DNA alkylating agent prodrug comprising aziridine structure in combination with Anti-allergic drug for treating tumor and cancer

Abstract

Use of a drug containing an AKR1C3 enzyme-activated DNA alkylating agent prodrug compound comprising an aziridine structure and a salt thereof, an ester thereof, a solvate thereof, and an isotopomer thereof in combination with an anti-allergic drug / non-steroidal analgesic drug for treating cancer and tumor patients, and use of the drug containing the AKR1C3 enzyme-activated DNA alkylating agent prodrug compound comprising the aziridine structure and the salt thereof, the ester thereof, the solvate thereof, and the isotopomer thereof in combination with the anti-allergic drug / non-steroidal analgesic drug in the preparation of a drug for treating a cancer and tumor.

Description

Combination of DNA alkylating agent prodrugs containing aziridine structure and antiallergic drugs for the treatment of tumors and cancer Technical Field

[0001] The present invention relates to a method for treating tumors and cancers, in particular to a method for treating cancers and tumor patients with specific gene expression, and belongs to the field of tumor treatment. Background Art

[0002] AKR1C3 enzyme-activated DNA alkylating agent prodrug compounds containing an aziridine structure, such as AST-3424, are currently undergoing clinical trials. According to patent applications PCT / CN2020 / 101870 and WO2021 / 008520, due to the presence of the aziridine structure, the AST-3424 compound will undergo ring-opening degradation when in contact with water, which will make the injection solution with water as the solvent lack long-term stability and cannot be directly developed into an aqueous solution injection. Therefore, a high-concentration concentrated solution is developed by mixing a C2-C8 monohydric alcohol with a C2-C8 diol or triol; and if necessary, in order to increase the concentration and solubility, other excipients may be added, such as high-molecular polymer surfactants: polyethylene glycol, polyvinyl alcohol, polyvinyl pyrrolidone, poloxamer, and polysorbate.

[0003] In particular, the applicant has discovered through research that DNA alkylating agent prodrug compounds containing an aziridine structure, such as AST-3424, have poor stability after being formulated into aqueous solutions due to their aziridine structure:

[0004] (AST-3424 compound),

[0005] In addition, although the aqueous solutions of some of the above six compounds have a certain stability under certain conditions (low temperature, avoid light), barely meeting the requirements for long-term storage of preparations, the solubility of the above compounds in water is not high. Therefore, whether using a non-aqueous concentrated solution similar to AST-3424 (such as using ethanol + propylene glycol organic solvent or using an injection oil solution, etc.) or a concentrated aqueous solution with the addition of a surfactant, it may involve the addition of organic solvents or surfactants to the injection solution, which may cause patients to have allergies or pain at the injection site.

[0006] Although cancer and tumor patients can be treated clinically with antiallergic drugs to prevent allergies, or with trace amounts of analgesics (usually opioids such as procaine, lidocaine, or NSAIDs such as flurbiprofen axetil) added to injections for pain relief, literature suggests that therapeutic drugs, including these analgesics and antiallergic drugs, may inhibit AKR1C3 (references 1-9), potentially rendering these AKR1C3-activated antitumor and cancer drugs ineffective. Therefore, it is necessary to select suitable antiallergic and analgesic drugs to alleviate the problems of allergies and injection site pain caused by the addition of organic solvents or surfactants to the injections, while also not affecting the activity of AKR1C3-activated DNA alkylating agent prodrug compounds containing an aziridine structure. Summary of the Invention

[0007] Through experiments, the applicant screened a series of anti-allergic drugs and NSAIDs analgesics and found that some drugs, when used in combination with AKR1C3 enzyme-activated DNA alkylating agent anti-tumor prodrugs, would not affect the activity of anti-tumor drugs in inhibiting tumor cell proliferation. Therefore, the combination of such drugs with AKR1C3 enzyme-activated DNA alkylating agent prodrugs is expected to alleviate patients' allergies and pain at the injection site caused by the addition of organic solvents or surfactants to the injection solution, while not affecting the anti-tumor therapeutic effect of AKR1C3 enzyme-activated DNA alkylating agent prodrugs containing an aziridine structure.

[0008] To this end, the following proposal is proposed.

[0009] The treatment method uses an AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, and isotopomers in combination with anti-allergic drugs to treat cancer and tumor patients.

[0010] The invention relates to a use of an AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, and isotope isomers in combination with an anti-allergic drug in the preparation of a drug for treating cancer and tumors.

[0011] The method comprises using an AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, and isotopomers in combination with non-steroidal analgesics to treat cancer and tumor patients.

[0012] Disclosed is a use of an AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, and isotope isomers in combination with a non-steroidal analgesic drug in the preparation of a drug for treating cancer and tumors.

[0013] A DNA alkylating agent prodrug compound refers to a prodrug compound that is metabolized and converted into a DNA alkylating agent, and the DNA alkylating agent eventually undergoes alkylation reaction with DNA in cancer cells to destroy the DNA structure of the cancer cells and ultimately lead to cell death.

[0014] Most prodrug compounds are metabolized and transformed under specific physiological conditions. For anti-tumor and anti-cancer drugs, these physiological conditions are typically microenvironments unique to tumor tissue or cancer cells, such as high expression of certain transport proteins on the cell membrane, high expression or concentration of certain enzymes or proteins in the intracellular or extracellular environment that are higher than normal cells due to enrichment, or conditions such as hypoxia or abnormal pH. Therefore, the existence of these microenvironments is generally believed to be caused by certain specific mechanisms of tumor tissue or cancer cells.

[0015] By activating the metabolism of prodrugs into effective anti-tumor drugs through these tumor microenvironments, targeted treatment of tumor / cancer cells in the above microenvironment is achieved.

[0016] AKR1C3 enzyme activates DNA alkylating agent prodrugs, that is, compounds in the form of prodrugs are catalyzed by AK R1C3 in the biochemical environment of cells to ultimately produce cytotoxic DNA alkylating agent toxins, thereby exerting a cancer cell-killing effect. The experimental test and determination method can be found in WO2022048492A1.

[0017] The AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure refers to a compound that meets the above definition and contains an aziridine in the compound structure. The compound of the structural fragment is specifically selected from the compounds of structural formula (4), (6)-(10).

[0018] Where T is X, Y, Z, R, A, and X 10 The definition is as described in the claims of patent application PCT / US2016 / 021581, publication number WO2016145092A1 (corresponding to Chinese application number 2016800150788, publication number CN107530556A), and the synthesis and preparation methods of specific compounds are also described in the above application, which are hereby incorporated into this application in their entirety, and are specifically defined as:

[0019] X 10 O, S, SO or SO2;

[0020] A is C6-C 10 Aryl, 5-15 membered heteroaryl or -N=CR 1 R 2 ;

[0021] R 1 and R2 are independently hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4-15 membered heterocycle, ether, -CONR 13 R 14 or -NR 13 COR 14 ;

[0022] X, Y and Z are each independently hydrogen, CN, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4-15 membered heterocycle, ether, -CONR 13 R 14 or -NR 13 COR 14 ;

[0023] R is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4-15 membered heterocycle, ether, -CONR 13 R 14 or -NR 13 COR 14 ;

[0024] R 13 and R 14 are independently hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4-15 membered heterocycle or ether;

[0025] and

[0026] These alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclic, heteroaryl, and ether groups are substituted or unsubstituted.

[0027] Among them, R1, R2, R3, R4, R5, R8, R9, R 10 The definition is as described in the claims of patent application PCT / CN2020 / 089692, publication number WO2020228685A9 (corresponding to Chinese application number 2020800358890, publication number CN113853379A), and the synthesis preparation method of the specific compound is also described in the above application, which is hereby incorporated into this application in its entirety, and is specifically defined as:

[0028] R1 is C6-C 10 Aryl or Z-substituted aryl, 4-15 membered heterocyclic or Z-substituted heterocyclic, 5-15 membered heteroaryl or Z-substituted heteroaryl, 7-15 membered fused ring or Z-substituted fused ring;

[0029] R2 is hydrogen, a halogen atom, a cyano group or an isocyano group, a hydroxyl group, a mercapto group, an amino group, OTs, OMS, a C1-C6 alkyl group or a Z-substituted alkyl group, a C2-C6 alkenyl group or a Z-substituted alkenyl group, a C2-C6 alkynyl group or a Z-substituted alkynyl group, a C3-C8 cycloalkyl group or a Z-substituted cycloalkyl group, a C6-C 10 Aryl or Z-substituted aryl, 4-15 membered heterocyclic or Z-substituted heterocyclic, 5-15 membered heteroaryl or Z-substituted heteroaryl, ether of 1-6 carbon atoms or Z-substituted alkoxy of 1-6 carbon atoms, -CONR 6 R 7 、-SO2NR 6 R 7 、-SO2R 6 、-OCOO-R 6 、-COOR 6 、-NR 6 COR 7 、-OCOR 6 、-NR 6 SO2R 7 、-NR 6 SO2NR 6 R 7 Or R2 and the atoms on the R1 group to which it is bonded together form a 7-15 membered fused ring or a Z-substituted fused ring;

[0030] R3 is hydrogen, halogen, cyano or isocyano, hydroxyl, thiol, amine, OTs, OMS, C1-C6 alkyl or Z substituted alkyl, C2-C6 alkenyl or Z substituted alkenyl, C2-C6 alkynyl or Z substituted alkynyl, C3-C8 cycloalkyl or Z substituted cycloalkyl, C6-C 10 Aryl or Z-substituted aryl, 4-15 membered heterocyclic or Z-substituted heterocyclic, 5-15 membered heteroaryl or Z-substituted heteroaryl, C1-C6 alkoxy or Z-substituted C1-C6 alkoxy, -CONR 6 R 7 、-SO2NR 6 R 7 、-SO2R 6 、-OCO-R 6 、-OCOO-R 6 、-COOR 6 、-NR 6 COR 7 , -OCOR 6 、-NR 6 SO2R 7 ;

[0031] R4 and R5 are each independently hydrogen, a halogen atom, a cyano group or an isocyano group, a hydroxyl group, a mercapto group, an amino group, OTs, OLCMS, a C1-C6 alkyl group or a Z-substituted alkyl group, a C2-C6 alkenyl group or a Z-substituted alkenyl group, a C2-C6 alkynyl group or a Z-substituted alkynyl group, a C3-C8 cycloalkyl group or a Z-substituted cycloalkyl group, a C6-C 10 Aryl or Z-substituted aryl, 4-15 membered heterocyclic or Z-substituted heterocyclic, 5-15 membered heteroaryl or Z-substituted heteroaryl, C1-C6 alkoxy or Z-substituted C1-C6 alkoxy, -CONR 6 R 7 、-SO2NR 6 R 7 、-SO2R 6 、-OCOO-R 6 、-COOR 6、 -NR 6 COR 6 、-OCOR 6 、-NR 6 SO2R 7 Or R4, R5 and the atoms on the benzene ring to which they are bonded together form a 7-15 membered fused ring or a Z-substituted fused ring;

[0032] R 6 and R 7 Each is independently hydrogen, cyano or isocyano, C1-C6 alkyl or Z substituted alkyl, C2-C6 alkenyl or Z substituted alkenyl, C2-C6 alkynyl or Z substituted alkynyl, C3-C8 cycloalkyl or Z substituted cycloalkyl, C6-C 10 Aryl or Z-substituted aryl, 4-15 membered heterocyclic or Z-substituted heterocyclic, 5-15 membered heteroaryl or Z-substituted heteroaryl, C1-C6 alkoxy or Z-substituted C1-C6 alkoxy, or R 6 、R 7 The group and the atoms to which it is bonded together form a 5-7 membered heterocyclic group or a Z-substituted 5-7 membered heterocyclic group;

[0033] R8, R 10 Each is independently hydrogen, deuterium, aryl or Z-substituted aryl, C1-C6 alkyl or Z-substituted alkyl, C2-C6 alkenyl or Z-substituted alkenyl, C2-C6 alkynyl or Z-substituted alkynyl, C3-C8 cycloalkyl or Z-substituted cycloalkyl, and at least one of them must be hydrogen or deuterium;

[0034] R9 is a substituted C6-C 10 aryl, substituted 4- to 15-membered heterocyclic ring having at least one fluorine atom or nitro group, substituted 5- to 15-membered heteroaryl group having at least one fluorine atom or nitro group.

[0035] The Z substituent is a halogen atom, a cyano group or an isocyano group, a hydroxyl group, a thiol group, an amine group, OTs, OMS, a C1-C3 alkyl group or a substituted alkyl group, a C1-C3 alkoxy group or a substituted alkoxy group, a C2-C3 alkenyl group or a substituted alkenyl group, a C2-C3 alkynyl group or a substituted alkynyl group, a C3-C8 cycloalkyl group or a substituted cycloalkyl group, an aromatic ring, a heterocyclic ring, a heteroaromatic ring and a condensed ring or a substituted aromatic ring, a heterocyclic ring, a heteroaromatic ring and a condensed ring, and the substitution is monosubstitution or geminal disubstitution;

[0036] Substitution of C6-C in R9 10 The substituents of the aryl group, the substituted 4-15 membered heterocyclic ring, and the substituted 5-15 membered heteroaryl group are halogen atoms, nitro groups, cyano groups or isocyano groups, hydroxyl groups, amino groups, C1-C3 alkyl groups or alkoxy groups, alkenyl groups, alkynyl groups, cycloalkyl groups or benzene rings, substituted benzene rings, C1-C3 alkoxy groups, or halogen atom-substituted alkoxy groups.

[0037] in:

[0038] A is a substituted or unsubstituted C6-C10 aryl, biaryl or substituted biaryl, 5-15 membered heteroaryl or -N=CR 1 R 2 , wherein the substituent when substituted is selected from the group consisting of: halogen, -CN, -NO2, –O-(CH2)-O-, -CO2H and its salts, -OR 100 、-CO2R 100 、-CONR 101 R 102 、-NR 101 R 102 、-NR 100 SO2R 100 、-SO2R 100 、-SO2NR 101 R 10 2 , C1-C6 alkyl, C3-C10 heterocyclic group;

[0039] Among them, R 100 、R 101 and R 102 are independently hydrogen, C1-C8 alkyl, C6-C12 aryl; or R 101 and R 102 together with the nitrogen atom to which it is attached, form a 5-7 membered heterocyclic ring;

[0040] wherein the alkyl group and the aryl group are each substituted with 1-3 halo groups or 1-3 C1-C6 alkyl groups;

[0041] R 1 and R 2 each independently phenyl or methyl;

[0042] X, Y and Z are each independently hydrogen or halo;

[0043] R is hydrogen or C1-C6 alkyl or halogen-substituted alkyl.

[0044] Wherein, Rw is defined as described in the claims of patent application PCT / CN2020 / 120281, publication number WO2021068952A1 (corresponding to Chinese application number 202080071652.8, publication number CN114555574A), and the synthesis and preparation methods of specific compounds are also described in the above-mentioned application, which are hereby incorporated into this application in their entirety, and are specifically defined as:

[0045] Rw is

[0046] R1 is H, C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocycloalkyl, 5-6 membered heteroaryl or phenyl, wherein the C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocycloalkyl, 5-6 membered heteroaryl and phenyl are optionally substituted by 1, 2 or 3 R a replaced by;

[0047] Each R a are independently H, F, Cl, Br, I, -CN, -OH, C 1-3 Alkoxy or C 1-3 alkyl;

[0048] R2 is H or C 1-6 alkyl;

[0049] Or R1 and R2 are linked together to form a 4-6 membered heterocycloalkyl group, wherein the 4-6 membered heterocycloalkyl group is optionally substituted by 1, 2 or 3 R b replaced by;

[0050] Each R b are independently H, F, Cl, Br, I, -CN, -OH, -NH2, -OCH3, -OCH2CH3, -CH3 or -CH2CH3;

[0051] R3 is H, F, Cl, Br, I, -OH, -NH2, C 1-3 Alkoxy or C 1-3 alkyl;

[0052] Or R2 and R3 are connected together to make the structural unit for

[0053] T1 is -(CR c R d ) m -or-(CR c R d ) n -O-;

[0054] m is 1, 2, or 3;

[0055] n is 1 or 2;

[0056] T2 is N or CH;

[0057] R c and R d Each independently is H, F, C 1-3 Alkyl or C 1-3 alkoxy;

[0058] R4, R5 and R6 are each independently H, F, Cl, Br, I, C 1-3 Alkyl or C 1-3 alkoxy;

[0059] T is N or CH;

[0060] R7 and R8 are each independently H, F, Cl, Br or I;

[0061] R9 and R 10 Each independently is H, F, Cl, Br, I, -CN or

[0062] The 4-6 membered heterocycloalkyl and 5-6 membered heteroaryl each contain 1, 2, 3 or 4 heteroatoms independently selected from N, -O- and -S-.

[0063] Wherein, the definitions of R1, R2, R3, R4, and T are as described in the claims of patent application PCT / CN2021 / 118597, publication number WO2022057838A1. The synthesis and preparation methods of specific compounds are also described in the above-mentioned application, which are hereby incorporated into this application in their entirety and are specifically defined as:

[0064] T is N or CH;

[0065] R1 and R2 are each independently H, F, Cl, Br, I or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 R a replaced by;

[0066] Each R a independently F, Cl, Br, I, -CN, -OH or -NH2;

[0067] R3 and R4 are each independently H, F, Cl, Br, I, CN, C 1-3 Alkyl, C 1-3 Alkoxy, Among them, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 R e replaced by;

[0068] R b and R c Each independently represents H, -CH3, -CH2CH3, -(CH2)2CH3, -CH(CH3)2;

[0069] R d It is -CH3, -CH2CH3, -(CH2)2CH3, -CH(CH3)2;

[0070] Each R e independently F, Cl, Br, I, -CN, -OH or -NH2.

[0071] The compound of structural formula (4) is selected from the following structural compounds:

[0072] (AST-3424 compound),

[0073] The synthetic preparation method of the compound is also described in patent application PCT / US2016 / 021581, publication number WO2016145092A1 (corresponding to Chinese application number 2016800150788, publication number CN107530556A), which is hereby incorporated into the present application in its entirety.

[0074] The compounds of structural formula (6) and (7) are selected from the following structural compounds:

[0075] (AST compound, compound A),

[0076] The synthetic preparation method of the compound is also recorded in patent application PCT / CN2020 / 089692, publication number WO2020228685A9 (corresponding to Chinese application number 2020800358890, publication number CN113853379A), the full text of which is hereby introduced into this application.

[0077] The compound of structural formula (8) is selected from the following structural compounds:

[0078] (AST compound, compound A). The compound of structural formula (9) is selected from the following structural compounds:

[0079] The synthetic preparation method of the compound is also recorded in patent application PCT / CN2020 / 120281, publication number WO2021068952A1 (corresponding to Chinese application number 202080071652.8, publication number CN114555574A), the full text of which is hereby introduced into this application.

[0080] The compound of structural formula (10) is selected from the following structural compounds:

[0081] The synthetic preparation method of the compound is also described in patent application PCT / CN2021 / 118597, publication number WO2022057838A1, the entire text of which is incorporated herein by reference.

[0082] The drugs described in this article refer to the compounds corresponding to the drugs, preparations or raw materials. When referring to drugs or preparations, the prepared drugs contain the active ingredient compounds (raw materials) or their salts, esters, solvates, isotope isomers within a specific dosage range, and / or the prepared drugs are in a specific dosage form and administered by a specific method.

[0083] Regarding the compounds described herein, if their chemical structures contain organic amine structures and P=O double bond structures, the compounds may also be administered in the form of salts. That is, the present invention provides pharmaceutically acceptable salts of the compounds, which may be basic salts, including salts formed with inorganic bases (e.g., alkali metal hydroxides, alkaline earth metal hydroxides, etc.) or with organic bases (e.g., monoethanolamine, diethanolamine, or triethanolamine, etc.). Alternatively, the salts may be acidic salts, including salts formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, perchloric acid, sulfuric acid, or phosphoric acid, etc.) or with organic acids (e.g., methanesulfonic acid, trifluoromethanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, fumaric acid, oxalic acid, maleic acid, citric acid, etc.). Similarly, they may react with certain acids or alcohols to form esters, and thus the compounds may also be administered in the form of esters.

[0084] Similarly, the above-mentioned compound concept also includes various crystal forms of the compound, such as PCT application PCT / CN2023 / 080261 and publication WO2023169462A1, which disclose a crystal form of an AKR1C3-activated anticancer prodrug compound:

[0085] For various reasons, these compounds may also form solvates with certain solvents, such as hydrates or alcoholates, and thus the compounds may also be administered in the form of solvates. The selection and preparation of acceptable salts, esters and solvates of compounds are well known in the art.

[0086] The term "isotopic variant" refers to a compound that contains unnatural proportions of isotopes at one or more of the atoms that constitute such compound. In certain embodiments, an "isotopic variant" of a compound contains unnatural proportions of one or more isotopes, including but not limited to hydrogen ( 1 H), deuterium ( 2 H), tritium ( 3 H), carbon-11 ( 11 C), carbon-12 ( 12 C), carbon-13 ( 13 C), carbon-14 ( 14 C), nitrogen-13 ( 13 N), nitrogen-14 ( 14 N), nitrogen-15 ( 15 N), oxygen-14 ( 14 O), oxygen-15 ( 15 O), oxygen-16 ( 16 O), oxygen-17 ( 17 O), oxygen-18 ( 18 O), fluorine-17 ( 17 F), fluorine-18 ( 18 F), phosphorus-31 ( 31 P), phosphorus-32 ( 32 P), phosphorus-33 ( 33 P), sulfur-32 ( 32 S), sulfur-33 ( 33 S), sulfur-34 ( 34 S), sulfur-35 ( 35 S), sulfur-36 ( 36 S), chlorine-35 ( 35 Cl), chlorine-36 ( 36 Cl), chlorine-37( 37 Cl), bromine-79( 79 Br), bromine-81( 81 Br), iodine-123 ( 123 I), iodine-125( 125 I), iodine-127( 127 I), iodine-129( 129 I) and iodine-131( 131 I). In certain embodiments, an "isotopic variant" of a compound is a stable form, i.e., non-radioactive. In certain embodiments, an "isotopic variant" of a compound contains unnatural proportions of one or more isotopes, including but not limited to hydrogen ( 1 H), deuterium ( 2 H), carbon-12 ( 12 C), carbon-13 ( 13C), nitrogen-14 ( 14 N), nitrogen-15 ( 15 N), oxygen-16 ( 16 O), oxygen-17 ( 17 O), oxygen-18 ( 18 O), fluorine-17 ( 17 F), phosphorus-31 ( 31 P), sulfur-32 ( 32 S), sulfur-33 ( 33 S), sulfur-34 ( 34 S), sulfur-36 ( 36 S), chlorine-35 ( 35 Cl), chlorine-37( 37 Cl), bromine-79( 79 Br), bromine-81( 81 Br) and iodine-127( 127 I). In certain embodiments, an "isotopic variant" of a compound is an unstable form, i.e., radioactive. In certain embodiments, an "isotopic variant" of a compound contains unnatural proportions of one or more isotopes, including but not limited to tritium ( 3 H), carbon-11 ( 11 C), carbon-14 ( 14 C), nitrogen-13 ( 13 N), oxygen-14 ( 14 O), oxygen-15 ( 15 O), fluorine-18 ( 18 F), phosphorus-32 ( 32 P), phosphorus-33 ( 33 P), sulfur-35( 35 S), chlorine-36 ( 36 Cl), iodine-123 ( 123 I), iodine-125( 125 I), iodine-129( 129 I) and iodine-131( 131 I) It is understood that in the compounds provided herein, any hydrogen may be replaced by, for example, 2 H is D, or any carbon can be, for example 13 C, or any nitrogen may be e.g. 15 N, and any oxygen may be 18 O. In certain embodiments, an "isotopic variant" of a compound contains unnatural proportions of deuterium (D).

[0087] In addition to containing the compounds of formula (4), (6)-(10), the above-mentioned drugs should also be added with pharmaceutically acceptable excipients or vehicles according to the specific characteristics of the drug, medicine, or preparation. The drug can be in any dosage form for clinical administration, such as tablets, suppositories, dispersible tablets, enteric-coated tablets, chewable tablets, orally disintegrating tablets, capsules, sugar-coated tablets, granules, dry powders, oral solutions, small needles for injection, freeze-dried powder for injection, or large infusions. Depending on the specific dosage form and administration method, the pharmaceutically acceptable excipients or vehicles in the drug can include one or more of the following: diluents, solubilizers, disintegrants, suspending agents, lubricants, adhesives, fillers, flavoring agents, sweeteners, antioxidants, surfactants, preservatives, encapsulating agents, and pigments.

[0088] "Cancer" refers to leukemias, lymphomas, carcinomas, and other malignant tumors (including solid tumors) or proliferations (malignant hyperplasias) with the potential for unrestricted growth that can spread locally by invasion and systemically by metastasis, and can include, but are not limited to, cancers of the adrenal glands, bones, brain, breast, bronchi, colon and / or rectum, gall bladder, head and neck, kidney, larynx, liver, lung, nervous tissue, pancreas, prostate, parathyroid gland, skin, stomach, and thyroid gland. Some other examples of cancer include acute and chronic lymphocytic and granulocytic tumors, adenocarcinomas, adenomas, basal cell carcinomas, dysplastic epithelial cells and carcinoma in situ of the cervix, Ewing's sarcoma, epidermoid carcinoma, giant cell tumors, glioblastoma multiforme, hair cell tumors, intestinal ganglioneuromas, proliferative corneal nerve tumors, pancreatic islet cell carcinoma, Kaposi's sarcoma, leiomyoma, leukemias, lymphomas, malignant carcinoid tumors, malignant melanomas, malignant hypercalcemia, Marfanoid bodies, tumors, medullary epithelial carcinoma, metastatic skin cancer, mucosal neuroma, myeloma, mycosis fungoides, neuroblastoma, osteosarcoma, osteogenic and other sarcomas, ovarian tumor, pheochromocytoma, polycythemia vera, primary brain tumor, small cell lung cancer, squamous cell carcinoma of both ulcerative and papillary types, hyperplasia, seminoma, soft tissue sarcoma, retinoblastoma, rhabdomyosarcoma, renal cell tumor, localized skin lesions, reticulum cell sarcoma, and Wilm's tumor.

[0089] The recommended dosage and dosage form of AST-3424 or this type of AKR1C3 enzyme-activated DNA alkylating agent prodrug for the treatment of cancer can refer to the patent application texts submitted by Threshold and Ascentawits, OBI, Vybio and other companies (such as WO2017087428A1, WO2017087428A1, WO2019062919A1, WO2021008520A1) and clinical trials registered with the FDA and NMPA (CTR20201915, CTR20201908, CTR20191399, CTR20191371, CTR20220957 and NCT04315324, NCT03592264).

[0090] Allergic reactions, also known as hypersensitivity reactions, are pathological immune responses to allergens in the body. Common diseases include anaphylactic shock, bronchial asthma, allergic rhinitis, urticaria, eczema, etc. Since the anticancer drug involved in this application, an aziridine-containing AKR1C3 enzyme-activating DNA alkylating agent prodrug, is administered as an injection, and the allergen is the surfactant added to the injection, it is necessary to administer an anti-allergy drug before administering the anticancer drug to prevent allergies.

[0091] Antiallergic drugs refer to drugs used to prevent and treat allergic reaction diseases or symptoms. Antiallergic drugs are generally divided into four categories based on their mechanism of action: antihistamines, allergic reaction mediator blockers, calcium supplements, and immunosuppressants.

[0092] Antihistamines include histamine H1 receptor blockers and histamine H2 receptor blockers. Histamine H1 receptor blockers include diphenhydramine, promethazine, cetirizine, loratadine, desloratadine, fexofenadine, efletirizine, chlorpheniramine, cyproheptadine, and terfenadine. Antihistamine H2 receptor antagonists include cimetidine, ranitidine, and famotidine.

[0093] Allergic reaction mediator blockers, also known as mast cell stabilizers, mainly include sodium cromoglycate, sodium cromolyn, ketotifen, etc.

[0094] Calcium supplements, including calcium gluconate and calcium chloride, increase the density of capillaries, reduce permeability, and thus reduce exudation, thereby alleviating or relieving allergic symptoms. They are usually injected intravenously and are effective quickly.

[0095] Immunosuppressants have nonspecific inhibitory effects on the body's immune function and are effective against all types of allergic reactions. They include adrenal cortical hormones such as Prednisone, Dexamethasone, Hydrocortisone, Methylprednisolone, as well as cyclophosphamide and thioguanine.

[0096] Combination therapy refers to the use of two or more drugs simultaneously or successively in one course of treatment.

[0097] Generally speaking, combination therapy requires exploring different dosages and dosing cycles based on the characteristics of the disease and the types of combined drugs. Only based on the above conditions can the combination drug treatment plan explored achieve better therapeutic effects than single drug treatment.

[0098] The drug dosage and administration cycle of the combination therapy need to be explored through clinical trials.

[0099] Through experiments, it was found that the following compounds, when used in combination with AKR1C3 enzyme-activated DNA alkylating agent prodrug compounds containing an aziridine structure, had no effect on the inhibition of cancer cell proliferation, or had negligible inhibition of proliferation:

[0100] Diphenhydramine, Promethazine, Cetirizine, Loratadine, Desloratadine, Fexofenadine, Cimetidine, Ranitidine, Famotidine, Hydrocortisone, Methylprednisolone.

[0101] Therefore, when using the above-mentioned AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, and isotopomers to treat cancer and tumors, drugs containing the above-mentioned anti-allergic drugs and their salts, esters, solvates, and isotopomers can be used to prevent or treat allergies that may be caused.

[0102] Obviously, in order to prevent allergies, the antiallergic drug is administered before the drug of the AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, isotope isomers.

[0103] Drugs containing AKR1C3 enzyme-activated DNA alkylator prodrug compounds and their salts, esters, solvates, and isotopomers contain surfactants, organic solvents, or oils for enhancing drug solubility. Such surfactants are highly likely to be the cause of allergic reactions when injecting AKR1C3 enzyme-activated DNA alkylator prodrug anticancer drugs.

[0104] Studies have shown that surfactants, organic solvents, and oils used in pharmaceuticals to increase the solubility of the API or system stability, particularly in oil-soluble injections, organic solvent injections, emulsion injections, and microsphere injections developed due to insufficient or unstable water solubility, can trigger allergic reactions after injection into the human body. Common allergic reactions in this setting include hypotension, bronchospasm, facial flushing, rash, dyspnea or even respiratory failure, tachycardia, fever, and chills. While this application proposes the use of the aforementioned specific antiallergic drugs for pre-administration to prevent or treat allergies caused by the surfactants, organic solvents, or oils contained in the formulation when administering the AKR1C3 enzyme-activating DNA alkylating agent prodrug, and while this does not affect the cancer and tumor treatment efficacy of the AKR1C3 enzyme-activating DNA alkylating agent prodrug, it cannot completely prevent allergic reactions in patients after the combined use of the aforementioned two drugs. The combination regimen proposed in this application can only reduce the probability of allergies or alleviate the severity of allergies. If allergic conditions or symptoms occur, medical institutions and clinical professionals should choose allergy drugs for treatment according to the situation, without having to stick to the above-mentioned specific drugs.

[0105] In order to improve the solubility and stability of the AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure in water or organic solvents, the surfactant used in the injection is a non-ionic surfactant, preferably a non-ionic surfactant with a high molecular weight structure, such as polyoxyethylene alkyl ethers alkylphenol ethoxylates, Tweens, Spans, Cremophor, Poloxamer, Polyvinylpyrrolidone, Polyethylene glycol PEG, and Polyvinyl alcohol PVA.

[0106] To further alleviate pain caused by injections containing surfactants, organic solvents, and oils, analgesics can be used in conjunction with injection. To achieve pain relief without compromising the efficacy of AKR1C3 enzyme-activated DNA alkylating agent prodrugs in treating cancer and tumors, nonsteroidal analgesics (NSAIDs) are commonly used and non-addictive pain relievers. However, studies (references 1-9) have shown that some nonsteroidal analgesics are AKR1C3 inhibitors and may affect the efficacy of AKR 1C3 enzyme-activated DNA alkylating agent prodrugs. Therefore, it is necessary to select an appropriate nonsteroidal analgesic and dosing regimen.

[0107] Experiments have shown that drugs containing diclofenac and its salts or esters, flurbiprofen and its salts or esters can be used in combination with drugs containing aziridine-structured AKR1C3 enzyme-activated DNA alkylating agent prodrug compounds and their salts, esters, solvates, and isotopomers to relieve pain. When used in combination, the drugs can be administered simultaneously, in advance, or delayed.

[0108] Experiments have shown that drugs containing Loxoprofen and its salts or esters, and Naproxen and its salts or esters can be used in combination with drugs containing aziridine structures, AKR1C3 enzyme-activated DNA alkylating agent prodrug compounds, and their salts, esters, solvates, and isotopomers to relieve pain. When used in combination, the drugs should be administered after a certain interval of time with the drugs containing aziridine structures, AKR1C3 enzyme-activated DNA alkylating agent prodrug compounds, and their salts, esters, solvates, and isotopomers.

[0109] If a drug containing loxoprofen and its salts or esters, or naproxen and its salts or esters is administered, the drug containing an aziridine structure-activated DNA alkylating agent prodrug compound and its salts, esters, solvates, or isotopomers should be administered after these drugs are metabolized and eliminated; or if a drug containing an aziridine structure-activated DNA alkylating agent prodrug compound and its salts, esters, solvates, or isotopomers is administered, the drug containing loxoprofen and its salts or esters, or naproxen and its salts or esters should be administered only after the aziridine structure-activated DNA alkylating agent prodrug compound and its salts, esters, solvates, or isotopomers are metabolized and eliminated.

[0110] The method comprises using an AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, and isotopomers in combination with non-steroidal analgesics to treat cancer and tumor patients.

[0111] Preferably, the non-steroidal analgesic is selected from diclofenac and its salts or esters, flurbiprofen and its salts or esters, and the non-steroidal analgesic can be administered simultaneously with the AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, isotopomers, or the like, or administered in advance or delayed.

[0112] Disclosed is a use of an AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, and isotope isomers in combination with a non-steroidal analgesic drug in the preparation of a drug for treating cancer and tumors.

[0113] Preferably, the non-steroidal analgesic drug is selected from Loxoprofen and its salts or esters, Naproxen and its salts or esters, and the non-steroidal analgesic drug should be administered at a certain interval with the drug containing an aziridine structure-containing AKR1C3 enzyme-activated DNA alkylating agent prodrug compound and its salts, esters, solvates, and isotopomers. BRIEF DESCRIPTION OF THE DRAWINGS

[0114] Figure 1 shows the in vitro proliferation inhibition curves and IC values ​​of different concentrations of diphenhydramine combined with compound A. 50 result;

[0115] Figure 2 shows the in vitro proliferation inhibition curves and IC values ​​of different concentrations of Cimetidine combined with Compound A. 50 result;

[0116] Figure 3 shows the in vitro proliferation inhibition curves and IC values ​​of different concentrations of Ranitidine combined with Compound A. 50 result;

[0117] Figure 4 shows the in vitro proliferation inhibition curves and IC of different concentrations of promethazine combined with compound A. 50 result;

[0118] Figure 5 shows the in vitro proliferation inhibition curves and IC values ​​of different concentrations of famotidine combined with compound A. 50 result;

[0119] Figure 6 shows the in vitro proliferation inhibition curves and IC values ​​of different concentrations of cetirizine dihydrochloride combined with compound A. 50 result;

[0120] Figure 7 shows the in vitro proliferation inhibition curves and IC values ​​of different concentrations of loratadine combined with compound A. 50 result;

[0121] Figure 8 shows the in vitro proliferation inhibition curves and IC values ​​of different concentrations of Desloratadine combined with Compound A. 50 result;

[0122] Figure 9 shows the in vitro proliferation inhibition curves and IC values ​​of different concentrations of Fexofenadine combined with Compound A. 50 result;

[0123] Figure 10 shows the in vitro proliferation inhibition curves and IC of different concentrations of hydrocortisone combined with compound A. 50 result;

[0124] Figure 11 shows the in vitro proliferation inhibition curves and IC values ​​of different concentrations of methylprednisolone combined with compound A. 50 result;

[0125] Figure 12 shows the in vitro proliferation inhibition curves and IC50 results of different combinations of diclofenac sodium and compound A. From top to bottom, the figure shows four different combination dosing regimens: pretreatment with diclofenac sodium or AST-3021 for 2 hours, followed by washing, and then adding compound A for 2 hours before washing away all the compounds; adding diclofenac sodium or AST-3021 and compound A simultaneously, and then washing away all the compounds after 2 hours; pretreatment with compound A for 2 hours, followed by washing away, and then adding diclofenac sodium or AST-3021 for 2 hours before washing away all the compounds; and the inhibition curves and results of pretreatment with diclofenac sodium or AST-3021 for 2 hours, followed by adding compound A for 2 hours before washing away all the compounds.

[0126] Figure 13 shows the in vitro proliferation inhibition curves and IC50 results of different combinations of loxoprofen sodium and compound A. From top to bottom, the figure shows four different combination dosing regimens: pretreatment with loxoprofen sodium or AST-3021 for 2 hours, followed by washing, and then adding compound A for 2 hours before washing away all the compounds; adding loxoprofen sodium or AST-3021 and compound A simultaneously, and then washing away all the compounds after 2 hours; pretreatment with compound A for 2 hours, followed by washing away, and then adding loxoprofen sodium or AST-3021 for 2 hours before washing away all the compounds; and the inhibition curves and results of pretreatment with loxoprofen sodium or AST-3021 for 2 hours, followed by adding compound A for 2 hours before washing away all the compounds.

[0127] Figure 14 shows the in vitro proliferation inhibition curves and IC50 results of different combinations of Flurbiprofen axetil and Compound A. From top to bottom in the figure are four different combination dosing regimens: Flurbiprofen axetil or AST-3021 pretreatment for 2 hours, followed by washing, and then adding Compound A for 2 hours before all compounds are washed away; Flurbiprofen axetil or AST-3021 and Compound A are added simultaneously, and then all compounds are washed away after 2 hours; Compound A is added first for pretreatment for 2 hours and then washed away, and Flur biprofen axetil or AST-3021 is added for 2 hours and then all compounds are washed away; Flurbiprofen axetil or AST-3021 is pretreated for 2 hours, and then Compound A is added for 2 hours and then all compounds are washed away; the inhibition curves and results are shown for Flurbiprofen axetil or AST-3021 pretreatment for 2 hours, followed by Compound A for 2 hours and then all compounds are washed away;

[0128] Figure 15 shows the in vitro proliferation inhibition curves and IC50 results of different combination orders of naproxen and compound A. From top to bottom in the figure are four different combination dosing regimens: pretreatment with naproxen or AST-3021 for 2 hours, washing off, adding compound A for 2 hours, and then washing away all compounds; adding naproxen or AST-3021 and compound A at the same time, and then washing away all compounds after 2 hours; adding compound A for pretreatment for 2 hours and then washing off, adding naproxen or AST-3021 for 2 hours, and then washing away all compounds; pretreatment with naproxen or AST-3021 for 2 hours, adding compound A for 2 hours, and then washing away all compounds. The inhibition curves and results. DETAILED DESCRIPTION

[0129] The present invention is described below with reference to specific examples. Those skilled in the art will appreciate that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention in any way.

[0130] The experimental methods in the following examples are conventional methods unless otherwise specified. The medicinal materials, reagents, etc. used are all commercially available products unless otherwise specified.

[0131] "Patient" and "subject" are used interchangeably to refer to a mammal in need of cancer treatment. Typically, the patient is a human. Typically, the patient is a human diagnosed with cancer. In certain embodiments, a "patient" or "subject" may refer to a non-human mammal, such as a non-human primate, dog, cat, rabbit, pig, mouse, or rat, used to screen, characterize, and evaluate drugs and therapies.

[0132] "Treatment" or "treating a patient" refers to administering, using or applying to a patient a therapeutically effective amount of a drug related to the present invention.

[0133] "Administering" or "applying" a drug to a patient refers to direct administration or administration (which may be administered or administered to a patient by a medical professional or may be self-administered or administered) and / or indirect administration or administration, which may be the act of prescribing a drug. For example, a physician who instructs a patient to self-administer or administer a drug and / or provides a prescription for a drug to a patient is administering or administering a drug to a patient.

[0134] A "therapeutically effective amount" of a drug refers to an amount of the drug that, when administered or used to a patient suffering from cancer, will have the desired therapeutic effect (e.g., alleviation, amelioration, remission, or elimination of the clinical manifestations of one or more cancers in the patient). The therapeutic effect does not necessarily occur by administering or applying a single dose and may only occur after administering or applying a series of doses. Thus, a therapeutically effective amount can be administered or applied in one or more doses.

[0135] "Treatment" of a condition or patient refers to taking steps to obtain beneficial or desired results (including clinical results). For the purposes of this invention, beneficial or desired clinical results include, but are not limited to, alleviation or amelioration of one or more symptoms of cancer; reduction in the extent of the disease; delay or slowing of disease progression; improvement, remission, or stabilization of the disease state; or other beneficial results. In some instances, treatment of cancer may result in a partial response or stabilization of the disease.

[0136] "Tumor cell" refers to a tumor cell of any appropriate species (eg, mammalian, such as murine, canine, feline, equine, or human).

[0137] The above description of the specific embodiments of the present invention does not limit the present invention. Those skilled in the art can make various changes or modifications based on the present invention. As long as they do not depart from the spirit of the present invention, they should fall within the scope of the claims attached to the present invention.

[0138] Example 1 Solubility and formulation of AST-3424

[0139] The following experiments illustrate the stability and solubility of AST-3424, a DNA alkylating agent prodrug containing an aziridine structure. These data demonstrate that the compound exhibits poor water solubility and poor stability in aqueous solutions, whereas it exhibits good solubility in organic solvents (alcoholic solvents) and excellent solution stability.

[0140] For specific experimental content, please refer to the patent application for stable AST-3424 injection preparation and preparation method, PCT / CN2020 / 101870, corresponding to Example 1 of publication number WO2021008520A1, study on the solubility and solution stability of AST-3424.

[0141] Example 2 Study on the Solubility and Solution Stability of AST-3424 Analog Compound A

[0142] Related substances and content determination: HPLC was used for detection. The detection conditions and parameters were similar to those for AST-3424. See the relevant section in patent publication WO2021 / 008520, corresponding to paragraphs 133-138 of CN 112469394 B. The related substances were determined using the integrated area normalization method, and the content was quantified using the external standard method.

[0143] Compound No. 16 (hereinafter referred to as Compound A) disclosed in patent application PCT / CN2020 / 089692 (publication number WO2020228685A9) and patent application PCT / CN2021 / 129077 was synthesized and prepared according to the descriptions in the above patent applications.

[0144] 2.1 Investigation of the solubility of compound A in different solvents

[0145] The solubility of compound A in single solvent water, acid and alkali aqueous solutions, isopropanol, and methanol was investigated. The test results are shown in Table 1.

[0146] Table 1: Solubility test results of compound A at 25°C (determined according to the Chinese Pharmacopoeia)

[0147] The above investigation results show that compound A has extremely low solubility in water and acid-base aqueous solutions, so it is chosen to be dissolved in a non-aqueous solvent to screen out a better solvent or co-solvent.

[0148] In order to increase the concentration of the API in the injection solution, the solubility of the API in a single excipient was investigated to screen for a better solvent, cosolvent, or solubilizer. The solubility test of Compound A in a single solvent was conducted as follows:

[0149] For each solvent, add a certain amount of API at 25°C and stir. Observe the dissolution of the API in the solvent. If dissolution is complete, continue adding the API until a supersaturated solution is obtained. Samples are taken every 2 hours for filtration and assay. The solubility of each solvent at 25°C is shown in Table 2.

[0150] Table 2: Solubility data of compound A in a single organic solvent at 25°C

[0151] From the above data, it can be seen that compound A has the highest solubility in N,N-dimethylacetamide, exceeding 200 mg / mL, its solubility in polyoxyethylene (35) castor oil is about 20 mg / mL, and its solubility in polyethylene glycol 400 is slightly higher than that in anhydrous ethanol.

[0152] When designing an injection formulation, appropriate excipients must be added. These excipients should not affect the drug's efficacy, interfere with testing, or cause toxicity or significant irritation at concentrations sufficient to identify optimal solvents, cosolvents, or solubilizers. The solubility of Compound A in various excipients was investigated to identify the optimal solvent, cosolvent, or solubilizer.

[0153] Given that Compound A and Compound AST-3424 are structurally similar, both are nitrobenzyl derivatives, and both are linked to an aziridine-structured DNA alkylating agent that can effectively kill tumor cells. Furthermore, both can target and treat tumor tissues or cells that overexpress aldehyde-keto reductase 1C3 (AKR1C3), releasing the aziridine-structured DNA alkylating agent in tumor tissues or cells that overexpress the AKR1C3 enzyme to kill the tumor tissues or cells. Therefore, based on the same formulation formulation development approach, the inventors designed and screened an injection formulation similar to Compound AST-3424 (the preferred formulation for AST-3424: 10 mg / mL Compound AST-3424 + 75% ethanol + 25% propylene glycol) for formulation research on Compound A.

[0154] In addition to the solvent used in the formulation of the reference compound AST-3424, anhydrous ethanol is used as a commonly used and relatively safe non-aqueous solvent injection excipient. In the formulation of the developed product, N,N-dimethylacetamide (DMA) or polyoxyethylene (35) castor oil (ELP) is considered to improve solubility and enhance process convenience. The formulation composition and solubility test results of different excipients for Compound A are shown in Table 3.

[0155] Table 3: Prescription solution without ELP added and its preparation process and phenomena

[0156] Note: F1 is the group that only added anhydrous ethanol, and the other groups added other excipients / solvents on the basis of anhydrous ethanol, among which F2 is the propylene glycol group, F3 is the glycerol group, and F4 is the DMA group.

[0157] According to the formulation design for compound AST-3424, the volume ratio of ethanol to propylene glycol is 3:1. For a 30 mL formulation solution containing 300 mg of Compound A, the amount of propylene glycol used (approximately 7.8 g) is between the amounts used in formulations F2-1 and F2-2. The above experimental observations indicate that Compound A cannot be completely dissolved according to the formulation design for compound AST-3424, necessitating the development of alternative formulations for the preparation of Compound A.

[0158] The above formula was processed to obtain the processed formula. The phenomena before and after the treatment indicate that the surfactant polyoxyethylene (35) castor oil (ELP) and the cosolvent N,N-dimethylacetamide (DMA) facilitate the dissolution of the API. Subsequent research will be conducted using anhydrous ethanol and the cosolvent DMA. Since surfactants can increase the solubility of the API in anhydrous ethanol, preliminary research was conducted using different surfactants.

[0159] 2.2 Stability study of compound A in different solvent systems

[0160] In the formula of Table 3 above, F1, F2-1, F2-2, F3-1, and F3-2 could not be completely dissolved, so they were stirred and treated with ELP respectively, and finally completely dissolved, and finally a new formula was obtained as shown in Table 4 below.

[0161] Table 4: New prescription after adding ELP

[0162] The stability of compound A in the above formulation was investigated. At predetermined time points (0 days, 5 days, and 10 days), 1 mL of sample was taken for HPLC analysis, and its content, total impurities, and moisture data were detected respectively. The different investigation conditions and experimental results are shown in Table 5.

[0163] Table 5: Stability of Compound A in different formulation excipients at 2-8°C

[0164] Note: NA means no placement conditions were involved on day 0.

[0165] The above data show that the content, total impurities and moisture content of compound A are relatively stable when placed at 2-8°C for 10 days in each solvent system.

[0166] The above investigation results show that compound A has extremely low solubility in water and acid-base aqueous solutions, but dissolves well in non-aqueous solvents. The cosolvent ELP can improve the solubility of the compound in the solvent.

[0167] Example 3 Preliminary investigation of the solubility of compound X in different solvents

[0168] Compound X The synthesis was carried out according to WO2021068952.

[0169] The solubility of the compound X in single solvent water, acid and alkali aqueous solutions, isopropanol, and methanol was investigated. The test results are shown in Table 6.

[0170] Table 6: Solubility test results of compound X at 25°C (determined according to the Chinese Pharmacopoeia)

[0171] Since the amount of compound X prepared in this synthesis is small, less than 1g, the 1g:1000mL ratio specified in the Chinese Pharmacopoeia was reduced to 1mg:1mL. That is, 1.00mg was accurately weighed and placed in a 2mL disposable Eppendorf microcentrifuge tube. The Eppendorf microcentrifuge tube was placed on a vibration table, and the solvent was added dropwise using a pipette for solubility testing.

[0172] Ethanol+ELP, isopropanol+ELP, DMF+ELP, and DMA+ELP respectively represent the dissolution of compound X that was not dissolved after 10 μl of ELP was added to the experimental tubes of ethanol, isopropanol, DMF, and DMA in the above table.

[0173] The above experimental results show that compound X has extremely low solubility in water and acid-base aqueous solutions, and slightly higher solubility in organic solvents such as ethanol, isopropanol, DMF, and DMA, but cannot meet the requirements of subsequent formulations. The use of cosolvent ELP can significantly improve the solubility of the drug in organic solvents.

[0174] As shown in Examples 1, 2, and 3, the DNA alkylating agent prodrugs AST-3424, Compound A, and Compound X containing an aziridine structure have poor solubility in water. However, the solubility and stability can be improved by using organic solvents or adding surfactants. Therefore, organic solvents or surfactants should be used as cosolvents to improve solubility when preparing injection preparations.

[0175] Example 4 In vitro cytotoxicity experiment of antiallergic drugs combined with compound A

[0176] Brief description of the experimental process

[0177] 1) Add H460 cell suspension to a 96-well plate, 100 μL per well, with a cell density of 2000 cells / well.

[0178] 2) The cells were cultured in a 37°C, 5% CO2 incubator overnight.

[0179] 3) Compound treatment

[0180] Single drug administration: 24 hours after cell plating, add 99.5 μL of growth medium to each well. Add 0.5 μL of test compound at different concentrations, shake gently to ensure uniform mixing, and then place in a 37°C, 5% CO2 incubator.

[0181] Combination therapy: 24 hours after plating cells, add 99 μL of growth medium to each well. Add 0.5 μL of the combination compound (antiallergic drug) at the desired concentration and gently shake to ensure uniform mixing. After incubation for 2 hours, add 0.5 μL of the test compound (Compound A) at varying concentrations and gently shake to ensure uniform mixing. Then, place the cells in a 37°C, 5% CO2 incubator.

[0182] 4) Place the cell plate in an incubator for 72 hours.

[0183] 5) The cell test plate was placed at room temperature for equilibration for 30 minutes, and 100 μL of culture medium was discarded from each well.

[0184] 6) Add 25 μL of CTG reagent to each well, place on a fast shaker for 2 minutes, and place at room temperature in the dark for 30 minutes.

[0185] 7) Read the chemiluminescent signal value using a multifunctional microplate reader with a reading time of 1000 ms.

[0186] 8) Calculate IC using GraphPad Prism 5 software 50 , using the nonlinear fitting formula to obtain the IC of the compound 50 (half inhibitory concentration).

[0187] During the above experiments, the dosage concentration of antiallergic drugs was designed with reference to the maximum blood drug concentration Cmax in humans reported in the literature, and several multiples of the corresponding molar concentration were used in combination.

[0188] The antiallergic drugs used and related information are shown in Table 7.

[0189] Table 7: Information about antiallergic drugs used in combination studies

[0190] The specific test results are shown in Figures 1 to 11.

[0191] Cancer cell proliferation inhibition experiment after different drugs and different doses combined IC 50 The results are summarized in Table 8 below.

[0192] Table 8: IC values ​​of cancer cell proliferation inhibition after different drugs and different doses of anti-allergic drugs were combined with Compound A 50 result

[0193] Comparison of IC of combination and single-drug compounds 50 The difference is not obvious, that is, each drug pretreated for 2 hours and then combined with compound A has no effect on the toxicity of compound A in H460 cells.

[0194] Example 5 In vitro cytotoxicity experiment of non-steroidal analgesics combined with compound A

[0195] Brief description of the experimental process

[0196] 1) Add H460 cell suspension to a 96-well plate, 100 μL per well, with a cell density of 2000 cells / well.

[0197] 2) The cells were cultured in a 37°C, 5% CO2 incubator overnight.

[0198] 3) Compound treatment

[0199] Single drug application: 24 hours after cell plating, add 99.5 μL of growth medium to each well. Add 0.5 μL of test compound at different concentrations, shake gently to ensure uniform mixing, and then place in a 37°C, 5% CO2 incubator.

[0200] Combination medication:

[0201] Each drug group was divided into 4 combination treatment groups corresponding to 4 plates. 24 hours after cell plating, each well was filled with 99 μL of growth medium. The treatment process for each plate was as follows:

[0202] Plate 1: Add 0.5 μL of AST-3021 / Naproxen / Diclofenac Sodium / Loxoprofen Sodium / Flurbiprofen Axetil to each well and gently shake to ensure uniform mixing. After placing in an incubator for 2 hours, wash away the compounds with culture medium. Then, add 0.5 μL of Compound A at different concentrations and place in an incubator for 2 hours. After that, wash away all the compounds with culture medium (pre-treat with the nonsteroidal anti-inflammatory drug and AKR1C3 inhibitor AST-3021 for 2 hours, wash away the compounds, then add Compound A for 2 hours before washing away all the compounds).

[0203] Plate 2: Simultaneously add 0.5 μL of AST-3021 / Naproxen / Diclofenac Sodium / Loxoprofen Sodium / Flurbiprofen Axetil to each well, then add 0.5 μL of Compound A at different concentrations. Place the plates in the incubator for 2 hours, then wash away all compounds with culture medium (the nonsteroidal anti-inflammatory drug and AKR1C3 inhibitor AST-3021 is added simultaneously with Compound A, and after 2 hours, all compounds are washed away).

[0204] Plate 3. Add 0.5 μL of Compound A at different concentrations to each well and shake gently to ensure uniform mixing. After placing in the incubator for 2 hours, wash away the compound with culture medium. Then add 0.5 μL of AST-3021 / Naproxen / Diclofenac Sodium / Loxoprofen Sodium / Flurbiprofen Axetil to each well and place in the incubator for 2 hours. Wash away the compound with culture medium (pre-treat with Compound A for 2 hours, then wash away the compound. Then add the non-steroidal anti-inflammatory drug and AKR1C3 inhibitor AST-3021 for 2 hours before washing away all compounds).

[0205] Plate 4. Add 0.5 μL of AST-3021 / Naproxen / Diclofenac Sodium / Loxoprofen sodium / Flurbiprofen axetil to each well and gently shake to ensure uniform mixing. Place in the incubator for 2 hours, then add 0.5 μL of Compound A at different concentrations, incubate in the incubator for 2 hours, and then wash away the compounds with culture medium (pre-treat with the non-steroidal anti-inflammatory drug and AKR1C3 inhibitor AST-3021 for 2 hours, add Compound A for 2 hours, and then wash away all compounds).

[0206] 4) Place the cell plate in an incubator for 72 hours.

[0207] 5) The cell test plate was placed at room temperature for 30 minutes to equilibrate, and 100 μL of culture medium was discarded from each well.

[0208] 6) Add 25 μL of CTG reagent to each well, place on a fast shaker for 2 minutes, and place at room temperature in the dark for 30 minutes.

[0209] 7) Read the chemiluminescent signal value using a multifunctional microplate reader with a reading time of 1000 ms.

[0210] 8) Calculate IC using GraphPad Prism 5 software 50 , using the nonlinear fitting formula to obtain the IC of the compound 50 (half inhibitory concentration).

[0211] During the above experiments, the dosing concentrations of AST-3021 / Naproxen / Diclofenac Sodium / Loxoprofen Sodium / Flurbiprofen axetil were designed with reference to the maximum blood drug concentration Cmax in humans reported in the literature, and were used in combination using several multiples of the corresponding molar concentrations.

[0212] The nonsteroidal analgesics used and related information are shown in Table 9.

[0213] Table 9: Information on nonsteroidal analgesics selected for combination studies

[0214] AST-3021 is a selective AKR1C3 inhibitor, which is compound 36 in Flanagan et al., Bioorganic and Medicinal Chemistry (2014) pp. 967-977.

[0215] The specific test results are shown in Figures 12 to 15.

[0216] The experimental data in the second graph corresponding to plate 2 in Figure 12 is abnormal due to incorrect dosing. Although there are no reliable experimental results for this plate, based on the results of the other three plates, especially the fourth plate (the fourth graph), it can be inferred that the combination of diclofenac sodium and Compound A in different drug treatment sequences has no significant effect on the in vitro cytotoxicity of Compound A and does not need to be repeated.

[0217] IC of different drugs and different administration sequences in combination for cancer cell proliferation inhibition experiments 50 The results are summarized in Table 10 below.

[0218] Table 10: IC values ​​of cancer cell proliferation inhibition after different drugs and different doses of anti-allergic drugs were combined with Compound A 50 result

[0219] Comparison of IC values ​​of combined and single compounds 50 It can be seen from the values ​​that for Diclofenac Sodium and Flurbiprofen axetil, although the order of adding drugs is different in different combination regimens, the IC 50 The difference in the values ​​is not obvious, so it can be determined that different dosing orders have no effect on the cytotoxicity of compound A, that is, simultaneous administration, early administration, or delayed administration will not affect the cancer cell proliferation inhibition effect of compound A when used in combination.

[0220] By comparing Loxoprofen sodium and Naproxen, we can see that:

[0221] Loxoprofen sodium combined with compound A or pretreated for 2 hours before combined with compound A inhibited the in vitro cytotoxicity of compound A, IC 50 The difference is 3-4 times; pretreatment with Loxoprofen sodium or compound A for 2 hours, followed by washing and then combined use, has no effect on the in vitro cytotoxicity of compound A; this suggests that the simultaneous presence of Loxoprofen sodium and compound A inhibits the in vitro cytotoxicity of compound A.

[0222] After pretreatment with naproxen for 2 hours, it was washed off and compound A was added again, which had no effect on the in vitro cytotoxicity of compound A; however, the simultaneous addition of naproxen and compound A inhibited the in vitro cytotoxicity of compound A; pretreatment with compound A for 2 hours, washed off and naproxen had no effect on the in vitro cytotoxicity of compound A; pretreatment with naproxen for 2 hours and then co-treatment with compound A for 2 hours inhibited the in vitro cytotoxicity of compound A, that is, when naproxen and compound A are present at the same time, the in vitro cytotoxicity of compound A is inhibited.

[0223] Examples 4 and 5 demonstrate that the combined use of Compound A, a DNA alkylating agent prodrug containing an aziridine structure, with certain specific antiallergic drugs does not inhibit the in vitro cancer cell proliferation activity of Compound A, regardless of the order of administration. However, when combined with certain non-steroidal analgesics, the in vitro cancer cell proliferation activity of Compound A may be significantly affected, and the administration time intervals of the respective drugs must be separated.

[0224] References:

[0225] 1. Higaki Y, Usami N, Shintani S, Ishikura S, El-Kabbani O, Hara A. Selective and potent inhibitors of human 20alpha-hydroxysteroid dehydrogenase (AKR1C1) that metabolizes ne urosteroids derived from progesterone.Chem Biol Interact.2003;143-144:503-513.doi:10.1016 / s0009-2797(02)00206-5

[0226] 2、Bydal P,Luu-The V,Labrie F,Poirier D.Steroidal lactones as inhibitors of 17beta-h ydroxysteroid dehydrogenase type 5:chemical synthesis,enzyme inhibitory activity,and asses sment of estrogenic and androgenic activities.Eur J Med Chem.2009;44(2):632-644.doi:10.1016 / j.ejmech.2008.03.020

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Claims

1. A method of treating cancer and tumor patients using AKR1C3 enzyme-activated DNA alkylating agent prodrug compounds containing an aziridine structure and their salts, esters, solvates, and isotope isomers in combination with antiallergic drugs.

2. Use of AKR1C3 enzyme-activated DNA alkylating agent prodrug compounds containing aziridine structures and their salts, esters, solvates, isotope isomers in combination with antiallergic drugs in the preparation of drugs for treating cancer and tumors.

3. Treatment method, using AKR1C3 enzyme-activated DNA alkylating agent prodrug compounds containing aziridine structure and their salts, esters, solvates, isotope isomers in combination with non-steroidal analgesics to treat cancer and tumor patients.

4. Use of AKR1C3 enzyme-activated DNA alkylating agent prodrug compounds containing aziridine structures and their salts, esters, solvates, isotope isomers in combination with non-steroidal analgesics in the preparation of drugs for treating cancer and tumors.

5. The method or use according to any one of claims 1 to 4, wherein: The AKR1C3 enzyme activated DNA alkylating agent prodrug compound is selected from the structural formulas (4), (6)-(10): Where T is X, Y, Z, R, A, and X 10 The definition is as set forth in the claims of patent application PCT / US2016 / 021581, publication number WO2016145092A1 (corresponding to Chinese application number 2016800150788, publication number CN107530556A); Among them, R1, R2, R3, R4, R5, R8, R9, R 10 The definition is as set forth in the claims of patent application PCT / CN2020 / 089692, publication number WO2020228685A9 (corresponding to Chinese application number 2020800358890, publication number CN113853379A); in: A is a substituted or unsubstituted C6-C10 aryl, biaryl or substituted biaryl, 5-15 membered heteroaryl or -N=CR 1 R 2 , wherein the substituent when substituted is selected from the group consisting of: halogen, -CN, -NO2, -O-(CH2)-O-, -CO2H and its salts, -OR 100 、-CO2R 100 、-CONR 101 R 102 、-NR 101 R 102 、-NR 100 S02R 100 、-SO2R 100 、-SO2NR 101 R 10 2 , C1-C6 alkyl, C3-C10 heterocyclic group; Among them, R 100 , R 101 and R 102 are independently hydrogen, C1-C8 alkyl, C6-C12 aryl; or R 101 and R 102 Together with the nitrogen atom to which it is attached, it forms a 5-7 membered heterocyclic ring; wherein the alkyl and aryl groups are each substituted with 1-3 halogen groups or 1-3 C1-C6 alkyl groups; R 1 and R 2 each independently is phenyl or methyl; X, Y and Z are each independently hydrogen or halo; R is hydrogen or C1-C6 alkyl or halogen-substituted alkyl; Wherein, the definition of Rw is as described in the claims of patent application PCT / CN2020 / 120281, publication number WO2021068952A1 (corresponding to Chinese application number 202080071652.8, publication number CN114555574A); Among them, the definitions of R1, R2, R3, R4, and T are as recorded in the claims of patent application PCT / CN2021 / 118597, publication number WO2022057838A1.

6. The method and use according to claim 5, The compound of structural formula (4) is selected from the following structural compounds: The compounds of structural formula (6) and (7) are selected from the following structural compounds: The compound of structural formula (8) is selected from the following structural compounds: The compound of structural formula (9) is selected from the following structural compounds: The compound of structural formula (10) is selected from the following structural compounds:

7. The method or use according to any one of claims 1 and 2, wherein: The antiallergic drug is selected from the following compounds and their salts, esters, solvates, and isotope isomers: Diphenhydramine, Promethazine, Cetirizine, Loratadine, Desloratadine, Fexofenadine, Cimetidine, Ranitidine, Famotidine, Hydrocortisone, Methylprednisolone, The anti-allergic drug is administered before the drug of the AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and the salt, ester, solvate, isotope isomer thereof to prevent allergies.

8. The method or use according to any one of claims 3 and 4, wherein: The non-steroidal analgesic drug is selected from diclofenac and its salts or esters, flurbiprofen and its salts or esters, The non-steroidal analgesic drug can be administered simultaneously with the AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, isotope isomers, or administered in advance or delayed.

9. The method or use according to any one of claims 3 and 4, wherein: The non-steroidal analgesic drug is selected from Loxoprofen and its salts or esters, Naproxen and its salts or esters, The non-steroidal analgesic drug should be administered at a certain interval from the drug of the AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salt, ester, solvate, and isotope isomer.

10. The method or use according to any one of claims 1 to 4, wherein: The AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salt, ester, solvate, and isotope isomer drug contain a surfactant for enhancing the solubility of the drug.

11. The method and use according to claim 10, wherein the surfactant is a nonionic surfactant, preferably selected from alkylphenol ethoxylates, Tweens, Span, Cremophor, Poloxamer, Polyvinylpyrrolidone, Polyethylene glycol PEG, and Polyvinyl alcohol PVA.

12. The method or use according to any one of claims 1 and 2, further used in combination with the following drugs to relieve pain: Diclofenac and its salts or esters, Flurbiprofen and its salts or esters, When used in combination, the drug can be administered simultaneously, in advance, or delayed with the AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, and isotope isomers.

13. The method and use according to any one of claims 1 and 2, further used in combination with the following drugs to relieve pain: Loxoprofen and its salts or esters, Naproxen and its salts or esters, When used in combination, the drug should be administered at a certain interval with the AKR1C3 enzyme-activated DNA alkylating agent prodrug compound containing an aziridine structure and its salts, esters, solvates, and isotope isomers.

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