Pharmaceutical composition for treating esophageal cancer and application thereof

Abstract

The invention discloses a pharmaceutical composition for treating esophageal cancer and application of the pharmaceutical composition. The pharmaceutical composition comprises an iminase I inhibitor and an ERK1 / 2 inhibitor. According to the application disclosed by the invention, the synergistic interaction effect of the combination of the topoisomerase I inhibitor SN-38 and the ERK1 / 2 inhibitor SCH772984 on the treatment of the esophageal cancer is found for the first time, the treatment effect on the esophageal cancer can be remarkably improved by combining the SN-38 and the SCH772984, and a brand new treatment scheme is provided for the technical field of esophageal cancer treatment; important scientific significance and clinical application value are realized.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically, relates to a pharmaceutical composition for treating esophageal cancer and an application thereof, and more specifically, relates to an application of a topoisomerase I inhibitor SN-38 and an ERK1 / 2 inhibitor SCH772984 in combination in the preparation of a drug for treating esophageal cancer. Background Art

[0002] Esophageal cancer has a poor prognosis and poses a serious threat to human health. The histological types of esophageal cancer are mainly divided into esophageal squamous cell carcinoma and esophageal adenocarcinoma. Since the 1970s, the incidence of esophageal squamous cell carcinoma in many Western countries has been declining. In contrast, the incidence of esophageal adenocarcinoma has increased rapidly, becoming one of the fastest growing malignant tumors.

[0003] At present, esophageal cancer is usually treated with surgery, radiotherapy, chemotherapy, and drug therapy. However, the effect of surgical treatment alone is poor. For patients with early esophageal cancer, the prognosis is good, and the 5-year survival rate is usually greater than 90%; but for patients with advanced esophageal cancer, the prognosis is poor, and the 5-year survival rate is about 17%. Chemotherapy, as an adjuvant treatment after esophageal cancer surgery, can improve the effect of surgical treatment and reduce the recurrence rate. However, the chemotherapy regimen currently used in clinical practice can only relieve symptoms and prolong survival, and the chemotherapy regimen has strong toxic side effects. Therefore, there is still a lack of effective drugs for the treatment of esophageal cancer in this field.

[0004] It can be seen that the chemotherapy regimen for the treatment of esophageal cancer has strong toxic side effects and the current lack of more effective esophageal cancer treatment drugs in this field has greatly affected the current clinical efficacy of esophageal cancer treatment. Therefore, finding new and more effective esophageal cancer treatment methods is of special importance and urgency. Summary of the invention

[0005] In view of this, in order to solve the above-mentioned technical problems currently faced in this field, the purpose of the present invention is to provide a combined pharmaceutical composition for treating esophageal cancer, thereby providing an effective drug combination strategy for the treatment of esophageal cancer.

[0006] In order to achieve the above object, the present invention adopts the following technical solution: A first aspect of the present invention provides a pharmaceutical composition for treating and / or preventing esophageal cancer.

[0007] Furthermore, the pharmaceutical composition comprises a topoisomerase I inhibitor and an ERK1 / 2 inhibitor.

[0008] Furthermore, the topoisomerase I inhibitor is SN-38, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof.

[0009] Furthermore, the ERK1 / 2 inhibitor is SCH772984, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof.

[0010] Furthermore, the concentration ratio of SN-38 to SCH772984 is 1:500 to 1:0.02.

[0011] Preferably, the concentration ratio of SN-38 to SCH772984 is 1:125 to 1:0.08.

[0012] Furthermore, the concentration ratio of SN-38 and SCH772984 is (0.01 μM~1 μM): (0.02 μM~5 μM).

[0013] Preferably, the concentration ratio of SN-38 and SCH772984 is (0.04 μM~1 μM): (0.08 μM~5 μM).

[0014] In a specific embodiment of the present invention, the inventors of the present invention discovered for the first time that the combination of the topoisomerase I inhibitor SN-38 and the ERK1 / 2 inhibitor SCH772984 has a synergistic effect in the treatment of esophageal cancer. The present invention has demonstrated through experiments that the combination of the two can significantly inhibit the growth of esophageal cancer cells and promote the apoptosis of esophageal cancer cells, showing a significant synergistic killing effect.

[0015] In the present invention, the topoisomerase I inhibitor is a class of compounds that can specifically inhibit the activity of DNA topoisomerase I, which can bind to topoisomerase I to form a stable DNA-topoisomerase I-inhibitor ternary complex, thereby preventing topoisomerase I from normal repair and relaxation of DNA, leading to DNA chain breakage and cell cycle arrest, and ultimately inducing tumor cell apoptosis.

[0016] In some embodiments, the topoisomerase I inhibitor comprises any agent capable of inhibiting the activity of DNA topoisomerase I. For example, the topoisomerase I inhibitor includes but is not limited to: SN-38, camptothecin, hydroxycamptothecin, irinotecan, topotecan, Genz 644282. In a specific embodiment of the present invention, the topoisomerase I inhibitor is SN-38.

[0017] In the present invention, the ERK1 / 2 inhibitor is a class of inhibitors targeting extracellular signal-regulated kinase (ERK) 1 and 2. ERK1 / 2 plays an important role in basic processes such as cell growth, proliferation and survival. Therefore, ERK1 / 2 inhibitors play a role in treating various cancers by inhibiting the activity of ERK1 / 2.

[0018] In some embodiments, the ERK1 / 2 inhibitor includes but is not limited to SCH772984, D3C-002, BPI-27336, AZD0364, LY3214996. In a specific embodiment of the present invention, the ERK1 / 2 inhibitor is SCH772984.

[0019] In some embodiments, the pharmaceutically acceptable salt refers to an acid salt formed with an inorganic and / or organic acid and a basic salt formed with an inorganic and / or organic base. In addition, when the compound contains a basic moiety (such as, but not limited to, pyridine or imidazole) and an acidic moiety (such as, but not limited to, a carboxylic acid), a zwitterion may be formed and the zwitterion is included in the pharmaceutically acceptable salts described herein. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, but other salts are also useful. A pharmaceutically acceptable salt of the compound can be formed, for example, by reacting the compound with an amount of an acid or a base in a medium, such as a medium in which the salt is precipitated or an aqueous medium (freeze-dried after the reaction).

[0020] Specific pharmaceutically acceptable salts include those salts which are suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic response, etc., and which are commensurate with a reasonable benefit or risk ratio, within the scope of sound medical judgment. Pharmaceutically acceptable salts (pharmaceutically acceptable salts) are well known in the art. Pharmaceutically acceptable salts of the compounds of the present invention include salts derived from suitable inorganic and organic acids and inorganic and organic bases.

[0021] The example of pharmaceutically acceptable nontoxic acid addition salt is the salt formed with inorganic acid, for example hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or the salt formed with organic acid, for example acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid. Also include the salt formed using the conventional methods in the art, for example, ion exchange method.Other pharmaceutically acceptable salts include: adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, gluconate, glycerophosphate, hemisulfate, enanthate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, mesylate. Pharmaceutically acceptable salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium salts and the like.

[0022] In some embodiments, the hydrate refers to a compound combined with water. It will be understood by those skilled in the art that an organic compound can form a complex with a solvent in which it reacts or precipitates or crystallizes, and these complexes are called solvates. When the solvent is water, the complex is called a hydrate.

[0023] In some embodiments, the solvate refers to a form of the compound or its salt that is combined with a solvent, usually formed by a solvolysis reaction. This physical association may include hydrogen bonding. Conventional solvents include water, methanol, ethanol, acetic acid, DMSO, THF, ether, etc. In some cases, the solvate will be able to separate, for example, when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid. Solvates include solvates in solution and separable solvates.

[0024] In some embodiments, the crystalline form refers to the crystalline form of a compound with a specific crystal packing arrangement. Different crystalline forms generally have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardnesses, crystal shapes, photoelectric properties, stability, and solubility. Recrystallization solvents, crystallization rates, storage temperatures, and other factors may lead to a dominant crystalline form. Various polymorphs of a compound can be prepared by crystallization under different conditions.

[0025] In some embodiments, the SN-38, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof; the SCH772984, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof, are all within the scope of protection of the present invention. In a specific embodiment of the present invention, the pharmaceutical composition is a pharmaceutical composition comprising SN-38 and SCH772984.

[0026] In the present invention, the treatment and / or prevention refers to delaying the development of a disease, preventing the development of a disease, and / or reducing the severity of the symptoms that will develop or are expected to develop. Therefore, these terms include improving existing disease symptoms, preventing additional symptoms, improving or preventing potential metabolic causes of symptoms, inhibiting a disorder or disease, for example, preventing the development of a disorder or disease, alleviating a disorder or disease, regressing a disorder or disease, alleviating a condition caused by a disease or disorder, or stopping symptoms of a disease or disorder.

[0027] Furthermore, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

[0028] In some embodiments, the pharmaceutically acceptable carrier and / or excipient include, but are not limited to, diluents, binders, surfactants, wetting agents, adsorption carriers, lubricants, fillers, and disintegrants.

[0029] In some embodiments, the diluent includes, but is not limited to, lactose, sodium chloride, glucose, urea, starch, water, and the like.

[0030] In some embodiments, the binder includes but is not limited to starch, pregelatinized starch, dextrin, maltodextrin, sucrose, gum arabic, gelatin, methyl cellulose, carboxymethyl cellulose, ethyl cellulose, polyvinyl alcohol, polyethylene glycol, polyvinyl pyrrolidone, alginic acid and alginate, xanthan gum, hydroxypropyl cellulose and hydroxypropyl methylcellulose, etc.

[0031] In some embodiments, the surfactant includes, but is not limited to, polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate, stearic acid monoglyceride, cetyl alcohol, and the like.

[0032] In some embodiments, the humectant includes but is not limited to glycerol, starch, and the like.

[0033] In some embodiments, the adsorption carrier includes, but is not limited to, starch, lactose, bentonite, silica gel, kaolin, and bentonite.

[0034] In some embodiments, the lubricant includes but is not limited to: zinc stearate, glyceryl monostearate, polyethylene glycol, talc, calcium and magnesium stearate, polyethylene glycol, boric acid powder, hydrogenated vegetable oil, sodium stearyl fumarate, polyoxyethylene monostearate, monolauric sucrose ester, sodium lauryl sulfate, magnesium lauryl sulfate, magnesium dodecyl sulfate, etc.

[0035] In some embodiments, the filler includes but is not limited to: mannitol (granular or powdered), xylitol, sorbitol, maltose, erythrose, microcrystalline cellulose, polymeric sugars, coupling sugars, glucose, lactose, sucrose, dextrin, starch, sodium alginate, laminarin powder, agar powder, calcium carbonate, sodium bicarbonate, etc.

[0036] In some embodiments, the disintegrant includes but is not limited to: cross-linked vinyl pyrrolidone, sodium carboxymethyl starch, low-substituted hydroxypropyl methyl, cross-linked sodium carboxymethyl cellulose, soybean polysaccharide, etc.

[0037] In some embodiments, the pharmaceutically acceptable carrier and / or excipient is used to help the stability of the formulation or to help improve its activity or its biological effectiveness or to produce an acceptable taste or smell in the case of oral administration as needed. The pharmaceutical composition thus prepared can be administered by any appropriate administration method known to those skilled in the art as needed. When the pharmaceutical composition is used, a safe and effective amount of the pharmaceutical composition of the present invention is administered to a human.

[0038] In some embodiments, the suitable dosage of the pharmaceutical composition described in the present invention can be prescribed in a variety of ways according to factors such as formulation method, administration method, patient's age, weight, gender, morbidity, diet, administration time, administration route, excretion rate and reaction sensitivity. A skilled physician can usually easily determine the prescription and the desired dosage that is effective for treatment.

[0039] Furthermore, the pharmaceutical composition of the present invention may also include additives such as stabilizers, bactericides, buffers, isotonic agents, chelating agents, pH control agents and surfactants.

[0040] Further, the pharmaceutical composition described in the present invention can also be used in combination with other drugs for treating esophageal cancer, and other therapeutic compounds can be administered simultaneously with the main active ingredient, or even administered simultaneously in the same composition. Other therapeutic compounds can also be administered separately in a separate composition or in a dosage form different from the main active ingredient. Partial doses of the main ingredient can be administered simultaneously with other therapeutic compounds, while other doses can be administered separately. During the treatment process, the dosage of the pharmaceutical composition of the present invention can be adjusted according to the severity of the symptoms, the frequency of recurrence, and the physiological response of the treatment regimen.

[0041] Furthermore, the pharmaceutical composition is a single compound preparation or a combination of two separate single preparations.

[0042] Furthermore, the compound preparation is a compound preparation comprising a topoisomerase I inhibitor and an ERK1 / 2 inhibitor.

[0043] Furthermore, the combination of single-ingredient preparations is a combination of a single-ingredient preparation comprising a topoisomerase I inhibitor and a single-ingredient preparation comprising an ERK1 / 2 inhibitor.

[0044] Furthermore, the two single-ingredient preparations in the combination of single-ingredient preparations are administered simultaneously or sequentially.

[0045] Furthermore, the topoisomerase I inhibitor is SN-38, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof.

[0046] Furthermore, the ERK1 / 2 inhibitor is SCH772984, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof.

[0047] In some embodiments, the compound preparation refers to a preparation made of two or more active pharmaceutical ingredients. For example, when the pharmaceutical composition of the present invention is a compound preparation, it may mean that it contains both the topoisomerase I inhibitor SN-38 and the ERK1 / 2 inhibitor SCH772984.

[0048] In some embodiments, the single-ingredient preparation refers to a preparation made with a single active pharmaceutical ingredient. For example, when the pharmaceutical composition of the present invention is a combination of single-ingredient preparations, it can represent a combination of a single-ingredient preparation containing the topoisomerase I inhibitor SN-38 and a single-ingredient preparation containing the ERK1 / 2 inhibitor SCH772984.

[0049] In some embodiments, the SN-38 and SCH772984 in the pharmaceutical composition can be administered simultaneously, separately, or sequentially. Wherein, simultaneously means that the two drugs are administered synchronously. If not administered simultaneously, they are administered sequentially within a time frame so that the two can act therapeutically in the same time frame. Therefore, sequential administration allows the administration of another drug within 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, or several hours after providing one drug, provided that the circulating half-life of the first administered drug allows the two to be present in therapeutically effective amounts at the same time. The time delay between the administration of each component will vary depending on the exact nature of the components, the interactions between them, and their respective half-lives. Unlike simultaneous or sequential, separately means that the interval between the administration of one drug and the other is significant, that is, when the second drug is administered, the first administered drug may no longer be present in the bloodstream in a therapeutically effective amount.

[0050] Furthermore, the dosage form of the pharmaceutical composition includes a parenteral dosage form and / or a parenteral dosage form.

[0051] Furthermore, the non-intestinal administration dosage form includes an injection dosage form, a cavity administration dosage form, a mucosal administration dosage form and / or a skin administration dosage form.

[0052] Furthermore, the dosage forms for administration via the gastrointestinal tract include tablets, granules, capsules, solutions, powders, sustained-release preparations, emulsions, suspensions, syrups and / or drops.

[0053] In some embodiments, the dosage form of the pharmaceutical composition of the present invention is a dosage form that is prepared by conventional methods and is conducive to administration, including but not limited to: parenteral dosage forms, gastrointestinal dosage forms, and specific examples include but are not limited to: aqueous solution injection, powder injection, pills, powders, tablets, patches, suppositories, emulsions, creams, gels, granules, capsules, aerosols, sprays, powder sprays, sustained-release agents and controlled-release agents, etc.

[0054] In some embodiments, the injection dosage forms include, but are not limited to, intravenous injections, intramuscular injections, subcutaneous injections, intradermal injections, intracavity injections and other injections.

[0055] In some embodiments, the cavity administration dosage form includes, but is not limited to, suppositories, aerosols, effervescent tablets, drops, pills, etc., and is used in the rectum, vagina, urethra, nasal cavity, ear canal, etc.

[0056] In some embodiments, the mucosal administration dosage form includes, but is not limited to, eye drops, nasal drops, eye ointments, gargles, sublingual tablets, adhesive tablets, patches, and the like.

[0057] In some embodiments, the skin administration dosage form includes, but is not limited to, topical solutions, lotions, liniments, ointments, plasters, pastes, patches, and the like.

[0058] In some embodiments, the pharmaceutical composition contains pharmaceutical excipients, which can be conventionally used in various preparations, including but not limited to isotonic agents, buffers, flavoring agents, excipients, fillers, adhesives, disintegrants and lubricants, etc.; or selected for use in order to adapt to the substance, including but not limited to emulsifiers, solubilizers, antibacterial agents, analgesics and antioxidants, etc. Such excipients can effectively improve the stability and solubility of the active ingredients contained in the composition or change the release rate and absorption rate of the active ingredients, thereby improving the metabolism of various active ingredients in the body, thereby enhancing the administration effect of the composition. In addition, excipients used to achieve specific administration purposes or methods, such as sustained-release administration, controlled-release administration and pulse administration, etc., include but are not limited to gelatin, albumin, chitosan, polyether and polyester polymer materials (for example: polyethylene glycol, polyurethane, polycarbonate and its copolymers, etc.). The main manifestations of the drug administration are: improving the therapeutic effect, improving the bioavailability, reducing toxic side effects and improving patient compliance, etc.

[0059] In some embodiments, the pharmaceutical composition or pharmaceutical preparation may also contain other drugs or agents that can be used for treatment and / or prevention, or auxiliary treatment and / or prevention. The other drugs or agents are not particularly limited. As long as they can produce the expected treatment and / or prevention, or auxiliary treatment and / or prevention effect on esophageal cancer, they are within the scope of protection of the present invention.

[0060] In some embodiments, the other drugs or agents that can be used for treatment and / or prevention, or adjuvant treatment and / or prevention include but are not limited to: chemotherapy drugs, specifically, the chemotherapy drugs include but are not limited to: taxane drugs (paclitaxel, docetaxel, cabazitaxel), antibiotic drugs (doxorubicin, epirubicin, daunorubicin, pirarubicin, etoposide, irinotecan, mitoxantrone), antimetabolites (cisplatin, carboplatin, lobaplatin, nedaplatin, oxaliplatin, gemcitabine, 5-fluorouracil), alkylating agents (cyclophosphamide, ifosfamide).

[0061] The second aspect of the present invention provides an in vitro method for inhibiting the growth of esophageal cancer cells or esophageal cancer organoids and / or promoting apoptosis of esophageal cancer cells or esophageal cancer organoids for non-therapeutic purposes.

[0062] Furthermore, the method comprises: treating esophageal cancer cells or esophageal cancer organoids with the pharmaceutical composition described in the first aspect of the present invention.

[0063] In a specific embodiment of the present invention, the present invention has experimentally demonstrated that the pharmaceutical composition has a significant synergistic inhibitory effect on the growth and proliferation of esophageal cancer cell lines. Therefore, the pharmaceutical composition can be used as an inhibitor for inhibiting the growth and proliferation of esophageal cancer cells or esophageal cancer organoids for non-therapeutic purposes, and can be used in the field of scientific research, such as more in-depth research on the growth and metabolic mechanism or behavior of esophageal cancer cell lines or esophageal cancer organoids, screening for potential drugs that can be used to treat esophageal cancer, etc.

[0064] The present invention also provides a method for treating and / or preventing esophageal cancer, the method comprising: administering a therapeutically and / or preventively effective amount of the pharmaceutical composition described in the first aspect of the present invention to a subject in need thereof.

[0065] In the present invention, the subject includes mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates such as chimpanzees and other apes and monkeys; farm animals such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs and cats; laboratory animals, including rodents such as rats, mice and guinea pigs, etc. Examples of non-mammals include, but are not limited to, birds, fish, etc. In one embodiment of the present invention, the subject is a human. The term subject includes confirmed patients.

[0066] In the present invention, the effective amount refers to the amount of the compound that effectively produces the desired preventive, alleviating or therapeutic effect. Depending on the compound, the symptom and its severity, the age of the treated mammal and other factors, the amount of the pharmaceutical composition of the present invention to achieve the effective amount will also vary accordingly, but the specific amount can be routinely determined by a person of ordinary skill in the art based on the knowledge in the art mastered by him / her and the content disclosed in the present invention, and the dosage that can produce the above-mentioned effects is within the scope of protection of the present invention.

[0067] In some embodiments, the administration of the pharmaceutical composition can be carried out in any form, including but not limited to: oral, topical, parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may include a penetration enhancer), oral and / or suppository administration, and other administration routes. In one embodiment of the invention, the active compound or compound combination as described above is provided in a solid dosage form known in the art. The most effective dosage form will depend on the bioavailability or pharmacokinetics of the selected specific agent and the severity of the patient's disease. Oral dosage forms are particularly preferred because they are easy to use and expected favorable patient compliance.

[0068] In some embodiments, the pharmaceutical composition or pharmaceutical preparation may be administered to the subject by oral administration, injection or topical administration. For example, the method may include administering the pharmaceutical composition or pharmaceutical preparation to the subject three times a day, once a day, once every two days, etc. In some embodiments, injection administration may include subcutaneous injection, intramuscular injection, intravenous injection, etc. In some embodiments, injection administration may include injecting the pharmaceutical composition directly into the lesion or in an area near the lesion. In some embodiments, a combination of different administration methods may be used.

[0069] The third aspect of the present invention provides any of the following applications: (1) Use of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor in combination in the preparation of a pharmaceutical composition for treating and / or preventing esophageal cancer; (2) Use of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor in combination in the preparation of a pharmaceutical preparation for the treatment and / or prevention of esophageal cancer; (3) The use of a combination of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor to inhibit the growth of esophageal cancer cells or esophageal cancer organoids and / or promote the apoptosis of esophageal cancer cells or esophageal cancer organoids in vitro for non-therapeutic purposes; (4) Use of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor in combination for the preparation of a reagent for inhibiting the growth of esophageal cancer cells or esophageal cancer organoids and / or promoting the apoptosis of esophageal cancer cells or esophageal cancer organoids in vitro; (5) Use of topoisomerase I inhibitors in the preparation of drugs for improving the therapeutic effect of ERK1 / 2 inhibitors in the treatment of esophageal cancer; (6) Use of ERK1 / 2 inhibitors in the preparation of drugs for improving the therapeutic effect of topoisomerase I inhibitors in the treatment of esophageal cancer.

[0070] Furthermore, the topoisomerase I inhibitor is SN-38, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof.

[0071] Furthermore, the ERK1 / 2 inhibitor is SCH772984, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof.

[0072] The present invention has the beneficial effects: The present invention discovers for the first time that the combination of SN-38 and SCH772984 has a synergistic effect in the treatment of esophageal cancer. The combination with SCH772984 can significantly improve the therapeutic effect of esophageal cancer and achieve better anti-tumor efficacy. The present invention provides a new treatment plan for the technical field of esophageal cancer treatment, which has important scientific significance and clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0073] Figure 1 It is a dose-response curve of SN-38 on KYSE150 cell line of esophageal squamous cell carcinoma; Figure 2 This is a dose-response curve of SCH772984 on esophageal squamous cell carcinoma KYSE150 cell line; Figure 3 It is a heat map of drug combination response of SN-38 and SCH772984 in esophageal squamous cell carcinoma KYSE150 cell line; Figure 4 The HSA model was used to quantify and analyze the synergy score and three-dimensional surface map of esophageal squamous cell carcinoma KYSE150 cell viability; Figure 5 The Loewe model was used to quantify and analyze the synergy score and three-dimensional surface map of esophageal squamous cell carcinoma KYSE150 cell viability; Figure 6 The Bliss model was used to quantify and analyze the synergy score and three-dimensional surface map of KYSE150 cell viability in esophageal squamous cell carcinoma cells; Figure 7 The ZIP model was used to quantify and analyze the synergy score and three-dimensional surface map of esophageal squamous cell carcinoma KYSE150 cell viability. DETAILED DESCRIPTION

[0074] The present invention is further described below in conjunction with specific embodiments, which are only used to explain the present invention and cannot be understood as limiting the present invention. It can be understood by those skilled in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and purpose of the present invention, and the scope of the present invention is defined by the claims and their equivalents.

[0075] The drugs, reagents and raw materials used in the present invention are easily obtained by ordinary technicians in the field and can be obtained from commercial channels unless otherwise specified. The experimental methods in the present invention that do not specify specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer. In particular, the following examples are only used to illustrate the present invention and should not limit the scope of the present invention in any way.

[0076] Example 1 In vitro proliferation inhibition experiment of esophageal cancer cells 1. Experimental methods The IC50 of SN-38 and SCH772984 on KYSE150 cells was detected by CCK8 assay. In order to more fully verify the killing ability of drugs on cells, we set 10 concentrations for each drug, and the specific drug concentrations are as follows: the drug concentration of SN-38 was set to: 0.0000μM, 0.0005μM, 0.0014μM, 0.0041μM, 0.0124μM, 0.0370μM, 0.1111μM, 0.3333μM, 1μM, 3μM, 9μM. The drug concentration of SCH772984 was set to 0.0003μM, 0.0012μM, 0.0049μM, 0.0195μM, 0.0781μM, 0.3125μM, 1.25μM, 5μM, 20μM, 80μM.

[0077] KYSE150 cells were counted, 3000 cells were plated in a 96-well plate, and after overnight attachment, the culture medium containing different concentrations of SN-38 and SCH772984 was replaced for 72 hours. The old culture medium was aspirated, and 100μl of new culture medium containing 10% CCK8 was added to each well. After two hours of incubation, the reading was taken at a wavelength of 450nm. Cell viability = (OD value of the experimental group - OD value of the blank control) / (OD value of the control group - OD value of the blank control) × 100%. Among them: The experimental group (Treatment group) refers to the cell group to which the drug to be tested or treatment is added. The control group (Control group) refers to the cell group without any treatment, which is used to establish a baseline. The blank control (Blank control) refers to the well with neither cells nor additives to the experimental group, which is used to correct the background absorbance.

[0078] 2. Experimental results The dose-response curves of SN-38 and SCH772984 on KYSE150 cell line of esophageal squamous cell carcinoma are shown in Figure 2. Figure 1 , 2 As shown, the results showed that the IC50 of SN-38 for KYSE150 was 3.018 μM, and the IC50 of SCH772984 for KYSE150 was 2.917 μM.

[0079] From the experimental results of single drug concentrations, we can see that low concentrations of SN-38 and SCH772984 have no obvious effect on the growth of KYSE150 cells. Next, we used a 6 x 6 method to select an intermediate concentration for a combined experiment to explore whether SN-38 and SCH772984 have a synergistic effect on the proliferation of esophageal cancer cells.

[0080] Example 2 Effect of SN-38 and SCH772984 on the proliferation of esophageal cancer cells 1. Experimental methods KYSE150 cells were counted and 3,000 cells were plated into a 96-well plate. After overnight attachment, the culture medium containing different concentrations of SN-38 and SCH772984 was replaced for 72 hours. The old culture medium was aspirated and 100 μl of new culture medium containing 10% CCK8 was added to each well. After incubation for two hours, the readings were taken at a wavelength of 450 nm.

[0081] 2. Experimental results Drug concentration settings see Figure 3 Heat map, SN-38 drug concentrations are 0μM, 0.01μM, 0.04μM, 0.11μM, 0.33μM and 1μM; SCH772984 drug concentrations are 0μM, 0.02μM, 0.08μM, 0.31μM, 1.25μM and 5μM. The results show that the combined use of the two has a better killing ability on KYSE150 cells than any single drug.

[0082] To determine whether SN-38 and SCH772984 have a synergistic effect on the growth of KYSE150 cells, we used the Synergy Fider online platform for computational analysis. First, we calculated the HSA (Highest Single Agent) synergy index, which is a mathematical model used to evaluate the effects of drug combinations. In drug combination experiments, the HSA model evaluates synergy by comparing the actual effect of the drug combination with the expected effect of the highest single drug effect. The calculation formula for the HSA synergy index is E HSA =E AB −max (E A , E B );Among them: E HSA represents the HSA synergy index, E AB represents the actual inhibitory effect of the drug combination (drug A and drug B), E A represents the inhibitory effect of drug A alone, E B Represents the inhibitory effect of drug B alone.

[0083] If the calculated HSA value is greater than 0, the drug combination is considered to have a synergistic effect; if the HSA value is less than 0, it may indicate an antagonistic effect between the drugs. The HSA model assumes that the expected effect of the drug combination is equal to the maximum effect of each individual drug. In practical applications, the HSA synergy index can help researchers understand the interaction characteristics of different drug combinations and optimize drug combination strategies. Our experimental results show that the HSA synergy score of SN-38 and SCH772984 is 19.054, indicating that the two have a strong synergistic effect ( Figure 4 ).

[0084] The Loewe additivity model is a mathematical model used to analyze drug interactions based on the assumption that no compound interacts with itself and that two doses from different compounds with the same effect are equivalent. This model can be used to predict the expected effects of two or more drugs when used in combination, if there is no interaction between these drugs.

[0085] In the Loewe model, if dose a' of drug A produces the same effect as dose b' of drug B, then any dose combination (a, b) that satisfies a * f_A^-1(E) + b * f_B^-1(E) = 1 must produce the same effect as a' and b', where f_A^-1(E) and f_B^-1(E) are doses that express effect E through the inverse function of the drug dose-response curve.

[0086] This model can be used to calculate the synergy score (S_LOEWE) of the drug combination, with the formula S_LOEWE = E_A,B,...,N - E_LOEWE, where E_LOEWE should satisfy Σ(x_k * f_k^-1(E)) = 1, where x_k represents the dose of the drug and f_k^-1(E) is the inverse function of the drug dose-response curve.

[0087] In practical applications, the Loewe model can help researchers evaluate the potential synergy of drug combinations, especially in drug development and clinical applications. Our experimental results show that the Loewe synergy score of SN-38 and SCH772984 is 10.632, indicating that the two have a strong synergistic effect ( Figure 5 ).

[0088] The Bliss synergy model is a mathematical model used to evaluate drug interactions. It is based on the assumption that two drugs act independently on the biological system without interaction. The synergy score (S_Bliss) of this model is calculated by the following formula: SBliss = E AB −(E A+E B −E A ×E B ). Among them, E AB is the actual effect of the combined action of the two drugs, E A and E B If SBliss is greater than 0, it is considered that there is a synergistic effect between the drugs; if it is equal to 0, it means that there is no interaction between the drugs; if it is less than 0, there may be an antagonistic effect.

[0089] A key feature of the Bliss model is that it takes into account the probability of independent drug effects and calculates the expected joint effect by multiplying the probabilities. This approach is very useful in drug combination studies, especially in high-throughput screening (HTS) experiments, where the potential synergistic effects of a large number of drug combinations can be quickly evaluated. Our experimental results show that the Bliss synergy score of SN-38 and SCH772984 is 13.94, indicating that the two have a strong synergistic effect ( Figure 6 ).

[0090] The ZIP (Zero Interaction Potency) model is a reference model for evaluating drug combination interactions. It is based on the assumption that if there is no interaction between two drugs, then their potency on the dose-response curve will not change. The ZIP model quantifies the strength of drug interactions by comparing the dose-response curves of single drugs and drug combinations.

[0091] In practical applications, the ZIP model is often used to analyze the synergistic effects of drug combinations, especially in high-throughput drug screening. It combines the advantages of the Loewe additivity and Bliss independence models and aims to systematically evaluate various drug interaction patterns that may occur. The synergy score (S_ZIP) of the ZIP model is calculated by the following formula: S ZIP =E A,B,....N −(E A +E B −E A ×E B +...+(−1) N+1 ×E A ×E B ×...×E N ). Among them, E A,B,....N is the actual effect of the drug combination, E A , E B , ...E N The effects of single drugs are respectively.

[0092] The characteristic of the ZIP model is that it takes into account the interaction of drugs at different doses and can be used to analyze the combination of two or more drugs. In the SynergyFinder tool, the ZIP model is used as the default synergy scoring model, which provides an automated outlier detection procedure, an extended curve fitting function, a statistical evaluation of repeated measures, and different summary reports and tables, allowing researchers to more accurately evaluate the synergy of drug combinations. Our experimental results show that the ZIP synergy score of SN-38 and SCH772984 is 13.879, indicating that the two have a strong synergistic effect ( Figure 7 ).

[0093] In summary, we used four different computational models to demonstrate that SN-38 and SCH772984 have a strong synergistic effect on the esophageal squamous cell carcinoma cell line KYSE150.

[0094] The description of the above embodiments is only used to understand the method and core idea of ​​the present invention. It should be pointed out that, for those skilled in the art, several improvements and modifications can be made to the present invention without departing from the principle of the present invention, and these improvements and modifications will also fall within the scope of protection of the claims of the present invention.

Claims

1. A pharmaceutical composition for treating and / or preventing esophageal cancer, characterized in that: The pharmaceutical composition comprises a topoisomerase I inhibitor and an ERK1 / 2 inhibitor.

2. The pharmaceutical composition according to claim 1, characterized in that The topoisomerase I inhibitor is SN-38, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof; The ERK1 / 2 inhibitor is SCH772984, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof.

3. The pharmaceutical composition according to claim 2, characterized in that The concentration ratio of SN-38 to SCH772984 is 1:500 to 1:0.02; Preferably, the concentration ratio of SN-38 to SCH772984 is 1:125 to 1:0.

08.

4. The pharmaceutical composition according to claim 3, characterized in that The concentration ratio of SN-38 and SCH772984 is (0.01 μM~1 μM): (0.02 μM~5 μM); Preferably, the concentration ratio of SN-38 and SCH772984 is (0.04 μM~1 μM): (0.08 μM~5 μM).

5. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

6. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition is a single compound preparation or a combination of two separate single preparations; The compound preparation is a compound preparation comprising a topoisomerase I inhibitor and an ERK1 / 2 inhibitor; The combination of single-ingredient preparations is a combination of a single-ingredient preparation comprising a topoisomerase I inhibitor and a single-ingredient preparation comprising an ERK1 / 2 inhibitor; The two single-ingredient preparations in the combination of single-ingredient preparations are administered simultaneously or sequentially; The topoisomerase I inhibitor is SN-38, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof; The ERK1 / 2 inhibitor is SCH772984, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof.

7. The pharmaceutical composition according to claim 1, characterized in that The dosage form of the pharmaceutical composition includes a non-gastrointestinal dosage form and / or a gastrointestinal dosage form; The non-gastrointestinal administration dosage forms include injection dosage forms, cavity administration dosage forms, mucosal administration dosage forms and / or skin administration dosage forms; The dosage forms for administration via the gastrointestinal tract include tablets, granules, capsules, solutions, powders, sustained-release preparations, emulsions, suspensions, syrups and / or drops.

8. A method for inhibiting the growth of esophageal cancer cells or esophageal cancer organoids and / or promoting apoptosis of esophageal cancer cells or esophageal cancer organoids in vitro for non-therapeutic purposes, characterized in that: The method comprises: treating esophageal cancer cells or esophageal cancer organoids with the pharmaceutical composition described in any one of claims 1 to 7.

9. Any one of the following applications, characterized in that: The applications include: (1) Use of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor in combination in the preparation of a pharmaceutical composition for treating and / or preventing esophageal cancer; (2) Use of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor in combination in the preparation of a pharmaceutical preparation for the treatment and / or prevention of esophageal cancer; (3) The use of a combination of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor to inhibit the growth of esophageal cancer cells or esophageal cancer organoids and / or promote the apoptosis of esophageal cancer cells or esophageal cancer organoids in vitro for non-therapeutic purposes; (4) Use of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor in combination for the preparation of a reagent for inhibiting the growth of esophageal cancer cells or esophageal cancer organoids and / or promoting the apoptosis of esophageal cancer cells or esophageal cancer organoids in vitro; (5) Use of topoisomerase I inhibitors in the preparation of drugs for improving the therapeutic effect of ERK1 / 2 inhibitors in the treatment of esophageal cancer; (6) Use of ERK1 / 2 inhibitors in the preparation of drugs for improving the therapeutic effect of topoisomerase I inhibitors in the treatment of esophageal cancer.

10. The use according to claim 9, characterized in that: The topoisomerase I inhibitor is SN-38, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof; The ERK1 / 2 inhibitor is SCH772984, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof.

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