WO2024007801A1 - Combination therapy of receptor tyrosine kinase inhibitor and biphenyl cyclooctadiene lignan and use thereof - Google Patents

Abstract

The present invention provides a combination therapy of a receptor tyrosine kinase inhibitor and biphenyl cyclooctadiene lignan and use thereof. The use comprises preparing a pharmaceutical composition. The pharmaceutical composition comprises a compound represented by formula (I) or a derivative thereof and biphenyl cyclooctadiene lignan. The pharmaceutical composition according to the embodiments of the present invention can effectively prolong the effect duration of the compound represented by formula (I) in vivo, significantly reduce the generation of a metabolite represented by formula (II), increase the blood-brain barrier permeability of the compound represented by formula (I), and improve the efficacy, thus providing more effective treatment and prevention against cancers.

Description

The combined use of receptor tyrosine kinase inhibitors and biphenylcyclooctadiene lignans and their uses Technical field

The present invention relates to the field of medicine. Specifically, the present invention relates to the combined use of receptor tyrosine kinase inhibitors and biphenylcyclooctadiene lignans and their uses. More specifically, the present invention relates to a pharmaceutical composition, Single dosage form, drug combination, kit, administration method of the compound represented by formula I or its salt, method of extending the half-life of the compound represented by formula I or its salt, reducing the compound represented by formula I or its salt to form the compound represented by formula II Methods.

Background technique

(R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate and its salts are not An inhibitor (EGFR-TKI) that is limited to oral administration and can inhibit the activity of epidermal growth factor receptor tyrosine kinase with activating mutations, initially developed for the treatment of non-small cell lung cancer (NSCLC) with central nervous system metastasis (CNS) ). (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate and its salts in The main CYP450 enzymes metabolized in the human body are CYP3A4 and CYP3A5 subtypes. Their drug efficacy increases with the increase in drug dosage. However, the occurrence of adverse reactions limits its further clinical application. The occurrence of adverse reactions may be related to the regular use. CYP450 enzymes, such as its subtype CYP3A metabolizing enzyme, are related to the generation of metabolites, and the generation of some metabolites increases at a higher rate than the administered dose. Existing research results show that (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylic The acid ester hydrochloride can penetrate the blood-brain barrier in the form of 100% concentration. The ability of the equally active metabolites generated during metabolism to penetrate the blood-brain barrier is significantly lower than that of the drug, resulting in reduced intracranial efficacy. Furthermore, different CYP450 enzyme inhibitors have been compared with (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine- Drug-drug interactions may occur when 1-carboxylic acid esters or their salts are combined. Therefore, research is still needed to explore a method that can improve (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxy. It is a safe and reliable method of medication.

Contents of the invention

This application is filed based on the inventor's discovery of the following issues and facts:

(R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate or its salt is An effective EGFR inhibitor that can penetrate the central nervous system, biphenylcyclooctadienol is a CYP450 enzyme inhibitor and can reduce serum alanine aminotransferase and play a role in protecting the liver. The inventor studied (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate or Its salt is used alone or in combination with various CYP450 enzyme inhibitors, such as schisandrin A, silibinin, ketoconazole, etc. It is found that schisandra Compared with other types of inhibitors, promethin has a significant inhibitory effect on human liver microsomal cytochrome CYP450 enzymes, and the degree of inhibition is moderate. Compared with (R)-4-(3-chloro-2-fluorophenyl) Amino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate or its salt is used alone. After combined use, the passage of biphenylcyclooctadiene lignans is inhibited. (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate by reducing CYP450 enzymes Or the metabolism of its salts, prolonging its residence time in the body to enhance the efficacy, and at the same time reducing the concentration or content of metabolites by inhibiting CYP450 cytochrome metabolic enzymes, thus improving both efficacy and safety.

In a first aspect of the invention, the invention provides a pharmaceutical composition. According to an embodiment of the present invention, the pharmaceutical composition includes: a compound represented by Formula I or a salt thereof and biphenylcyclooctadienolignan. The pharmaceutical composition according to the embodiment of the present invention can reduce the content of alanine aminotransferase in serum, play a protective role on the liver, and compared with the single use of the compound represented by formula I or its salt, the pharmaceutical composition can effectively prolong the The duration of drug efficacy of the compound represented by formula I or its salt in the body, and significantly reducing the concentration of the metabolite represented by formula II during the metabolic process, increasing the exposure of the compound represented by formula I or its salt in the body and the dose in brain tissue, Reducing the adverse reactions caused by the metabolites represented by formula II improves the efficacy and safety of the compounds represented by formula I or salts thereof, thereby effectively treating or preventing cancer.

According to embodiments of the present invention, the above-mentioned pharmaceutical composition may further include at least one of the following additional technical features:

According to an embodiment of the present invention, the salt of the compound represented by formula I includes at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate, benzoate, adipic acid Salts, succinates, methanesulfonates and maleates.

According to an embodiment of the present invention, the salt of the compound represented by Formula I is a hydrochloride.

According to embodiments of the present invention, the salt of the compound represented by Formula I is (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2, 4-Dimethylpiperazine-1-carboxylate hydrochloride.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignans include at least one of the following: schisandrin A, schisandrin ester A, schisandrol hae, schisandrin alcohol A, schisandrin B, and schisandrol.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignan is schisandrin A.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4:3). When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4:3), the half-life of the compound represented by formula I or its salt is extended. , the content of the metabolite shown in Formula II during the metabolic process is lower than that of the compound shown in Formula I or its salt alone, and it has strong anti-tumor ability and high safety. According to the description of the embodiments of this application, the dosages for mice and humans can be converted, and adding the above two substances using the converted values can effectively protect humans from or alleviate tumors.

According to an embodiment of the present invention, the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (1:10) to (5:6). When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (1:10) to (5:6), the half-life of the compound represented by formula I or its salt is significantly improved. Prolonged, the content of the metabolite shown in formula II during the metabolic process is significantly lower than that of the compound shown in formula I or its salt alone, and it has strong anti-tumor ability and high safety. According to the description of the embodiments of this application, the dosages for mice and humans can be converted, and adding the above two substances using the converted values can effectively protect humans from or alleviate tumors.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (7:50) to (4:5). When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (7:50) to (4:5), the half-life of the compound represented by formula I or its salt is significantly improved. Prolonged, the content of the metabolite represented by formula II during the metabolic process is significantly lower than that of the compound represented by formula I or its salt alone, and the pharmaceutical composition has strong anti-tumor ability and high safety.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is 3:20, 1:5, 81:250 or 81:125. When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is 3:20, 1:5, 81:250 or 81:125, the compound represented by formula I or its salt is The half-life is significantly extended, and the content of the metabolite represented by formula II during metabolism is significantly lower than when the compound represented by formula I or its salt is used alone. The pharmaceutical composition has strong anti-tumor ability and high safety.

According to an embodiment of the present invention, the pharmaceutical composition is an oral preparation. The pharmaceutical composition may be in the form of taking preparations, injection preparations, pills, sustained-release preparations, implants or aerosols, etc., and is not particularly limited. According to specific embodiments of the present invention, when the pharmaceutical composition When it is an oral preparation, the half-life of the single dosage form is longer, and the content of the metabolite shown in Formula II during metabolism is significantly lower than when the compound shown in Formula I or its salt is used alone, which is beneficial to improving the penetration of the compound shown in Formula I or its salt. The brain dose further improves the drug efficacy. Therefore, the pharmaceutical composition has stronger anti-tumor ability and higher safety.

According to some specific embodiments of the present invention, the pharmaceutical composition includes the compound represented by Formula I or a salt thereof provided herein, and Schisandrin A, as well as a pharmaceutically acceptable carrier. "Pharmaceutically acceptable carrier" may include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Specific examples may be one or more of water, saline, phosphate buffered saline, glucose, glycerin, ethanol, etc., and combinations thereof. In many cases, pharmaceutical compositions include isotonic agents, such as sugars, polyols (such as mannitol, sorbitol) or sodium chloride. Of course, pharmaceutically acceptable carriers may also include trace amounts of auxiliary substances, such as wetting agents or emulsifiers, preservatives or buffers, to extend the shelf life or effectiveness of the antibody.

For example, the compound represented by formula I of the present invention or a salt thereof and biphenylcyclooctadienolignan can be incorporated into a pharmaceutical composition suitable for parenteral administration (such as intravenous, subcutaneous, intraperitoneal, intramuscular) . These pharmaceutical compositions can be prepared in various forms. Examples include liquid, semi-solid, and solid dosage forms, including, but not limited to, liquid solutions (eg, injection solutions and infusion solutions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. Typical pharmaceutical compositions are in the form of oral preparations, injection solutions, or infusion solutions. The pharmaceutical composition may be administered orally, intravenously, or by injection, or intramuscularly or subcutaneously.

In a second aspect of the invention, a single dosage form is provided. According to the embodiment of the present invention, it includes: the compound represented by formula I or its salt and biphenylcyclooctadiene lignan. The inventor found that biphenylcyclooctadiene lignans, such as Schisandrin A, not only have the function of protecting the liver, but also have a significant inhibitory effect on liver microsomal cytochrome CYP450 enzymes. Furthermore, by inhibiting CYP450 enzymes, they reduce The metabolism of the compound represented by formula I or its salt prolongs its residence time in the body and enhances the therapeutic effect. At the same time, it reduces the concentration or content of the metabolite represented by formula II produced by the metabolic reaction in the serum at the same time, and increases the concentration or content of the compound represented by formula I or its salt. The exposure of its salt in the body and the dose in brain tissue reduce the adverse reactions caused by the metabolites shown in Formula II, so that the efficacy and safety of the compound shown in Formula I or its salt are improved. Therefore, the single The dosage form has strong anti-tumor ability and high safety. According to the description of the embodiments of this application, through the exchange of dosage between mice and humans, Calculation, using this ratio for conversion can effectively protect humans from or alleviate tumors.

According to embodiments of the present invention, the above-mentioned single dosage form may further include at least one of the following additional technical features:

According to embodiments of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4:3).

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (1:10) to (5:6).

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (7:50) to (4:5). When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (7:50) to (4:5), the half-life of the single dosage form is significantly extended, and the metabolism process of formula II The content of the metabolites shown is significantly lower than that of the compound shown in formula I or its salt alone, thereby increasing the brain-penetrating dose of the compound shown in formula I or its salt and improving the efficacy. Therefore, the single dosage form has strong anti-bacterial activity. tumor capability and higher safety.

According to an embodiment of the present invention, the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is 3:20, 1:5, 81:250 or 81:125. When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is 3:20, 1:5, 81:250 or 81:125, the half-life of the single dosage form is significantly extended, and the metabolism In the process, the content of the metabolite represented by formula II is significantly lower than that of the compound represented by formula I or its salt alone, thereby increasing the brain-penetrating dose of the compound represented by formula I or its salt and improving the drug efficacy. Therefore, the single dosage form has very high Strong anti-tumor ability and high safety.

According to an embodiment of the present invention, the single dosage form includes 200 to 300 mg of the compound represented by Formula I or a salt thereof. According to the description of the embodiments of this application, the above-mentioned single dosage form can be used 2 times, 3 times or 4 times a day, preferably 2 times. Through the conversion of dosages for mice and humans, this dosage can effectively protect humans after conversion. Avoid or alleviate tumors. Those skilled in the art can understand that the specifications of the single dosage form are not particularly limited. For example, the specifications of the single dosage form can be set according to the characteristics of different target groups. For example, the specifications of the single dosage form can be set according to the weight of the target group, such as a body weight of 40kg. , 60kg, 70kg, 80kg, 90kg, 100kg, etc., or set according to the mass ratio of the compound represented by Formula I or its salt described in this application to biphenylcyclooctadiene lignan, such as 3:20, 1:6, 5:6, 4:5, 81:250 or 81:125, etc. Or, the compound of formula I or its salt and biphenylcyclooctadienyl lignin can be used according to the characteristics of the target population and the Set the mass ratio of elements.

According to an embodiment of the present invention, the single dosage form is an oral preparation. The single dosage form may be an oral preparation, an injection preparation, a pill, a sustained-release preparation, an implant or an aerosol, and is not particularly limited. According to specific embodiments of the present invention, when the single dosage form is oral The half-life of the single dosage form is longer during preparation, and the content of the metabolite represented by formula II during the metabolism process is significantly lower than that of the compound represented by formula I or its salt alone, thereby increasing the penetration of the compound represented by formula I or its salt. Brain dose improves drug efficacy. Therefore, the single dosage form has stronger anti-tumor ability and higher safety.

In a third aspect of the invention, the invention proposes a drug combination. According to an embodiment of the present invention, the compound of formula I or a salt thereof is included as the first active ingredient and biphenylcyclooctadiene lignan as the second active ingredient. As mentioned above, the inventor found that Schisandrin A not only has the function of protecting the liver, but also can reduce the metabolism of the compound represented by Formula I or its salt by inhibiting CYP450 enzymes, such as subtype CYP3A, and prolong its retention in the body. time to enhance the therapeutic effect, and at the same time reduce the metabolism of formula II produced by the demethylation reaction in the serum at the same time It can increase the exposure of the compound represented by formula I or its salt in the body and the dose in brain tissue, reduce the adverse reactions caused by the metabolite represented by formula II, and improve the efficacy and safety of the compound represented by formula I or its salt. Therefore, the drug combination according to the embodiment of the present invention can significantly extend the half-life and brain-penetrating dose of the compound represented by Formula I or its salt, can effectively inhibit tumor growth, and has high safety.

According to embodiments of the present invention, the above-mentioned drug combination may further include at least one of the following additional technical features:

According to an embodiment of the present invention, the salt of the compound represented by formula I includes at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate, benzoate, adipic acid Salts, succinates, methanesulfonates and maleates.

According to an embodiment of the present invention, the salt of the compound represented by Formula I is a hydrochloride.

According to embodiments of the present invention, the salt of the compound represented by Formula I is (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2, 4-Dimethylpiperazine-1-carboxylate hydrochloride.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignans include at least one of the following: schisandrin A, schisandrin ester A, schisandrol hae, schisandrin alcohol A, schisandrin B, and schisandrol.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignan is schisandrin A.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4:3). When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4:3), the half-life of the compound represented by formula I or its salt is extended. , the content of the metabolite shown in Formula II during the metabolic process is lower than that of the compound shown in Formula I or its salt alone, and it has strong anti-tumor ability and high safety. According to the description of the embodiments of this application, the dosages for mice and humans can be converted, and adding the above two substances using the converted values can effectively protect humans from or alleviate tumors.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (1:10) to (5:6). When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (1:10) to (5:6), the half-life of the compound represented by formula I or its salt is significantly improved. Prolonged, the content of the metabolite shown in formula II during the metabolic process is significantly lower than that of the compound shown in formula I or its salt alone, thereby increasing the brain-penetrating dose of the compound shown in formula I or its salt, and improving the drug efficacy, as described after drug combination. The compound represented by formula I or its salt has stronger anti-tumor ability and higher safety. According to the description of the embodiments of the present application, by converting the dosage between mice and humans, using this ratio can effectively protect humans from or alleviate tumors.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (7:50) to (4:5). When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (7:50) to (4:5), the half-life of the compound represented by formula I or its salt is significantly improved. Prolonged, the content of the metabolite represented by formula II is significantly lower than that of the compound represented by formula I or its salt alone, thereby increasing the brain-penetrating dose of the compound represented by formula I or its salt, and improving the drug efficacy. After drug combination, formula I is described The indicated compound or its salt has stronger anti-tumor ability and higher safety.

According to an embodiment of the present invention, the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is 3:20, 1:5, 81:250 or 81:125. The mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is 3:20, 1:5, 81:250 or 81: When 125, the half-life of the compound represented by formula I or its salt is significantly extended, and the content of the metabolite represented by formula II is significantly lower than that of the compound represented by formula I or its salt alone, thereby increasing the concentration of the compound represented by formula I or its salt. The brain-penetrating dose of the salt can improve the drug efficacy. After drug combination, the compound represented by formula I or its salt has stronger anti-tumor ability and higher safety.

According to an embodiment of the present invention, the compound represented by Formula I or its salt and the biphenylcyclooctadiene lignan are formulated together or separately.

According to an embodiment of the present invention, the compound represented by formula I or its salt and the biphenylcyclooctadiene lignan are used simultaneously or separately.

It should be noted that the drug combination includes combinations that are separated in time and/or space, as long as the compound represented by formula I or its salt and biphenylcyclooctadienolignan can cooperate to achieve the purpose of the present invention. For example, the components contained in the pharmaceutical combination can be administered to the subject or the sample to be tested as a whole, or separately. When the components contained in the pharmaceutical combination are administered to the subject or sample to be tested separately, the individual components may be administered to the subject or sample to be tested simultaneously or sequentially.

According to an embodiment of the present invention, the biphenylcyclooctadienolignan is administered prior to the compound represented by Formula I or a salt thereof. Those skilled in the art can understand that the order of administration of the biphenylcyclooctadienol and the compound represented by formula I or its salt is not particularly limited, as long as the diphenylcyclooctadienol It is sufficient that the compound can play a role in the metabolic cycle of the compound represented by formula I or its salt. For example, the biphenylcyclooctadienolignan is administered before the compound represented by formula I or its salt, Or the biphenylcyclooctadiene lignan is administered after the compound represented by formula I or its salt, or the biphenylcyclooctadienolignan and the compound represented by formula I or its salt are administered Simultaneous administration is within the scope of protection of this application. According to some specific embodiments of the present invention, when the biphenylcyclooctadienolignan precedes the (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquin When oxazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride is administered, the half-life of the compound represented by formula I or its salt can be significantly extended, and during its metabolism The content of the metabolite shown in the formula II produced is significantly reduced, so that the compound shown in the formula or its salt has a higher brain penetration rate, which increases the brain penetration dose and further improves the drug efficacy. The drug combination has a good anti-tumor effect and security.

According to an embodiment of the invention, the second active ingredient is in an oral dosage form. The second active ingredient may be in the form of taking preparations, injection preparations, pills, sustained-release preparations, implants or aerosols, etc., and is not particularly limited. According to specific embodiments of the present invention, when the second active ingredient When the active ingredient is an oral preparation, the half-life of the single dosage form is longer, and the content of the metabolite shown in Formula II during metabolism is significantly lower than when the compound shown in Formula I or its salt is used alone, and the compound shown in Formula I or its salt has Stronger anti-tumor ability and higher safety.

In a fourth aspect of the invention, the invention provides a medicine box. According to an embodiment of the present invention, the pharmaceutical kit includes: a compound represented by Formula I or a salt thereof as the first active ingredient and biphenylcyclooctadienolignan as the second active ingredient. As mentioned above, the inventor found that biphenylcyclooctadienolignan reduces the metabolism of the compound represented by formula I or its salt by inhibiting CYP450 enzymes, such as its subtype CYP3A enzyme, prolongs its residence time in the body and enhances the therapeutic effect. , while reducing the concentration or content of the metabolite represented by formula II in the serum at the same time, increasing the exposure of the compound represented by formula I or its salt in the body and the dose in brain tissue, and reducing the adverse reactions caused by the metabolite represented by formula II, The efficacy and safety of the compound represented by formula I or its salt are improved, and the compound can effectively inhibit tumor growth and has high safety.

According to an embodiment of the present invention, the above-mentioned medicine box may further include at least one of the following additional technical features:

According to an embodiment of the present invention, the salt of the compound represented by formula I includes at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate, benzoate, adipic acid Salts, succinates, methanesulfonates and maleates.

According to an embodiment of the present invention, the salt of the compound represented by Formula I is a hydrochloride.

According to embodiments of the present invention, the salt of the compound represented by Formula I is (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2, 4-Dimethylpiperazine-1-carboxylate hydrochloride.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignans include at least one of the following: schisandrin A, schisandrin ester A, schisandrol hae, schisandrin alcohol A, schisandrin B, and schisandrol.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignan is schisandrin A.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4:3). When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4:3), the half-life of the compound represented by formula I or its salt is extended. , the content of the metabolite shown in Formula II during the metabolic process is lower than that of the compound shown in Formula I or its salt alone, and it has strong anti-tumor ability and high safety. According to the description of the embodiments of this application, the dosages for mice and humans can be converted, and adding the above two substances using the converted values can effectively protect humans from or alleviate tumors.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (1:10) to (5:6). When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (1:10) to (5:6), the half-life of the compound represented by formula I or its salt is significantly improved. Prolonged, the content of the metabolite of the metabolite shown in formula II during the metabolic process is significantly lower than that of the compound shown in formula I or its salt alone, and it has strong anti-tumor ability and high safety. According to the description of the embodiments of this application, the dosages for mice and humans can be converted, and adding the above two substances using the converted values can effectively protect humans from or alleviate tumors.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (7:50) to (4:5). When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (7:50) to (4:5), the half-life of the compound represented by formula I or its salt is significantly improved. Prolonged, the content of the metabolite of the metabolite shown in Formula II during the metabolic process is significantly lower than that of the compound shown in Formula I or its salt alone, and the kit has strong anti-tumor ability and high safety.

According to an embodiment of the present invention, the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is 3:20, 1:5, 81:250 or 81:125. When the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is 3:20, 1:5, 81:250 or 81:125, the compound represented by formula I or its salt is The half-life is significantly extended, and the content of the metabolite represented by formula II during metabolism is significantly lower than when the compound represented by formula I or its salt is used alone. The pharmaceutical composition has strong anti-tumor ability and high safety.

According to an embodiment of the present invention, the compound represented by Formula I or its salt and the biphenylcyclooctadiene lignan are formulated together or separately. Those skilled in the art can understand that the preparation method of the compound represented by formula I or its salt and the biphenylcyclooctadiene lignan is not particularly limited, and they can be prepared separately or together.

According to an embodiment of the present invention, the compound represented by formula I or its salt and the biphenylcyclooctadiene lignan are used simultaneously or separately. It should be noted that in the method, the compound represented by formula I or its salt and the biphenylcyclooctadiene lignan are used separately in time and/or space, as long as the compound represented by formula I or its salt is used separately in time and/or space. The salt and biphenylcyclooctadiene lignan salt can cooperate to achieve the purpose of the present invention. For example, the compound represented by Formula I or a salt thereof and the biphenylcyclooctadienolignan can be administered to the subject or the sample to be tested as a whole, or separately to the subject or the sample to be tested. When the compound represented by Formula I or a salt thereof and the biphenylcyclooctadienolignan are administered to a subject or a sample to be tested separately, each component may be administered to the subject or sample to be tested simultaneously or sequentially. sample.

According to an embodiment of the present invention, the biphenylcyclooctadienolignan is administered prior to the compound represented by Formula I or a salt thereof. Those skilled in the art can understand that the order of administration of the biphenylcyclooctadienol and the compound represented by formula I or its salt is not particularly limited, as long as the diphenylcyclooctadienol It is sufficient that the compound can play a role in the metabolic cycle of the compound represented by formula I or its salt. For example, the biphenylcyclooctadienolignan is administered before the compound represented by formula I or its salt, Or the biphenylcyclooctadiene lignan is administered after the compound represented by formula I or its salt, or the biphenylcyclooctadienolignan and the compound represented by formula I or its salt are administered Simultaneous administration is within the scope of protection of this application. According to some specific embodiments of the present invention, when the biphenylcyclooctadienolignan is administered before the compound represented by formula I or its salt, the half-life of the compound represented by formula I or its salt can be It is significantly prolonged, and the content of the metabolite shown in formula II produced during its metabolism is significantly reduced, and it has good anti-tumor effect and safety.

According to an embodiment of the invention, the second active ingredient is in an oral dosage form. The second active ingredient may be in the form of taking preparations, injection preparations, pills, sustained-release preparations, implants or aerosols, etc., and is not particularly limited. According to specific embodiments of the present invention, when the second active ingredient When the active ingredient is an oral preparation, the half-life of the kit is longer, and the content of the metabolite shown in Formula II during metabolism is significantly lower than when the compound shown in Formula I or its salt is used alone, and the kit has strong anti-tumor ability. and higher security.

In the fifth aspect of the present invention, the present invention provides a method of administering the compound represented by Formula I or a salt thereof. According to an embodiment of the present invention, the method includes combining the compound represented by Formula I or a salt thereof with biphenylcyclooctadienolignan.

According to an embodiment of the present invention, the above method further includes at least one of the following additional technical features:

According to an embodiment of the present invention, the salt of the compound represented by formula I includes at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate, benzoate, adipic acid Salts, succinates, methanesulfonates and maleates.

According to an embodiment of the present invention, the salt of the compound represented by Formula I is a hydrochloride.

According to embodiments of the present invention, the salt of the compound represented by Formula I is (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2, 4-Dimethylpiperazine-1-carboxylate hydrochloride.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignans include at least one of the following: schisandrin A, schisandrin ester A, schisandrol hae, schisandrin alcohol A, schisandrin B, and schisandrol.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignan is schisandrin A.

According to an embodiment of the present invention, the combined medication includes administering the compound represented by formula I or its salt and biphenylcyclooctadienolignan simultaneously or separately.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4:3). According to the description of the embodiments of the present application, by converting the dosage between mice and humans, this ratio conversion can effectively protect humans from or alleviate tumors.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan during the medication process is (1:10) to (5:6).

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan during the medication process is (7:50) to (4:5).

According to an embodiment of the present invention, the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan during the medication process is 3:20, 1:5, 81:250 or 81:125.

According to an embodiment of the present invention, the biphenylcyclooctadienolignan is administered prior to the compound represented by Formula I or a salt thereof. Those skilled in the art can understand that the order of administration of the biphenylcyclooctadienol and the compound represented by formula I or its salt is not particularly limited, as long as the diphenylcyclooctadienol It is sufficient that the compound can play a role in the metabolic cycle of the compound represented by formula I or its salt. For example, the biphenylcyclooctadienolignan is administered before the compound represented by formula I or its salt, Or the biphenylcyclooctadiene lignan is administered after the compound represented by formula I or its salt, or the biphenylcyclooctadienolignan and the compound represented by formula I or its salt are administered Simultaneous administration is within the scope of protection of this application.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignan is administered orally. Those skilled in the art can understand that the biphenylcyclooctadiene lignans can be in the form of oral preparations, injection preparations, pills, sustained-release preparations, implants or aerosols, etc., and are not particularly limited. According to In specific embodiments of the present invention, when the biphenylcyclooctadiene lignan is an oral preparation, the half-life of the compound represented by formula I or its salt is longer, and the content of metabolite 1 during metabolism is significantly lower than that of the compound alone. Using the compound represented by formula I or its salt, the administration method has stronger anti-tumor ability and higher safety.

According to an embodiment of the present invention, the dosage of the biphenylcyclooctadiene lignan is 0 to 25 mg/kg.

According to an embodiment of the present invention, the dosage of the biphenylcyclooctadiene lignan is 5 to 25 mg/kg. According to specific embodiments of the present invention, when the dosage of the biphenylcyclooctadiene lignan is 5 to 25 mg/kg, the half-life of the compound represented by formula I or its salt can be significantly extended, and the The content of the metabolites shown in formula II produced during the metabolism process is significantly reduced, which increases the exposure of the compound shown in formula I or its salt in the body and the dose in brain tissue, and reduces the adverse reactions caused by the metabolites shown in formula II, so that The efficacy and safety of the compound represented by formula I or its salt are improved, and the administration method can more effectively inhibit tumor growth and has higher safety.

In the sixth aspect of the present invention, the present invention proposes a method for extending the half-life of the compound represented by formula I or its salt. According to an embodiment of the present invention, the method includes combining the compound represented by Formula I or a salt thereof with biphenylcyclooctadiene lignan. As mentioned above, the inventor found that biphenylcyclooctadienolignans reduce the metabolism of the compound represented by formula I or its salt by inhibiting CYP450 enzymes, such as its subtype CYP3A enzyme, prolonging its residence time in the body and enhancing the therapeutic effect. , Therefore, the method according to the embodiment of the present invention can effectively extend the half-life of the compound represented by formula I or its salt.

According to an embodiment of the present invention, the above method further includes at least one of the following additional technical features:

According to an embodiment of the present invention, the salt of the compound represented by formula I includes at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate, benzoate, adipic acid Salts, succinates, methanesulfonates and maleates.

According to an embodiment of the present invention, the salt of the compound represented by Formula I is a hydrochloride.

According to embodiments of the present invention, the salt of the compound represented by Formula I is (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2, 4-Dimethylpiperazine-1-carboxylate hydrochloride.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignans include at least one of the following: schisandrin A, schisandrin ester A, schisandrol hae, schisandrin alcohol A, schisandrin B, and schisandrol.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignan is schisandrin A.

According to an embodiment of the present invention, the combined medication includes administering the compound represented by formula I or its salt and biphenylcyclooctadienolignan simultaneously or separately. The method of combined administration will not be described in detail, and reference may be made to the method of combined administration in the administration method of the compound represented by formula I or its salt described in the fifth aspect.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4:3). According to the description of the embodiments of the present application, by converting the dosage between mice and humans, using this ratio conversion can effectively extend the efficacy time of protecting humans from or alleviating tumors.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan during the medication process is (1:10) to (5:6).

According to an embodiment of the present invention, the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan during the medication process is (7:10) to (4:5).

According to an embodiment of the present invention, the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan during the medication process is 3:20, 1:5, 81:250 or 81:125.

According to an embodiment of the present invention, the biphenylcyclooctadienolignan is administered prior to the compound represented by Formula I or a salt thereof. As mentioned above, the order of administration of the biphenylcyclooctadienolignan and the compound represented by Formula I or its salt is not particularly limited, as long as the biphenylcyclooctadienolignan can It only needs to be effective within the metabolic cycle of the compound represented by formula I or its salt. For example, the biphenylcyclooctadienolignan is administered before the compound represented by formula I or its salt, or the The biphenylcyclooctadiene lignan is administered after the compound represented by the formula I or its salt, or the biphenylcyclooctadiene lignan and the compound represented by the formula I or its salt are administered simultaneously. All drugs are within the protection scope of this application.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignan is administered orally.

According to an embodiment of the present invention, the dosage of the biphenylcyclooctadiene lignan is 0 to 25 mg/kg.

According to an embodiment of the present invention, the dosage of the biphenylcyclooctadiene lignan is 5 to 25 mg/kg. According to specific embodiments of the present invention, when the dosage of the biphenylcyclooctadiene lignan is 5 to 25 mg/kg, the half-life of the compound represented by formula I or its salt can be significantly extended, and the The content of the metabolites shown in formula II produced during the metabolic process is significantly reduced, and the method can more effectively extend the The half-life of the compound represented by formula I or its salt is increased, and its brain penetration rate is increased to improve the drug efficacy.

In the seventh aspect of the present invention, the present invention proposes a method for reducing the compound represented by formula I or its salt to produce the compound represented by formula II. According to an embodiment of the present invention, the method includes combining the compound represented by Formula I or a salt thereof with biphenylcyclooctadienolignan. As mentioned above, the inventor found that biphenylcyclooctadienolignan inhibits CYP450 enzymes, such as its subtype CYP3A enzyme, thereby reducing the metabolism of the compound represented by formula I or its salt, and reducing the concentration of the metabolite represented by formula II in the serum. or content, increase the exposure of the compound represented by formula I or its salt in the body and the dose in brain tissue, reduce the adverse reactions caused by the metabolite represented by formula II, and make the compound represented by formula I or its salt more effective and safe sex has been improved.

According to an embodiment of the present invention, the above method further includes at least one of the following additional technical features:

According to an embodiment of the present invention, the salt of the compound represented by formula I includes at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate, benzoate, adipic acid Salts, succinates, methanesulfonates and maleates.

According to an embodiment of the present invention, the salt of the compound represented by Formula I is a hydrochloride.

According to embodiments of the present invention, the salt of the compound represented by Formula I is (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2, 4-Dimethylpiperazine-1-carboxylate hydrochloride.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignans include at least one of the following: schisandrin A, schisandrin ester A, schisandrol hae, schisandrin alcohol A, schisandrin B, and schisandrol.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignan is schisandrin A.

According to an embodiment of the present invention, the combined medication includes administering the compound represented by formula I or its salt and biphenylcyclooctadienolignan simultaneously or separately.

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4:3).

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan during the medication process is (1:10) to (5:6).

According to an embodiment of the present invention, the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan during the medication process is (7:50) to (4:5).

According to an embodiment of the present invention, the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan during the medication process is 3:20, 1:5, 81:250 or 81:125.

According to an embodiment of the present invention, the biphenylcyclooctadienolignan is administered prior to the compound represented by Formula I or a salt thereof. As mentioned above, the order of administration of the biphenylcyclooctadienolignan and the compound represented by Formula I or its salt is not particularly limited, as long as the biphenylcyclooctadienolignan can It only needs to be effective within the metabolic cycle of the compound represented by formula I or its salt. For example, the biphenylcyclooctadienolignan is administered before the compound represented by formula I or its salt, or the The biphenylcyclooctadiene lignan is administered after the compound represented by the formula I or its salt, or the biphenylcyclooctadiene lignan and the compound represented by the formula I or its salt are administered simultaneously. All drugs are within the protection scope of this application.

According to an embodiment of the present invention, the biphenylcyclooctadiene lignan is administered orally.

According to an embodiment of the present invention, the dosage of the biphenylcyclooctadiene lignan is 0 to 25 mg/kg.

According to an embodiment of the present invention, the dosage of the biphenylcyclooctadiene lignan is 5 to 25 mg/kg. According to specific embodiments of the present invention, when the dosage of Schisandrin A is 5 to 25 mg/kg, the compound represented by Formula I or its salt can more effectively reduce the amount of Formula II produced in the serum during the metabolism process. Therefore, the method according to the embodiment of the present invention can more significantly reduce the content of the metabolite shown in Formula II during the metabolism of the compound shown in Formula I or its salt.

Detailed ways

The embodiments of the present invention are described in detail below. The following embodiments are illustrative and are intended to explain the present invention, but should not be understood as limiting the present invention.

In the process of describing the present invention, the relevant terms in this article have been explained and explained. These explanations and explanations are only for the convenience of understanding of the scheme and cannot be regarded as limiting the protection scheme of the present invention.

In this application, "compounds represented by formula I" all refer to compounds with the following structures:

In this application, the "hydrochloride of the compound represented by formula I", "(R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl The molecular formula of "2,4-dimethylpiperazine-1-carboxylate hydrochloride", "test drug" and "original drug" is C ₂₂ H ₂₃ ClFN ₅ O _3. The chemical structure is:

In this application, the molecular formula of "metabolite represented by formula II" and "metabolite 1" is C ₂₁ H ₂₁ ClFN ₅ O ₃ , and its chemical structure is:

In this application, "Schisandrin A" refers to a substance with a molecular formula of C ₂₄ H ₃₂ O ₆ and a molecular weight of 416.507. Its chemical structure is:

The molecular formula of "Schisandrin A" is C ₃₀ H ₃₂ O ₉ , the molecular weight is 536.577, and the chemical structure is:

The molecular formula of "Schizandrol" is C ₂₃ H ₂₈ O ₇ , the molecular weight is 416.47, and the chemical structure is:

The molecular formula of "Schizandrol Methyl" is C ₂₄ H ₃₂ O ₇ , the molecular weight is 432.51, and the chemical structure is:

The molecular formula of "Schisandrin B" is C ₂₃ H ₂₈ O ₆ , the molecular weight is 400.46, and the chemical structure is:

The molecular formula of "Schizandrol" is C ₂₃ H ₃₀ O ₆ , the molecular weight is 402.48, and the chemical structure is:

In this application, the "single dosage form" refers to a dosage form that is used up after a single administration of a preparation.

In this application, the "drug combination" refers to the simultaneous or sequential application of two or more drugs to achieve therapeutic purposes. The result is mainly to increase the efficacy of the drug or to reduce the toxic and side effects of the drug.

The solutions of the present invention will be explained below with reference to examples. Those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention. If specific techniques or conditions are not specified in the examples, the techniques or conditions described in literature in the field or product instructions will be followed. If the manufacturer of the reagents or instruments used is not indicated, they are all conventional products that can be purchased commercially.

Example 1 Detection of CYP450 Enzyme Inhibitory Ability of CYP450 Enzyme Inhibitors

In this application, the inventor is interested in the drug (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1- Carboxylate Hydrochloride was studied alone and in combination with CYP450 enzyme inhibitors, wherein the (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazoline-6- The chemical structure of 2,4-dimethylpiperazine-1-carboxylate hydrochloride is:

The specific experimental operations are as follows:

The experimental system uses human liver microsomes. There are 4 groups of liver microsomes each, namely 1) (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl. 2,4-Dimethylpiperazine-1-carboxylate hydrochloride group (test drug alone group); 2) (R)-4-(3-chloro-2-fluorophenylamino)-7 -Methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride combined with Schizandrin A (WWZ) group (test drug + WWZ group); 3) ( R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride with water Silybin group (test drug + silibinin group), 4) (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2 , 4-dimethylpiperazine-1-carboxylate hydrochloride combined with ketoconazole group (test drug + ketoconazole group). The specific experimental steps are as follows:

Precisely measure (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate salt The stock solution of acid salt (Stock0009) is 1 mg/mL, and diluted with PBS buffer solution to a standard working solution with a molar concentration of 10 μM. Precisely measure (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate salt The stock solution of acid salt (Stock0009) of 1 mg/mL was diluted into a standard working solution with a molar concentration of 20 μM with a PBS buffer solution containing 5 μM of Schisandrin A, silibinin, or ketoconazole, and then 10 μM was obtained. Mixed working solution 2 containing 5 μM of various CYP450 enzyme inhibitors. Weigh about 7mg of NADPH (reduced form) and dilute it with PBS buffer solution to a working solution with a molar concentration of 10mM. The initial concentration of liver microsomes (HLM) was 20 mg/mL, and it was diluted with PBS buffer solution to a liver microsome working solution with a molar concentration of 2 mg/mL. Take each of the above working solutions and prepare according to Table 1.

Table 1: Preparation method of liver microsome incubation system sample

Mix the four test working solutions, HLM, and PBS buffer solutions in incubation tubes according to the volumes in the table (in triplicate), and preheat in a 37°C water bath for 5 minutes. At the same time, 10mM NADPH was also preheated in a 37°C water bath. After sufficient preheating, add 30 μL NADPH to the liver microsome incubation tube, shake the incubation tube gently, and start incubation. Take 40 μL of incubation tubes at 2 minutes, 15 minutes, 1 hour, 2 hours, and 4 hours after starting the incubation. Add 160 μL of acetonitrile solution containing internal standard (propranolol 20 ng/mL). Each sample was shaken and centrifuged for 10 min (centrifugal force 20200g), and the supernatant was taken and injected for LC/MS/MS analysis.

The experimental results are shown in Table 2. By treating the drug (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperidine Comparing the clearance effect of oxazine-1-carboxylate hydrochloride alone and in combination with various CYP450 enzyme inhibitors, the inventor found that compared with other high and low intensity CYP450 enzyme inhibitors, Schisandrin A has a higher The degree of inhibition of CYP450 enzymes is moderate. In this study, the moderate inhibitor WWZ) was selected for follow-up research.

Table 2:

Example 2 (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate salt Study on the metabolic stability of acid salt in liver microparticles in vitro

On the basis of Example 1, this experiment mainly investigated the original drug (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-di Differences in metabolic stability of methylpiperazine-1-carboxylate hydrochloride in liver microsome incubation system, and changes in metabolic stability after combined use with the inhibitor WWZ. The experimental system uses human liver microsomes. There are three groups of liver microsomes, each of which is 1) (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl. 2,4-Dimethylpiperazine-1-carboxylate hydrochloride group (test drug alone group); 2) (R)-4-(3-chloro-2-fluorophenylamino)-7 -Methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride combined with WWZ group (test drug + WWZ group); 3) Positive control drug (Vera Pami, VPL) group. The specific experimental steps are as follows:

Precisely measure (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate salt The stock solution of acid salt (Stock0009) is 1 mg/mL, and diluted with PBS buffer solution to a standard working solution with a molar concentration of 10 μM. Precisely measure (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate salt The stock solution (Stock0009) of acid salt 1 mg/mL was diluted with PBS buffer solution dissolved with Schisandrin A (5 μM) to a standard working solution with a molar concentration of 20 μM, and then a mixed working solution containing 5 μM Schisandrin A was obtained as 10 μM. 2. Measure 100 μL of verapamil 1 mg/mL stock solution (StockVPL) and dilute it with PBS buffer solution to a working solution with a molar concentration of 10 μM as a positive control. Weigh about 7mg of NADPH (reduced form) and dilute it with PBS buffer solution to a working solution with a molar concentration of 10mM. The initial concentration of liver microsomes (HLM) was 20 mg/mL in PBS. The buffer solution was diluted to a liver microsomal working solution with a molar concentration of 2 mg/mL. Take each of the above working solutions and prepare according to Table 3.

Table 3: Preparation method of liver microsome incubation system sample

Mix the three test working solutions, HLM, and PBS buffer solutions in incubation tubes according to the volumes in the table (in triplicate), and preheat in a 37°C water bath for 5 minutes. At the same time, 10mM NADPH was also preheated in a 37°C water bath. After sufficient preheating, add 30 μL NADPH to the liver microsome incubation tube, shake the incubation tube gently, and start incubation. At 2 min, 15 min, 1 h, 2 h, and 4 h after starting the incubation, add 160 μL of acetonitrile solution containing the internal standard (propranolol 20 ng/mL) into a 40 μL incubation tube. Each sample was shaken and centrifuged for 10 min (centrifugal force 20200g), and the supernatant was taken and injected for LC/MS/MS analysis.

Verapamil was used as a positive control (sampling time points were 30 min and 2 h after incubation), and verapamil without NADPH (replaced by PBS) was used as a negative control. The drug-free (replaced by PBS) group served as a blank control.

Human liver microsomal (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate The results of the in vitro metabolic stability test of hydrochloride are shown in Table 4. Among them, for verapamil, which was used as a positive control, 10% of the prototype drug remained after 2 hours of incubation, indicating that the enzyme metabolic activity of this batch of incubation system was normal. The test drug was metabolized after incubation in human liver microsomes, and the prototype concentration decreased with increasing incubation time. At the same time, (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride Metabolic rate slows down after taking WWZ.

Table 4:

Example 3 (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate salt Study on the metabolic kinetics in SD rats after combined administration of acid salt and Schisandrin A

This example is carried out on the basis of Examples 1 and 2, mainly investigating the test drug ((R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazoline- Changes in metabolic kinetics in SD rats after combined administration of 6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride and schizandrin. In this study, 9 healthy growing male rats aged 6-8 weeks and weighing 220-300g were divided into 3 groups: the test drug (5mg/kg) group was administered by gavage; the group was administered 25 mg of Schisandrin A by gavage. /kg and then intragastric administration of the test drug (5mg/kg) group (A0009+WWZ(PO)); intravenous injection of Schisandrin A (10mg/kg) followed by intragastric administration of the test drug (5mg/kg) group ( A0009+WWZ(IV)).

The whole blood of the rats in the above three groups was collected at the following time points: before administration and 5min, 15min, 30min, 1h, 2h, 4h, 8h, 12h, 24h, and 36h after administration. Blood collection site and volume: About 0.3 mL of blood is collected from the retroocular venous plexus. The obtained whole blood sample was centrifuged (centrifugal force 2000g, 4°C) for 10 min, and the upper plasma was taken. Plasma samples were processed using organic solvent protein precipitation. Take 100 μL of plasma, add 400 μL of internal standard acetonitrile solution (propranolol, 20 ng/mL), vortex and mix evenly, centrifuge for 10 min (centrifugal force 20200g, 4°C), and take the supernatant and inject it for LC/MS/MS analysis. The Winnonlin pharmacokinetic program was used to analyze the measured data and calculate the main pharmacokinetic parameters.

The blood drug concentration-time change data of the test drugs in each group of animals are shown in Table 5, and the main pharmacokinetic parameters are shown in Tables 6 and 7. Based on the in vivo metabolism results, it can be seen that the in vivo PK behavior of the test drug compounds is affected after taking WWZ. This effect is related to the administration method of co-administration of WWZ. Intravenous co-administration did not change the clearance rate of the test drug, while oral co-administration improved the bioavailability of the test drug. From this, the inventor speculates that drug interactions with WWZ cannot simply be explained by the inhibitory effect of WWZ on liver metabolic enzymes, but are mainly manifested in the absorption process. WWZ mainly acts on the intestines and liver in the absorption phase, improving bioavailability by increasing the absorption of the test drug or reducing the first pass of the liver and intestines. After oral administration of WWZ, (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylic The average exposure AUCinf and half-life t _1/2 h of acid ester hydrochloride were significantly higher than those of (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazoline alone -6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride group.

table 5:

Table 6: Pharmacokinetic parameters of the test drugs in single administration and combined WWZ (gastric administration, intravenous injection) trials

Example 4 (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate salt Study on metabolic transformation in SD rats after combined use of acid salt and WWZ

In order to further observe the kinetic characteristics of metabolite 1 (compound of formula II) in the metabolism process of the test drug after taking WWZ in vivo, this part of the study used the SD rat plasma samples obtained in Example 3 to detect Concentration-time changes of metabolite 1, and pharmacokinetic parameters were calculated. For specific experimental operations, refer to Example 2.

The concentration-time change results of metabolite 1 in the plasma of SD rats in each group are shown in Table 7. Among them, the data in Table 7 and Table 8 show that the metabolic production pattern of metabolite 1 has changed after taking WWZ; whether it is taken orally or When WWZ was administered intravenously together, within 8 hours after its administration, the concentration of metabolite 1 was lower than that of the test drug alone group, indicating that the combined administration of WWZ can effectively inhibit the production of metabolite 1. for testing The blood exposure Cmax of metabolite 1 in the drug alone group was as high as 261ng/mL 4 hours after administration. After oral or intravenous administration of WWZ, it first reduced the exposure of metabolite 1 in the blood, with Cmax of 190ng/mL and 118ng/mL respectively; secondly, it prolonged the peak time of metabolite 1, which was 8h or 12h after administration. Reach Cmax. Data from the exposure amount and peak time of metabolite 1 show that the use of WWZ has the characteristics of reducing metabolite 1.

Table 7: Plasma concentration of metabolite 1 in SD rat plasma samples

Table 8:

Example 5 (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate salt The growth inhibitory effect of combined use of acid salt and WWZ on the growth of human lung cancer PC9 transplanted tumors in nude mice

Based on Examples 1-4, this example continues to study the test drugs and WWZ. The PC9 tumor tissue is taken out under sterile conditions and cut into tumor pieces of 2.0mm×2.0mm×2.0mm, uniformly Inoculate into children weighing 16.0-18.0g BALB/cAnSlacNifdc-nu male nude mice were injected subcutaneously on the left axillary back, and the day of inoculation was recorded as D0. When the tumor volume reaches about 200.0 mm ³ (D6), the tumors are randomly divided into groups according to the tumor volume: solvent control group 1 (25% PEG400 + pH 5.5 water), solvent control group 2 (pH 5.5 water), WWZ ( 25.0 mg/kg) group, test drug [(R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yl 2,4-dimethylpiper Azine-1-carboxylate hydrochloride] group (8.1 mg/kg) group, test drug [(R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazole Phenolin-6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride] group (16.2 mg/kg), WWZ (25.0 mg/kg) + test drug group (8.1 mg/kg) WWZ (25.0mg/kg) + test drug group (16.2mg/kg), a total of 7 groups, 10 animals in each group; on the day of grouping, animals in all groups began to be given intragastric administration, once a day, for a total of 28 days. Dosed 28 times.

Instructions for dosage: Studies have shown that in the in vivo anti-tumor study of the test drug in mouse xenograft tumor model, the dosage calculated based on the free base is 7.5 mg/kg and 15 mg/kg, and has been proven to have better anti-tumor effect, this dose was continued to be used in this study. According to CoA, the free base content of this batch of compounds (batch number: CPo121798-04-05-02-34-01-RS) is 91.85%, the molecular weight of the free base is 459.9, and the molecular weight of the hydrochloride is 496.4. Calculated based on the hydrochloride content in CoA, the doses used in mice in this study were 8.1 mg/kg and 16.2 mg/kg respectively. The effective doses were converted into human doses of 0.7 mg/kg and 1.4 mg/kg based on the body surface area dose. . Instructions on the dosage of WWZ: The current drug name of WWZ in human clinical use is Wuester Capsule (TCM), and its preparation is a capsule, each capsule containing WWZ 11.25mg. The clinical usage and dosage is two capsules each time, three times a day, and the daily dosage is 67.5mg. This dose is the highest clinically used daily dose for adults (60kg). This highest dose is converted into a dose for mice in this study by body surface area dose. In this study, mice were given a dose group of WWZ. , that is, the highest effective dose is 25.0mg/kg.

During the experiment, tumor volume and body weight were measured and recorded twice a week. At the end of the experiment, the tumor volume was counted to calculate the tumor growth inhibition rate TGI and relative tumor proliferation rate T/C%. The tumor volume was measured twice a week using a vernier caliper to measure the long and short diameters of the tumor. The volume calculation formula is: Tumor volume = 0.5 × long diameter × short diameter ² .
Tumor growth inhibition rate (TGI, %) = (1-T (tumor volume of treatment group)/C (tumor volume of solvent control group)) × 100.
Relative tumor proliferation rate (T/C, %) = RTV of treatment group/RTV of solvent control group.

The test results are shown in Table 9.

Table 9:


Note: Evaluation criteria: T/C (%) > 40% is invalid; T/C (%) ≤ 40%; TGI: tumor inhibition rate; T/C%: relative tumor growth rate.

The data in Table 9 shows that the groups of WWZ (25.0mg/kg), test drug (8.1mg/kg), test drug (16.2mg/kg), WWZ (25.0mg/kg) + test drug were administered intragastrically. (8.1mg/kg), WWZ (25.0mg/kg) + test drug (16.2mg/kg) group, on D34, except for the WWZ (25.0mg/kg) group, the tumor growth inhibition rates of the other treatment groups (according to tumor Volume and tumor weight) were both greater than 60%, and T/C were both less than 40%. Prompt to administer test drug (8.1mg/kg), test drug (16.2mg/kg), WWZ (25.0mg/kg) + test drug (8.1mg/kg), WWZ (25.0mg/kg) by gavage +The test drug (16.2 mg/kg) has a significant inhibitory effect on the tumor growth of human lung cancer PC9 transplanted tumors in nude mice.

In the description of this specification, reference to the terms "one embodiment," "some embodiments," "an example," "specific examples," or "some examples" or the like means that specific features are described in connection with the embodiment or example. , structures, materials or features are included in at least one embodiment or example of the invention. In this specification, the schematic expressions of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the specific features, structures, materials or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, those skilled in the art may combine and combine different embodiments or examples and features of different embodiments or examples described in this specification unless they are inconsistent with each other.

Although the embodiments of the present invention have been shown and described above, it can be understood that the above-mentioned embodiments are illustrative and should not be construed as limitations of the present invention. Those of ordinary skill in the art can make modifications to the above-mentioned embodiments within the scope of the present invention. The embodiments are subject to changes, modifications, substitutions and variations.

Claims (54)

A pharmaceutical composition, characterized by comprising: a compound represented by formula I or a salt thereof and biphenylcyclooctadienolignan
The pharmaceutical composition according to claim 1, wherein the salt of the compound represented by formula I includes at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate, Benzoate, adipate, succinate, methanesulfonate and maleate. The pharmaceutical composition according to claim 1, characterized in that the salt of the compound represented by Formula I is a hydrochloride. The pharmaceutical composition according to claim 1, characterized in that the salt of the compound represented by formula I is (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquin Zozolin-6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride. The pharmaceutical composition according to claim 1, wherein the biphenylcyclooctadiene lignans include at least one of the following: schisandrin A, schisandrin ester A, schisandrol acetate, schisandrin ester A, Schisandrin B and Schisandrol. The pharmaceutical composition according to claim 1, wherein the biphenylcyclooctadienolignan is schisandrin A. The pharmaceutical composition according to claim 1, characterized in that the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadienolignan is (1:31) to (4:3) . The pharmaceutical composition according to claim 1, characterized in that the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadienolignan is (1:10) to (5:6) . The pharmaceutical composition according to claim 1, characterized in that the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadienolignan is (7:50) ~ (4:5) . The pharmaceutical composition according to claim 1, characterized in that the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is 3:20, 1:5, 81:250 Or 81:125. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is an oral preparation. A single dosage form, characterized by comprising: a compound represented by formula I or a salt thereof and biphenylcyclooctadienolignan
The single dosage form according to claim 12, characterized in that the salt of the compound represented by formula I includes at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate, benzene Formate, adipate, succinate, methanesulfonate and maleate. The single dosage form according to claim 12, characterized in that the salt of the compound represented by Formula I is a hydrochloride. The single dosage form according to claim 12, characterized in that the salt of the compound represented by formula I is (R)-4-(3-chloro-2-fluorophenylamino)-7-methoxyquinazole Phin-6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride. The single dosage form according to claim 12, wherein the biphenylcyclooctadiene lignans include at least one of the following: schisandrin A, schisandrin ester A, schisandrol acetate, schisandrin ester A, schisandrin Bactin and Schizandrol. The single dosage form according to claim 12, characterized in that the biphenylcyclooctadiene lignan is schisandrin A. The single-dose form according to claim 12, characterized in that the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4:3). The single-dose form according to claim 12, characterized in that the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (1:10) to (5:6). The single-dose form according to claim 12, characterized in that the mass ratio of the compound represented by Formula I or its salt to biphenylcyclooctadiene lignan is (7:50) to (4:5). The single dosage form according to claim 12, characterized in that the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is 3:20, 1:5, 81:250 or 81:125. The single dosage form according to claim 12, characterized in that the single dosage form includes 200 to 300 mg of the compound represented by Formula I or a salt thereof. The single dosage form according to claim 12, wherein the single dosage form is an oral preparation. A drug combination, characterized by comprising: a compound represented by formula I or a salt thereof as the first active ingredient and biphenylcyclooctadienolignan as the second active ingredient
A pharmaceutical kit, characterized by comprising: a compound represented by Formula I or a salt thereof as the first active ingredient and biphenylcyclooctadienolignan as the second active ingredient
The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, wherein the salt of the compound represented by formula I includes at least one of the following: sulfate, phosphate, citrate, Tartrate, fumarate, benzoate, adipate, succinate, methanesulfonate and maleate. The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, wherein the salt of the compound represented by formula I is a hydrochloride. The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, characterized in that the salt of the compound represented by formula I is (R)-4-(3-chloro-2-fluorophenylamino) )-7-methoxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride. The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, wherein the biphenylcyclooctadiene lignans include at least one of the following: schisandrin A, schisandrin A, Schisandrin alcohol, Schisandra alcohol A, Schisandrin B and Schisandra phenol. The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, wherein the biphenylcyclooctadienol is Schisandrin A. The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, wherein the compound represented by formula I The mass ratio of the substance or its salt to biphenylcyclooctadiene lignan is (1:31) ~ (4:3). The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, characterized in that the mass ratio of the compound represented by formula I or its salt and biphenylcyclooctadienolignan is (1: 10)～(5:6). The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, characterized in that the mass ratio of the compound represented by formula I or its salt and biphenylcyclooctadienolignan is (7: 50)～(4:5). The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, characterized in that the mass ratio of the compound represented by formula I or its salt and biphenylcyclooctadienolignan is 3:20 , 1:5, 81:250 or 81:125. The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, characterized in that the compound represented by formula I or its salt and biphenylcyclooctadiene lignan are prepared together or separately. The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, characterized in that the compound represented by formula I or its salt and biphenylcyclooctadienolignan are used simultaneously or separately. The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, wherein the biphenylcyclooctadienolignan is administered prior to the compound represented by formula I or its salt. The pharmaceutical combination according to claim 24 or the pharmaceutical kit according to claim 25, wherein the second active ingredient is in an oral dosage form. A method of administering a compound represented by formula I or a salt thereof, which is characterized in that the compound represented by formula I or a salt thereof is combined with biphenylcyclooctadienolignans
A method for extending the half-life of a compound represented by Formula I or a salt thereof, which is characterized in that the compound represented by Formula I or a salt thereof is combined with biphenylcyclooctadiene lignan
A method for reducing the compound represented by formula I or a salt thereof to produce a compound represented by formula II, characterized in that the compound represented by formula I or a salt thereof is combined with biphenylcyclooctadienolignans
The method according to any one of claims 39 to 41, wherein the combined medication includes administering the compound represented by formula I or its salt and biphenylcyclooctadiene lignan simultaneously or separately. The method according to any one of claims 39 to 41, characterized in that the salt of the compound represented by formula I includes at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate acid salts, benzoates, adipates, succinates, mesylates and maleates. The method according to any one of claims 39 to 41, characterized in that the salt of the compound represented by formula I is a hydrochloride. The method according to any one of claims 39 to 41, characterized in that the salt of the compound represented by formula I is (R)-4-(3-chloro-2-fluorophenylamino)-7-methyl Oxyquinazolin-6-yl 2,4-dimethylpiperazine-1-carboxylate hydrochloride. The method according to any one of claims 39 to 41, characterized in that the biphenylcyclooctadiene lignans include at least one of the following: schisandrin A, schisandrin ester A, schisandra alcohol, schisandrin Alcohol A, Schisandrin B and Schisandrol. The method according to any one of claims 39 to 41, characterized in that the biphenylcyclooctadienolignan is Schisandrin A. The method according to any one of claims 39 to 41, characterized in that the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (1:31) to (4 :3). The method according to any one of claims 39 to 41, characterized in that the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (1:10) to (5 :6). The method according to any one of claims 39 to 41, characterized in that the mass ratio of the compound represented by formula I or its salt to biphenylcyclooctadiene lignan is (7:50) to (4 :5). The method according to any one of claims 39 to 41, characterized in that the mass ratio of the compound represented by formula I or its salt and biphenylcyclooctadiene lignan is 3:20, 1:5, 81:250 or 81:125. The method according to any one of claims 39 to 41, wherein the biphenylcyclooctadienolignan is administered prior to the compound represented by Formula I or a salt thereof. The method according to any one of claims 39 to 41, wherein the biphenylcyclooctadiene lignan is administered orally. The method according to any one of claims 39 to 41, characterized in that the dosage of biphenylcyclooctadiene lignan is 0 to 25 mg/kg, preferably 5 to 25 mg/kg.