapeutic effects against cancer, such as the effect of inhibiting tumor formation or tumor size, become more pronounced than when the above pharmaceutical composition is administered alone.

[0038] A pharmaceutical composition according to one aspect may exhibit a synergistic effect when administered in combination with an anticancer drug, compared to when the anticancer drug is administered alone or when the pharmaceutical composition is administered alone.

[0039] In one embodiment, a synergistic effect was confirmed by co-administering paclitaxel, a microtubule inhibitor, and the inflammatory macrophages (M1-BMDMs). In an animal model with ovarian cancer, paclitaxel 10 mg / kg and inflammatory macrophages (M1-BMDMs) 1 x 10 6 As a result of injecting cells / mousses separately and administering them in combination to determine the degree of tumor size reduction, it was confirmed that the most superior tumor size reduction effect was observed when paclitaxel and inflammatory macrophages were administered in combination (see Example 7).

[0040] In one embodiment, when the pharmaceutical composition is administered in combination with another anticancer agent, the pharmaceutical composition may be administered after the anticancer agent has been administered.

[0041] In one embodiment, when the pharmaceutical composition is administered before an anticancer drug is administered, that is, when an anticancer drug is administered after the pharmaceutical composition is administered, the preventive or therapeutic effect of the pharmaceutical composition against cancer may not be excellent due to the effect of the anticancer drug. Accordingly, the above pharmaceutical composition may be administered after an anticancer drug has been administered, and specifically, the above pharmaceutical composition may be administered 6 to 24 hours after the anticancer drug has been administered, 9 to 24 hours after, 12 to 24 hours after, 15 to 24 hours after, 18 to 24 hours after, 21 to 24 hours after, 6 to 21 hours after, 9 to 21 hours after, 12 to 21 hours after, 15 to 21 hours after, 18 to 21 hours after, 6 to 18 hours after, 9 to 18 hours after, 12 to 18 hours after, 15 to 18 hours after, 6 to 15 hours after, 9 to 15 hours after, 12 to 15 hours after, 6 to 12 hours after, 9 to 12 hours after, or 6 to 9 hours after It may be administered. More specifically, the above pharmaceutical composition may be administered 9 to 24 hours after the anticancer drug is administered, 12 to 24 hours after, or 15 to 24 hours after.

[0042] The above pharmaceutical composition may be administered orally or parenterally. When the above pharmaceutical composition is administered parenterally, it may be administered via 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, intranasal injection, intra-intestinal injection, local injection, sublingual injection, rectal injection, ventricular injection, intracerebral micro-injection, intraconjunctival injection, or thoracic injection. Specifically, the above pharmaceutical composition may be administered intravenously.

[0043] The term "cancer" above refers to a disease caused by cells that possess aggressive characteristics, such as dividing and growing beyond normal growth limits; invasive characteristics, such as infiltrating surrounding tissues; and metastatic characteristics, such as spreading to other parts of the body or outside.

[0044] In one embodiment, the cancer may be a solid cancer.

[0045] The above “solid tumor” refers to a tumor that forms in a solid form, meaning a cancer that is localized to a specific organ or tissue within the body and forms a mass. Unlike blood cancer, which occurs in a liquid state such as blood or the lymphatic system, solid tumors are cancers formed by the aggregation of tumor cells; they arise from the excessive proliferation of cells in tissues and are treated in various ways, such as surgery, radiation therapy, and chemotherapy.

[0046] In one embodiment, M1 macrophages play a role in directly killing cancer cells by secreting cytokines such as IFN-γ within the tumor to produce reactive oxygen species and nitric oxide. Additionally, they play a role in maintaining an anti-tumor environment by presenting cancer cell antigens to activate immune cells such as T cells to eliminate cancer cells, interfering with tumor cell angiogenesis, and suppressing immunosuppressive components. Accordingly, when the proportion of M1 macrophages within a solid tumor increases, it is possible to effectively suppress the tumor to reduce its size and simultaneously prevent or treat cancer.

[0047] In one embodiment, the solid tumor may be one or more selected from the group consisting of brain tumor, melanoma, multiple myeloma, non-small cell lung cancer, oral cancer, liver cancer, stomach cancer, colon cancer, breast cancer, triple negative breast cancer (TNBC), lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cervical cancer, ovarian cancer, colorectal cancer, small intestine cancer, rectal cancer, fallopian tube carcinoma, pro-anal cancer, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, lymphoma, bladder cancer, gallbladder cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, kidney cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system tumor, spinal cord tumor, brainstem glioma and pituitary adenoma, and specifically, It may be one or more selected from the group consisting of ovarian cancer, esophageal cancer, breast cancer, lung cancer, Kaposi's sarcoma, cervical cancer, head and neck cancer, gastric cancer, prostate cancer, non-small cell lung cancer, small cell lung cancer, bladder cancer, brain cancer, melanoma, testicular cancer, and pancreatic cancer, and more specifically, it may be one or more selected from the group consisting of ovarian cancer, esophageal cancer, breast cancer, lung cancer, Kaposi's sarcoma, cervical cancer, and pancreatic cancer.

[0048] The term "prevention" above refers to any act of suppressing cancer in an individual or delaying its onset by administering a pharmaceutical composition according to one aspect.

[0049] The term "treatment" above refers to any act in which the symptoms of cancer in an individual are improved or beneficially altered by the administration of a pharmaceutical composition according to one aspect.

[0050] The term "administration" above means introducing a specific substance to an individual in an appropriate manner.

[0051] Another aspect provides a method for the prevention or treatment of cancer comprising the step of administering inflammatory macrophages to an individual in need.

[0052] The above "inflammatory macrophages," "administration," "cancer," "prevention," and "treatment" may be within the aforementioned scope.

[0053] The term “individual” above refers to all living organisms capable of carrying cancer, including humans, rats, mice, and livestock. Specific examples may include mammals, including humans.

[0054] In one embodiment, the step of administering the inflammatory macrophages may be a step of administering the macrophages in combination with a pharmaceutical composition having a preventive or therapeutic effect against other cancers, namely an anticancer agent.

[0055] The term “concurrent administration” above refers to the administration of two or more drugs simultaneously or during the same treatment period.

[0056] In one embodiment, the method may further include a step of administering a pharmaceutical composition for the prevention or treatment of other cancers, i.e., an anticancer agent, prior to the step of administering inflammatory macrophages.

[0057] In one embodiment, the inflammatory macrophages may be administered in combination with other known anticancer agents having a preventive or therapeutic effect against cancer, 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, taking all of the above factors into consideration, and this can be easily determined by a person skilled in the art.

[0058] When the above-mentioned inflammatory macrophages are administered in combination with other anticancer agents, a synergistic effect may occur in which the preventive or therapeutic effects against cancer, such as the tumor formation inhibitory effect or tumor size inhibitory effect, become more pronounced than when the above-mentioned pharmaceutical composition is administered alone.

[0059] Inflammatory macrophages according to one aspect can exhibit a synergistic effect when administered in combination with an anticancer drug, compared to when the anticancer drug is administered alone or when the inflammatory macrophages are administered alone.

[0060] In one embodiment, when the inflammatory macrophages are administered in combination with another anticancer agent, the inflammatory macrophages may be administered after the anticancer agent has been administered.

[0061] In one embodiment, when the inflammatory macrophages are administered before the anticancer agent is administered, that is, when the anticancer agent is administered after the inflammatory macrophages are administered, the effect of the anticancer agent may result in the prevention or treatment of cancer by the inflammatory macrophages not being excellent. Accordingly, the above-mentioned inflammatory macrophages may be administered after the anticancer drug is administered, and specifically, the above-mentioned inflammatory macrophages may be administered 6 to 24 hours after the anticancer drug is administered, 9 to 24 hours after, 12 to 24 hours after, 15 to 24 hours after, 18 to 24 hours after, 21 to 24 hours after, 6 to 21 hours after, 9 to 21 hours after, 12 to 21 hours after, 15 to 21 hours after, 18 to 21 hours after, 6 to 18 hours after, 9 to 18 hours after, 12 to 18 hours after, 15 to 18 hours after, 6 to 15 hours after, 9 to 15 hours after, 12 to 15 hours after, 6 to 12 hours after, 9 to 12 hours after, or 6 to It may be administered after 9 hours. More specifically, the inflammatory macrophages may be administered after 9 to 24 hours, 12 to 24 hours, or 15 to 24 hours after the anticancer drug is administered.

[0062] In one embodiment, the inflammatory macrophages are 1 x 10 7 Up to 3 x 10 9cells / kg may be administered, specifically, the inflammatory macrophages are 1 x 10 7 Up to 3 x 10 9 cells / kg, 1.5 x 10 7 Up to 3 x 10 9 cells / kg, 2 x 10 7 Up to 3 x 10 9 cells / kg, 2.5 x 10 7 Up to 3 x 10 9 cells / kg, 1 x 10 8 Up to 3 x 10 9 cells / kg, 1.5 x 10 8 Up to 3 x 10 9 cells / kg, 2 x 10 8 Up to 3 x 10 9 cells / kg, 2.5 x 10 8 Up to 3 x 10 9 cells / kg, 1 x 10 8 Up to 3 x 10 8 cells / kg, 1.5 x 10 8 Up to 3 x 10 8 cells / kg, 2 x 10 8 Up to 3 x 10 8 cells / kg, 2.5 x 10 8 Up to 3 x 10 8 cells / kg, 1 x 10 8 Up to 2.5 x 10 8 cells / kg, 1.5 x 10 8 Up to 2.5 x 10 8 cells / kg, 2 x 10 8 Up to 2.5 x 10 8 cells / kg, 1 x 10 9 Up to 3 x 10 9 cells / kg, 1.5 x 10 9 Up to 3 x 10 9 cells / kg, 2 x 10 9 Up to 3 x 10 9 cells / kg or 2.5 x 10⁶ 9Up to 3 x 10 9 cells / kg may be administered. More specifically, the inflammatory macrophages are 1 x 10 8 Up to 3 x 10 8 cells / kg, 1.5 x 10 8 Up to 3 x 10 8 cells / kg, 2 x 10 8 Up to 3 x 10 8 cells / kg, 2.5 x 10 8 Up to 3 x 10 8 cells / kg, 1 x 10 8 Up to 2.5 x 10 8 cells / kg, 1.5 x 10 8 Up to 2.5 x 10 8 cells / kg or 2 x 10 8 Up to 2.5 x 10 8 cells / kg, can be administered.

[0063] The above inflammatory macrophages may be administered orally or parenterally, and when the above inflammatory macrophages are administered parenterally, they may be administered via topical 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, intranasal injection, intra-intestinal injection, local injection, sublingual injection, rectal injection, ventricular injection, intracerebral micro-injection, intraconjunctival injection, or thoracic injection. Specifically, the above inflammatory macrophages may be injected intravenously.

[0064] Another aspect provides a health functional food for the prevention or improvement of cancer containing inflammatory macrophages.

[0065] The above "inflammatory macrophages," "cancer," and "prevention" may be within the aforementioned range.

[0066] The term "improvement" above refers to any action that at least reduces parameters related to the condition being treated, such as the severity of symptoms.

[0067] The term “health functional food” above includes health functional foods, health foods, and health supplement foods. “Health food” refers to a food that has active effects on maintaining or promoting health compared to general foods, and “health supplement food” refers to a food intended for the purpose of health support.

[0068] The above-mentioned health functional food may be formulated into one selected from the group consisting of tablets, pills, powders, granules, powders, capsules, and liquid formulations, by further including one or more of a carrier, a diluent, an excipient, and an additive. Foods to which glabridin may be added according to one aspect include various food products, powders, granules, tablets, capsules, syrups, beverages, gum, tea, vitamin complexes, health functional foods, etc.

[0069] Specific examples of the above carrier, excipient, diluent, and additive may be one or more 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.

[0070] 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, when manufacturing food or beverages, the above-mentioned health functional food may be added in an amount of about 15% by weight or less, specifically about 10% by weight or less, with respect to the raw materials. However, in the case of long-term consumption for the purpose of health and hygiene or health control, the above amount may be less than the above range.

[0071] The above-mentioned health functional food may include other ingredients as essential components in addition to the active ingredients mentioned above. For example, it may include various flavoring agents or natural carbohydrates as additional ingredients, such as in ordinary beverages. Examples of the above-mentioned natural carbohydrates 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 mentioned above, natural flavoring agents (taumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) may be advantageously used. The proportion of the above-mentioned natural carbohydrates may be appropriately determined by the choice of a person skilled in the art.

[0072] In addition to the above, a health functional food according to one aspect may include 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. These ingredients may be used independently or in combination, and the proportion of these additives may also be appropriately selected by a person skilled in the art.

[0073] In one embodiment, the health functional food may further include a health functional food for the prevention or improvement of other cancers in addition to the inflammatory macrophages.

[0074] The above-mentioned health functional food for the prevention or improvement of other cancers may be provided in combination with other health functional foods for the prevention or improvement of other cancers that are conventionally known or newly developed health functional foods for the prevention or improvement of other cancers.

[0075] If the above-mentioned health functional food further includes other health functional foods for the prevention or improvement of cancer, it is important that the amounts mixed allow for maximum effect to be obtained with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.

[0076] If the above-mentioned health functional food further includes another health functional food for the prevention or improvement of cancer, a synergistic effect may occur in which the cancer prevention or improvement effect is more pronounced than when only the other health functional food for the prevention or improvement of cancer containing the above-mentioned inflammatory macrophages is included as an active ingredient.

[0077] In addition, in one embodiment, the health functional food may be consumed alone or in combination with the other health functional food for the prevention or improvement of cancer. That is, the health functional food may be consumed in conjunction with other known health functional foods having a preventive or improving effect on other cancers, or newly developed health functional foods for the prevention or improvement of other cancers; it 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 considering all of the above factors, and this can be easily determined by a person skilled in the art.

[0078] When the above-mentioned health functional food is consumed in combination with other health functional foods for the prevention or improvement of cancer, a synergistic effect may occur in which the cancer prevention or improvement effect becomes more pronounced than when the above-mentioned health functional food is consumed alone.

[0079] In one embodiment, the inflammatory macrophages are 1 x 10 7 Up to 3 x 10 9 cells / kg can be ingested, and specifically, the above-mentioned inflammatory macrophages are 1 x 10 7 Up to 3 x 10 9 cells / kg, 1.5 x 10 7 Up to 3 x 10 9 cells / kg, 2 x 10 7 Up to 3 x 10 9 cells / kg, 2.5 x 10 7 Up to 3 x 10 9 cells / kg, 1 x 10 8 Up to 3 x 10 9 cells / kg, 1.5 x 10 8 Up to 3 x 10 9 cells / kg, 2 x 10 8 Up to 3 x 10 9 cells / kg, 2.5 x 10 8 Up to 3 x 10 9cells / kg, 1 x 10 8 Up to 3 x 10 8 cells / kg, 1.5 x 10 8 Up to 3 x 10 8 cells / kg, 2 x 10 8 Up to 3 x 10 8 cells / kg, 2.5 x 10 8 Up to 3 x 10 8 cells / kg, 1 x 10 8 Up to 2.5 x 10 8 cells / kg, 1.5 x 10 8 Up to 2.5 x 10 8 cells / kg, 2 x 10 8 Up to 2.5 x 10 8 cells / kg, 1 x 10 9 Up to 3 x 10 9 cells / kg, 1.5 x 10 9 Up to 3 x 10 9 cells / kg, 2 x 10 9 Up to 3 x 10 9 cells / kg or 2.5 x 10⁶ 9 Up to 3 x 10 9 cells / kg can be ingested. More specifically, the above-mentioned inflammatory macrophages are 1 x 10 8 Up to 3 x 10 8 cells / kg, 1.5 x 10 8 Up to 3 x 10 8 cells / kg, 2 x 10 8 Up to 3 x 10 8 cells / kg, 2.5 x 10 8 Up to 3 x 10 8 cells / kg, 1 x 10 8 Up to 2.5 x 10 8 cells / kg, 1.5 x 10 8 Up to 2.5 x 10 8 cells / kg or 2 x 10 8 Up to 2.5 x 10 8 cells / kg, can be consumed.

[0080] Another aspect provides the use of the inflammatory macrophages for the manufacture of drugs for cancer prevention or treatment.

[0081] The above "cancer," "prevention," "treatment," and "inflammatory macrophages" may be within the aforementioned range.

[0082] In one embodiment, the inflammatory macrophages may be administered in combination with a pharmaceutical composition, namely an anticancer agent, that has a preventive or therapeutic effect against other cancers.

[0083] The term “concurrent administration” above refers to the administration of two or more drugs simultaneously or during the same treatment period.

[0084] In one embodiment, the inflammatory macrophages may be administered in combination with other known anticancer agents having a preventive or therapeutic effect against cancer, 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, taking all of the above factors into consideration, and this can be easily determined by a person skilled in the art.

[0085] When the above-mentioned inflammatory macrophages are administered in combination with other anticancer agents, a synergistic effect may occur in which the preventive or therapeutic effects against cancer, such as the effect of inhibiting tumor formation or tumor size, become more pronounced than when the above-mentioned inflammatory macrophages are administered alone.

[0086] Inflammatory macrophages according to one aspect can exhibit a synergistic effect when administered in combination with an anticancer drug, compared to when the anticancer drug is administered alone or when the inflammatory macrophages are administered alone.

[0087] In one embodiment, when the inflammatory macrophages are administered in combination with another anticancer agent, the inflammatory macrophages may be administered after the anticancer agent has been administered.

[0088] In one embodiment, when the inflammatory macrophages are administered before the anticancer agent is administered, that is, when the anticancer agent is administered after the inflammatory macrophages are administered, the effect of the anticancer agent may result in the prevention or treatment of cancer by the inflammatory macrophages not being excellent. Accordingly, the above-mentioned inflammatory macrophages may be administered after the anticancer drug is administered, and specifically, the above-mentioned inflammatory macrophages may be administered 6 to 24 hours after the anticancer drug is administered, 9 to 24 hours after, 12 to 24 hours after, 15 to 24 hours after, 18 to 24 hours after, 21 to 24 hours after, 6 to 21 hours after, 9 to 21 hours after, 12 to 21 hours after, 15 to 21 hours after, 18 to 21 hours after, 6 to 18 hours after, 9 to 18 hours after, 12 to 18 hours after, 15 to 18 hours after, 6 to 15 hours after, 9 to 15 hours after, 12 to 15 hours after, 6 to 12 hours after, 9 to 12 hours after, or 6 to It may be administered after 9 hours. More specifically, the inflammatory macrophages may be administered after 9 to 24 hours, 12 to 24 hours, or 15 to 24 hours after the anticancer drug is administered.

[0089] In one embodiment, the inflammatory macrophages are 1 x 10 7 Up to 3 x 10 9 cells / kg may be administered, specifically, the inflammatory macrophages are 1 x 10 7 Up to 3 x 10 9 cells / kg, 1.5 x 10 7 Up to 3 x 10 9cells / kg, 2 x 10 7 Up to 3 x 10 9 cells / kg, 2.5 x 10 7 Up to 3 x 10 9 cells / kg, 1 x 10 8 Up to 3 x 10 9 cells / kg, 1.5 x 10 8 Up to 3 x 10 9 cells / kg, 2 x 10 8 Up to 3 x 10 9 cells / kg, 2.5 x 10 8 Up to 3 x 10 9 cells / kg, 1 x 10 8 Up to 3 x 10 8 cells / kg, 1.5 x 10 8 Up to 3 x 10 8 cells / kg, 2 x 10 8 Up to 3 x 10 8 cells / kg, 2.5 x 10 8 Up to 3 x 10 8 cells / kg, 1 x 10 8 Up to 2.5 x 10 8 cells / kg, 1.5 x 10 8 Up to 2.5 x 10 8 cells / kg, 2 x 10 8 Up to 2.5 x 10 8 cells / kg, 1 x 10 9 Up to 3 x 10 9 cells / kg, 1.5 x 10 9 Up to 3 x 10 9 cells / kg, 2 x 10 9 Up to 3 x 10 9 cells / kg or 2.5 x 10⁶ 9 Up to 3 x 10 9 cells / kg may be administered. More specifically, the inflammatory macrophages are 1 x 10 8 Up to 3 x 10 8 cells / kg, 1.5 x 108 Up to 3 x 10 8 cells / kg, 2 x 10 8 Up to 3 x 10 8 cells / kg, 2.5 x 10 8 Up to 3 x 10 8 cells / kg, 1 x 10 8 Up to 2.5 x 10 8 cells / kg, 1.5 x 10 8 Up to 2.5 x 10 8 cells / kg or 2 x 10 8 Up to 2.5 x 10 8 cells / kg, can be administered.

[0090] The above inflammatory macrophages may be administered orally or parenterally, and when the above inflammatory macrophages are administered parenterally, they may be administered via topical application to the skin or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, intra-arterial injection, intramedullary injection, intracardiac injection, intradural injection, transdermal injection, intranasal injection, intra-intestinal injection, local injection, sublingual injection, rectal injection, ventricular injection, intracerebral micro-injection, intraconjugation injection, or intrathoracic injection. Specifically, the above inflammatory macrophages may be injected intravenously.

[0091]

[0092] Inflammatory macrophages according to one aspect possess excellent preventive or therapeutic effects against cancer, such as increasing the proportion of inflammatory macrophages, including M1-type macrophages, within the tumor while simultaneously significantly reducing tumor size. Furthermore, when administered in combination with an anticancer drug or when an anticancer drug is additionally included in the pharmaceutical composition, even if the anticancer drug is included at a relatively low dose, it exhibits significantly greater preventive or therapeutic effects against cancer compared to using each substance alone, while simultaneously avoiding the side effects of the anticancer drug. Therefore, the pharmaceutical composition according to one aspect can be utilized in various industrial fields as a monotherapy or combination therapy for anticancer treatment.

[0093]

[0094] Figure 1 is a schematic diagram showing the process of preparing an ovarian cancer animal model by xenotransplanting an A2780 cell line.

[0095] Figure 2 shows the tumor size of an animal model according to the number of xenografted A2780 cells.

[0096] Figure 3 shows the appearance of the animal model on days 14 and 21 according to the number of xenotransplanted A2780 cells.

[0097] Figure 4 shows the tumor growth pattern and pathological characteristics according to the number of xenografted A2780 cells.

[0098] Figure 5 is a schematic diagram showing the process of manufacturing bone marrow-derived macrophages used in the treatment of ovarian cancer.

[0099] Figure 6 shows the results of confirming whether the prepared macrophages express markers using a flow cytometer.

[0100] Figure 7 shows the results of confirming cell morphology after treating bone marrow-derived macrophages and RAW 256.7 cells with LPS and IL-4.

[0101] Figure 8 shows the results of identifying the types of mRNA and proteins expressed in bone marrow-derived macrophages upon treatment with LPS and IL-4.

[0102] Figure 9 is a schematic diagram showing the experimental process of treating ovarian cancer in an animal model of ovarian cancer by treating it with M1-BMDMs.

[0103] Figure 10 shows the change in tumor size of an animal model as a result of treatment with M1-BMDMs.

[0104] Figure 11 shows the appearance of an animal model on days 14 and 21 following treatment with M1-BMDMs.

[0105] Figure 12 is a schematic diagram showing the experimental process for isolating single cells from tumor tissues of an ovarian cancer animal model and a treatment group animal model for the analysis of the polarity of macrophages within the tumor.

[0106] Figures 13 and 14 show the change in the proportion of macrophages in an animal model over time following treatment with M1-BMDMs.

[0107] Figures 15 and 16 show the results of confirming the ratio of M1 and M2 macrophages through flow cytometry of single cells isolated from tumor tissue.

[0108] Figure 17 is a schematic diagram showing the experimental process of treating ovarian cancer in an animal model of ovarian cancer by treating it with paclitaxel.

[0109] Figure 18 shows the change in tumor size of an animal model according to the paclitaxel treatment dose.

[0110] Figure 19 shows the appearance and H&E staining results of an animal model on days 14 and 21 according to the paclitaxel treatment dose.

[0111] Figure 20 is a schematic diagram showing the experimental process of treating ovarian cancer in an animal model of ovarian cancer by combining M1-BMDMs and paclitaxel.

[0112] Figure 21 shows the change in tumor size in an animal model treated with M1-BMDMs and paclitaxel.

[0113] Figure 22 shows the appearance and H&E staining results on days 14 and 21 of an animal model treated with M1-BMDMs and paclitaxel.

[0114]

[0115] The present invention will be explained in more detail below through examples. However, these examples are intended to illustrate the invention and the scope of the invention is not limited to these examples.

[0116]

[0117] Reference Example - Preparation of an Ovarian Cancer Animal Model

[0118] Animal models were prepared to confirm the therapeutic effect of inflammatory macrophages on ovarian cancer in vivo (Examples 1 to 7). An ovarian cancer xenograft mouse model was prepared by injecting human ovarian cancer cell line A2780 cells (ECACC, Wiltshire, UK) into 8-week-old female mice (BALB / c-Foxn1 nu / Arc-Gem nude), and the overall experimental process is shown in Figure 1.

[0119] Specifically, 1 x 10 A2780 cells 3 , 1 x 10 4 , 1 x 10 5 and 1 x 10 6 After injecting the cells into the dorsal subcutaneous (sc) of mice, they were observed for up to 28 days.

[0120] Observations showed that the tumor size in the mouse model increased over time in proportion to the number of injected A2780 cells. A2780 cells 1 x 10 3 In the group transplanted according to the number of cells, no tumors were observed until day 21, but A2780 cells 1 x 10 5 , 1 x 10 6In the group transplanted according to the number of cells, it was observed that the tumor size increased exponentially starting from the 14th day. On the other hand, A2780 cells at 1 x 10 4 It was confirmed that in the group transplanted with a cell count of [amount], tumors formed of an appropriate size for conducting experiments to verify the therapeutic effect of inflammatory macrophages on ovarian cancer; thus, the aforementioned A2780 cells [1 x 10] 4 Subsequent experiments were conducted with a group transplanted with the same number of cells (Fig. 2).

[0121] The tumor images of the animal model according to the number of transplanted A2780 cells are shown in Fig. 3, and Fig. 4 is data for analyzing the tumor tissue, in which various locations of the tumor were confirmed by performing HE staining.

[0122]

[0123] Examples

[0124] Example 1. Preparation and verification of bone marrow-derived macrophages (BMDMs)

[0125] To obtain bone marrow-derived macrophages (BMDMs) used in the treatment of ovarian cancer, the tibia and femur of 8-9 week old C57BL6 male mice were used. The specific procedure is as shown in Figure 5.

[0126] Bone marrow-derived cells (BMDCs) were obtained by washing the bone marrow of a tibia and femur with both ends severed using DPBS supplemented with 2% FBS (Gibco) via a 26 G syringe. The samples were centrifuged at 500 xg and washed with DPBS supplemented with 2% FBS, then incubated in 1 X erythrocyte lysis solution (BioLegend, San Diego, CA, USA) at 4°C for 5 minutes to isolate the BMDCs. The isolated BMDCs were differentiated into macrophages (Mφs) by culturing them for 7 days in RPMI medium (Gibco) containing M-CSF (Macrophage Colony-Stimulating Factor) (50 ng / ml).

[0127] To confirm whether the bone marrow-derived macrophages prepared above exhibit macrophage characteristics, the expression levels of macrophage markers CD45, CD11b, and F4 / 80 proteins were checked using a flow cytometer. As a result, it was confirmed that CD45, CD11b, and F4 / 80 were all expressed positively at a rate of over 99%, indicating that the hematopoietic stem cells had completely differentiated into macrophages (Fig. 6).

[0128]

[0129] Example 2. Induction of polarization in bone marrow-derived macrophages (BMDMs) and confirmation of cell morphology

[0130] Bone marrow-derived macrophages (BMDMs) prepared in Example 1 above and RAW 264.7 mouse monocyte cell line (ATCC, Manassas, VA, USA) were treated with LPS and IL-4 to induce polarization of the macrophages into M1 or M2 macrophages, and the changes in cell morphology were observed using an optical microscope.

[0131] It was confirmed that both types of cells differentiated into M1 macrophages upon treatment with LPS (100 ng / ml) for 24 hours, and into M2 macrophages upon treatment with IL-4 (20 ng / ml) for 24 hours. M1-BMDMs exhibited an oval shape with elongated pseudopodia, whereas M2-BMDMs were round with short pseudopodia. RAW 264.7 cells maintained an overall round or oval shape even after differentiation into M1 / M2, and morphological variation between cells was less than that of BMDMs. It was confirmed that the morphology of the differentiated cells was distinctly different between the two types (Fig. 7).

[0132]

[0133] Example 3. Confirmation of marker expression in bone marrow-derived macrophages (BMDMs)

[0134] The markers expressed in the macrophages that induced polarization in Example 2 above were analyzed at the mRNA level and the protein level.

[0135] At the mRNA level, in the LPS-treated group, the M2 macrophage markers Fizz1, Mgl2, and Pd-l2 were not expressed, while the expression of the M1 macrophage markers Scos3, iNOS, and Tnf-α was increased. Additionally, in the IL-4-treated group, it was confirmed that the macrophage markers Scos3, iNOS, and Tnf-α were not expressed, while the expression of the M2 macrophage markers Fizz1, Mgl2, and Pd-l2 was increased.

[0136] Similarly, at the protein level, in the LPS-treated group, M2 macrophage markers CD206 and ARG1 were not expressed, while the expression of M1 macrophage markers INOS and COX2 proteins increased. In addition, in the IL-4-treated group, M1 macrophage markers INOS and COX2 were expressed in small amounts, but it was confirmed that the expression of M2 macrophage markers CD206 and ARG1 proteins significantly increased. Therefore, it was found that BMDMs were polarized into M1 macrophages upon treatment with LPS (Fig. 8).

[0137]

[0138] Example 4. Confirmation of tumor size reduction effect by M1-BMDMs treatment

[0139] In the animal model prepared in Reference Example 1 above, 1 x 10 M1-type bone marrow-derived macrophages (M1-BMDMs) differentiated in Example 2 above were added. 6 or 2.5 x 10 6 It was injected at a concentration of cells / mouse. M1-BMDMs were intravenously injected a total of two times on days 7 and 14 after A2780 cells were transplanted. Subsequently, the degree of tumor size reduction in mice was checked using a caliper until day 21. The overall experimental procedure is shown in Figure 9.

[0140] Experimental results showed that the tumor size of the control group not treated with M1-BMDMs was 1848.41 mm 3 On the other hand, M1-BMDMs 1 x 10 6 The tumor size of the experimental group treated with cells / mouse was 1477.67 mm 3 and 2.5 x 10 M1-BMDMs 6 The tumor size of the experimental group treated with cells / mouse was 1057.46 mm 3It was confirmed that the tumor size of mice decreased over time in proportion to the concentration of treated M1-BMDMs (Figs. 10 and 11).

[0141]

[0142] Example 5. Confirmation of changes in the ratio of macrophages within tumors by M1-BMDMs treatment

[0143] The ratio of macrophages within the tumor and their polarity in the animal model prepared in Reference Example 1 above were analyzed. M1-BMDMs were injected into the animal model in the same manner as in Example 4 above, and tumor tissues were collected at 1, 3, 6, 12, 24, 48, and 72 hours, after which single cells were isolated (Fig. 12).

[0144] The isolated single cells were stained with F4 / 80 antibody, and the proportion of the macrophage population within the tumor was analyzed using a flow cytometer. The results confirmed that F4 / 80 within the tumor + The proportion of cells was 12.16% in the control A2780 tumor, and when M1-BMDMs were injected, it increased to 17.39% at 1 hour but decreased to 8.83% after 6 hours. Subsequently, it showed the highest proportion at 20.88% at 12 hours, and exhibited a gradual decreasing trend to 11.38% after 24 hours, 9.91% at 48 hours, and 7.92% at 72 hours. Consequently, it was confirmed that the injection of M1-BMDMs temporarily increased macrophage infiltration within the tumor, with the proportion of macrophages being highest particularly 12 hours after injection (Figs. 13 and 14).

[0145] Next, the single cells were stained with CD80 antibodies (used as M1 macrophage markers) and CD206 antibodies (used as M2 macrophage markers), and the ratio of M1 to M2 macrophages in the final single cells was determined using a flow cytometer. The results showed that in the control group, M1 cells accounted for 31%, M2 cells for 3%, and double-positive cells for 44% of the macrophages, indicating that double-positive cells accounted for a higher proportion than M2 alone. However, at 1 hour after injecting M1-BMDMs, the proportion of M1 increased to 32%, and the proportion of double-positive cells increased significantly to 61%. Furthermore, at 6 hours, the proportion of M1 increased even more significantly to 59%. At 12 hours after injection, the proportion of M1 cells remained dominant at 54%, while double-positive cells accounted for 39%. However, starting from 24 hours, the proportion of M1 cells gradually decreased to 35%, 46% at 48 hours, and 38% at 72 hours. Conversely, the proportion of double-positive cells showed a recovery trend, reaching 53% at 24 hours, 23% at 48 hours, and 41% at 72 hours.

[0146] That is, when M1-BMDMs are injected, the subtype of macrophages within the tumor rapidly switches to the anti-tumor M1 type, and it was confirmed that the effect was most pronounced, particularly at 6 to 12 hours (Figs. 15 and 16).

[0147]

[0148] Example 6. Setting of paclitaxel-treated dose for co-administration with M1-BMDMs

[0149] An experiment was conducted to determine the appropriate treatment dose of paclitaxel for co-administration with the M1-BMDMs differentiated in Example 2 above.

[0150] Paclitaxel 10 mg / kg or 20 mg / kg was injected into an animal model in the same manner as in Example 4 above. Subsequently, the degree of tumor size reduction in mice was checked using a caliper up to day 21. The overall experimental procedure is shown in Fig. 17.

[0151] As a result of checking the tumor size of mice on day 21, the tumor size of the control group not treated with paclitaxel was approximately 2000 mm 3 In contrast, the experimental group treated with 10 mg / kg of paclitaxel had a tumor size of 1500 mm 3 And, the experimental group treated with 20 mg / kg of paclitaxel had a tumor size of 900 mm 3 It was confirmed that (Fig. 18). In addition, H&E staining confirmed that tumor necrosis was prominently observed in the experimental group treated with 20 mg / kg of paclitaxel, and that the tumor reduction effect was the most excellent. That is, it was confirmed that the tumor size of mice decreased over time in proportion to the dose of paclitaxel administered (Fig. 19).

[0152] Therefore, in order to determine whether the therapeutic effect is maximized when M1-BMDMs, whose effects were confirmed in Example 4 above, are administered in combination with paclitaxel, the appropriate treatment dose of paclitaxel in the subsequent combination administration experiment was set to 10 mg / kg.

[0153]

[0154] Example 7. Confirmation of tumor size reduction effect by co-administration of M1-BMDMs and paclitaxel

[0155] 1 x 10 M1-BMDMs in the same manner as Example 4 above 6cells / mouse and 10 mg / kg of paclitaxel were injected. However, to avoid interference with the therapeutic effect of M1-BMDMs, paclitaxel was injected 12 hours before injecting M1-BMDMs. Subsequently, the degree of tumor size reduction in mice was checked using a caliper until day 21, and the overall experimental procedure is shown in Figure 20.

[0156] As a result of checking the tumor size of the mice on day 21, the tumor size of the control group was approximately 1250 mm 3 , M1-BMDMs 1 x 10 6 Approximately 600 mm when cells / mouse are treated alone 3 , when paclitaxel 10 mg / kg was administered alone, approximately 900 mm 3 , when M1-BMDMs are administered in combination with paclitaxel, approximately 400 mm 3 As a result, it was confirmed that the tumor size decreased by more than three times compared to the control group when administered in combination, compared to when each substance was treated alone (p≤0.05) (Fig. 21). In addition, when paclitaxel was treated alone at a high concentration (20 mg / kg) or M1-BMDMs at 2.5 x 10 6 It was confirmed that it has a superior tumor reduction effect compared to treatment with cells / mouse alone.

[0157] Similarly, the results confirmed by H&E staining showed that tumor necrosis was prominently observed when M1-BMDMs and paclitaxel were administered in combination, confirming that the tumor reduction effect was the best (Fig. 22).

Claims

1. A pharmaceutical composition for the prevention or treatment of cancer comprising inflammatory macrophages.

2. A pharmaceutical composition according to claim 1, wherein the inflammatory macrophages are one or more selected from the group consisting of M1-type macrophages and M1-like macrophages.

3. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises an anticancer agent.

4. A pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is administered in combination with an anticancer agent.

5. A pharmaceutical composition according to claim 3 or 4, wherein the anticancer agent is a microtubule-targeting agent.

6. A pharmaceutical composition according to claim 3 or 4, wherein the anticancer agent is one or more selected from the group consisting of cabazitaxel, paclitaxel, docetaxel, vinblastine, vincristine, and vinorelbine.

7. The pharmaceutical composition of claim 4, wherein the pharmaceutical composition is administered after an anticancer agent has been administered.

8. The pharmaceutical composition of claim 4, wherein the pharmaceutical composition is administered 12 to 24 hours after the administration of an anticancer agent.

9. In Claim 1, the inflammatory macrophages are 1 x 10 7 Up to 3 x 10 9 A pharmaceutical composition administered in cells / kg.

10. A pharmaceutical composition according to claim 1, wherein the cancer is a solid tumor.

11. A pharmaceutical composition according to claim 1, wherein the cancer is one or more selected from the group consisting of ovarian cancer, esophageal cancer, breast cancer, lung cancer, Kaposi's sarcoma, cervical cancer, head and neck cancer, gastric cancer, prostate cancer, non-small cell lung cancer, small cell lung cancer, bladder cancer, brain cancer, melanoma, testicular cancer, and pancreatic cancer.

12. A pharmaceutical composition according to claim 1, wherein the pharmaceutical composition increases the proportion of inflammatory macrophages within a tumor.

13. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is administered intravenously.

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