A pharmaceutical composition for treating esophageal cancer and its application

Abstract

The present invention discloses a pharmaceutical composition for treating esophageal cancer and its application. The pharmaceutical composition contains a topoisomerase I inhibitor and an ERK1 / 2 inhibitor. The present invention discovers for the first time that the combination of the topoisomerase I inhibitor SN-38 and the ERK1 / 2 inhibitor SCH772984 has a synergistic effect on the treatment of esophageal cancer. By combining SN-38 and SCH772984, the treatment effect on esophageal cancer can be significantly improved. The present invention provides a brand-new treatment plan for the technical field of esophageal cancer treatment, which has important scientific significance and clinical application value.

Description

Technical Field

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

[0002] The prognosis of esophageal cancer is poor, which seriously endangers 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 has shown a downward trend in many Western countries. On the contrary, the incidence of esophageal adenocarcinoma has increased rapidly, becoming one of the malignant tumors with the fastest growth rate.

[0003] Currently, esophageal cancer is usually treated by surgery, radiotherapy, chemotherapy, drug therapy, etc. However, the effect of simple surgical treatment is not good. For patients with early esophageal cancer, the prognosis is relatively 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 surgical treatment effect and reduce the recurrence rate. However, the current chemotherapy regimens used clinically can only relieve symptoms and prolong the survival period, and the chemotherapy regimens have strong toxic and side effects. Therefore, there is still a lack of effective drugs for treating esophageal cancer in the current field.

[0004] It can be seen that the chemotherapy regimens for treating esophageal cancer have strong toxic and side effects and the current situation that there is still a lack of more effective drugs for treating esophageal cancer in this field greatly affects the curative effect of treating esophageal cancer clinically at present. Therefore, it is of special importance and urgency to find new and more effective treatment means for esophageal cancer. Summary of the Invention

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

[0006] In order to achieve the above purpose, the present invention adopts the following technical solutions:

[0007] The first aspect of the present invention provides a pharmaceutical composition for treating and / or preventing esophageal cancer.

[0008] Furthermore, the pharmaceutical composition comprises a topoisomerase I inhibitor and an ERK1 / 2 inhibitor.

[0009] Furthermore, the topoisomerase I inhibitor is SN-38, or its pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form.

[0010] Furthermore, the ERK1 / 2 inhibitor is SCH772984, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate or crystalline form thereof.

[0011] Furthermore, the concentration ratio of SN-38 to SCH772984 is 1:500 to 1:0.02.

[0012] Preferably, the concentration ratio of SN-38 to SCH772984 is 1:125 to 1:0.08.

[0013] Furthermore, the concentration ratio of SN-38 to SCH772984 is (0.01 μM to 1 μM):(0.02 μM to 5 μM).

[0014] Preferably, the concentration ratio of SN-38 to SCH772984 is (0.04 μM to 1 μM):(0.08 μM to 5 μM).

[0015] In a specific embodiment of the present invention, the inventors of the present invention first discovered that the combination of the topoisomerase I inhibitor SN-38 and the ERK1 / 2 inhibitor SCH772984 has a synergistic effect on 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.

[0016] In the present invention, the topoisomerase I inhibitor is a class of compounds that can specifically inhibit the activity of DNA topoisomerase I. It can bind to topoisomerase I to form a stable DNA-topoisomerase I-inhibitor ternary complex, thereby preventing the normal repair and relaxation process of topoisomerase I on DNA, resulting in DNA strand breaks and cell cycle arrest, and ultimately inducing apoptosis of tumor cells.

[0017] In some embodiments, the topoisomerase I inhibitor comprises any reagent capable of inhibiting the activity of DNA topoisomerase I. Exemplarily, the topoisomerase I inhibitor includes but is not limited to: SN-38, camptothecin, hydroxycamptothecin, irinotecan, topotecan, Genz644282. In a specific embodiment of the present invention, the topoisomerase I inhibitor is SN-38.

[0018] In the present invention, the ERK1 / 2 inhibitor is a class of inhibitors targeting extracellular signal-regulated kinases (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.

[0019] 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.

[0020] In some embodiments, the pharmaceutically acceptable salt refers to acid addition salts formed with inorganic and / or organic acids and base salts formed with inorganic and / or organic bases. Additionally, when the compound contains a basic moiety (e.g., but not limited to, pyridine or imidazole) and an acidic moiety (e.g., but not limited to, carboxylic acid), zwitterions can be formed and the zwitterions are included in the pharmaceutically acceptable salts described in this application. Preferably, they are pharmaceutical (i.e., non-toxic, physiologically acceptable) salts, but other salts are also useful. The pharmaceutically acceptable salts of the compound can be formed, for example, by reacting the compound with an appropriate amount of acid or base in a medium, such as a medium in which the salt precipitates or an aqueous medium (lyophilized after the reaction).

[0021] Specific pharmaceutically acceptable salts include those salts that are within the scope of sound medical judgment, suitable for contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. The pharmaceutically acceptable salts of the compounds described in the present invention include salts derived from suitable inorganic and organic acids and inorganic and organic bases.

[0022] Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or salts formed with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid. Also included are salts formed using conventional methods in the art, such as ion exchange methods. Other pharmaceutically acceptable salts include: adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, gluconate, glycerophosphate, hemisulfate, heptanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate. Pharmaceutically acceptable salts derived from suitable bases include alkali metal, alkaline earth metal, and ammonium salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium salts, etc.

[0023] In some embodiments, the hydrate refers to a compound combined with water. Those skilled in the art will understand that an organic compound can form a complex with a solvent, which reacts in the solvent or precipitates or crystallizes out from the solvent, and these complexes are called solvates. When the solvent is water, the complex is called a hydrate.

[0024] In some embodiments, the solvate refers to a compound combined with a solvent or its salt form, which is usually formed by a solvent decomposition reaction. This physical association may include hydrogen bonding. Conventional solvents include water, methanol, ethanol, acetic acid, DMSO, THF, diethyl ether, etc. In some cases, the solvate will be able to be separated, for example, when one or more solvent molecules are incorporated into the lattice of the crystalline solid. Solvates include solvates in solution state and separable solvates.

[0025] In some embodiments, the crystalline form refers to the crystalline form of a compound with a specific crystal packing arrangement. Different crystalline forms usually have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardnesses, crystal shapes, optoelectronic properties, stabilities, and solubilities. Recrystallization solvents, crystallization rates, storage temperatures, and other factors can lead to the dominance of one crystalline form. Various polymorphs of a compound can be prepared by crystallization under different conditions.

[0026] In some embodiments, the SN-38, or its pharmaceutically acceptable salts, hydrates, enantiomers, diastereomers, solvates, or crystalline forms; the SCH772984, or its pharmaceutically acceptable salts, hydrates, enantiomers, diastereomers, solvates, or crystalline forms, are all within the scope of the present invention. In a specific embodiment of the present invention, the pharmaceutical composition is a pharmaceutical composition comprising SN-38 and SCH772984.

[0027] 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 the potential metabolic causes of symptoms, inhibiting disorders or diseases, for example, preventing the development of disorders or diseases, alleviating disorders or diseases, regressing disorders or diseases, alleviating the conditions caused by diseases or disorders, or stopping the symptoms of diseases or disorders.

[0028] Furthermore, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

[0029] 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, disintegrants.

[0030] In some embodiments, the diluent includes, but is not limited to: lactose, sodium chloride, glucose, urea, starch, water, etc.

[0031] In some embodiments, the binder includes, but is not limited to: starch, pregelatinized starch, dextrin, maltodextrin, sucrose, gum arabic, gelatin, methylcellulose, carboxymethylcellulose, ethylcellulose, polyvinyl alcohol, polyethylene glycol, polyvinylpyrrolidone, alginic acid and its salts, xanthan gum, hydroxypropylcellulose, and hydroxypropylmethylcellulose, etc.

[0032] In some embodiments, the surfactant includes, but is not limited to: polyoxyethylene sorbitan fatty acid esters, sodium dodecyl sulfate, monoglyceryl stearate, cetyl alcohol, etc.

[0033] In some embodiments, the humectant includes, but is not limited to: glycerol, starch, etc.

[0034] In some embodiments, the adsorption carrier includes, but is not limited to: starch, lactose, bentonite, silica gel, kaolin, and saponite, etc.

[0035] In some embodiments, the lubricant includes, but is not limited to: zinc stearate, monoglyceryl stearate, polyethylene glycol, talc powder, calcium and magnesium stearate, polyethylene glycol, boric acid powder, hydrogenated vegetable oil, sodium stearyl fumarate, polyoxyethylene monostearate, sucrose monolaurate, sodium lauryl sulfate, magnesium lauryl sulfate, sodium dodecyl magnesium sulfate, etc.

[0036] In some embodiments, the filler includes, but is not limited to: mannitol (granular or powdered), xylitol, sorbitol, maltose, erythritol, microcrystalline cellulose, polydextrose, coupled sugar, glucose, lactose, sucrose, dextrin, starch, sodium alginate, laminarin powder, agar powder, calcium carbonate, and sodium bicarbonate, etc.

[0037] In some embodiments, the disintegrant includes, but is not limited to: cross-linked vinylpyrrolidone, sodium carboxymethyl starch, low-substituted hydroxypropylmethyl, cross-linked sodium carboxymethyl cellulose, soy polysaccharide, etc.

[0038] In some embodiments, the pharmaceutically acceptable carrier and / or excipient are used as needed to assist in the stability of the formulation or to help improve its activity or its bioavailability or to produce an acceptable taste or odor in the case of oral administration. The pharmaceutical composition thus formulated can be administered with the drug by any suitable administration method known to those skilled in the art as needed. When using the pharmaceutical composition, a safe and effective amount of the pharmaceutical composition of the present invention is administered to a human.

[0039] In some embodiments, the suitable dosage of the pharmaceutical composition of the present invention can be prescribed in various ways according to factors such as the formulation method, administration method, age, weight, sex, morbidity, diet, administration time, administration route, excretion rate, and sensitivity of the patient. Skilled doctors can usually easily determine the prescription and the desired therapeutically effective dosage.

[0040] Furthermore, the pharmaceutical composition of the present invention may further include additives such as stabilizers, bactericides, buffers, isotonic agents, chelating agents, pH control agents, and surfactants.

[0041] Furthermore, the pharmaceutical composition described in the present invention can also be used in combination with other drugs for treating esophageal cancer. Other therapeutic compounds can be administered simultaneously with the main active ingredient, or even in the same composition. Other therapeutic compounds can also be administered separately in a separate composition or in a different dosage form 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 to the treatment regimen.

[0042] Furthermore, the pharmaceutical composition is a single compound preparation or a combination of two separate single-agent preparations.

[0043] Furthermore, the compound preparation is a compound preparation containing a topoisomerase I inhibitor and an ERK1 / 2 inhibitor.

[0044] Furthermore, the combination of single-agent preparations is a combination of a single-agent preparation containing a topoisomerase I inhibitor and a single-agent preparation containing an ERK1 / 2 inhibitor.

[0045] Furthermore, the administration methods of the two single-agent preparations in the combination of single-agent preparations are simultaneous administration or sequential administration.

[0046] Furthermore, the topoisomerase I inhibitor is SN-38, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate, or crystalline form thereof.

[0047] Furthermore, the ERK1 / 2 inhibitor is SCH772984, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereomer, solvate, or crystalline form thereof.

[0048] In some embodiments, the compound preparation refers to a preparation made of two or more active pharmaceutical ingredients. For example, when the pharmaceutical composition in the present invention is a compound preparation, it can represent that it simultaneously contains the topoisomerase I inhibitor SN-38 and the ERK1 / 2 inhibitor SCH772984.

[0049] In some embodiments, the single-agent formulation refers to a formulation made of a single active pharmaceutical ingredient. For example, when the pharmaceutical composition in the present invention is a combination of single-agent formulations, it can represent a combination of single-agent formulations respectively containing the topoisomerase I inhibitor SN-38 and a single-agent formulation containing the ERK1 / 2 inhibitor SCH772984.

[0050] In some embodiments, SN-38 and SCH772984 in the pharmaceutical composition can be administered simultaneously, separately, or sequentially. Among them, simultaneous administration means that the two drugs are administered synchronously. If not administered simultaneously, they are administered sequentially within a time range such that both can be therapeutically effective within the same time range. Therefore, sequential administration can allow the administration of the other drug 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, or several hours after the administration of one drug, provided that the circulating half-life of the first-administered drug allows for a therapeutically effective amount of both to be present simultaneously. The time delay between the administrations of the components will vary depending on the exact nature of the components, the interactions between them, and their respective half-lives. Different from simultaneous or sequential administration, separate administration 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.

[0051] Furthermore, the dosage forms of the pharmaceutical composition include parenteral dosage forms and / or enteral dosage forms.

[0052] Furthermore, the parenteral dosage forms include injection dosage forms, cavity dosage forms, mucosal dosage forms, and / or cutaneous dosage forms.

[0053] Furthermore, the enteral dosage forms include tablets, granules, capsules, solutions, powders, sustained-release agents, emulsions, suspensions, syrups, and / or drops.

[0054] In some embodiments, the dosage forms of the pharmaceutical composition of the present invention are dosage forms that are prepared by conventional methods and are conducive to administration, including but not limited to: parenteral dosage forms, enteral dosage forms. Specific examples include but are not limited to: aqueous injection solutions, powder for injection, pills, powders, tablets, patches, suppositories, emulsions, creams, gels, granules, capsules, aerosols, sprays, powder inhalations, sustained-release agents, and controlled-release agents, etc.

[0055] In some embodiments, the injection dosage forms include but are not limited to: intravenous injections, intramuscular injections, subcutaneous injections, intradermal injections, and intra-cavity injections, and other various injections.

[0056] In some embodiments, the intravaginal drug dosage forms include, but are not limited to: suppositories, aerosols, effervescent tablets, drops, dripping pills, etc., for use in the rectum, vagina, urethra, nasal cavity, ear canal, etc.

[0057] In some embodiments, the mucosal drug dosage forms include, but are not limited to: eye drops, nasal drops, ophthalmic ointments, gargles, sublingual tablets, adhesive tablets, film dressings, etc.

[0058] In some embodiments, the topical drug dosage forms include, but are not limited to: topical solutions, lotions, liniments, ointments, plasters, pastes, patches, etc.

[0059] In some embodiments, the pharmaceutical composition contains pharmaceutical excipients. The pharmaceutical excipients can be those conventionally used in various preparations, including but not limited to: isotonic agents, buffers, flavoring agents, excipients, fillers, binders, disintegrants, and lubricants, etc.; or those selected to be compatible with the substance, including but not limited to: emulsifying agents, solubilizing agents, bacteriostatic 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, etc., thereby improving the metabolism of various active ingredients in the body and further enhancing the administration effect of the composition. In addition, excipients can also be used to achieve specific administration purposes or methods, such as: sustained-release administration, controlled-release administration, and pulsed administration, etc., including but not limited to: gelatin, albumin, chitosan, polyethers, and polyester polymers (for example: polyethylene glycol, polyurethane, polycarbonate, and their copolymers, etc.). The main manifestations beneficial to administration include: improving the therapeutic effect, increasing the bioavailability, reducing the toxicity and side effects, and improving the patient compliance, etc.

[0060] In some embodiments, the pharmaceutical composition or pharmaceutical preparation may further contain other drugs or reagents that can be used for treatment and / or prevention, or adjuvant treatment and / or prevention. There is no particular limitation on the other drugs or reagents, as long as they can produce the expected treatment and / or prevention, or adjuvant treatment and / or prevention effects on esophageal cancer, they are all within the protection scope of the present invention.

[0061] In some embodiments, the other drugs or reagents that can be used for treatment and / or prevention, or adjuvant treatment and / or prevention include but are not limited to: chemotherapeutic drugs. Specifically, the chemotherapeutic drugs include but are not limited to: taxane drugs (paclitaxel, docetaxel, cabazitaxel), antibiotic drugs (doxorubicin, epirubicin, daunorubicin, pirarubicin, etoposide, irinotecan, mitoxantrone), antimetabolic drugs (cisplatin, carboplatin, lobaplatin, nedaplatin, oxaliplatin, gemcitabine, 5-fluorouracil), alkylating agents (cyclophosphamide, ifosfamide).

[0062] The second aspect of the present invention provides a method for non-therapeutically 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.

[0063] Further, the method includes: treating esophageal cancer cells or esophageal cancer organoids with the pharmaceutical composition described in the first aspect of the present invention.

[0064] In a specific embodiment of the present invention, the present invention has experimentally demonstrated that the pharmaceutical composition has a significant synergistic effect in inhibiting the growth and proliferation of esophageal cancer cell lines. Therefore, the pharmaceutical composition can be used as an inhibitor for non-therapeutically inhibiting the growth and proliferation of esophageal cancer cells or esophageal cancer organoids and can be used in the scientific research field, such as more in-depth research on the growth, metabolic mechanisms or behaviors of esophageal cancer cell lines or esophageal cancer organoids, screening for potential drugs that can be used to treat esophageal cancer, and the like.

[0065] The present invention also provides a method for treating and / or preventing esophageal cancer, the method including: administering to a subject in need a therapeutically and / or prophylactically effective amount of the pharmaceutical composition described in the first aspect of the present invention.

[0066] 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 cows, 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 diagnosed patients.

[0067] In the present invention, the effective amount refers to the amount of the compound that effectively produces the desired prophylactic, alleviating, or therapeutic effect. Depending on factors such as the compound, the symptoms and their severity, and the age of the mammal being treated, the amount of the pharmaceutical composition of the present invention that reaches the effective amount will also vary, but the specific dosage can be routinely determined by those of ordinary skill in the art based on the knowledge in this field combined with the content disclosed in the present invention. Doses that can produce the above effects are all within the protection scope of the present invention.

[0068] 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 penetration enhancers), buccal, and / or suppository administration, and other administration routes. In one embodiment of the present invention, the active compound or compound combination as described above is provided in solid dosage forms well known in the art. The most effective dosage form will depend on the bioavailability or pharmacokinetics of the specific agent selected and the severity of the patient's disease. Oral dosage forms are particularly preferred because of their ease of administration and expected favorable patient compliance.

[0069] In some embodiments, the pharmaceutical composition or pharmaceutical preparation can be administered to a subject by oral administration, injection administration, or topical administration. For example, the method can 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 can include subcutaneous injection, intramuscular injection, intravenous injection, etc. In some embodiments, injection administration can include directly injecting the pharmaceutical composition into the lesion or an area near the lesion. In some embodiments, a combination of different administration methods can be used.

[0070] The third aspect of the present invention provides an application in any of the following aspects:

[0071] (1) The application of the combination of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor in the preparation of a pharmaceutical composition for the treatment and / or prevention of esophageal cancer;

[0072] (2) The application of the combination of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor in the preparation of a pharmaceutical preparation for the treatment and / or prevention of esophageal cancer;

[0073] (3) The application of the combination of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor in the in vitro non-therapeutic inhibition of the growth of esophageal cancer cells or esophageal cancer organoids and / or the promotion of the apoptosis of esophageal cancer cells or esophageal cancer organoids;

[0074] (4) The application of the combination of a topoisomerase I inhibitor and an ERK1 / 2 inhibitor in the preparation of a reagent for in vitro inhibition of the growth of esophageal cancer cells or esophageal cancer organoids and / or the promotion of the apoptosis of esophageal cancer cells or esophageal cancer organoids;

[0075] (5) The application of a topoisomerase I inhibitor in the preparation of a drug for enhancing the therapeutic effect of an ERK1 / 2 inhibitor in the treatment of esophageal cancer;

[0076] (6) The application of an ERK1 / 2 inhibitor in the preparation of a drug for enhancing the therapeutic effect of a topoisomerase I inhibitor in the treatment of esophageal cancer.

[0077] Further, the topoisomerase I inhibitor is SN-38, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereoisomer, solvate or crystalline form thereof.

[0078] Further, the ERK1 / 2 inhibitor is SCH772984, or a pharmaceutically acceptable salt, hydrate, enantiomer, diastereoisomer, solvate or crystalline form thereof.

[0079] Advantages of the present invention:

[0080] The present invention for the first time discovers that the combination of SN-38 and SCH772984 has a synergistic effect on the treatment of esophageal cancer. By combining with SCH772984, the treatment effect on esophageal cancer can be significantly improved, and better anti-tumor efficacy can be achieved. 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. Description of the Drawings

[0081] Figure 1 It is a dose-response curve graph of SN-38 acting on esophageal squamous cell carcinoma KYSE150 cell line;

[0082] Figure 2 It is a dose-response curve graph of SCH772984 acting on esophageal squamous cell carcinoma KYSE150 cell line;

[0083] Figure 3 It is a drug combination response heat map of SN-38 and SCH772984 acting jointly on esophageal squamous cell carcinoma KYSE150 cell line;

[0084] Figure 4 It is a synergistic effect score and three-dimensional surface graph of quantifying and analyzing the viability of esophageal squamous cell carcinoma KYSE150 cells using the HSA model;

[0085] Figure 5 It is a synergistic effect score and three-dimensional surface graph of quantifying and analyzing the viability of esophageal squamous cell carcinoma KYSE150 cells using the Loewe model;

[0086] Figure 6 It is a synergistic effect score and three-dimensional surface graph of quantifying and analyzing the viability of esophageal squamous cell carcinoma KYSE150 cells using the Bliss model;

[0087] Figure 7 It is a synergistic effect score and three-dimensional surface graph of quantifying and analyzing the viability of esophageal squamous cell carcinoma KYSE150 cells using the ZIP model. Detailed Embodiments

[0088] The present invention will be further described below in conjunction with specific embodiments. The specific embodiments are only used to explain the present invention and should not be construed as a limitation of the present invention. Those of ordinary skill in the art can understand that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and purposes of the present invention. The scope of the present invention is defined by the claims and their equivalents.

[0089] The drugs, reagents, and raw materials used in the present invention are easily obtained by those of ordinary skill in the art. Without special instructions, they can all be obtained from commercial channels. The experimental methods without specific conditions noted in the present invention are usually carried out under conventional conditions or according to the conditions recommended by the manufacturers. In particular, the following embodiments are only used to illustrate the present invention and should not limit the scope of the present invention in any way.

[0090] Example 1 In vitro proliferation inhibition experiment of esophageal cancer cells

[0091] 1. Experimental method

[0092] The IC50 values of SN-38 and SCH772984 on KYSE150 cells were detected by CCK8 experiment. In order to more fully verify the killing ability of the drugs on cells, 10 concentrations were set for each drug. The specific drug concentrations are as follows: The drug concentrations of SN-38 were set as: 0.0000 μM, 0.0005 μM, 0.0014 μM, 0.0041 μM, 0.0124 μM, 0.0370 μM, 0.1111 μM, 0.3333 μM, 1 μM, 3 μM, 9 μM. The drug concentrations of SCH772984 were set as 0.0003 μM, 0.0012 μM, 0.0049 μM, 0.0195 μM, 0.0781 μM, 0.3125 μM, 1.25 μM, 5 μM, 20 μM, 80 μM.

[0093] Count KYSE150 cells, seed 3000 cells into a 96-well plate. After overnight attachment, replace the culture medium with different concentrations of SN-38 and SCH772984 and culture for 72 hours. Aspirate the old culture medium, add 100 μl of new culture medium containing 10% CCK8 to each well, and read the absorbance at a wavelength of 450 nm after incubation for two hours. 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%. Wherein: The experimental group (Treatment group) refers to the cell group to which the drug or treatment to be tested 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 without cells and without the additives in the experimental group, which is used to correct the background absorbance.

[0094] 2. Experimental Results

[0095] The dose-response curves of SN-38 and SCH772984 on esophageal squamous cell carcinoma KYSE150 cell line are respectively as Figure 1 、 2 shown. The results show that the IC50 of SN-38 for KYSE150 is 3.018 μM, and the IC50 of SCH772984 for KYSE150 is 2.917 μM.

[0096] From the experimental results of the single drug concentration, we can see that low concentrations of SN-38 and SCH772984 have no obvious effect on the growth of KYSE150 cells. Next, we selected intermediate concentrations in a 6x6 manner to conduct a combined experiment to explore whether SN-38 and SCH772984 have a synergistic effect on the proliferation of esophageal cancer cells.

[0097] Example 2 Proliferation effect of SN-38 and SCH772984 in combination on esophageal cancer cells

[0098] 1. Experimental method

[0099] Count KYSE150 cells, seed 3000 cells into a 96-well plate. After overnight attachment, replace the culture medium with different concentrations of SN-38 and SCH772984 and culture for 72 hours. Aspirate the old culture medium, add 100 μl of new culture medium containing 10% CCK8 to each well, incubate for two hours, and then take readings at a wavelength of 450 nm.

[0100] 2. Experimental results

[0101] The drug concentration settings are shown in Figure 3 the heat map. The drug concentrations of SN-38 are 0 μM, 0.01 μM, 0.04 μM, 0.11 μM, 0.33 μM and 1 μM respectively; the drug concentrations of SCH772984 are 0 μM, 0.02 μM, 0.08 μM, 0.31 μM, 1.25 μM and 5 μM respectively. The results show that the combined application of the two has a better killing ability on KYSE150 cells than any single drug.

[0102] To clarify whether SN-38 and SCH772984 have a synergistic effect on the growth of KYSE150 cells, we used the Synergy Fider online platform for calculation and analysis. First, we calculated the HSA (Highest SingleAgent) synergy index. HSA is a mathematical model used to evaluate the effect of drug combinations. In drug combination experiments, the HSA model evaluates the synergistic effect by comparing the actual effect of the drug combination with the expected effect of the highest single drug effect. The calculation formula of the HSA synergy index is EHSA = E AB − max(E A , E B ) ; where: 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.

[0103] If the calculated HSA value is greater than 0, it is considered that the drug combination has 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 of the individual drug effects. In practical applications, the HSA synergy index can help researchers understand the interaction characteristics of different drug combinations, and then optimize the drug combination strategy. Our experimental results show that the HSA synergy score of SN-38 and SCH772984 is 19.054, indicating a strong synergistic effect ( Figure 4 ).

[0104] The Loewe additivity model is a mathematical model for analyzing drug interactions. It is based on the assumptions that no compound interacts with itself and that two doses from different compounds with the same action are equivalent. This model can be used to predict the expected effect when two or more drugs are used in combination, if there is no interaction between these drugs.

[0105] In the Loewe model, if the dose a' of drug A produces the same effect as the 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 the doses representing the effect E through the inverse functions of the drug dose-response curves.

[0106] This model can be used to calculate the synergy score (S_LOEWE) of the drug combination, and the formula is 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.

[0107] In practical applications, the Loewe model can help researchers evaluate the potential synergistic effects of drug combinations, especially in drug development and clinical applications. Our experimental results show that the Loewe synergyscore of SN-38 and SCH772984 is 10.632, indicating a strong synergistic effect between the two ( Figure 5 ).

[0108] 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 a 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, and E A and E B are the effects of the two drugs acting alone, respectively. 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.

[0109] A key feature of the Bliss model is that it takes into account the probability of independent drug action and calculates the expected combined effect through the product of probabilities. This method is very useful in drug combination research, 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 a strong synergistic effect between the two ( Figure 6 ).

[0110] The ZIP (Zero Interaction Potency) model is a reference model used to evaluate the interaction of drug combinations. 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 intensity of drug interaction by comparing the dose-response curves of single drugs and drug combinations.

[0111] In practical applications, the ZIP model is usually 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 possible drug interaction patterns. The synergy score (S_ZIP) of the ZIP model is calculated by the following formula: S ZIP =EA,B,....N −(E A +E B −E A ×E B +...+(−1) N+1 ×E A ×E B ×...×E N ). Where E A,B,....N is the actual effect of the drug combination, and E A , E B ,... E N are the effects of the individual drugs respectively.

[0112] The ZIP model is characterized by taking into account the interactions of drugs at different doses and can be used to analyze combinations 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, extended curve fitting capabilities, statistical evaluation of repeated measurements, and different summary reports and tables, enabling 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 a strong synergistic effect between the two ( Figure 7 ).

[0113] In summary, we have shown using four different computational models that SN-38 and SCH772984 have a strong synergistic effect on the esophageal squamous cell carcinoma cell line KYSE150.

[0114] The description of the above embodiments is only for understanding the method of the present invention and its core idea. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications will also fall within the protection scope 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 SN-38 and SCH772984.

2. The pharmaceutical composition according to claim 1, characterized in that The concentration ratio of SN-38 and SCH772984 is (0.01 μM~1 μM): (0.02 μM~5 μM).

3. The pharmaceutical composition according to claim 2, characterized in that The concentration ratio of SN-38 and SCH772984 is (0.04 μM~1 μM): (0.08 μM~5 μM).

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

5. 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 SN-38 and SCH772984; The combination of single-ingredient preparations is a combination of a single-ingredient preparation containing SN-38 and a single-ingredient preparation containing SCH772984; The two single-ingredient preparations in the combination of the single-ingredient preparations are administered simultaneously or sequentially.

6. 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.

7. Use of SN-38 and SCH772984 in combination for preparing a pharmaceutical composition for treating and / or preventing esophageal cancer.

8. Use of SN-38 and SCH772984 in combination for preparing a pharmaceutical preparation for treating and / or preventing esophageal cancer.

9. Use of SN-38 in the preparation of a drug for improving the therapeutic effect of SCH772984 in treating esophageal cancer.

10. Use of SCH772984 in the preparation of a drug for improving the therapeutic effect of SN-38 in treating esophageal cancer.

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