Application of cinnamyl alcohol and derivatives thereof in preparation of medicines for preventing or treating tumors

Abstract

The invention discloses application of cinnamyl alcohol and derivatives thereof in preparation of medicines for preventing or treating tumors, and relates to the technical field of biological medicines. Wherein the tumor is a solid tumor and comprises at least one of epithelial tumor, sarcoma and lymphoma. The research shows that the cinnamyl alcohol medicine can inhibit the growth and metastasis of melanoma, prevent and treat the recurrence of the melanoma after operation, and prolong the survival time of individuals. In addition, the cinnamyl alcohol drugs can also significantly inhibit proliferation of other epithelial tumors such as breast cancer, glioma, lung cancer and the like. Therefore, the invention develops a new field of cinnamyl alcohol drugs in anti-tumor treatment; the important guiding significance is realized on the clinical application of tumor prevention and treatment.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to the application of cinnamyl alcohol and its derivatives in preparing medicines for preventing or treating tumors. Background Art

[0002] Tumor is one of the most common fatal diseases, which has brought great economic pressure and heavy medical burden to the society. Clinically, there are solid tumors and non-solid tumors. Among them, solid tumors are tangible tumors, including epithelial tumors, sarcomas and lymphomas. Epithelial tumors account for 50%-70% of primary ovarian tumors, and their malignant types account for 85%-90% of ovarian malignant tumors. Epithelial tumors include melanoma, breast cancer, lung cancer, liver cancer and glioma. At present, the treatment of tumors is a comprehensive treatment with surgery as the main treatment and radiotherapy and chemotherapy as the auxiliary treatment. However, despite the application of these treatments, many patients still have recurrence and metastasis, resulting in poor prognosis. Therefore, seeking a more effective drug for tumor treatment is an urgent clinical requirement.

[0003] Cinnamyl alcohol is an organic compound with the molecular formula C9H 10 O. Its natural products exist in the form of esters in cinnamon leaves, hyacinth oil and styrax esters, etc., and are often used as spices, fragrances, preservatives, deodorants, etc. In recent years, some studies have begun to focus on the medicinal value of cinnamyl alcohol and explore its potential in anti-inflammatory, antiviral, anti-tumor and other diseases. However, the application of cinnamyl alcohol drugs in preventing and treating tumor growth, metastasis and postoperative recurrence is imperfect and needs further development to provide new application areas and scientific basis for clinical application in treating tumors.

[0004] In view of this, the present invention is proposed. Summary of the invention

[0005] The purpose of the present invention is to provide the use of cinnamyl alcohol and its derivatives in the preparation of drugs for preventing or treating tumors, so as to solve the above technical problems.

[0006] The present invention is achieved in that:

[0007] In a first aspect, an embodiment of the present invention provides a use of cinnamyl alcohol and its derivatives in preparing a drug for preventing or treating a tumor, wherein the tumor is a solid tumor; the solid tumor includes at least one of an epithelial tumor, a sarcoma, and a lymphoma.

[0008] In a second aspect, an embodiment of the present invention provides an anti-epithelial tumor drug, which includes cinnamyl alcohol and its derivatives and pharmaceutical excipients; wherein the epithelial tumor includes at least one of melanoma, breast cancer, lung cancer and glioma.

[0009] In a third aspect, an embodiment of the present invention provides a use of cinnamyl alcohol and its derivatives in the preparation of a combined therapeutic drug for treating tumors, wherein the combined therapeutic drug is selected from any one of a surgical treatment drug, a chemotherapy drug and an immunotherapy drug.

[0010] The present invention has the following beneficial effects:

[0011] The cinnamyl alcohol and its derivatives provided by the embodiment of the present invention can inhibit the growth and metastasis of tumors and prevent and treat the recurrence of tumors after surgery in the preparation of drugs for preventing or treating tumors, and can prolong the survival time of individuals. Tumors are solid tumors, which include at least one of epithelial tumors, sarcomas and lymphomas. Therefore, the present invention provides a new application field and scientific basis for cinnamyl alcohol drugs, which has important guiding significance for their clinical application in preventing and treating epithelial tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0013] Figure 1 In Example 1 of the present invention, when cinnamyl alcohol is used to prevent and treat melanoma in vitro, it can significantly inhibit tumor proliferation;

[0014] Figure 2 In Example 1 of the present invention, when cinnamyl alcohol is used to prevent and treat melanoma in vitro, it can significantly inhibit tumor migration;

[0015] Figure 3 In Example 1 of the present invention, when cinnamyl alcohol is used to prevent and treat melanoma in vitro, it can significantly inhibit the migration, invasion and metastasis of the tumor;

[0016] Figure 4 In Example 2 of the present invention, when cinnamyl alcohol is used in vivo to prevent and treat subcutaneous in situ melanoma, it can significantly inhibit the growth of tumors;

[0017] Figure 5 In Example 3 of the present invention, cinnamyl alcohol is used in vivo in combination with surgery to treat melanoma, which can significantly inhibit the recurrence of melanoma and significantly prolong the survival time of individuals;

[0018] Figure 6 In Example 4 of the present invention, when cinnamyl alcohol is used to prevent and treat breast cancer, glioma, and lung cancer in vitro, it can significantly inhibit the proliferation of tumors. DETAILED DESCRIPTION

[0019] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0020] Traditional Chinese medicine culture has a long history. Since "Shennong tasted hundreds of herbs", "medicine" and "food" have been inseparable. In the process of the development of food and medicine, a unique "medicine and food are of the same origin" culture has been formed, which believes that many foods are both food and medicine, and food and medicine can also prevent and treat diseases. In recent years, the National Health Commission has successively issued relevant policies to regulate products of medicine and food of the same origin. Now there are more than 100 food and drug substances that are traditionally used as food and listed in the "Pharmacopoeia of the People's Republic of China" (hereinafter referred to as the "Chinese Pharmacopoeia"). Cinnamon is one of them.

[0021] Cinnamon bark, also known as male cinnamon, big cinnamon, mushroom cinnamon, tube cinnamon, spicy cinnamon, etc., is derived from the dried bark of the cinnamon bark of the Lauraceae plant. It was first recorded in "Shennong's Herbal Classic" and listed as a top-grade medicine. It is a commonly used Chinese medicine in clinical practice of traditional Chinese medicine. "Pharmacopoeia" records that cinnamon bark is spicy, sweet, and very hot. It enters the kidney, spleen, heart, and liver meridians. It replenishes fire and helps yang, draws fire back to the origin, dispels cold and relieves pain, and warms and dredges the meridians. It is used for impotence, cold uterus, cold pain in the waist and knees, asthma caused by kidney deficiency, floating of virtual yang, dizziness and red eyes, cold pain in the heart and abdomen, vomiting and diarrhea caused by deficiency and cold, abdominal pain caused by cold hernia, dysmenorrhea and amenorrhea. "Shennong's Herbal Classic" records that cinnamon bark is used to treat cough and reverse breathing, vomiting and breathing caused by qi stagnation in the throat, benefit joints, and replenish the middle and replenish qi. "Bielu" records that cinnamon bark is used to treat heart pain, side wind, side pain, warm tendons, dredge meridians, stop restlessness, and sweat. It warms the middle part of the body, benefits the liver and lungs, treats cold and heat in the heart and abdomen, cold diseases, cholera cramps, headaches, back pain, stops saliva, coughs, and nasal congestion; it can induce abortion, strengthen bones and joints, dredge blood vessels, and regulate deficiency; it promotes hundreds of medicines and is fearless. The Materia Medica records that cinnamon is mainly used to treat nine kinds of heartache, kill three insects, break blood, dredge menstruation, treat soft feet, numbness, numbness, retained placenta, eliminate cough, qi stagnation, and numbness, stop cold air in the abdomen, unbearable pain, and treat diarrhea and nasal polyps. Kill plant poisons. The Rihuazi Materia Medica records that cinnamon can treat all kinds of wind, replenish five kinds of labor and seven kinds of injuries, open nine orifices, benefit joints, benefit essence, improve eyesight, warm the waist and knees, break up lumps, eliminate blood stasis, treat wind arthritis, joint contracture, connect tendons and bones, and grow muscles. The Origin of Medicine records that cinnamon replenishes the deficiency of the lower jiao, treats deep cold and chronic cold, and spontaneous sweating due to superficial deficiency. "Compendium of Materia Medica" records that cinnamon can treat cold arthralgia, aphonia, blood loss due to yin excess, diarrhea, epilepsy, and blood loss due to yang deficiency, and can support carbuncles and pox, induce blood to turn into sweat and pus, and detoxify snake venom.

[0022] Cinnamyl alcohol, also known as cinnamic acid alcohol, is a natural compound extracted from the bark of the cinnamon tree. It has a fragrant and spicy taste. Cinnamyl alcohol is widely used in the food, pharmaceutical and spice industries. It is expected to be prepared into a product that can be used as both medicine and food, which can supplement nutrition and treat diseases at the same time.

[0023] At present, the medicinal value of cinnamyl alcohol is mainly reflected in anti-inflammatory, antiviral, anti-tumor and other diseases, and its applicability in preventing or treating tumors is rarely studied. The present invention studies the new uses of cinnamyl alcohol drugs, as follows:

[0024] In a first aspect, an embodiment of the present invention provides a use of cinnamyl alcohol and its derivatives in the preparation of a drug for preventing or treating a tumor, wherein the tumor is a solid tumor, and the solid tumor includes at least one of an epithelial tumor, a sarcoma, and a lymphoma.

[0025] It should be noted that in the optional embodiments of the present invention, epithelial tumors are used as representatives to study the applicability of cinnamyl alcohol and its derivatives in the preparation of drugs for preventing or treating tumors. In other embodiments of the present invention, sarcomas and lymphomas can be selected for research. Malignant tumors that occur in the epithelium are called cancers, which are the most common type of malignant tumors in humans. Cancers often grow mainly in an invasive manner, so they are not clearly demarcated from surrounding tissues. Cancers that occur on the surface of the skin and mucous membranes often appear polyp-like, mushroom-like or cauliflower-like in appearance, and often have necrosis and ulceration on the surface; those that occur in organs are often irregular nodules, infiltrating into surrounding tissues in a tree root-like or crab-like shape, with a hard texture, and the cut surface is often off-white and dry. Under the microscope, cancer cells can be arranged in glandular, nested or cord-like shapes, with clear boundaries from the stroma. They can also grow diffusely and infiltratingly in the stroma, with unclear boundaries from the stroma.

[0026] Sarcoma is a malignant tumor originating from mesenchymal tissue (including connective tissue and muscle), which often occurs in the skin, subcutaneous tissue, periosteum and both ends of long bones.

[0027] Lymphoma is a malignant tumor originating from lymphatic tissue, including two major types: Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL).

[0028] In an alternative embodiment, the epithelial tumor comprises at least one of melanoma, breast cancer, lung cancer and glioma.

[0029] Melanoma, usually refers to malignant melanoma, is a highly malignant tumor derived from melanocytes, referred to as melanoma. It mostly occurs in the skin, and can also be seen in mucous membranes and internal organs, accounting for about 3% of all tumors. Cutaneous malignant melanoma ranks third among skin malignancies (accounting for about 6.8%-20%), and its 5-year survival rate is even lower than 5%. It is more common in adults, with a high incidence in white people with fair skin, and a lower incidence in Asians and Africans with dark skin, and it is rarely seen in children. Some patients have familial multiple phenomena. Malignant melanoma can evolve from congenital or acquired benign melanocytic nevi, or from the malignant transformation of dysplastic nevi, or it can be a new occurrence. In recent years, the incidence and mortality of malignant melanoma have increased year by year, and its age of death is lower than that of other solid tumors. In addition to early surgical resection, malignant melanoma lacks specific treatment and has a poor prognosis.

[0030] Breast Cancer, also known as breast cancer, is a common malignant tumor in women, arising from the uncontrolled proliferation of mammary epithelial cells. High-risk factors for cancer include genetics and estradiol exposure, such as early menarche, late menopause, infertility, and breast radiation exposure during childhood. In addition to high-risk groups, late development, long lactation, and early childbearing can reduce the chance of breast cancer. Tumor types are divided into non-invasive cancer and invasive cancer, and can develop to many organs throughout the body. The development of breast cancer can be divided using the TNM (Tumor Nodes Metastasis) system. The treatment options for breast cancer are diverse, such as surgery, chemotherapy, radiotherapy, hormone therapy, and targeted therapy, and the chance of recurrence after treatment is high.

[0031] Lung cancer, also known as primary bronchial lung cancer, is the most common lung malignancy originating from the trachea, bronchial mucosa or glands. According to the histopathological characteristics, it is mainly divided into non-small cell carcinoma (including adenocarcinoma and squamous cell carcinoma) and small cell carcinoma. Globally, the incidence and mortality of lung cancer are extremely high and on the rise. The incidence and mortality of lung cancer in men rank first among malignant tumors. For the treatment of lung cancer, commonly used methods include surgery, radiotherapy and chemotherapy. The prognosis of the disease varies depending on the patient's condition, such as tumor type, tumor stage, age, gender, etc., but overall, the five-year survival rate of lung cancer is not high. In terms of prevention and treatment, prevention is the key, including reducing smoking, avoiding occupational and environmental exposure, and regular physical examinations.

[0032] Glioma, also known as neuroglioma or glioma cell tumor, refers to a tumor derived from the neuroepithelium and is the most common type of intracranial tumor. According to the different sources, glioma can be divided into astrocytoma, oligodendroglioma, ependymoma, mixed glioma, glioblastoma, etc. The treatment of glioma is a comprehensive treatment based on surgery, combined with chemotherapy, targeted therapy and other therapies.

[0033] In an optional embodiment, the drug has at least one of the following uses:

[0034] (1) Used to inhibit tumor growth; (2) Used to inhibit tumor migration; (3) Used to inhibit tumor invasion; (4) Used to inhibit tumor metastasis.

[0035] Among them, in an optional embodiment of the present invention, the above-mentioned use of the drug is studied in an in vitro melanoma model, a breast cancer model, a glioma model and a lung cancer model. Specifically (taking melanoma as an example for explanation), the study on inhibiting tumor growth includes the following implementation steps:

[0036] (1) Cultivation of melanoma cell lines.

[0037] B16 and A375 melanoma cell lines were selected for in vitro culture, generally using DMEM medium containing 10% fetal bovine serum, and the culture conditions were 37° C. in a humidified environment containing 5% carbon dioxide.

[0038] (2) Cinnamyl alcohol drug administration and proliferation detection test.

[0039] The cells were digested with pancreatic enzymes for 2-5 minutes to keep the cells in suspension; then the effect of pancreatic enzymes was neutralized with fresh serum-containing medium, and the tumor cells were collected into a centrifuge tube. After centrifugation at 1000 rpm / min for 3-5 minutes, a cell pellet was obtained. After resuspending the precipitated cells with serum-containing medium, an appropriate amount (about 5×10^3 cells / well) of cells were seeded into a 96-well plate. After 24 hours, the cells were treated with drugs. Different groups of cells were treated with 0μM-120μM, and cell viability was detected by MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide) colorimetric method 48 hours after drug administration.

[0040] Further, in an optional embodiment of the present invention, the specific administration concentration of cinnamyl alcohol drugs in the in vitro melanoma model is 0μM, 3.75μM, 7.5μM, 15μM, 30μM, 60μM, 120μM, and the dosage of different cell groups is reasonably adjusted according to the actual situation. Among them, the preferred administration concentration is 0μM-120μM.

[0041] It should be noted that in the optional embodiments of the present invention, the breast cancer cell line, glioma cell line and lung cancer cell line are respectively selected from BT474 breast cancer cell line, A172 glioma cell line and A549 lung cancer cell line, which can be adjusted to other types of cell lines according to actual conditions.

[0042] The specific dosage concentrations of cinnamyl alcohol drugs in in vitro breast cancer models, glioma models and lung cancer models were 0 μM, 10 μM, 20 μM, 40 μM, 80 μM, 160 μM and 160 μM, and the dosage of different cell groups was reasonably adjusted according to the actual situation.

[0043] In other embodiments of the present invention, the cinnamyl alcohol drug is selected from any one of cinnamyl alcohol, dihydrocinnamyl alcohol and cinnamyl alcohol glycoside, which can be reasonably selected according to actual conditions.

[0044] In other embodiments of the present invention, the culture medium, the type of enzyme, the number of cells inoculated, etc. can be reasonably adjusted according to the actual experimental conditions.

[0045] In an optional embodiment of the present invention, the experiment for inhibiting tumor migration adopts a scratch experiment, specifically (taking melanoma as an example for explanation), comprising the following implementation steps:

[0046] (1) Cultivation of melanoma cell lines. The relevant implementation steps are the same as those for culturing melanoma cell lines for inhibiting tumor growth.

[0047] (2) Administration and migration detection test of cinnamyl alcohol drugs.

[0048] After the cells were seeded into a 96-well plate, a wound marker was used to scratch a wound in the middle of the well, and the width of the scratch was recorded by taking a photo. The cells were then cultured with serum-free medium, and each group was given medication, divided into a control group and a cinnamyl alcohol administration group. After 48 hours of medication, the width of the scratch was recorded again by taking a photo, and the migration distance of the cells was obtained by subtracting the two widths.

[0049] Furthermore, in the embodiment of the present invention, the preferred administration concentration for the migration detection test is 30 μM-60 μM, and the specific administration concentration can be reasonably adjusted according to the actual cell type cultured.

[0050] In addition, in an optional embodiment of the present invention, the experiment for inhibiting tumor migration, invasion and metastasis adopts a transwell migration and invasion experiment, specifically (taking melanoma as an example for explanation), comprising the following implementation steps:

[0051] Melanoma cells were seeded into six-well plates and divided into control group and cinnamyl alcohol group. For transwell migration experiment, after digestion, centrifugation and resuspension of cells in each group, appropriate amount of cells (about 2×10^4 cells / well) were seeded into transwell chamber (8.0μm, SPL, Japan), and 200μL serum-free culture medium was added into the chamber as the upper chamber of transwell system. At the same time, the transwell chamber was placed into a well of 24-well plate as the lower chamber of the system, and 500μL culture medium containing 10% fetal bovine serum was added. After 36 hours, the chamber was removed, the upper cells of the membrane (cells that did not penetrate the membrane) were gently wiped off, and the cells of the lower layer of the membrane (cells that had penetrated the membrane) were fixed with 4% paraformaldehyde, followed by staining and observation. For transwell invasion experiment, the basic steps were the same as transwell migration experiment, but a layer of matrix gel (Matrigel matrix, BD, USA) was pre-laid on the transwell chamber.

[0052] It should be noted that, in other embodiments of the present invention, the specifications of the transwell chamber, the specifications of the matrix gel, the culture medium and the number of cells to be inoculated can be reasonably adjusted according to actual conditions.

[0053] In an optional embodiment of the present invention, the study of the drug inhibiting the in situ growth of tumors in vivo, specifically (taking melanoma as an example for explanation), includes the following implementation steps:

[0054] (1) Establishment of subcutaneous tumor implant model. First, culture the B16 melanoma cell line in vitro, generally using a 15 cm culture dish. The rest of the digestion, centrifugation, and collection of tumor cell suspension are the same as before. Finally, resuspend the cells with PBS and bring the cell suspension into the animal room. Use a 1 mL syringe to inject the tumor cell suspension subcutaneously into the nude mouse (right buttocks), with an injection volume of 10^6 cells / 100 μL.

[0055] (2) Treatment with cinnamyl alcohol drugs. A cinnamyl alcohol solution was prepared with 10% dimethyl sulfoxide (DMSO) solvent and administered by intraperitoneal injection. The drug was administered every day for 14 consecutive days. A control group (intraperitoneal injection of the corresponding amount of solvent) and a cinnamyl alcohol administration group were set up.

[0056] (3) Three weeks after subcutaneous tumor implantation, the nude mice were euthanized and the tumors were removed for recording and observation of tumor size.

[0057] It should be noted that the mice used in the present invention for the study of inhibiting the in situ growth of melanoma in vivo are balb / c nude mice, a total of 18 mice (6 mice in each of the control group, the low-dose cinnamyl alcohol group and the high-dose cinnamyl alcohol group), half of which are male and half are female, 4-5 weeks old, and the average weight is 25 grams.

[0058] In an alternative embodiment, the drug is used to reduce the recurrence rate of tumors after surgery.

[0059] Tumor recurrence refers to the reappearance of the same tumor after a period of clinical cure or remission after surgery or radiotherapy. Recurrent tumors can occur at the original site (such as recurrence of the anastomosis after gastric cancer surgery), adjacent sites, or distant sites. There are many reasons for recurrence:

[0060] (1) Residual tumor cells: Due to incomplete tumor removal, there are residual tumor cells. It may also be due to the limitation of radiation dose during radiotherapy (due to the limited tolerance of the body), which makes it impossible to kill all tumor cells.

[0061] (2) The existence of latent metastatic lesions: When the body's immune function declines, tumor cells can regain their growth vitality. It is worth noting that sometimes the occurrence of multiple malignant tumors can be mistaken for tumor recurrence. For this so-called "recurrent tumor", surgical resection is still worthwhile.

[0062] In an optional embodiment of the present invention, the drug is used to inhibit at least one of the following parameters of a tumor: size, mass and area.

[0063] In an optional embodiment of the present invention, the drug is used to prolong the survival time of a subject;

[0064] Furthermore, the survival time of the subjects after surgical resection is prolonged.

[0065] In an optional embodiment of the present invention, the study of the drug inhibiting tumor recurrence after tumor resection in vivo, specifically (using melanoma as an example for explanation), includes the following implementation steps:

[0066] (1) Establishment of an in situ melanoma model. The procedure for subcutaneous melanoma implantation is the same as the study of drugs inhibiting in situ melanoma growth in vivo. After the subcutaneous tumor model is established, the volume of the subcutaneous tumor is measured daily. When the long diameter of the subcutaneous tumor reaches about 1.5 cm, the tumor can be surgically removed. Generally, it takes about 1-2 weeks for a B16-derived subcutaneous tumor to grow to 1.5 cm (long diameter).

[0067] It should be noted that the size of the tumor and the time required for growth will vary with the type of cells inoculated and the size of the nude mice, and can be reasonably adjusted based on actual conditions.

[0068] (2) Surgical removal of subcutaneous tumors.

[0069] The first step is preoperative preparation. You need to prepare avertin and isoflurane gas anesthesia, prepare the instruments and equipment required for sterile surgery, and disinfect the mice before surgery.

[0070] The second step is anesthesia. The combination of intraperitoneal infiltration anesthesia and gas anesthesia can be used. About 200 μL of the above-mentioned avertin is injected intraperitoneally to achieve intraperitoneal infiltration anesthesia, and isoflurane is used to maintain anesthesia.

[0071] The third step is tumor removal. The mouse is fixed on the operating table and fully exposed. The skin is then cut open, the tumor is separated along the edge of the healthy skin, and finally the tumor is removed in one piece.

[0072] Step 4: Suturing: After hemostasis, use No. 1 thread (diameter 0.20 mm) to perform simple interrupted sutures.

[0073] Step 5: Perioperative observation: If any nude mice died during the perioperative period (before surgery to 1 day after surgery), they were eliminated.

[0074] (3) Treatment with cinnamyl alcohol drugs. A cinnamyl alcohol solution was prepared with 10% dimethyl sulfoxide (DMSO) solvent and administered by intraperitoneal injection. The drug was administered every day for 14 consecutive days. A control group (intraperitoneal injection of the corresponding amount of solvent) and a cinnamyl alcohol administration group were set up.

[0075] It should be noted that the dosage concentration here is determined comprehensively based on the body weight of nude mice and the dosage concentration obtained from previous in vitro melanoma model studies.

[0076] (4) Follow-up. Regularly observe and record the tumor recurrence of each group of mice. Record and make statistics on the size of the recurrent tumors in the two groups of mice 3 weeks after surgery. Follow up for two months, record the survival status of each group of mice, and finally perform survival analysis.

[0077] In an optional embodiment of the present invention, the administration method of the drug is selected from any one of intravenous injection, intramuscular injection, intraperitoneal injection, intrathecal administration and oral administration.

[0078] In an optional embodiment of the present invention, the dosage form of the drug is selected from any one of an injection, a tablet, an implant, a capsule and a sustained-release controlled agent.

[0079] The administration method of the injection dosage form is selected from any one of intraperitoneal injection, intravenous injection and intramuscular injection. In an optional embodiment of the present invention, intraperitoneal injection is used. In other embodiments of the present invention, other injection methods can be selected according to actual needs.

[0080] In a second aspect, an embodiment of the present invention provides an anti-epithelial tumor drug, which includes cinnamyl alcohol and its derivatives and pharmaceutical excipients; wherein the epithelial tumor includes at least one of melanoma, breast cancer, lung cancer and glioma.

[0081] In an optional embodiment of the present invention, the pharmaceutical excipients include at least one of a pH regulator, a surfactant, a suspending agent, an isotonicity regulator, and a local analgesic.

[0082] It should be noted that the pharmaceutical excipients here mainly involve injection dosage forms. If it is an oral dosage form, the pharmaceutical excipients should be reasonably adjusted according to actual needs.

[0083] In a third aspect, an embodiment of the present invention provides a use of cinnamyl alcohol and its derivatives in the preparation of a combined therapeutic drug for treating tumors, wherein the combined therapeutic drug is selected from any one of a surgical treatment drug, a chemotherapy drug, and an immunotherapy drug.

[0084] Among them, surgical treatment is one of the most common cancer treatments, especially for early-stage tumors. In addition to completely removing the tumor, surgery can also be an important method for staging and histological examination of the lesion, determining the extent of the lesion and selecting subsequent treatment plans. In an optional embodiment of the present invention, the surgical treatment method selected is to remove the tumor. In other embodiments of the present invention, other forms of surgical methods can be selected according to actual needs.

[0085] Chemotherapy is a conventional treatment method that mainly uses drugs to kill tumor cells or inhibit their growth and spread to achieve the purpose of treatment. The specific chemotherapy plan depends on factors such as the type of cancer, the patient's health status, and the stage of the tumor.

[0086] Immunotherapy activates and enhances the body's immune system's own defenses against cancer to increase resistance to tumors or directly kill tumor cells.

[0087] In an optional embodiment of the present invention, the immunotherapy method is selected from any one of antibodies, vaccines, CAR-T cell therapy and TCR-T cell therapy.

[0088] Among them, "CAR-T cell therapy", or "chimeric antigen receptor T cell", is a cellular immunotherapy based on T cell-mediated CAR. It uses gene transduction technology to introduce antigen-specific CAR molecules into T cells, giving T cells the ability to accurately attack tumors, as if a "navigation head" is installed on the T cells, producing immediate and long-term anti-cancer effects.

[0089] "TCR-T cell therapy" or "T cell receptor engineered T cell therapy" uses genetic engineering technology to transfer antigen-specific TCR gene sequences (α and β chains) into T cells, enabling them to specifically recognize tumor antigens and accurately kill cancer cells.

[0090] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0091] Example 1

[0092] This example provides a cinnamyl alcohol drug that inhibits the proliferation, migration, invasion and metastasis of melanoma in an in vitro melanoma model. The relevant implementation steps are as follows:

[0093] (1) Cultivation of melanoma cell lines B16 and A375 melanoma cell lines were selected for in vitro culture, generally using DMEM medium containing 10% fetal bovine serum, and the culture conditions were 37° C. in a humidified environment containing 5% carbon dioxide.

[0094] (2) Cinnamyl alcohol drug administration and proliferation detection test. Melanoma grows in an adherent state in vitro. Trypsin can be used to digest the cells for 2-5 minutes to suspend them. Then, the effect of trypsin is neutralized with fresh serum-containing culture medium. The tumor cells are collected into a centrifuge tube and centrifuged at a speed of 1000 rpm / min for 3 minutes to obtain a cell pellet. After resuspending the precipitated cells with serum-containing culture medium, an appropriate amount of cells (about 5×10^3 cells / well) are seeded into a 96-well plate. After 24 hours, the cells are treated with medication. The specific cinnamyl alcohol drug used is cinnamyl alcohol.

[0095] Different groups of cells were given different doses. B16 type melanoma cell lines were given 0μM, 7.5μM, 15μM, 30μM, 60μM, and 120μM, respectively; A375 type melanoma cell lines were given 0μM, 3.75μM, 7.5μM, 15μM, 30μM, and 60μM, respectively; 48 hours after administration, MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide) colorimetric method was used to detect cell viability. Related results are shown in Figure 1 .

[0096] from Figure 1 It can be seen that cinnamyl alcohol can significantly inhibit the proliferation of B16 and A375 melanoma cells in vitro when the concentration is greater than 15 μM, and the inhibitory effect on the proliferation of B16 melanoma cells is better than that on the proliferation of A375 melanoma cells.

[0097] (3) Administration of cinnamyl alcohol drugs and scratch experiment. The culture, collection and plating of melanoma cell lines were performed in the same manner as above. In this experiment, B16 melanoma cells were used. After the cells were seeded into a 96-well plate, a wound marker was used to scratch a scratch in the middle of the well, and a photo was taken to record the width of the scratch. The cells were then cultured in serum-free culture medium, and each group was drugged and divided into a control group (0 μM) and a cinnamyl alcohol drug group (30 μM). 48 hours after drug administration, another photo was taken to record the width of the scratch, and the two widths were subtracted to obtain the migration distance of the cells. For related results, see Figure 2 .

[0098] from Figure 2 It can be seen that cinnamyl alcohol can significantly inhibit the migration ability of B16 melanoma cells in vitro.

[0099] (4) Administration of cinnamyl alcohol drugs and transwell migration and invasion experiments. B16 melanoma cells were seeded into six-well plates and divided into control group (0 μM) and cinnamyl alcohol administration group (60 μM, 120 μM). For the transwell migration experiment, after digestion, centrifugation and resuspension of cells in each group, an appropriate amount of cells (about 2 × 10^4 cells / well) were seeded into a transwell chamber (8.0 μm, SPL, Japan), and 200 μL of serum-free culture medium was added to the chamber as the upper chamber of the transwell system. At the same time, the transwell chamber was placed in a well of a 24-well plate as the lower chamber of the system, and 500 μL of culture medium containing 10% fetal bovine serum was added. After 36 hours, the chamber was removed, the upper layer of cells on the membrane (cells that did not penetrate the membrane) were gently wiped off, and the cells on the lower layer of the membrane (cells that had penetrated the membrane) were fixed with 4% paraformaldehyde, followed by staining and observation. For the transwell invasion assay, the basic steps are the same as the transwell migration assay, but a layer of Matrigel matrix (BD, USA) is pre-painted on the transwell chamber. Figure 3 .

[0100] from Figure 3 It can be seen that cinnamyl alcohol can significantly inhibit the migration and invasion ability of B16 melanoma cells through the transwell membrane in vitro, suggesting that cinnamyl alcohol can inhibit the migration, invasion and metastasis of melanoma.

[0101] Example 2

[0102] This embodiment provides a cinnamyl alcohol drug for inhibiting the growth of melanoma in situ tumors in vivo, and the relevant implementation steps are as follows:

[0103] (1) Establishment of subcutaneous tumor implant model. First, culture the B16 melanoma cell line in vitro, generally using a 15 cm culture dish. The rest of the digestion, centrifugation, and collection of tumor cell suspension are the same as before. Finally, resuspend the cells with PBS and bring the cell suspension into the animal room. Use a 1 ml syringe to inject the tumor cell suspension subcutaneously into the nude mouse (right buttocks), with an injection volume of 10^6 cells / 100 μL.

[0104] The mice were balb / c nude mice, 18 in total (6 in each of the control group, the low-dose cinnamyl alcohol group and the high-dose cinnamyl alcohol group), half of which were male and half were female, 4-5 weeks old, and had an average body weight of 25 grams.

[0105] (2) Treatment with cinnamyl alcohol drugs.

[0106] A cinnamyl alcohol solution was prepared with a 10% dimethyl sulfoxide (DMSO) solvent and administered by intraperitoneal injection. The drug was administered every day for 14 consecutive days. A control group (intraperitoneal injection of a corresponding amount of solvent), a cinnamyl alcohol-10 mg / kg group and a cinnamyl alcohol-30 mg / kg group were set up. The specific cinnamyl alcohol drug used was cinnamyl alcohol.

[0107] (3) Three weeks after subcutaneous tumor implantation, the nude mice were euthanized and the tumors were removed for recording and observation of tumor size. Figure 4 It was shown that 30 mg / kg of cinnamyl alcohol could significantly inhibit the growth of subcutaneous in situ melanoma.

[0108] Example 3

[0109] This embodiment provides a method for administering a cinnamyl alcohol drug after tumor resection in vivo, and the relevant implementation steps are as follows:

[0110] (1) Establishment of in situ melanoma model. The operation of subcutaneous melanoma implantation is the same as that in Example 2. After the subcutaneous tumor model is established, the volume of the subcutaneous tumor is observed and measured every day. When the long diameter of the subcutaneous tumor reaches about 1.5 cm, the tumor can be surgically removed. Generally, it takes about 1-2 weeks for B16-derived subcutaneous tumors to grow to 1.5 cm (long diameter).

[0111] (2) Surgical removal of subcutaneous tumors.

[0112] The first step is preoperative preparation. You need to prepare avertin and isoflurane gas anesthesia, prepare the instruments and equipment required for sterile surgery, and disinfect the mice before surgery.

[0113] The second step is anesthesia. The combination of intraperitoneal infiltration anesthesia and gas anesthesia can be used. About 200 μL of the above-mentioned avertin is injected intraperitoneally to achieve intraperitoneal infiltration anesthesia, and isoflurane is used for maintenance anesthesia.

[0114] The third step is tumor removal. The mouse is fixed on the operating table and fully exposed. The skin is then cut open, the tumor is separated along the edge of the healthy skin, and finally the tumor is removed in one piece.

[0115] Step 4: Suturing: After hemostasis, use No. 1 thread (diameter 0.20 mm) to perform simple interrupted sutures.

[0116] Step 5: Perioperative observation: If any nude mice died during the perioperative period (before surgery to 1 day after surgery), they were eliminated.

[0117] (3) Treatment with cinnamyl alcohol drugs.

[0118] A cinnamyl alcohol solution was prepared with 10% dimethyl sulfoxide (DMSO) solvent and administered by intraperitoneal injection. The administration was performed every day for 14 consecutive days. A control group (intraperitoneal injection of a corresponding amount of solvent) and a cinnamyl alcohol administration group (30 mg / kg) were set up. The specific cinnamyl alcohol drug used was cinnamyl alcohol.

[0119] (4) Follow-up.

[0120] The tumor recurrence of each group of mice was observed and recorded regularly. The size of the recurrent tumors in the two groups of mice 3 weeks after surgery was recorded and statistically analyzed. The mice were followed up for two months, and the survival status of each group of mice was recorded. Finally, survival analysis was performed. Figure 5 .

[0121] from Figure 5 It can be seen that cinnamyl alcohol drugs can significantly inhibit the recurrence of melanoma in vivo and prolong the survival time of individuals after surgical resection.

[0122] Example 4

[0123] This embodiment provides a cinnamyl alcohol drug that inhibits tumor proliferation in an in vitro breast cancer model, a glioma model, and a lung cancer model. The relevant implementation steps are as follows:

[0124] (1) Cultivation of breast cancer cell lines, glioma cell lines and lung cancer cell lines. BT474 breast cancer cell lines, A172 glioma cell lines and A549 lung cancer cell lines were selected for in vitro culture, generally using DMEM medium containing 10% fetal bovine serum, and the culture conditions were 37°C in a humidified environment containing 5% carbon dioxide.

[0125] (2) Cinnamyl alcohol drug administration and proliferation detection test. Breast cancer, glioma and lung cancer all grow in an adherent state in vitro. Trypsin can be used to digest the cells for 2-5 minutes to suspend them, and then the effect of trypsin can be neutralized with fresh serum-containing culture medium. The tumor cells are then collected into a centrifuge tube and centrifuged at a speed of 1000 rpm / min for 3 minutes to obtain a cell pellet. After resuspending the precipitated cells with serum-containing culture medium, an appropriate amount of cells (about 5×10^3 cells / well) are seeded into a 96-well plate. After 24 hours, the cells are treated with medication. The specific cinnamyl alcohol drug used is cinnamyl alcohol.

[0126] Three tumor cell lines were administered in vitro, with doses of 0μM, 10μM, 20μM, 40μM, 80μM, 160μM, and 160μM, respectively; 48 hours after administration, cell viability was detected using MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide) colorimetric method. Figure 6 .

[0127] from Figure 6 It can be seen that cinnamyl alcohol can significantly inhibit the proliferation of BT474 breast cancer cell line and A549 lung cancer cells in vitro when the concentration is greater than 10 μM; and cinnamyl alcohol can significantly inhibit the proliferation of A172 glioma cells in vitro when the concentration is greater than 40 μM.

[0128] In summary, the use of cinnamyl alcohol and its derivatives provided by the present invention in the preparation of drugs for preventing or treating tumors has the following characteristics:

[0129] 1) The present invention has created a new application field for cinnamyl alcohol drugs, and adopted a variety of cell and animal models to highly simulate the clinical scenario of melanoma, and to detect the therapeutic effect of cinnamyl alcohol drugs on the model.

[0130] 2) Cinnamyl alcohol drugs can significantly inhibit the proliferation of melanoma.

[0131] 3) Cinnamyl alcohol drugs can significantly inhibit the metastasis of melanoma.

[0132] 4) Cinnamyl alcohol drugs can significantly inhibit the recurrence of melanoma and prolong the survival time of individuals after surgical resection.

[0133] 5) Cinnamyl alcohol drugs can also significantly inhibit the proliferation of other epithelial tumors such as breast cancer, glioma, and lung cancer.

[0134] The invention provides a new application field and scientific basis for cinnamyl alcohol drugs, and has important guiding significance for the clinical application of cinnamyl alcohol drugs in preventing and treating solid tumors.

[0135] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Use of cinnamyl alcohol and its derivatives in the preparation of drugs for preventing or treating tumors, characterized in that: The tumor is a solid tumor; The solid tumor includes at least one of an epithelial tumor, a sarcoma, and a lymphoma.

2. The use according to claim 1, characterized in that: The epithelial tumor includes at least one of melanoma, breast cancer, lung cancer and glioma.

3. The use according to claim 1, characterized in that: The drug has at least one of the following uses: (1) Used to inhibit tumor growth; (2) Used to inhibit tumor migration; (3) Used to inhibit tumor invasion; (4) Used to inhibit tumor metastasis.

4. The use according to claim 1, characterized in that: The drug is used to reduce the postoperative recurrence rate of tumors.

5. The use according to claim 1, characterized in that: The administration method of the drug is selected from any one of intravenous injection, intramuscular injection, intraperitoneal injection, intrathecal administration and oral administration.

6. The use according to claim 1, characterized in that: The dosage form of the drug is selected from any one of injection, tablet, implant, capsule and sustained-release controlled agent.

7. A drug for treating epithelial tumors, characterized in that: The medicine comprises cinnamyl alcohol and its derivatives and pharmaceutical excipients; the epithelial tumor comprises at least one of melanoma, breast cancer, lung cancer and glioma.

8. The drug according to claim 7, characterized in that The pharmaceutical excipients include at least one of a pH regulator, a surfactant, a suspending agent, an isotonicity regulator, and a local analgesic.

9. Use of cinnamyl alcohol and its derivatives in the preparation of combined therapeutic drugs for treating tumors, characterized in that: The combined therapeutic drug is selected from any one of surgical treatment drugs, chemotherapy drugs, and immunotherapy drugs.

10. The use according to claim 9, characterized in that: The immunotherapy method is selected from any one of antibodies, vaccines, CAR-T cell therapy and TCR-T cell therapy.

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