Correlation between AKR1C3 enzyme expression levels and KRAS mutations, and their medical use.

Abstract

Correlation between AKR1C3 enzyme expression levels and KRAS mutations, and their medical applications. In the case of AKR1C3 enzyme-activated anticancer prodrugs, KRAS gene mutations can directly function as a test target before drug administration; that is, patients with KRAS mutations have high AKR1C3 enzyme expression levels, and AKR1C3 enzyme expression level or AKR1C3 RNA testing is unnecessary. In other words, a positive KRAS mutation test result can be used as a screening indicator to screen for patients with high AKR1C3 enzyme expression levels.

Description

[Technical Field]

[0001] The present invention relates to a method for treating cancer, and more particularly to a method for treating cancer having a specific gene. [Background technology]

[0002] AST-3424 (OBI-3424 / TH3424) is an AKR1C3 enzyme-activated DNA alkylating agent prodrug that has entered Phase II clinical trials in both China and the United States (clinical registrations in China are CTR20201915, CTR20201908, CTR20191399, and CTR20191371; clinical registrations in the United States are NCT04315324 and NCT03592264). It is specifically activated by the AKR1C3 enzyme, which is highly expressed in various tumors, and releases the DNA alkylating agent to exert an antitumor effect.

[0003] Preclinical studies have shown that the efficacy of AST-3424 is highly correlated with AKR1C3 enzyme expression, indicating that even slight differences in enzyme expression significantly impact the drug's ultimate therapeutic efficacy (AACR 2016, Abstract # 1369: In vitro and in vivo antitumor activity of TH3424: Preclinical rationale for a highly selective AKR1C3 prodrug for treating hepatocellular carcinomas; AACR-NCI-EORTC Annual Meeting, 2017, Abstract: LB-B16: The AKR1C3-Activated Prodrug OBI-3424 Exerts Profound In Vivo Efficacy Against Preclinical Models of T-Cell Acute Lymphoblastic Leukemia (T-ALL); a Pediatric Preclinical Testing Consortium Study; Clin Cancer Res 2019; 25: 4493-503: OBI-3424, a Novel AKR1C3-Activated Prodrug,Exhibits Potent Efficacy against Preclinical Models of T-ALL;Am J Cancer Res 2021;11(7):3645-3659,A novel selective AKR1C3-activated prodrug AST-3424 / OBI-3424 exhibits broad anti-tumor activity).

[0004] The aforementioned Phase II clinical trials in China and the United States will investigate the relationship between AKR1C3 enzyme expression levels and therapeutic efficacy in cancer patients and determine quantitative AKR1C3 expression levels to screen patients who will benefit from the experimental drug.

[0005] In other words, it is necessary to detect the expression level of the AKR1C3 enzyme in patients before administering the aforementioned AKR1C3 enzyme-activated anticancer prodrug, and the drug can only provide a better therapeutic effect in patients whose AKR1C3 enzyme expression level reaches a predetermined level.

[0006] Based on the Phase I clinical results of the drug, researchers determined that patients with AKR1C3 enzyme expression levels of H-score ≥ 135 (IHC) could be enrolled in Phase II clinical trials (Safety, Pharmacokinetics, and Clinical Activity of OBI-3424, an AKR1C3-Activated Prodrug, in Patients with Advanced or Metastatic Solid Tumors: A Phase 1 Dose-Escalation Study, J Clin Oncol 40, 2022, suppl 16; abstract 3030, DOI:10.1200 / JCO.2022.40.16_suppl.3030; Tsimberidou, AM, Verschraegen, CF, Wesolowski, R. et al. Phase 1 dose-escalation study evaluating the safety, pharmacokinetics, and clinical activity of OBI-3424 in patients with advanced or metastatic solid tumors. Br J Cancer(2023).https: / / doi.org / 10.1038 / s41416-023-02280-4).

[0007] In addition to AST-3424, a similar AKR1C3 enzyme-activated DNA alkylating anticancer prodrug, TFX05-01, is in Phase I clinical trials (Clinical registration in China: CTR20220957; Clinical registration in the US: NCT05434299. Charles Z Ding, Zhe Cai, Wei Sha. Preclinical evaluation of TFX05-01, a selective AKR1C3-targeted prodrug for solid tumor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022;2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5691), and this too needs to be investigated to examine the relationship between AKR1C3 enzyme expression levels and the therapeutic effect of the drug in clinical trials.

[0008] To quantitatively detect the expression level of the AKR1C3 enzyme, International Publication No. 2022048492 discloses an IHC detection method using FFPE tumor tissue sections, and International Publication No. 2022183483 discloses a PCR detection method using blood samples. The former requires the use of FFPE sections of the patient's tumor tissue as the detection sample and is therefore suitable for detecting patients with solid tumors. The latter requires the use of the patient's blood sample and is therefore suitable for hematological malignancies, particularly leukemia.

[0009] However, in practice, for some patients with malignant solid tumors, such as most pancreatic cancers, some liver cancers, almost all intrahepatic cholangiocarcinomas, primary brain cancers, and metastatic brain cancers, it is not possible to obtain viable tissue from these tumor tissues and prepare FFPE sections as IHC detection samples.

[0010] Therefore, it is necessary to identify alternative detection techniques and targets, screen patients who can benefit from AKR1C3 enzyme-activated antitumor (cancer) prodrug therapy based on the detection results, and complete the drug administration treatment. [Overview of the project]

[0011] In further preclinical efficacy studies, the inventors found that the AKR1C3 enzyme-activating prodrugs AST-3424 and AST had significant inhibitory effects on PDX tumor models and cells with Kras-G12D / G12C mutations (Examples 1-9 of PCT / CN2022 / 120817, publication number WO2023 / 046060). The inventors hypothesized whether Kras-G12D / G12C mutations are related in some way to high expression of the AKR1C3 enzyme: whether patients with Kras-G12D / G12C mutations also have high expression of the AKR1C3 enzyme.

[0012] Based on the above hypothesis, the inventors performed AKR1C3 RNA detection and AKR1C3 protein expression detection in the corresponding tissue of the PDX model as described above (Example 10 of PCT / CN2022 / 120817, Publication No. WO2023 / 046060). The results demonstrated high levels of AKR1C3 RNA and protein expression in the above model with the Kras-G12D / G12C mutation, thereby confirming the above hypothesis: the Kras-G12D / G12C mutation is associated with high expression of the AKR1C3 enzyme. The Kras-G12D / G12C mutation is accompanied by high expression of the AKR1C3 enzyme.

[0013] To verify the conclusion that the Kras-G12D mutation is associated with high expression of AKR1C3, we collected AKR1C3 expression data from 179 PDX models carrying the Kras-G12D mutation and found the following: The distribution of AKR1C3 expression in the KRAS G12D PDX model is mainly concentrated in moderate and high expression levels, accounting for 90.4%.

[0014] To verify the conclusion that the Kras-G12C mutation is associated with high expression of the AKR1C3 enzyme, we collected AKR1C3 expression data from 51 PDX models with the Kras-G12C mutation and found the following: In the KRAS G12C PDX model, AKR1C3 expression levels are equally distributed across low, moderate, and high expression levels, with moderate expression levels accounting for 66.7%.

[0015] To verify the conclusion that the Kras-G13D mutation is associated with high expression of the AKR1C3 enzyme, we collected AKR1C3 expression data from 48 PDX models carrying the Kras-G13D mutation and found the following: In the KRAS G13D PDX model, the distribution of AKR1C3 expression levels is mainly concentrated in moderate and high expression levels, with expression levels above moderate accounting for 83.3%.

[0016] The inventors further learned from the literature that KRAS G12D mutant tumor cells can directly upregulate and activate NRF2 (Nature, 2011, 475:106; Cancer Res, 2014, 74:7430) via the RAF-MEK-ERK-Jun signaling pathway (Nature, 2011, 475:106). Activated NRF2 can further upregulate and activate a series of downstream genes, including AKR1C3 (Chem Res Toxicol, 2017, 30:162; Cancer, 2019, 11:1715). Based on the above regulatory pathway, it is understood that gene mutations that can upregulate or activate NRF2, such as the G12D mutation, can ultimately upregulate the AKR1C3 gene, resulting in increased expression of the AKR1C3 enzyme in cancer cells.

[0017] Based on the above experiments and literature review, the inventors proposed that for AKR1C3 enzyme-activated anticancer prodrugs, KRAS gene mutations can be directly used as a detection target before drug administration. In other words, patients with KRAS mutations are patients with high expression of the AKR1C3 enzyme, and detection of AKR1C3 enzyme expression levels or AKR1C3 RNA is unnecessary. That is, a positive detection result for KRAS mutations can be used as a screening indicator for screening patients with high expression of the AKR1C3 enzyme.

[0018] For this purpose, this application provides the following technical solutions.

[0019] Solution 1 A method for administering cancer, tumors, or disorders or cell proliferation disorders resulting from cancer or tumors, comprising administering an AKR1C3 enzyme-activated prodrug to a patient if, without detecting the AKR1C3 protein expression level or AKR1C3 RNA content in the patient's tumor or cancer tissue, the patient's tumor or cancer tissue is detected to have a KRAS gene mutation, or the patient is detected to have a KRAS gene mutation.

[0020] Solution 2 A method for treating cancer, tumors, or disorders or proliferative diseases resulting from cancer or tumors, comprising the steps of administering a drug or formulation containing an AKR1C3 enzyme-activating prodrug, and detecting KRAS gene mutations, but not comprising the step of detecting the amount of AKR1C3 protein expression or AKR1C3 RNA content in the patient's tumor or cancerous tissue, A method for administering a drug or formulation containing an AKR1C3 enzyme-activated prodrug to a patient if the patient's tumor or cancerous tissue is detected to have a KRAS gene mutation, or if the patient is detected to have a KRAS gene mutation.

[0021] Solution 3 A pharmaceutically acceptable use of an AKR1C3 enzyme-activated prodrug in the manufacture of a drug for the treatment of cancer, tumors, or disorders or proliferative disorders resulting from cancer or tumors, characterized in that the drug is formulated for administration to a patient in whom a tumor or cancerous tissue has been detected to have a KRAS gene mutation, or to a patient in whom a KRAS gene mutation has been detected, and the detection of AKR1C3 protein expression levels or AKR1C3 RNA content in the patient's tumor or cancerous tissue is not required.

[0022] Solution 4 A drug formulation unit package comprising a separate packaging container for holding a drug formulation, an outer packaging component for housing the separate packaging container, and a drug usage instruction manual, wherein the drug formulation contains an AKR1C3 enzyme-activated prodrug, and the drug usage instruction manual is The drugs are used to treat patients with cancer, tumors, or disorders or proliferative diseases resulting from cancer or tumors. KRAS gene mutation detection is performed on patients without detecting AKR1C3 protein expression levels or AKR1C3 RNA content in tumor or cancerous tissue. A drug unit package that records that if the patient's tumor or cancerous tissue is detected to have a KRAS gene mutation, or if the patient is detected to have a KRAS gene mutation, a drug or formulation containing an AKR1C3 enzyme-activated prodrug will be administered to the patient.

[0023] An AKR1C3 enzyme-activating prodrug refers to a compound that acts as a prodrug. The prodrug molecule reacts with the AKR1C3 enzyme and, after the reaction, releases a cytotoxic antitumor compound.

[0024] Broadly speaking, an AKR1C3 enzyme-activated prodrug is one that satisfies at least one of the following conditions, but is not limited to these:

[0025] A. AKR1C3 inhibitors (e.g., compound 36, i.e., Bioorganic and Medicinal Chemistry, 2014:962-977) [ka] The inhibitory effect of compounds on cancer cell proliferation detected in the presence of AKR1C3 inhibitors is smaller than the inhibitory effect detected in the absence of AKR1C3 inhibitors. 50 When quantified, the IC of compounds against cancer cell lines detected in the presence of an AKR1C3 inhibitor is used. 50 However, IC was detected in the absence of the AKR1C3 inhibitor. 50 When the value is greater than the given value, the compound can be determined to be an AKR1C3-activating anticancer drug (lysis-prodrug). Specific compounds are described in the following patent document. PCT / US2016 / 021581, publication number WO2016145092A1, corresponding to Chinese application No. 2016800150788, publication number CN107530556A. PCT / US2016 / 062114, publication number WO2017087428, corresponding to Chinese application No. 2016800446081, publication number CN108290911A. PCT / US2016 / 025665, publication number WO2016161342, corresponding to Chinese application No. 2016800200132, publication number CN108136214A, and This is similar to the compound disclosed in PCT / NZ2019 / 050030, publication number WO2019190331, corresponding to Chinese application No. 2019800234236, publication number CN111918864A. The entire contents of the above-mentioned patent document are incorporated herein by reference.

[0026] Among these, the compounds disclosed in patent application PCT / US2016 / 021581, publication number WO2016145092A1, corresponding to Chinese application No. 2016800150788, publication number CN107530556A; patent application PCT / US2016 / 062114, publication number WO2017087428, corresponding to Chinese application No. 2016800446081, publication number CN108290911A; and patent application PCT / US2016 / 025665, publication number WO2016161342, corresponding to Chinese application No. 2016800200132, publication number CN108136214A, ultimately undergo cleavage. [ka] It is a cleavage prodrug that is metabolized to primary drugs, paclitaxel, camptothecin, and other drugs, which play a role as a primary drug. The compound disclosed in patent application PCT / NZ2019 / 050030, publication number WO2019190331, corresponding to Chinese application CN2019800234236, publication number CN111918864A, is a cleavage prodrug that is ultimately cleaved and metabolized to primary drugs that play a role as a nitrogen mustard structure drug.

[0027] The inhibitory effects of compounds on the proliferation of cancer cells with different levels of B.AKR1C3 enzyme expression differed significantly; the inhibitory effect on the proliferation of cancer cells with high AKR1C3 enzyme expression was far greater than the inhibitory effect on the proliferation of cancer cells with low AKR1C3 enzyme expression. 50 When quantified, the IC of compounds against cancer cells with high AKR1C3 enzyme expression 50 The value is an IC for cancer cells with low AKR1C3 enzyme expression. 50 When the value is lower than the specified value, the compound can be identified. The specific compound is disclosed in patent application PCT / CN2020 / 120281, publication number WO2021068952A1, the entire contents of which are incorporated herein by reference.

[0028] In the prior art, all AKR1C3 enzyme activating drugs (such as AST-3424 and AST-001, which are in clinical trials) require patients with solid tumors to provide pathological wax blocks or sections (including stored pathological wax blocks and sections) for AKR1C3 expression analysis, preferably recent samples. Only patients whose AKR1C3 expression in tumor tissue has been confirmed to reach a certain level by IHC detection can be enrolled in clinical trials.

[0029] In fact, for various reasons, some patients may not be able to obtain qualified tissue samples during tumor resection surgery, and some patients may not have previously undergone tumor resection surgery. Therefore, a large number of patients cannot provide samples for IHC detection to detect the expression level of the AKR1C3 enzyme.

[0030] The four solutions described above employ relatively mature KRAS gene mutation detection positive results (detecting the presence of KRAS gene mutations) to replace detection results of high levels of AKR1C3 protein expression or high AKR1C3 RNA content. In one embodiment, for patients whose previous detection results were positive for KRAS gene mutations, it can be directly determined that they have high expression of the AKR1C3 enzyme without further detection. In another embodiment, for patients who have not previously been detected for KRAS gene mutations, a mature KRAS gene mutation kit can be used for detection, which is convenient and readily available. Furthermore, while the sample for gene mutation detection is blood, which is easy to collect and has high patient compliance, tumor tissue samples used for IHC detection must be obtained by biopsy as an invasive procedure or during tumor resection surgery.

[0031] KRAS gene mutations are selected from KRAS-G12D, KRAS-G12V, KRAS-G12C, KRAS-G12A, and KRAS-G13D mutations.

[0032] Mutations in either or both of the genes corresponding to KRAS are approved in China. Amoy Dx, National Equipment Registration Number 20153401126, Human KRAS Gene Mutation Detection Kit (Fluorescence PCR Method) It can be detected and diagnosed using commercially available (companion) diagnostic kits such as Tellgen, National Equipment Registration No. 20163401341, Human K-RAS Gene 7 Mutation Detection Kit (PCR Fluorescence Method).

[0033] Clearly, NGS sequencing (YS450 gene NGS large panel) can also be used to determine specific KRAS mutation subtypes.

[0034] The specific information corresponding to this is shown in the table below.

[0035] [Table 1] [Table 2]

[0036] Generally speaking, KRAS gene mutations are detected using complete samples from a patient's tumor or cancerous tissue. However, there may be situations where it is necessary to detect or diagnose a patient's own KRAS gene mutation status for various reasons, such as the patient being unable to provide a qualified tumor or cancerous tissue sample, the current detection method being inaccurate, or the tumor or cancerous tissue no longer reflecting the patient's recent condition. For example, blood can be used for gene mutation detection, and the detection results can represent the state of the patient's tumor or cancerous tissue. Circulating tumor cell (CTC) detection can detect the KRAS gene mutation status of specific cancer cells (all tumor cells released into the peripheral blood) using a complete detection sample from the patient using peripheral blood. In some cases, the gene mutation status of parents in a biological parentage relationship can be used to diagnose or assist in diagnosing the gene mutation status. This is a situation where a patient has been detected to have a KRAS gene mutation, or where a patient with a KRAS gene mutation has been detected.

[0037] In this application, gene mutation refers to pathogenic gene mutation.

[0038] The TMB (Tumor Mutation Load) level of the described gene mutations is moderate.

[0039] Tumor Mutation Load (TMB) varies in height between different tumor types; generally, a TMB greater than 20 mutations / Mb (Mb represents bases per million) is considered high, a TMB less than 10 mutations / Mb is considered low, and a median TMB is considered moderate. At the 2017 World Congress on Lung Cancer, Squibb Inc. presented the results of a clinical trial called CheckMate-032. This was a Phase II clinical trial that enrolled 401 patients with advanced lung cancer who had failed first-line treatment and were treated with PD-1 inhibitors alone or in combination with ipilimumab. Patients were divided into three categories according to their TMB level: high TMB, moderate TMB, and low TMB. Among patients who received combination therapy, the efficacy rates for the three groups were 62%, 20%, and 23%, respectively. The efficacy rate in the high TMB group was three times higher than in the other two groups. The median overall survival for the three groups was 22.0 months, 3.6 months, and 3.4 months, respectively, with a six-fold difference between 22.0 months and 3.4 months. This trial demonstrated that different TMB levels significantly affect the efficacy of different cancer treatments.

[0040] The AKR1C3 enzyme-activating prodrug is selected from the compounds of formulas (1) to (12) or their pharmaceutically acceptable salts, solvates, or isotopic isomers.

[0041] [ka] X, Y, Z, R, T, A, and X 10The definition is described in the claims of patent application PCT / US2016 / 021581 with publication number WO2016145092A1 (corresponding to Chinese application No. 2016800150788 with publication number CN107530556A). The methods for synthesizing and preparing specific compounds are also described in the above patent application, and the whole is incorporated herein by reference. The specific definition is as follows.

[0042] X 10 is O, S, SO or SO2, A is C6 - C 10 aryl, 5 - to 15 - member heteroaryl or -N = CR 1 R 2 and R 1 and R 2 are each independently hydrogen, C1 - C6 alkyl, C3 - C8 cycloalkyl, C6 - C 10 aryl, 4 - to 15 - member heterocycle, ether, -CONR 13 R 14 or -NR 13 COR 14 and 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 - to 15 - member heterocycle, ether, -CONR 13 R 14 or -NR 13 COR 14 and R is hydrogen, C1 - C6 alkyl, C2 - C6 alkenyl, C2 - C6 alkynyl, C3 - C8 cycloalkyl, C6 - C 10 aryl, 4 - to 15 - member heterocycle, ether, -CONR 13 R 14 or -NR 13 COR 14 and R 13 and R 14 are each independently hydrogen, C1 - C6 alkyl, C3 - C8 cycloalkyl, C6 - C 10Aryl, 4-15 membered heterocycle, ether, T is -OP(Z 1 ) One or more Z joined to the part 5 -X 5 -Y 5 Contains a phosphoramidate alkylating agent including a portion, Z 5 X is a heteroatom containing nitrogen, sulfur, or oxygen. 5 Y is substituted or unsubstituted ethylene. 5 is a halogen or another leaving group, or Z 5 -X 5 -Y 5 They both form an aziridinyl (NCH2CH2) moiety, Z 1 is either O or S, Alkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, aryl groups, heterocyclyl groups, heteroaryl groups, and ether groups can be substituted or unsubstituted.

[0043] The AKR1C3 enzyme-activating prodrug compound of formula (1) is selected from compounds having the following structural formulas.

[0044] [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0045] The specific synthesis method of the compound of formula (1) and the corresponding spectral data are disclosed in International Publication No. 2016145092 (corresponding to Chinese Publication Text CN107530556A), which is incorporated herein by reference in its entirety.

[0046] [ka] X, Y, Z, R, D, L 1 , A and X 10 The definition is described in the claims of PCT / US2016 / 025665, publication number WO2016 / 161342A1 (corresponding to Chinese application No. 2016800200132, publication number CN108136214A), and the methods for the synthesis and preparation of specific compounds are also described in the above patent application, which are incorporated herein by reference in their entirety. The specific definition is as follows:

[0047] X 10 These are O, S, SO, or SO2. A is C6~C 10 Aryl, 5-15 member heteroaryl, or -N=CR 1 R 2 And, R 1 and R 2 These are, independently, hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, and C6-C 10 Aryl, 4-15 membered heterocyclic rings, 5-15 membered heteroaryls, ethers, -CONR 13 R 14 or -NR 13 COR 14 And, X, Y, and Z are independently hydrogen, CN, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, and C6-C 10 Aryl, 4-15 membered heterocyclic rings, 5-15 membered heteroaryls, ethers, -CONR 13 R 14 or -NR 13 COR 14 And, Each R independently consists of hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, and C6-C 10 Aryl, 4-15 membered heterocyclic rings, 5-15 membered heteroaryls, ethers, -CONR 13 R 14 or -NR 13 COR 14 And, R 13 and R 14 These are, independently, hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, and C6-C 10 They are aryl groups, 4-15 membered heterocyclic rings, 5-15 membered heteroaryl groups, or ethers. L 1 And D is defined as follows: L 1 The following can be selected: [ka] R 40 and R 41 These are, independently, hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, and C6-C 10 It is an aryl, a 4- to 15-membered heterocyclic ring, or a 5- to 15-membered heteroaryl. R 42 This is a C2-C3 alkylene or heteroalkylene which may be optionally substituted with 1-3 C1-C6 alkyl groups. V(-) is any anion, preferably a pharmaceutically acceptable anion. D is the part that makes D-OH an anticancer agent, where OH is an aliphatic hydroxyl or a phenolic hydroxyl, or an OH part bonded to a phosphorus atom as provided herein, or L 1 teeth, [ka] And, R 40 As defined above, R43 is hydrogen or forms a heterocyclic ring together with D, and the phenyl moiety is optionally substituted, D is D-NR 43 H is a moiety that makes an anticancer agent, or L 1 is a bond, -O-C(R 40 R 41 )2-, -O-C(R 40 R 41 )-NR 40 R 41 (+)-C(R 40 R 41 )- or

Chemical formula

[0048] The AKR1C3 enzyme-activated prodrug compound of formula (2) is selected from compounds having the following structural formulas.

[0049]

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0050] The specific synthesis method of the compound of formula (2) and the corresponding spectral data are disclosed in International Publication No. 2016161342 (corresponding to Chinese publication texts CN108136214A and CN112142692A), which are incorporated herein by reference in their entirety.

[0051] [ka] R1, R2, R3, R4, R5, R8, R9 and R 10 The definition is described in the claims of patent application PCT / CN2020 / 089692, publication number WO2020228685A1 (corresponding to Chinese application No. 2020800358890, publication number CN113853379A), and the methods for the synthesis and preparation of specific compounds are also described in the above patent application, which are incorporated herein by reference in their entirety. The specific definition is as follows:

[0052] R1 is C6~C 10 These are aryl or Z-substituted aryl rings, 4-15 member heterocyclic rings or Z-substituted heterocyclic rings, 5-15 member heteroaryl or Z-substituted heteroaryl rings, and 7-15 member fused rings or Z-substituted fused rings. R2 is hydrogen, halogen atom, cyano or isocyano, hydroxy, sulfhydryl, amino, 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 10Aryl or Z-substituted aryl, 4- to 15-membered heterocyclic ring or Z-substituted heterocyclic ring, 5- to 15-membered heteroaryl or Z-substituted heteroaryl, ether having 1 to 6 carbon atoms or Z-substituted alkoxy having 1 to 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 または-NR 6 SO2NR 6 R 7 、またはR 2 は、それが結合している基R 1 中の原子と一緒になって、7~15員の縮合環またはZ置換縮合環を形成し、 R3 is hydrogen, halogen, cyano or isocyano, hydroxy, sulfhydryl, amino, 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- to 15-membered heterocyclic ring or Z-substituted heterocyclic ring, 5- to 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 、または-NR 6 SO2R 7 であり、 R4 and R5 are, independently, hydrogen, halogen atoms, cyano or isocyano, hydroxy, sulfhydryl, amino, 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 heterocycle or Z-substituted heterocycle, 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 or -NR 6 SO2R 7 And, or, R 4 and R 5 These atoms, together with the atoms in the benzene ring to which they are bonded, form a 7-15 member fused ring or a Z-substituted fused ring. R6 and R7 are 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 It is an aryl or Z-substituted aryl, a 4-15 member heterocycle or Z-substituted heterocycle, a 5-15 member heteroaryl or Z-substituted heteroaryl, a C1-C6 alkoxy or Z-substituted C1-C6 alkoxy, or R 6 and R 7 These, together with the atoms to which they are bonded, form 5-7 membered heterocyclines or Z-substituted 5-7 membered heterocyclines. R8 and R 10Each of these 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 R 8 and R 10 At least one of them must be hydrogen or deuterium. R9 is a substituted C6-C atom substituted with at least one fluorine atom or nitro group. 10 The aryl group is a substituted 4- to 15-membered heterocyclic ring substituted with at least one fluorine atom or nitro group, or a substituted 5- to 15-membered heteroaryl group substituted with at least one fluorine atom or nitro group. Substituent Z is a halogen atom, cyano or isocyano, hydroxy, sulfhydryl, amino, OTs, OMS, C1-C3 alkyl or substituted alkyl, C1-C3 alkoxy or substituted alkoxy, C2-C3 alkenyl or substituted alkenyl, C2-C3 alkynyl or substituted alkynyl, C3-C8 cycloalkyl or substituted cycloalkyl, aromatic ring, heterocycle, heteroaromatic ring and fused ring or substituted aromatic ring, heterocycle, heteroaromatic ring and fused ring, and the substitution pattern is mono or geminal disubstituted. In R9, substitution C6~C 10 Substitutions in aryls, substituted 4-15 membered heterocycles, or substituted 5-15 membered heteroaryls include halogen atoms, nitro, cyano or isocyano, hydroxy, amino, C1-C3 alkyl or alkoxy, alkenyl, alkynyl, cycloalkyl or benzene rings, substituted benzene rings, C1-C3 alkoxy, or halogen atom-substituted alkoxy.

[0053] The AKR1C3 enzyme-activating prodrug compound of formula (3) is selected from compounds having the following structural formulas.

[0054] [ka] [ka]

[0055] The AKR1C3 enzyme-activating prodrug compound of formula (4) is selected from compounds having the following structural formulas.

[0056] [ka]

[0057] Specific synthesis methods for the compounds of formulas (3) and (4), as well as the corresponding spectral data, are disclosed in International Publication No. 2020228685 (corresponding to Chinese Publication Text CN113853379A), which is incorporated herein by reference in its entirety.

[0058] [ka] A is either substituted or non-substituted C6~C 10 Aryl, biaryl or substituted biaryl, 5-15 member heteroaryl, or -N=CR 1 R 2 Here, the substituents are halogen, -CN, -NO2, -O-(CH2)-O-, -CO2H and their salts, -OR 100 , -CO2R 100 ,-CONR 101 R 102 , -NR 101 R 102 , -NR 100 SO2R 100 , -SO2R 100 -SO2NR 101 R 102 , C1-C6 alkyl and C3-C 10 Selected from a group consisting of heterocyclines, R 100 , R 101 and R 102 These are, independently, hydrogen, C1-C8 alkyl or C6-C 12 It is either aryl or R 101 and R 102These, together with the nitrogen atoms to which they are bonded, form a 5-7 membered heterocycle. The alkyl and aryl groups are each substituted with 1 to 3 halogen groups or 1 to 3 C1-C6 alkyl groups. R 1 and R 2 These are, independently, phenyl or methyl, X, Y, and Z are each independently either hydrogen or a halogen. R is hydrogen, a C1-C6 alkyl group, or a halogen-substituted alkyl group.

[0059] The AKR1C3 enzyme-activating prodrug compound of formula (5) is selected from compounds having the following structural formulas.

[0060] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry]

[0061] Specific synthetic methods of the compound of formula (5) and the corresponding spectral data are disclosed in International Publication No. 2020228685 (corresponding to Chinese Publication Text CN113853379A) and International Publication No. 2016145092 (corresponding to Chinese Publication Text CN107530556A), which are hereby incorporated by reference in their entirety into this specification.

[0062] [Chemistry] The definition of Rw is described in the claims of the patent application PCT / CN2020 / 120281 (corresponding to Chinese Application No. 202080071652.8 with Publication No. CN11455574A) of Publication No. WO2021068952A1. The synthetic and preparation methods of specific compounds are also described in the above patent application, which are hereby incorporated by reference in their entirety into this specification. The specific definition is as follows.

[0063] Rw is [Chemistry] and R1 is H, C 1~6 alkyl, C 3~6 cycloalkyl, 4-6 member heterocycloalkyl, 5-6 member heteroaryl or phenyl, where C 1~6 alkyl, C 3~6Cycloalkyl, 4-6 member heterocycloalkyl, 5-6 member heteroaryl, and phenyl compounds have 1, 2, or 3 R a It is sometimes substituted in the base, Each R a These are independently H, F, Cl, Br, I, -CN, -OH, C 1~3 Alkoxy or C 1~3 It is alkyl, R2 is H or C 1~6 It is alkyl, Alternatively, R1 and R2, together with the N atom to which they are bonded, form a 4-6 member heterocycloalkyl group, where the 4-6 member heterocycloalkyl group has 1, 2, or 3 R b It is sometimes substituted in the base, Each R b These are independently H, F, Cl, Br, I, -CN, -OH, -NH2, -OCH3, -OCH2CH3, -CH3 or -CH2CH3, R3 consists of H, F, Cl, Br, I, -OH, -NH2, and C. 1~3 Alkoxy or C 1~3 It is alkyl, Alternatively, R2 and R3 may combine together to form a structural unit. [ka] of, [ka] west, T1 is -(CR c R d ) m - or - (CR c R d ) n -O-, m is 1, 2, or 3. n is either 1 or 2. T2 is N or CH. R c and R d These are H, F, and C, respectively, independently. 1~3 Alkyl or C 1~3 It is an alkoxy, R4, R5, and R6 are each independently H, F, Cl, Br, I, C 1~3 alkyl or C 1~3 alkoxy, T is N or CH, R7 and R8 are each independently H, F, Cl, Br, or I, R9 and R 10 are each independently H, F, Cl, Br, I, -CN, or 4- to 6-membered heterocycloalkyl and 5- to 6-membered heteroaryl each contain 1, 2, 3, or 4 heteroatoms independently selected from N, -O-, and -S-.

[0064] The AKR1C3 enzyme-activating prodrug compound of formula (6) is selected from compounds having the following structural formulas.

[0065]

Chemical formula

[0066] A specific synthesis method of the compound of formula (6) and the corresponding spectral data are disclosed in International Publication No. 2021068952 (corresponding to Chinese Publication Text CN114555574A), the whole of which is incorporated herein by reference.

[0067]

Chemical formula

[0068] T is N or CH, R1 and R2 are each independently H, F, Cl, Br, I, or C 1~3It is alkyl, and here, C 1~3 Alkyl groups consist of 1, 2, or 3 R groups. a It is sometimes substituted in the base, Each R a These are independently F, Cl, Br, I, -CN, -OH, or -NH2. R3 and R4 are independently H, F, Cl, Br, I, CN, and C, respectively. 1~3 Alkyl, C 1~3 Alkoxy, [ka] And here, C 1~3 Alkyl groups consist of 1, 2, or 3 R groups. e It is sometimes substituted in the base, R b and R c These are, independently, H, -CH3, -CH2CH3, -(CH2)2CH3, or -CH(CH3)2, R d These are -CH3, -CH2CH3, -(CH2)2CH3, or -CH(CH3)2. Each R e These are independently F, Cl, Br, I, -CN, -OH, or -NH2.

[0069] The AKR1C3 enzyme-activating prodrug compound of formula (7) is selected from compounds having the following structural formulas.

[0070] [ka]

[0071] The specific synthesis method of the compound of formula (7) and the corresponding spectral data are disclosed in International Publication No. 2022057838, which is incorporated herein by reference in its entirety.

[0072] [ka] The definitions of A, E, G, X, and Y are described in the claims of patent application PCT / NZ2019 / 050030, publication number WO2019190331A1 (corresponding to Chinese patent application No. 2019800234236, publication number CN111918864A), and the methods for the synthesis and preparation of specific compounds are also described in the above patent application, which is incorporated herein by reference in its entirety. The specific definitions are as follows:

[0073] A is H, C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, CFH2, CF2H, CF3, F, Cl, Br, I, OCF3, COR, or CON(R)2. E is SO or SO2, X is Cl, Br, I, or OSO2R. Y is Cl, Br, I, or OSO2R. Each R is independently H or C1-C6 alkyl. G is a group selected from the group consisting of equations (B) to (AA), [ka] R1 is H, C1-C6 alkyl, CH2(CH2) n R1 is OH, CH2CH(OH)CH2OH, phenyl, pyridyl, benzyl, or pyridylmethyl, provided that if R1 is phenyl, pyridyl, benzyl, or pyridylmethyl, R1 may be substituted at any available position with C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, OR6, N(R6)(R7), CFH2, CF2H, CF3, F, Cl, Br, I, OCF3, COR6, CON(R6)(R7), SOR6, SON(R6)(R7), SO2R6, SO2N(R6)(R7), CN, or NO2. R2 and R3 are independently H, C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, OR6, N(R6)(R7), CFH2, CF2H, CF3, F, Cl, Br, I, OCF3, COR6, CON(R6)(R7), SOR6, SON(R6)(R7), SO2R6, SO2N(R6)(R7), CN, or NO2. R4 is N(R6)(R7), OH, OCH2(CH2) n N(R6)(R7) or CH2(CH2) n N(R6)(R7), R5 is H or a C1-C6 alkyl group. R6 and R7 are either independently H or C1-6 alkyl, or together R6 and R7 form a substituted or unsubstituted 5-membered or 6-membered heterocycle. Z is either CH or N. W is CH2, O, S, SO, or SO2. n is between 0 and 6. * indicates a connection point with equation (I).

[0074] The AKR1C3 enzyme-activating prodrug compound of formula (8) is selected from compounds having the following structural formulas.

[0075] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0076] The specific synthesis method of the compound of formula (8) and the corresponding spectral data are disclosed in International Publication No. 2019190331 (corresponding to the Chinese Publication Text CN111918864A), which is incorporated herein by reference in its entirety.

[0077] [ka] R w X, R4, R 10 , R 13 , and R 14 The definition is described in the claims of patent application PCT / CN2022 / 098082, publication number WO2022258043A1, and the methods for the synthesis and preparation of specific compounds are also described in the above patent application, which are incorporated herein by reference in their entirety. The specific definition is as follows:

[0078] Each of the two X groups independently performs CR 15 or N, R 13 and R 14 Each of these can independently be hydrogen, C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, C6-C 20Aryl, 5-20 member heterocyclyl, halogen-substituted C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, halogen-substituted C6-C 20 It is an aryl or halogen-substituted 5-20 member heterocycline, R 13 and R 14 Neither of them is hydrogen, R 10 These are hydrogen, C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, C6-C 20 Aryl, 5-20 member heterocyclyl, halogen-substituted C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, halogen-substituted C6-C 20 These are aryl or halogen-substituted 5-20 member heterocyclines. Alternatively, R 10 R 10 , R 13 and R 14 Under conditions consistent with the aforementioned definition, R 13 or R 14 It can bond to form a 5-9 membered ring, R4 and R 15 Each of these can independently be hydrogen, halogen, C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, cyano, 5-20 member heterocyclyl, C6-C 20 Aryl, or halogen-substituted C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, 5-20 membered heterocyclyl, or C6-C 20 It is Ariel, Alternatively, R 10 and R 15 The above R 10 and R 15 Under conditions consistent with the definition, it can form a 4-12 membered cyclic hydrocarbon or heterocycle. R W teeth [ka] And, A is CR 16 Or N, and the position of A can be different in the ring. R 16These are hydrogen, C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, C6-C 20 Aryl, 5-20 member heterocyclyl, halogen-substituted C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl; halogen-substituted C6-C 20 These are aryl or halogen-substituted 5-20 member heterocyclines. R6 and R7 meet the following conditions: Each of R6 and R7 is independently hydrogen, halogen, cyano, hydroxyl, C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, 5-20 member heterocyclyl, C6-C 20 Aryl, or halogen-substituted C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, 5-20 membered heterocyclyl, C6-C 20 Aryl, or cyano-substituted C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, 5-20 membered heterocyclyl, C6-C 20 Aryl or hydroxyl-substituted C1-C6 alkyl, cycloalkyl, alkoxy, 5-20 membered heterocyclyl, C6-C 20 aryl or -CONR 11 R 12 Alternatively, -CH2NR 11 R 12 And, Alternatively, R6 and R7 combine to form a 5- to 8-membered monocyclic or fused heterocyclic ring containing at least one of N, S, and O, or two or three of N, S, and O. Alternatively, R6 and R7 are bonded to form a 5-8 member monocyclic or fused heterocycle containing at least one N, S, and O, or two or three N, S, and O, and the monocyclic or fused heterocycle is substituted with C1-C6 alkyl groups. R6 is CR 16 It can bond to form a 5- to 9-membered ring, a heterocycle, or an aromatic heterocycle. R 11 and R 12 The following conditions must be met: R 11 and R12 Each of them is independently a C1-C6 alkyl or halogen-substituted C1-C6 alkyl, or R 11 and R 12 Under conditions that match the above definition, -CONR 11 R 12 N or -CH2NR 11 R 12 It forms a 5-7 membered ring with N.

[0079] The AKR1C3 enzyme-activating prodrug compound of formula (9) is selected from compounds having the following structural formulas.

[0080] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0081] The specific synthesis method of the compound of formula (9) and the corresponding spectral data are disclosed in International Publication No. 2022258043, which is incorporated herein by reference in its entirety.

[0082] [ka] The definitions of R1, R2, R3, R4, G1, G2, G3, G4, E, T, Y, Z, m, n, s, t, v, w and ring A are described in the claims of patent application CN202210585771.6, publication number CN115403579A, and the methods for the synthesis and preparation of certain compounds are also described in the aforementioned patent application, which is incorporated herein by reference in its entirety. The specific definitions are as follows:

[0083] G 1 , G 2 , G 3 or G 4 They are identical or different, and each is independently CR 5 or N atom, Each R 5 These are the same or different, and each independently consists of a hydrogen atom, halogen, alkyl, alkenyl, alkynyl, alkoxy, hydroxyl, cyano, amino, nitro, and -NR. a R b -C(O)NR a R b, selected from cycloalkyl, heterocyclyl, aryl and heteroaryl, where alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, hydroxyl, oxo, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl. Y is -(C(R y2 R y3 )) f -NR y1 -,-(C(R y2 R y3 )) g -O-, -(C(R y2 R y3 )) h -S-, -(C(R y2 R y3 )) h -S(O)-, -(C(R y2 R y3 )) h -S(O)2-, -C(R) y2 R y3 )-, -NR y1 -(C(R y2 R y3 )) f -, -O-(C(R y2 R y3 )) g -, -S-(C(R y2 R y3 )) h -, -S(O)-(C(R y2 R y3 )) h -, and -S(O)2-(C(R y2 R y3 )) h - Selected from, R y1 These are selected from hydrogen atoms, alkyl, haloalkyl, hydroxyalkyl, cycloalkyl and heterocyclyl, R y2 and R y3They are the same or different, and each is independently selected from hydrogen atoms, halogens, alkyls, haloalkyls, hydroxyalkyls, cycloalkyls, and heterocyclines. Alternatively, R y2 and R y3 Together they form =O, Z is either O or OH. [ka] These are single or double bonds, [ka] If it is a single bond, then Z is OH, [ka] If it is a double bond, then Z is O, E is selected from NH, O atoms, and S atoms. T is -C(R T1 R T2 )-, -NR T3 -, or -O- are selected, R T1 and R T2 Same or different, R T1 and R T2 Each of these is independently selected from hydrogen atoms, deuterium atoms, halogens, alkyls, haloalkyls, hydroxyalkyls, cycloalkyls, and heterocyclines. Alternatively, R T1 and R T2 These, together with the carbon atoms to which they are bonded, form a cycloalkyl or heterocycline, where each cycloalkyl or heterocycline is optionally substituted with one or more substituents selected independently from halogens, alkyls, and hydroxyls. R T3 These are selected from hydrogen atoms, alkyl, haloalkyl, hydroxyalkyl, cycloalkyl and heterocyclyl, Ring A is a 6- to 10-membered aryl or a 5- to 10-membered heteroaryl. Each R 1 The same or different, each R 1 These are independently hydrogen atoms, deuterium atoms, halogens, alkyls, alkenyls, alkynyls, alkoxys, hydroxyls, cyanos, and -NRs. a R b -C(O)NR a R b -S(O)NR a R b -S(O)2NR a R b ,-S(O)R c -S(O)2R c , -B(OR d )2, selected from nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl, where alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, hydroxyl, oxo, cyano, -NR a R b -C(O)NR a R b -S(O)NR a R b -S(O)2NR a R b ,-S(O)R c -S(O)2R c , -B(OR d )2, optionally substituted with one or more substituents selected from nitro, cycloalkyl, heterocyclyl, aryl, and heteroaryl, R a and R b Same or different, R a and R b Each of these independently consists of a hydrogen atom, alkyl, haloalkyl, hydroxyl, hydroxyalkyl, and -C(O)R e Selected from cycloalkyl and heterocyclyl, or R a and R bThese, together with the nitrogen atom to which they are bonded, form a cycloalkyl or heterocycline, where the cycloalkyl or heterocycline is optionally substituted with one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, hydroxyl, cyano, amino, nitro, cycloalkyl, heterocycline, aryl, and heteroaryl. R c is selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, where alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are optionally substituted with one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, hydroxyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, and heteroaryl. R d is a hydrogen atom or C 1~6 It is alkyl, R e is selected from alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl and heteroaryl, where alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally substituted with one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, hydroxyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl. Each R 2 The same or different, each R 2 These are independently selected from hydrogen atoms, deuterium atoms, halogens, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, oxo, hydroxyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl, Each R 3 The same or different, each R 3These are independently selected from hydrogen atoms, deuterium atoms, halogens, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, oxo, hydroxyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl, R 4 This is selected from hydrogen atoms, alkyl, haloalkyl, hydroxyl, and hydroxyalkyl. n is 0, 1, 2, or 3. v is 0, 1, or 2, w is 0, 1, or 2. f is 0, 1, or 2, g is 0, 1, or 2. h is 0, 1, or 2. m is 0, 1, 2, 3, 4, or 5. s is 0, 1, 2, 3, 4, 5, 6, 7, or 8. t is 0, 1, 2, 3, 4, 5, or 6. however, If Y is an O- atom and E is an O atom, then ring A is phenyl or a 5-6 member heteroaryl, and G 3 CR 5 or N atom, R 5 It is not a hydrogen atom, If Y is an O- atom and E is an S atom, then ring A is a phenyl or 5-6 member heteroaryl. G 1 , G 2 , G 3 , and G 4 All CR 5 And Y is NR y1 If n, v, and w are all 1, and E is an O atom, then 1) T is not CH2 or CD2, and 2) R 2 or R 3 At least one of them is a deuterium atom, and 3)R 4 4)R is selected from alkyl, haloalkyl, hydroxyl, and hydroxyalkyl. 1One of them is a 3- to 8-membered cycloalkyl or a 5- to 8-membered heterocycline, where the 3- to 8-membered cycloalkyl or 5- to 8-membered heterocycline is optionally substituted with one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, hydroxyl, oxo, cyano, amino, nitro, cycloalkyl, heterocycline, aryl and heteroaryl, 5) Ring A is a hydrogen atom or C 1~6 R is an alkyl group d That is the case.

[0084] The AKR1C3 enzyme-activated KARS inhibitor prodrug compound of formula (10) is selected from compounds having the following structural formulas.

[0085] [ka] [ka] [ka] [ka] [ka] [ka]

[0086] The specific synthesis method of the compound of formula (10) and the corresponding spectral data are disclosed in the Chinese Public Text CN115403579A, which is incorporated herein by reference in its entirety.

[0087] [ka] R 1 , R 2a , R 2b , R 3 , R 4, R 5 The definitions of n and Z are described in the claims of patent application PCT / IB2020 / 057285, publication number WO2021005586A1 (corresponding to Chinese application No. CN202080053804.1, publication number CN114206870A), and the methods for the synthesis and preparation of specific compounds are also described in the above patent application, which is incorporated herein by reference in its entirety. The specific definitions are as follows:

[0088] [ka] These are single or double bonds, Z is [ka] If it is a single bond, it is OH, or [ka] If it is a double bond, then it is O. Each R 1 These are independently (C1-C6) alkyl, (C1-C6) alkoxy, and (C0-C4) alkyl N(R) 8 Selected from the group consisting of )2 and halogeno, R 2a and R 2b Each of these is independently selected from the group consisting of H, (C1-C6) alkyl, and halogeno. Each R 3 It was independently selected from the group consisting of H and Halogeno, R 4 R is selected from the group consisting of aryls containing 1, 2, 3, or 4 heteroatoms independently selected from N, O, and S, 5-6 membered heteroaryls, and 9-10 membered condensed bicyclic heteroaryls containing 1, 2, 3, or 4 heteroatoms independently selected from N, O, and S, and any of the above is one or more R 6 It is sometimes replaced by, R 5H; (C1~C6) alkyl; (C2~C6) alkenyl; (C0~C4) alkyl OR 8 ;(C1~C4)alkyl(C3~C 10 )Cycloalkyl; Halo(C1~C6)alkyl; (C2~C3)alkynyl; (C1~C4)alkylN(R 10 Selected from the group consisting of )2, Each R 6 These are: halogen; (C1~C6)alkyl; (C1~C6)alkoxy; halo(C1~C6)alkyl; OH; aryl; 3~6 member heterocyclic; 5~6 member heteroaryl; (C0~C4)alkylS(O) m (C1-C6) alkyl; halo(C1-C6) alkoxy; (C0-C4) alkyl S(O) m N(R 8 )2;(C0~C4)alkylN(R 8 )2;(C0~C4)alkyl(CO)OR 7 ;N(R 8 )S(O) m (C1~C6) alkyl; N(R) 8 )S(O) m (C3~C6)Cycloalkyl; OP(O)(OH)2; (C0~C3)Alkyl(CO)NHR 11 (C0~C3) alkyl OR 7 ; and (C3~C 10 ) Independently selected from the group consisting of cycloalkyls, each R 6 If it is not a halogen, OH, or OP(O)(OH)2, then 1 to 3 R 9 In some cases, it is replaced by two adjacent R 6 These, together with the atoms to which they are bonded, form a 5-7 membered heterocycle or (C5-C8) cycloalkyl group. R 7 and R 8 Each of these independently has H, or 1 to 3 R 9 Selected from the group consisting of alkyl groups (C1-C6) which may be substituted in some cases, Each R 9Halogeno;-OH;amino;(C1~C4)alkylamino;di(C1~C4)alkylamino;OP(O)(OH)2;(C1~C6)alkyl;(C1~C3)alkynyl;(C1~C6)alkoxy;halo(C1~C6)alkyl;(C0~C4)alkylS(O) m (C1~C6) alkyl; halo(C1~C6) alkoxy; 3-6 membered heterocycles optionally substituted with oxo(=O); (C0~C4) alkylS(O) m N(R 10 )2;(C0~C4)alkyl(CO)R 10 ;(C0~C4)alkyl(CO)OR 10 (C0~C4) alkyl NR 10 S(O) m (C1~C6) alkyl; (C0~C4) alkyl OR 10 (C0~C4) alkyl N(R) 10 )2;(C0~C4)alkylCN;(C0~C4)alkylN(R 10 )2; and (C0~C4)alkyl(CO)N(R 10 ) Independently selected from the group consisting of 2, Each R 10 The group is independently selected from the group consisting of H; (C1-C6) alkyl; or 3-6 membered heterocycles, and the 3-6 membered heterocycle is optionally substituted with one or more substituents selected from (C1-C6) alkyl and oxo (=O). Each R 11 H; 1 to 4 R 12 A 4-6 member complex ring that is substituted in some cases; 1-4 R 12 (C3~C6) cycloalkyl groups that may be substituted with a halogen; (C0~C3) alkyl groups (C3~C6) cycloalkyl groups (C1~C3) alkyl groups that may be substituted with a halogen; 1 to 3 R groups 12 Independently selected from the group consisting of CH2-aryl;(C1~C6)alkyl;(C2~C6)alkenyl; or (C2~C6)alkynyl, each of which is optionally substituted, 13 It may be replaced in some cases. Each R 12This is independently selected from the group consisting of OH; (C1-C3) alkoxy; NH2; or (C1-C3) alkyl substituted with one or more OH groups. Each R 13 These are independently selected from the group consisting of halogens; OH; aminos; (C1-C4) alkylaminos; di(C1-C4) alkylaminos; (C1-C3) alkoxys; and C(O)-(C3-C8) cycloalkyls. m is 0, 1, or 2. n is 0, 1, or 2.

[0089] The AKR1C3 enzyme-activated KARS inhibitor prodrug compound of formula (11) is selected from compounds having the following structural formulas.

[0090] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0091] The specific synthesis method of the compound of formula (11) and the corresponding spectral data are disclosed in International Publication No. 2021005586 (corresponding to Chinese Publication Text CN114206870A), which is incorporated herein by reference in its entirety.

[0092] [ka] or a pharmaceutically acceptable salt thereof R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R a , R b The definitions of n1 and n2 are described in the claims of patent application PCT / CN2023 / 123253, publication number WO2024078392A1, and the methods for the synthesis and preparation of certain compounds are also described in the aforementioned patent application, which are incorporated herein by reference in their entirety. The specific definitions are as follows:

[0093] (2)R 1 and R 2 It bonds with the inserted atom to form a 4- to 8-membered carbon ring or heterocycle, which may be substituted, R 4 and R 5 This is as defined in (1), or (3)R 1 and R 5 It bonds with the inserted atom to form a 4- to 8-membered carbon ring or heterocycle, which may be substituted, R 2 and R 4 This is as defined in (1), or (4)R 4 and R 5 It bonds with the inserted atom to form a 4- to 8-membered carbon ring or heterocycle, which may be substituted, R 1 and R 2 is as defined in (1) or (2), or (1)R 1 C is substituted with hydrogen, deuterium, and optionally C 1~4 Alkyl, and possibly substituted C 2~4 Alkenyl or optionally substituted C 2~4 It is alkinyl, R 2 , R 4, and R 5 Each of these can be independently hydrogen, a halogen (e.g., F), or optionally substituted C. 1~4 Alkyl, and possibly substituted C 2~4 Alkenyl, possibly substituted C 2~4 Alkinyl, and C may be substituted. 1~4 An alkoxy, or optionally substituted, 3- to 5-membered ring, X is O, S, NR 10 C may be replaced depending on the circumstances. 1~4 Alkylene, or optionally substituted C 1~4 It is a heteroalkylene, and here, R 10 C is a hydrogen atom, which may be substituted in some cases. 1~4 Alkyl groups, optionally substituted 3- to 6-membered rings, or nitrogen protecting groups. R 3 C is a hydrogen atom, which may be substituted in some cases. 1~4 Alkyl, or optionally substituted, 3- to 10-membered rings, R 6 C is substituted with hydrogen, deuterium, and optionally C 1~4 Alkyl, and possibly substituted C 2~4 Alkenyl or optionally substituted C 2~4 It is alkinyl, Each of the integers n1 and n2 is independently 0, 1, 2, 3, or 4. R a and R b Each existence of C is independently and, depending on the case, substituted. 1~4 Alkyl or optionally substituted C 1~4 It is a heteroalkylene, or two R a or two R's b However, it bonds with the inserted atom to form a 3-6 membered ring which may be substituted, and any remaining R a and / or R b It is defined as above.

[0094] The AKR1C3 enzyme-activating compounds of formula (12) were selected from compounds 1 to 569 in Tables 1 to 15 of patent application PCT / CN2023 / 123253, publication number WO2024078392A1, and the structures of some of these compounds are listed below.

[0095] [ka] [ka] [ka]

[0096] The specific synthesis methods and corresponding spectral data for compounds 1 to 569 of formula (12) are disclosed in International Publication No. 2024078392, which is incorporated herein by reference in its entirety.

[0097] When the AKR1C3 enzyme-activating prodrug is selected from the compounds of formulas (1), (3) to (8), (12) or their pharmaceutically acceptable salts, solvates, or isotopic isomers, The patient's tumor or cancerous tissue is detected to have homology-dependent recombination repair deficiency (HRD), or the patient is detected to have homology-dependent recombination repair deficiency (HRD).

[0098] Having a homology-dependent recombination repair defect means that one or both of the following conditions must be met: Mutations occur in one or more genes involved in the homology-dependent recombination repair (HRR) process. The genomic scarring (GS) score reaches a predetermined value.

[0099] Cellular DNA damage can be divided into single-strand breaks (SSBs) and double-strand breaks (DSBs). Of these, SSBs are mainly repaired by poly-ADP-ribose polymerase (PARP) via nucleotide excision repair (NER), base excision repair (BER), and mismatch repair (MMR). DSBs are repaired via pathways such as homologous recombination repair (HRR), non-homologous end joining (NHEJ), and microhomologous intervening end joining (MMEJ).

[0100] Homology-dependent recombination repair (HRR) is a high-fidelity repair method and one of the core DNA damage repair methods. It primarily occurs during the S and G2 phases of the cell cycle and is a DNA repair mechanism that maintains genomic integrity and ensures high-fidelity transmission of genetic information. Many genes are involved in this process, including ATM, ATR, BARD1, BLM, BRCA1, BRCA2, BRIP1, CDK12, CHEK1, CHEK2, FAAP20, FAN1, FANCE, FANCL, FANCM, MRE11A, NBN, PALB2, POLQ, PPP2R2A, RAD51B, RAD51C, RAD51D, RAD54L, RBBP8, SLX4, XRCC2, and others.

[0101] Homologous recombination repair deficiency (HRD) is a biological event caused by various factors, including genetic and epigenetic inactivation of genes in the homology-dependent recombination repair pathway, and is widely present in various tumors. When DNA is damaged, if it cannot be properly repaired via the homology-dependent recombination repair (HRR) pathway, it is called homology-dependent recombination repair deficiency (HRD).

[0102] HRD can lead to phenomena of "genomic scarring," including loss of heterozygosity (LOH), telomere allele mismatch (TAI), and large-scale state transitions (LST).

[0103] When double-strand damage and homology-dependent recombination repair deficiency (HRD) occur simultaneously, cells initiate low-fidelity repair methods for double-strand damage, such as non-homologous end joining (NHEJ) and microhomonymous intercalation end joining (MMEJ). This leads to gene insertions / deletions and other errors, resulting in high genomic instability, which is known as "genomic scarring (GS)." Thus, the phenomenon of genomic instability caused by HRD is called the "genomic scarring (GS)" phenomenon.

[0104] Genomic scarring primarily includes three types: loss of heterozygosity (LOH), telomere allele mismatch (TAI), and large-scale state transition (LST). Detection of combinations of these three types can maximize the state of genomic instability, thereby allowing for inverse prediction of whether HRD is present within the cell.

[0105] In summary, homology-dependent recombination repair mutations (HRRm) cause homology-dependent recombination repair deficiencies (HRD), while genomic scarring (GS) is a symptom of HRD. Homologous-dependent recombination repair mutations (HRRm) result in homology-dependent recombination repair deficiencies (HRD), which in turn lead to genomic scarring (GS).

[0106] Therefore, homology-dependent recombination repair deficiency (HRD) can be determined based on either or both of the following conditions: detection of mutations in one or more genes associated with the homology-dependent recombination repair (HRR) process, and / or reaching a predetermined genomic scar (GS) score.

[0107] However, since DNA double-strand breaks (DSBs) primarily rely on homology-dependent recombination repair (HRR) and can also be repaired by non-homologous end joining (NHEJ) and microhomologous intervening end joining (MMEJ) pathways, detection of HRRm alone does not necessarily lead to 100% HRD. Mutations in genes affecting repair pathways such as non-homologous end joining (NHEJ) and microhomologous intervening end joining (MMEJ) can also lead to GS development, so GS detection alone does not necessarily determine with 100% certainty that GS is related to HRRm.

[0108] Therefore, combined detection of HRRm and GS provides a more comprehensive assessment of the HRD status.

[0109] Two detection kits, FoundationFocus CDx BRCA LOH Assay and Myriad Genetics myChoice® HRD, have been approved. Designed based on Western populations, these two HRD detection products have been clinically validated and approved by the U.S. Food and Drug Administration (FDA), but no HRD companion diagnostic products are currently available in China.

[0110] Details of these two HRD detection kits are shown in Figure 21 (this figure is taken from the academic report "From BRCA to HRD Detection" by Professor Ouyang Nengtai at the academic conference of the Gynecologic Oncology Molecular Diagnosis and Treatment Committee of the Southern China Cancer Clinical Research Association on January 27, 2021).

[0111] Detecting HRD requires simultaneously determining whether mutations exist in one or more genes involved in homology-dependent recombination repair (HRR) and whether the genomic scarring (GS) score has reached a predetermined value. HRD positivity is determined only when both conditions are met.

[0112] The HRRm gene detected by two HRD detection kits approved by the US FDA is BRCA. In fact, other genes associated with homology-dependent recombination repair processes can also be selected from ATM, ATR, BARD1, BLM, BRCA1, BRCA2, BRIP1, CDK12, CHEK1, CHEK2, FAAP20, FAN1, FANCE, FANCL, FANCM, MRE11A, NBN, PALB2, POLQ, PPP2R2A, RAD51B, RAD51C, RAD51D, RAD54L, RBBP8, SLX4, and XRCC2.

[0113] The methods for detecting genomic scarring vary. FoundationFocus CDx BRCA LOH Assay detects the LOH state, while Myriad Genetics myChoice® HRD detects the comprehensive state of LOH, TAI, and LST.

[0114] The aforementioned compounds (1), (3)-(8), and (12) are activated in vivo by the AKR1C3 enzyme, which is highly expressed in tumor cells, and release DNA alkylating agents to exert their effects.

[0115] The DNA alkylating agents released after AKR1C3 enzyme activation of compounds (1), (3)-(7), and (12) are [ka] That is the case.

[0116] The DNA alkylating agent released after activation of the AKR1C3 enzyme of compound (8) [ka] That is the case.

[0117] All of the cytotoxic compounds of these drugs after metabolic activation are DNA alkylating agents that act on the DNA double helix to cause crosslinking, thereby resulting in DNA double-strand breaks (DSBs). In other words, the ultimate effect of the aforementioned compounds (1)-(8) and (12) is to induce DNA double-strand damage and, furthermore, to kill cancer cells.

[0118] DSBs are primarily repaired via the homology-dependent recombination repair (HRR) pathway. When DNA is damaged and cannot be properly repaired via the HRR pathway, it is called a homology-dependent recombination repair deficiency (HRD).

[0119] Therefore, the compounds described above should exhibit superior therapeutic effects in tumor models with homology-dependent recombination repair deficiency (HRD) compared to non-mutated tumor models; that is, AKR1C3-activated DNA alkylating prodrugs should have more pronounced therapeutic effects in HRD-positive cancer and tumor patients. In vitro cell proliferation inhibition experiments and animal model experiments also support this.

[0120] With respect to the drugs or formulations described herein, the prepared drugs contain the indicated compound or its salt or solvate within a specific dosage range, and / or the prepared drugs are administered in a specific dosage form and a specific mode of administration.

[0121] The drugs or formulations described herein may further contain pharmaceutically acceptable excipients or carriers. The drugs may be in any dosage form for clinical administration, e.g., tablets, suppositories, dispersible tablets, enteric-coated tablets, chewable tablets, orally disintegrating tablets, capsules, sugar-coated tablets, granules, dried powders, oral solutions, small injections for injection, lyophilized powder injections for injection, or large injections. Depending on the specific dosage form and mode of administration, the pharmaceutically acceptable excipients or carriers of the drug may include one or more of the following: diluents, solubilizers, disintegrants, suspending agents, lubricants, binders, fillers, flavoring agents, sweeteners, antioxidants, surfactants, preservatives, coating agents, and pigments.

[0122] The “administration” of a drug or the “giveness” of a drug shall be defined as achieving a “therapeutic effective dose.” A “therapeutic effective dose” of a drug refers to the amount of drug administered to a patient with cancer that, when given, produces the intended therapeutic effect (e.g., relief, improvement, reduction, or elimination of one or more clinical symptoms of cancer in the patient). Therapeutic effects do not necessarily occur with a single dose, but may only occur after a series of doses. Therefore, a therapeutic effective dose may be administered in one or more doses.

[0123] "Administering" or "using" a drug to a patient refers to direct administration, which may be administered to or self-administered by a healthcare professional, and / or indirect administration, which may be the act of prescribing a drug. For example, a physician who instructs a patient to self-administer a drug and / or gives a patient a prescription for a drug administers a drug to the patient.

[0124] "Treatment" or "treatment of a patient" means administering, using, or giving a patient a therapeutically effective amount of the drug in relation to the present invention.

[0125] "Treating" a condition or patient, "treatment" or "treatment of" a condition or patient means taking measures to obtain a beneficial or desired outcome (including clinical outcomes). For the purposes of this invention, beneficial or desired clinical outcomes include, but are not limited to, relief or improvement of one or more symptoms of cancer, reduction of disease severity, delay or slowing of disease progression, improvement of the disease state, remission or stabilization, or other beneficial outcomes. In some cases, treatment of cancer may result in a partial response or stable disease progression.

[0126] "Tumor cells" or "cancer cells" refer to tumor / cancer cells of any appropriate species, such as mammals like mice, dogs, cats, horses, or humans.

[0127] The subject of “treating cancer or tumors” refers to any suitable species, i.e., mammals, fish, etc., such as mice, dogs, cats, horses, or humans. Preferably, the patient is a mammal, more preferably a human.

[0128] The terms “patient” and “individual” are used interchangeably and refer to mammals requiring treatment for cancer. Typically, the patient is a human. In certain embodiments, “patient” or “individual” may refer to non-human mammals, such as non-human primates, dogs, cats, rabbits, pigs, mice, or rats, used to screen, characterize, and evaluate drugs and therapies.

[0129] A “prodrug” refers to a compound that, after administration, is metabolized or converted into a compound (or drug) that is biologically active, or otherwise more active, in respect of at least one property. Prodrugs are chemically modified to be less active or inactive compared to the drug, but the chemical modification is such that the corresponding drug is produced by metabolism or other biological processes after the prodrug is administered. Compared to the active drug, prodrugs may have altered metabolic stability or transport properties, fewer side effects or lower toxicity, or improved flavor. Prodrugs may be synthesized using reactants other than the corresponding drug.

[0130] Salts are basic salts or acidic salts, and solvates are hydrates or alkoxides.

[0131] The compounds also include salt forms of structures of formulas I and II; that is, the present invention provides pharmaceutically acceptable salts of the compounds. The salts may be basic salts, including salts formed by compounds having an inorganic base (e.g., alkali metal hydroxides, alkaline earth metal hydroxides, etc.) or an organic base (e.g., monoethanolamine, diethanolamine, triethanolamine, etc.). Alternatively, the salts may be acidic salts, including salts formed by compounds having an inorganic acid (e.g., hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, perchloric acid, sulfuric acid, phosphoric acid, etc.) or an organic acid (e.g., methanesulfonic acid, trifluoromethanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, fumaric acid, oxalic acid, maleic acid, citric acid, etc.). The selection and preparation of acceptable salts, solvates, etc. of the compounds are well known in the art.

[0132] The compounds described herein may also be used in the form of solvates, such as hydrates and alkoxides, the alkoxides including ethanolates.

[0133] The concept of isotopic variants or isomers in this invention is based on the concept of isotopic variants and chiral isomers in patent application PCT / US2016 / 062114, publication number WO2017087428, which corresponds to Chinese application No. 2016800446081, publication number CN108290911A.

[0134] Terms not specifically described or illustrated in the text of this application should be referred to textbooks on medicinal chemistry, organic chemistry, biochemistry, and pharmacology. [Brief explanation of the drawing]

[0135] [Figure 1] This figure shows the correspondence between AKR1C3 RNA expression levels and NRF2 RNA expression levels in the KRAS-G12D mutation model. [Figure 2] This figure shows the correspondence between the RNA expression levels of AKR1C3 and NRF2 in the KRAS-G12C mutation model. [Figure 3] This figure shows the correspondence between AKR1C3 RNA expression levels and NRF2 RNA expression levels in the KRAS-G13D mutation model. [Figure 4] The results of Western blot assays of target proteins after cell lysis with different concentrations of SFN are shown. The left panel shows images of Western blots with different concentrations of SFN, while the right panel shows bar graphs of protein band density analysis for NRF2 and AKR1C3. In each concentration group, the left bar represents NRF2 and the right bar represents AKR1C3. [Figure 5] The results of Western blot assays of target proteins after cell lysis with different concentrations of AST are shown. The left panel shows images of the Western blots with different concentrations of AST, while the right panel shows bar graphs of protein band density analysis for p-ERK2 and ERK2 with different concentrations of AST. In each AST concentration group, the bars on the left represent p-ERK2, and the bars on the right represent ERK2. [Figure 6] The results of Western blot assays of target proteins after cell lysis treated with 1 μM AST for different time intervals are shown. The left panel shows Western blot images from treatment with 1 μM AST at different time intervals, while the right panel shows bar graphs of protein band density analysis for p-ERK2 and ERK2 from treatment with 1 μM AST at different time intervals. In each treatment time group, the left bar represents p-ERK2 and the right bar represents ERK2. [Figure 7] The results of Western blot assays of target proteins after AKR1C3-dependent cell lysis are shown. The left panel shows images from the AKR1C3-dependent Western blot assay, while the right panel shows bar graphs of AKR1C3-dependent protein band density analysis for p-ERK2 and ERK2. In each treatment group, the left bar represents p-ERK2, and the right bar represents ERK2. [Figure 8]This document shows the results of Western blot assays and cell apoptosis assays of target proteins after cell lysis in ERK2 inhibitor, AST monotherapy, and combination therapy groups. The upper panel shows Western blot images of the ERK2 inhibitor group (PD98059, suramin) as monotherapy or combination therapy, while the lower left panel shows bar graphs of protein band density analysis for the ERK2 inhibitor group (PD98059, suramin) as monotherapy or combination therapy. In each treatment group, the left bar represents p-ERK2 and the right bar represents ERK2. The lower right panel shows bar graphs of cell apoptosis for the ERK2 inhibitor group (PD98059, suramin) as monotherapy or combination therapy, where RLU (relative luminescence unit) indicates the relative luminescence unit of the apoptosis signal. [Figure 9] The results of Western blot assays and cell apoptosis assays of target proteins after cell lysis in ERK2 activators, AST monotherapy, and combination therapy groups are shown. The top panel shows Western blot images of ERK2 activators (TPAs) as monotherapy or combination therapy. The bottom left panel is a bar graph of density analysis for ERK2 activators (TPAs) as monotherapy or combination therapy. In each treatment group, the left bar represents p-ERK2 and the right bar represents ERK2. The bottom right panel is a bar graph of cell apoptosis for ERK2 activators (TPAs) as monotherapy or combination therapy, where RLU (relative luminescence unit) indicates the relative luminescence unit of the apoptosis signal. [Figure 10]This is a schematic diagram illustrating how AST induces apoptosis in cancer cells with the KRAS G12D pathogenic mutation via the ERK2 signaling pathway. (1), (3), and (4) refer to relevant and supporting literature. (1)Prior IA,Hood FE and Hartley JL.The Frequency of Ras Mutations in Cancer.Cancer Res 2020;80:2969-2974; (3)Park HE, Yoo SY, Cho NY, Bae JM, Han SW, Lee HS, Park KJ, Kim TY and Kang GH.Tumor microenvironment-adjusted prognostic implications of the KRAS mutation subtype in patients with stage III colorectal cancer treated with adjuvant FOLFOX.Sci Rep 2021;11:14609; (4)Patricelli MP,Janes MR,Li LS,Hansen R,Peters U,Kessler LV,Chen Y,Kucharski JM,Feng J,Ely T,Chen JH,Firdaus SJ,Babbar A,Ren P and Liu Y.Selective Inhibition of Oncogenic KRAS Output with Small Molecules Targeting the Inactive State. Cancer Discov 2016;6:316-329; 1) shows that AST compounds act on and activate the MEK-ERK signaling pathway. 2) The AKR1C3 enzyme activates AST prodrug compounds, ultimately releasing the DNA alkylating agent AST-2660. 3) Tumor cell proliferation is inhibited by AST-2660. [Figure 11] The results of AKR1C3 expression levels measured by Western blot assay in Capan-1, PLC / PRF / 5, HT29, and H460 cell lines are shown. [Figure 12]The results of AKR1C3 expression levels measured by Western blot assay in Capan-1, HepG2, and H460 cell lines are shown. [Figure 13] These are growth curves of tumor volume in mice from each group during in vivo efficacy experiments of AST-3424 in a Capan-1 CDX model with BRCA pathogenic mutations. [Figure 14] These are curves showing the change in body weight of mice in each group during an in vivo efficacy experiment of AST-3424 in a Capan-1 CDX model with a BRCA pathogenic mutation. [Figure 15] These are growth curves of tumor volume in mice from each group during in vivo efficacy experiments of AST-3424 in the non-BRCA pathogenic mutation model HepG2-GFP. [Figure 16] These are curves showing the change in body weight of mice in each group during in vivo efficacy experiments of AST-3424 in the non-BRCA pathogenic mutant model HepG2-GFP. [Figure 17] These are the tumor volume growth curves in mice from each group during in vivo efficacy experiments of compound S in the BRCA pathogenic mutation model HuPrime® gastric cancer GA6201. [Figure 18] These are curves showing the percentage change in body weight of mice in each group during in vivo efficacy experiments of compound S in the BRCA pathogenic mutation model HuPrime® gastric cancer GA6201. [Figure 19] These are the tumor volume growth curves in mice from each group during in vivo efficacy experiments of compound S in the non-BRCA pathogenic mutation model HuPrime® lung cancer LU5173. [Figure 20] These are curves showing the percentage change in body weight of mice in each group during in vivo efficacy experiments of compound S in the non-BRCA pathogenic mutation model HuPrime® lung cancer LU5173. [Figure 21] This is a schematic diagram illustrating the detection characteristics and positive test criteria for two FDA-approved HRD detection kits, FoundationFocus CDx BRCA LOH Assay and Myriad Genetics myChoice®. [Modes for carrying out the invention]

[0136] The present invention will be described below with specific examples. Those skilled in the art will understand that these examples are used solely to illustrate the present invention and are not intended to limit its scope.

[0137] The experimental methods in the following examples are conventional methods unless otherwise specified. Unless otherwise noted, the pharmaceutical raw materials and reagents used are commercially available products.

[0138] The above description of specific embodiments of the present invention is not intended to limit the invention. Those skilled in the art can make various changes or modifications in accordance with the present invention without departing from the spirit of the invention, all of which shall fall within the scope of the appended claims of the present invention.

[0139] Specific experiments / examples related to the present invention are presented below to illustrate the invention.

[0140] AST-3424 [ka] AST-3424 analog compound AST (referred to as compound S in some examples) [ka]

[0141] 1. Distribution of AKR1C3 in KRAS G12D PDX and correlation statistics with NRF2 A total of 179 model data sets containing the KRAS-G12D mutation were obtained from the public tumor PDX model gene database at CrownBio (https: / / www.crownbio.cn / model-systems / in-vivo / pdx-models / ), and the RNA expression levels of AKR1C3 in these models were counted. The results are shown in Table 1.

[0142] [Table 3]

[0143] Furthermore, the NRF2 expression levels of these KRAS-G12D mutation models were statistically analyzed. The RNA expression levels of AKR1C3 and NRF2 in these 179 PDX models were plotted on an XY coordinate system to obtain Figure 1.

[0144] The distribution of AKR1C3 expression levels in the KRAS G12D PDX model was mainly concentrated in moderate and high expression levels (LOG2(FPKM)≧4), which accounted for 90.4%.

[0145] In the KRAS G12D PDX model, the distribution of NRF2 expression levels was also concentrated in moderate and high expression levels, which was consistent with the trend in AKR1C3 protein expression levels and showed a certain correlation.

[0146] 2. Distribution of AKR1C3 in KRAS G12C PDX and correlation statistics with NRF2 A total of 51 model data sets containing the KRAS-G12C mutation were obtained from the public tumor PDX model gene database at CrownBio (https: / / www.crownbio.cn / model-systems / in-vivo / pdx-models / ), and the RNA expression levels of AKR1C3 in these models were counted. The results are shown in Table 2.

[0147] [Table 4]

[0148] Furthermore, the NRF2 expression levels of these KRAS-G12C mutation models were statistically analyzed. The RNA expression levels of AKR1C3 and NRF2 in these 51 PDX models were plotted on an XY coordinate system, resulting in Figure 2.

[0149] In the KRAS G12C PDX model, AKR1C3 expression levels are evenly distributed across low, moderate, and high levels, with moderate expression levels (LOG2(FPKM)≧4) accounting for 66.7%.

[0150] In the KRAS G12C PDX model, the distribution trend of NRF2 expression levels showed a weak correlation with the distribution of AKR1C3 protein expression levels.

[0151] 3. Distribution of AKR1C3 in KRAS G13D PDX and correlation statistics with NRF2 A total of 41 model data sets containing the KRAS-G13D mutation were obtained from the public tumor PDX model gene database at CrownBio (https: / / www.crownbio.cn / model-systems / in-vivo / pdx-models / ), and the RNA expression levels of AKR1C3 in these models were counted. The results are shown in Table 3.

[0152] [Table 5]

[0153] Furthermore, the NRF2 expression levels of these KRAS-G13D mutant models were statistically analyzed. The RNA expression levels of AKR1C3 and NRF2 in 48 PDX models were plotted on an XY coordinate system, resulting in Figure 3.

[0154] In the KRAS G13D PDX model, AKR1C3 expression levels are evenly distributed across low, moderate, and high levels, with moderate or higher expression levels accounting for 83.3%.

[0155] In the KRAS G13D PDX model, the distribution trend of NRF2 expression levels showed a certain correlation with the distribution of AKR1C3 protein expression levels.

[0156] The statistical results above revealed a positive correlation between NRF2 expression and AKR1C3 expression. High NRF2 expression frequently accompanied high AKR1C3 expression. Cells and tissues with KRAS mutations frequently showed moderate and high AKR1C3 expression. In summary, KRAS mutations (including KRAS-G13D, KRAS G12C, and KRAS G12D) were associated with moderate and high expression of NRF2 and AKR1C3. KRAS mutations (including KRAS-G13D, KRAS G12C, and KRAS G12D) frequently accompanied moderate and high expression of NRF2 and AKR1C3.

[0157] To further explain the relationship between KRAS mutations and NRF2 and AKR1C3, the inventors conducted related mechanistic research experiments.

[0158] 4. AKR1C3 expression levels in HPAF II tumor cells with KRAS G12D mutations were regulated by NRF2. An SFN compound (sulforaphane, NRF2 activator) solution was added to HPAF II cell suspension and incubated overnight at 37°C in a 5% CO2 incubator. Cell protein lysates were collected for Western blotting (WB) detection.

[0159] The experiment was divided into five groups: 0.5% DMSO, 0.875 μM SFN, 1.75 μM SFN, 3.5 μM SFN, and 7 μM SFN.

[0160] HPAF II cells were treated with different concentrations of NRF2 activator SFN, and then the expression levels of NRF2 and AKR1C3 were detected by Western blotting. The results of Western blotting and protein band density analysis of NRF2 and AKR1C3 at different SFN concentrations are shown in Figure 4.

[0161] The experimental results showed that as the SFN treatment concentration increased, the expression level of NRF2 significantly increased, and the expression of AKR1C3 also increased simultaneously.

[0162] The experimental results showed that the NRF2 activator SFN upregulated NRF2 expression in a concentration-dependent manner. Under the same experimental conditions, AKR1C3 expression was upregulated simultaneously with the increase in SFN concentration. This result clearly demonstrated that AKR1C3 expression is regulated by NRF2. Furthermore, when combined with the significant correlations between NRF2 and AKR1C3 mRNA levels in the moderate and high expression ranges in Examples 1-3, it was shown that AKR1C3 expression levels were positively correlated with NRF2 expression levels in PDX models with the KRAS G12D mutation.

[0163] The AKR1C3 enzyme-activating prodrugs AST-3424 and AST exhibited significant antitumor effects and were well-tolerated in PDX and CDX models with high AKR1C3 enzyme expression (Examples 1-9 of PCT / CN2022 / 120817, publication number WO2023 / 046060).

[0164] Combined with the experimental conclusions in Examples 1-3 that AKR1C3 expression levels are positively correlated with NRF2 expression levels, those skilled in the art can reasonably surmise that KRAS gene mutations upregulate or activate NRF2, thereby promoting high AKR1C3 expression, and further enabling the AKR1C3 enzyme-activating prodrugs AST-3424 and AST to exert significant antitumor effects.

[0165] All compounds of general formulas (1) to (12) in this application are AKR1C3 enzyme-activated anticancer prodrugs, which are cleaved under the action of the AKR1C3 enzyme to produce anticancer active drugs such as AST-2660, paclitaxel, SN-38, gemcitabine, and KARS inhibitors. Therefore, those skilled in the art can reasonably deduce that KRAS gene mutations result in high expression of the AKR1C3 enzyme by upregulating or activating NRF2, thereby enabling the compounds of general formulas (1) to (12) to be activated and metabolized into anticancer active drugs. In other words, patients detected to have a KRAS gene mutation may benefit from treatment with compounds of general formulas (1) to (12), particularly treatment with AST-3424 or AST.

[0166] Using the AKR1C3 enzyme-activating prodrug AST as an example, we will investigate its mechanism of action in tumor cells with KRAS mutations, particularly those with the KRAS G12D mutation.

[0167] 5. AST activates the MEK / ERK signaling pathway. AST-induced ERK phosphorylation experiments were conducted. Three sets of variables were used to perform experiments in three groups: AST concentration, AST action duration, and AKR1C3 enzyme dependence. Of these, the AKR1C3 enzyme-dependent variable was based on the AKR1C3 enzyme-specific inhibitor AST-3021, and experiments were performed in or without the presence of the inhibitor. The inhibitor used was AST-3021 (also known as TH-3021, whose structural formula was reported by Flanagan et al. in Bioorganic and Medicinal Chemistry (2014), pp. 962-977). [ka] This is compound 36.

[0168] The experiment was divided into three groups, each treated with either different concentrations of AST (Group 1), 1 μM AST at different time intervals (Group 2), or with / without the AKR1C3 inhibitor AST-3021 (Group 3).

[0169] The specific compounds to be treated in each of the three groups were as follows: - Group 1 (concentration groups): 0.5% DMSO treatment group (24 hours), 0.5 μM AST treatment group (24 hours), 1 μM AST treatment group (24 hours), 10 μM AST treatment group (24 hours), - Group 2 (Time Group): 0.5% DMSO treatment group (24 hours), 1 μM AST treatment group (2 hours), 1 μM AST treatment group (8 hours), 1 μM AST treatment group (24 hours), - Group 3 (AKR1C3-dependent group): 0.5% DMSO treatment group (24 hours), 1 μM AST treatment group (24 hours), 3 μM AST-3021 treatment group (24 hours), 1 μM AST + 3 μM AST-3021 treatment group (24 hours).

[0170] Each compound solution was added to an HPAF II cell suspension and incubated overnight in a 37°C, 5% CO2 incubator. Cell protein lysates were collected for Western blotting (WB) detection according to the experimental group.

[0171] Western blotting was used to detect the expression levels of NRF2 and AKR1C3. Figure 5 shows Western blot images and protein band density analysis results for p-NRF2 and NRF2 from group 1, treated with different concentrations of AST. Figure 6 shows Western blot images and protein band density analysis results for p-NRF2 and NRF2 from group 2, treated with 1 μM AST for different durations. Figure 7 shows Western blot images and protein band density analysis results for p-NRF2 and NRF2 from group 3, treated with / untreated with the AKR1C3 inhibitor AST-3021.

[0172] The experimental results showed the following: In Figure 5, p-NRF2 expression increased in proportion to the increase in AST treatment concentration. Treatment with 10 μM AST for 24 hours significantly increased p-NRF2 expression, but AST treatment at all concentrations did not affect NRF2 expression. In Figure 6, p-NRF2 expression remained essentially unchanged after treatment with 1 μM AST for 2 hours and 8 hours, but significantly increased after 24 hours compared to the DMSO control group. In Figure 7, only the 1 μM AST mono-drug treatment group (24 hours) showed a significant increase in p-NRF2 expression, while p-NRF2 expression remained essentially unchanged in the other groups (0.5% DMSO treatment group (24 hours), 3 μM AST-3021 treatment group (24 hours), and 1 μM AST + 3 μM AST-3021 treatment group (24 hours)). In the experiments involving the three groups described above, total ERK2 expression remained unchanged across all treatment groups.

[0173] The above experiments demonstrated that AST induces ERK2 phosphorylation in a concentration-dependent manner without affecting ERK2 expression levels. After treatment with 1 μmol / L AST for 24 hours, ERK2 phosphorylation levels were significantly increased compared to the DMSO control group. As shown in Figure 7, HPAF II cells were pretreated with the AKR1C3-specific inhibitor AST-3021 2 hours prior to the addition of 1 μmol / L AST, and 3 μmol / L AST-3021 strongly inhibited the AST-mediated increase in ERK2 phosphorylation. Treatment with AST-3021 alone did not affect p-ERK2 levels. Under all test conditions, total ERK2 expression remained unchanged. Actin Western blotting confirmed the consistency of loading in all samples. AST activated the MEK / ERK signaling pathway in a concentration, time, and AKR1C3-dependent manner.

[0174] 6. AST induces apoptosis in KRAS G12D cells via the ERK2 signaling pathway. This experiment used three MEK / ERK modulators, including two inhibitors and one activator. The inhibitors were PD98059, a specific inhibitor of MEK1 / 2, and suramin, a broad-spectrum growth factor receptor inhibitor. The activator of the ERK signaling pathway was phorbol ester (TPA). The chemical structures of the inhibitor PD98059, the inhibitor suramin, and the activator TPA are shown below.

[0175] [ka]

[0176] The experiment was divided into an inhibitor group and an activator group, and both were subjected to Western blot assays and cell apoptosis assays.

[0177] In the Western blot assay, HPAF II cell suspensions were treated with each compound, incubated overnight at 37°C in a 5% CO2 incubator, and cell protein lysates were collected for Western blot (WB) detection.

[0178] The specific compounds that should be treated with inhibitors and activators, as determined by Western blotting, were as follows: - Inhibitor groups: 1% DMSO treatment group, 1 μM AST treatment group, 30 μM PD98059 treatment group, 1 mM suramin treatment group, 1 μM AST + 30 μM PD98059 treatment group, 1 μM AST + 1 mM suramin treatment group, -Activator group: 1% DMSO treatment group, 0.5 μM AST treatment group, 25 nM TPA treatment group, 0.5 μM AST + 25 nM TPA treatment group.

[0179] Furthermore, we performed apoptosis detection of cells. 1) Prepare the Caspase Glo® 3 / 7 reagent and equilibrate it to room temperature. Mix the reagent buffer and substrate in a 1:1 ratio to prepare the Caspase Glo® 3 / 7 reagent. It can be stored at 4°C for 3 days.

[0180] 2) 100 μL of Caspase Glo® 3 / 7 reagent was added to each well containing 100 μL of blank (cell-free) cells, negative control cells, or treated cells. Due to the sensitivity of the assay, care should be taken to avoid cross-contamination by ensuring that the pipette tip does not touch the wells containing the samples. The plate was covered with a plate sealer or lid.

[0181] 3) Gently mix the reagent solution on a shaker at 300-500 rpm for 30 seconds, then leave it at room temperature in the dark for 1 hour.

[0182] 4) The luminescence of each sample was measured using a plate-reading luminometer. The luminescence values ​​for each group were calculated and analyzed using bar graphs.

[0183] The specific compounds that should be treated with inhibitors and activators in cell apoptosis are as follows: - Inhibitor groups: 1% DMSO treatment group, 1 μM AST treatment group, 30 μM PD98059 treatment group, 1 mM suramin treatment group, 1 μM AST + 30 μM PD98059 treatment group, 1 μM AST + 1 mM suramin treatment group, -Activator group: 1% DMSO treatment group, 1 μM AST treatment group, 50 nM TPA treatment group, 1 μM AST + 50 nM TPA treatment group.

[0184] Figure 8 shows the results of Western blotting, protein band density analysis, and cell apoptosis experiments for the ERK2 inhibitor, AST monotherapy, and combination therapy groups. Figure 9 shows the results of Western blotting, protein band density analysis, and cell apoptosis experiments for the ERK2 activator, AST monotherapy, and combination therapy groups.

[0185] The experimental results showed the following: In the inhibitor groups in Figure 8, the upper and lower left panels showed that treatment with PD98059 and suramin alone slightly decreased p-ERK2 expression, while the combination therapy group of PD98059 and AST significantly decreased p-ERK2 expression, the combination therapy group of suramin and AST slightly increased p-ERK2 expression, but the AST monotherapy group significantly increased p-ERK2 expression. The lower right panel showed that the AST monotherapy group significantly affected cell apoptosis, while the PD98059, suramin monotherapy groups, and their respective combination therapies with AST had little effect on cell apoptosis. In the activator groups in Figure 9, the upper and lower left panels showed that both the AST and TPA monotherapy groups increased p-ERK2 expression compared to the DMSO control group, while the combination therapy group of AST and TPA significantly increased p-ERK2 expression. The lower right panel shows that AST monotherapy induced cell apoptosis, while combination therapy with TPA showed a more significant effect on cell apoptosis.

[0186] Experimental results showed that treatment with the two ERK inhibitors, PD98059 and suramin alone, slightly reduced the level of endogenous ERK phosphorylation, while treatment with AST alone significantly increased the level of intracellular p-ERK2. Conversely, the combination of AST and the two ERK inhibitors almost completely counteracted the AST-induced increase in p-ERK2. Using the caspase 3 / 7 kit, the effects of AST, ERK2 inhibitors alone, and in combination on cellular apoptosis were detected, and the results showed that the ERK2 inhibitor significantly inhibited AST-induced cellular apoptosis. In contrast, treatment with the ERK2 activator TPA alone could also increase the level of intracellular p-ERK2, and combined treatment with TPA and AST further enhanced the levels of AST-induced ERK2 phosphorylation and cellular apoptosis. Treatment of cells with any of the above monotherapy or combination therapies did not affect total ERK2 expression. These results clearly demonstrate that AST induces apoptosis in KRAS G12D mutant cells via the ERK2 signaling pathway.

[0187] Combining Examples 4, 5, and 6 allows us to estimate the mechanism shown in Figure 10, namely AST-induced apoptosis in cancer cells with the KRAS G12D pathogenic mutation via the ERK2 signaling pathway. Further combination with studies of related literature shows that the MEK / ERK signaling pathway can be activated by the action of DNA alkylating agents (Wang X, Martindale JL and Holbrook NJ. Requirement for ERK activation in cisplatin-induced apoptosis. J Biol Chem 2000;275:39435-39443). Examples 11 and 12 confirmed that AST, a DNA alkylating agent prodrug, has the effect of activating the MEK / ERK signaling pathway, which can induce apoptosis in KRAS G12D mutant cancer cells via the ERK2 signaling pathway, while the AST compound is an AKR1C3 enzyme-activated DNA alkylating agent prodrug that exerts an anticancer effect by being activated by the AKR1C3 enzyme to produce AST-2660.

[0188] Considering the relevant regulatory pathways of AKR1C3 and ERK2 as illustrated in Figure 10, we hypothesize that AST compounds activate and phosphorylate ERK2, thereby activating the MEK-ERK pathway, upregulating Nrf2, and ultimately leading to high expression of the AKR1C3 enzyme, which then induces cancer cell apoptosis. In other words, the KRAS-G12D mutation can more effectively activate AST drugs by upregulating the RAF / MEK / ERK signaling pathway, upregulating Nrf2 protein expression, and thereby upregulating AKR1C3 protein expression.

[0189] In fact, studies have shown that KRAS gene mutations can upregulate the protein expression of Nrf2, and subsequently the protein expression of AKR1C3. These genes include various subtypes under KRAS mutations, namely KRAS-G12D, KRAS-G12V, KRAS-G12C, KRAS-G12A, and KRAS-G13D.

[0190] Further extending the mechanism of the AST compound illustrated in Figure 10 to other similar AKR1C3 enzyme-activated prodrugs, general formulas (1) to (12), KRAS mutations can more effectively activate AKR1C3 enzyme-activated prodrugs, thereby upregulating the RAF / MEK / ERK signaling pathway, upregulating Nrf2 protein expression, and thus upregulating AKR1C3 protein expression, thereby inducing apoptosis or inhibiting cell proliferation.

[0191] Furthermore, the inventors have experimentally found that the AKR1C3 enzyme-activated DNA alkylating agents AST-3424 and AST have superior therapeutic effects against tumor models with BRCA mutations.

[0192] Based on two approved HRD detection kits, FoundationFocus CDx BRCA LOH Assay and Myriad Genetics myChoice® HRD, cell lines with BRCA gene mutations were selected for the relevant experiments. In some cases, cell lines or models with BRCA pathogenic mutations were used in the experiments. According to the judgment rules of the above kits, BRCA mutation positivity can be determined as HRD positivity.

[0193] 7. In vitro cytotoxicity comparison of compound AST-3424 in tumor cells with non-pathogenic and pathogenic BRCA mutations. In vitro cytotoxicity (IC) of human-derived tumor cell lines 50Using the st(I) value, we compared whether the compound AST-3424 showed a significant difference in tumor cells with BRCA non-pathogenic and pathogenic mutations, thereby detecting whether BRCA pathogenic mutations enhance the susceptibility of tumor cells to AST-3424 at the in vitro cell level. The following cell lines were used as subjects in the experiment: the BRCA pathogenic mutant cell line Capan-1 (human pancreatic cancer cell line, BRCA2 pathogenic mutation site: p.S1982RfsTer22), the BRCA non-pathogenic mutant cell line PLC / PRF / 5 (human hepatocellular carcinoma cell line), and HT29 (human colorectal cancer cell line).

[0194] The activation mechanism of the prodrug AST-3424 and its analog compound S (i.e., the aforementioned compound AST) involves the release of a DNA alkylating agent via the action of the AKR1C3 enzyme, thereby inhibiting and killing tumor cells. Therefore, in the presence or absence of pathogenic gene mutations, the higher the expression level of the AKR1C3 enzyme, the stronger the expected cytotoxicity of the compound to tumor cells, and IC 50 The values ​​become lower. The inventors chose to detect the effect of BRCA pathogenic mutations on the sensitivity of cells to AST-3424 and its analog compound S when the AKR1C3 protein expression levels of each cell line are similar, or when the AKR1C3 enzyme protein expression level of BRCA pathogenic mutant cell lines is relatively lower than that of non-BRCA pathogenic mutant cell lines. The inventors identified and quantitatively analyzed the AKR1C3 protein expression levels of the above three cell lines by Western blot assay. Meanwhile, the AKR1C3 protein expression levels of the three cell lines were compared using the human lung cancer cell line H460, which has high AKR1C3 protein expression, as a control. The Western blot results showed that the AKR1C3 protein expression levels of the Capan-1 and HT29 cell lines were similar, with the PLC / PRF / 5 cell line showing slightly higher expression (Table 4, Figure 11).

[0195] [Table 6]

[0196] Using cell proliferation experiments, we investigated the sensitivity of various cells to AST-3424, i.e., IC. 50 Value detected. Unless otherwise specified, IC 50 The values ​​were tested by a CTG assay.

[0197] IC2C of test compound AST-3424 in different cells as measured by the experimental method described above. 50 The values ​​are listed in Table 5 below.

[0198] [Table 7]

[0199] The test results showed that IC of AST-3424 in the BRCA non-pathogenic mutant cell line HT29 compared to the BRCA pathogenic mutant cell line Capan-1. 50 This showed that the IC50 ratio of AST-3424 was 67 times higher than that of the Capan-1 cell line. Therefore, the IC50 ratio of AST-3424 was observed between the BRCA pathogenic mutant cell line Capan-1 and the BRCA non-pathogenic mutant cell line HT29, both of which have similar AKR1C3 protein expression. 50 There was a significant difference in the values. This indicates that tumor cells with BRCA pathogenic mutations are more sensitive to AST-3424 under similar AKR1C3 protein expression conditions. Despite the relatively lower AKR1C3 protein expression level in the BRCA pathogenic mutant cell line Capan-1 compared to the BRCA non-pathogenic mutant cell line PLC / PRF / 5, the IC of AST-3424 in the BRCA non-pathogenic mutant cell line PLC / PRF / 5 was significantly lower. 50 This remained 22-fold compared to Capan-1 cell lines. Therefore, tumor cells with BRCA pathogenic mutations were more sensitive to AST-3424; in other words, HRD-positive tumor cells were more sensitive to AST-3424.

[0200] 8. In vitro cytotoxicity comparison of AST-3424 analog compound S in tumor cells with non-pathogenic and pathogenic BRCA mutations. Using the same experimental methods and cell lines as above, we further tested whether compound S showed a significant difference in tumor cells with BRCA non-pathogenic and pathogenic mutations, thereby detecting whether BRCA pathogenic mutations enhance the susceptibility of tumor cells to compound S. The in vitro cytotoxicity results of compound S for the three cell lines are shown in Table 6 below.

[0201] [Table 8]

[0202] The test results showed that the IC50 of AST-3424 analog compound S in the BRCA non-pathogenic mutant cell line HT29 was higher compared to the BRCA pathogenic mutant cell line Capan-1. 50 This showed that the IC50 of AST-3424 analog compound S was 12 times higher than that of the Capan-1 cell line. Therefore, the IC50 of AST-3424 analog compound S was observed between the BRCA pathogenic mutant cell line Capan-1 and the BRCA non-pathogenic mutant cell line HT29, both of which have similar AKR1C3 protein expression. 50 There was another significant difference in the values. Despite the relatively lower expression level of AKR1C3 protein in the BRCA pathogenic mutant cell line Capan-1 compared to the BRCA non-pathogenic mutant cell line PLC / PRF / 5, the IC50 of AST-3424 analog compound S in the BRCA non-pathogenic mutant cell line PLC / PRF / 5 was significantly lower. 50 This was still five times higher than that of the Capan-1 cell line. Therefore, tumor cell models with BRCA pathogenic mutations were more sensitive to AST-3424 analog compound S.

[0203] The results of both experiments above showed that, under similar AKR1C3 protein expression conditions, AST-3424 and its analog compound S exhibited a more pronounced in vitro killing effect against tumor models with BRCA pathogenic mutations. Even when the AKR1C3 protein expression level in BRCA pathogenic mutant cell lines was relatively lower than that in BRCA non-pathogenic mutant cell lines, AST-3424 and its analog compound S still exhibited a more pronounced in vitro killing effect against tumor models with BRCA pathogenic mutations.

[0204] 9. Comparison of in vivo efficacy of compound AST-3424 in BRCA non-pathogenic and pathogenic mutation tumor models. To investigate whether there is a difference in the in vivo efficacy of AST-3424 between BRCA pathogenic and non-pathogenic tumor cells, the inventors selected two CDX models: human pancreatic cancer Capan-1 (BRCA2 pathogenic mutation model, mutation site: p.S1982RfsTer22) and BRCA non-pathogenic mutation model HepG2 (human hepatocellular carcinoma model). The mRNA expression levels of AKR1C3 detected in the Capan-1 and HepG2 cell lines were LOG2(FPKM)=6.6363 and LOG2(FPKM)=6.8572, respectively, indicating that the mRNA expression levels of AKR1C3 in the two models are similar. A comparison of AKR1C3 protein expression levels with those in the H460 cell line is shown in Table 7 and Figure 12. Protein expression results showed that AKR1C3 protein expression levels in Capan-1 cells with BRCA pathogenic mutations were lower than those in HepG2 cells with non-pathogenic BRCA mutations. Based on this, the inventors compared the effect of BRCA pathogenic mutations on the in vivo efficacy of AST-3424.

[0205] [Table 9]

[0206] 10. In vivo efficacy experiment of AST-3424 in a Capan-1 CDX model with BRCA pathogenic mutations. A subcutaneous xenograft model of human pancreatic cancer was established by subcutaneous inoculation of human pancreatic cancer Capan-1 cells into BALB / c nude mice. The experiment was divided into three groups: an olaparib 100 mg / kg monotherapy group, an AST-3424 1.5 mg / kg monotherapy group, and a 10% anhydrous ethanol + 10% polyoxyethylene (35) castor oil + 80% glucose injection D5W (pH 7.4) vehicle control group, with 6 mice in each group. The olaparib monotherapy group was administered once daily via oral gastric tube feeding for 30 consecutive days. Both the vehicle control group and the AST-3424 monotherapy group were administered via tail vein injection. The vehicle control group received the drug once weekly for 3 weeks. The AST-3424 monotherapy group received the drug once weekly for 2 weeks. Specific administration regimens and TGIs are shown in Table 8. The group treated with the study drug olaparib 100 mg / kg showed a slight tumor inhibitory effect 43 days after tumor cell inoculation, with a relative tumor growth inhibition rate (TGI) of 37.43%, which was not statistically significant compared to the control group (p>0.05). The group treated with the study drug AST-3424 1.5 mg / kg showed a very significant tumor inhibitory effect 43 days after tumor cell inoculation, with a relative tumor growth inhibition rate (TGI) of 97.97%. Tumors in four mice completely disappeared, and the complete tumor inhibition rate was 66.7%, which was statistically significant compared to the control group (p<0.001).

[0207] [Table 10]

[0208] The corresponding curves of tumor volume and mouse body weight percentage over time are shown in Figures 13 and 14.

[0209] The experimental results showed that in a Capan-1 CDX model with BRCA pathogenic mutations, the AST-3424 1.5 mg / kg monotherapy group demonstrated a significant tumor inhibitory effect, achieving a 97.97% TGI at the end of the experiment, and that tumors were completely eliminated in four mice in this group. The body weight percentage data from the above animal experiment showed that the body weight of the experimental animals in the AST-3424 administration group did not decrease, demonstrating that the drug was well tolerable.

[0210] 11. In vivo efficacy experiment of AST-3424 in the non-BRCA pathogenic mutation model HepG2-GFP. A human hepatocellular carcinoma (HCP) orthotopic xenograft model was established by orthotopic inoculation of human hepatocellular carcinoma (HCP) HepG2-GFP cells into the liver tissue of BALB / c nude mice, and tumor size was determined using the fluorescence region of green fluorescent protein (GFP). The experiment was divided into three groups: a sorafenib monotherapy group, an AST-3424 1.25 mg / kg monotherapy group, and a saline injection vehicle control group, with 10 mice in each group. The sorafenib monotherapy group received oral gastric tube feeding once daily for 35 consecutive days. Both the vehicle control group and the AST-3424 monotherapy group received the drug by tail vein injection, with weekly administration for two weeks, followed by a one-week break, and then weekly administration for another two consecutive weeks. Specific administration regimens and TGIs are shown in Table 9.

[0211] [Table 11]

[0212] Note: TGI was calculated using the following formula (based on tumor weight): TGI% = (1 - Treatment (T) / Control (C)) × 100% (T and C are the tumor weights of the treatment and control groups at the end of the experiment, respectively).

[0213] The corresponding curves of tumor volume and mouse body weight over time are shown in Figures 15 and 16.

[0214] The experimental results showed that in the BRCA non-pathogenic mutation model, the tumor fluorescence area measurement in the AST-3424 1.25 mg / kg group was significantly smaller than that in the vehicle control group, but the difference was not statistically significant (p>0.05), and the TGI was 60%. This suggests that AST-3424 1.25 mg / kg has a tendency to inhibit proliferation in the human hepatocellular carcinoma HepG2-GFP orthotopic model, but the inhibitory effect was not statistically significant, and the mouse tumors were not completely eliminated. The body weight data from the above animal experiments showed that the body weight of the experimental animals in the AST-3424 administration group did not decrease, demonstrating that the drug is well tolerable.

[0215] Summary of the experiment From the experimental examples 9-11 above, the inventors found a significant difference in the in vivo efficacy of AST-3424. AST-3424 1.5 mg / kg (administered once a week for two weeks) showed very significant in vivo efficacy against a Capan-1 CDX model with a BRCA pathogenic mutation, with a TGI of 97.97%. Tumors were completely removed in 4 out of 6 mice in the AST-3424-treated group, resulting in a removal rate of 67%. However, AST-3424 1.25 mg / kg (administered once a week for two weeks, followed by a one-week discontinuation, then once a week for another two consecutive weeks) did not show significant in vivo efficacy against the BRCA non-pathogenic mutation model HepG2-GFP when the total dose exceeded the dose for the Capan-1 model. The TGI was 60%, which was not significantly different from the control group, and tumors were not completely removed in the mice in the treated group. The results above showed that although the AKR1C3 protein expression level in the Capan-1 model was lower than that of the HepG2-GFP model, the in vivo efficacy of AST-3424 differed significantly because the Capan-1 model had BRCA pathogenic mutations while the HepG2-GFP model did not. The BRCA pathogenic mutations increased the sensitivity of the Capan-1 model to AST-3424 and promoted tumor tissue death by AST-3424.

[0216] 12. Comparison of in vivo efficacy of compound S in BRCA non-pathogenic and pathogenic mutation tumor models. To investigate the difference in in vivo efficacy of AST-3424 analog compound S between BRCA pathogenic mutation models and non-pathogenic mutation models, we selected two PDX models: human gastric cancer PDX model GA6201 (BRCA2 pathogenic mutation model, mutation site: L1732PfsTer9) and BRCA non-pathogenic mutation PDX model LU5173 (human lung cancer PDX model). Results detected by existing methods showed that the mRNA expression levels of AKR1C3 in GA6201 and LU5173 tumor tissues were LOG2(FPKM)=6.78 and LOG2(FPKM)=8.88, respectively, indicating that the RNA expression levels of AKR1C3 in these two models are similar. The AKR1C3 protein expression levels in these two models were detected by immunohistochemistry (IHC), and the results are shown in Table 10. Immunohistochemistry and databases suggested that the AKR1C3 protein expression levels in these two PDX models were similar. In this part of the study, we compared the effect of BRCA pathogenicity mutations on the in vivo efficacy of AST-3424 analog compound S using these two models with similar AKR1C3 protein expression levels.

[0217] [Table 12] Note: Standard scoring criteria for IHC staining of AKR1C3 protein expression are mentioned in the literature (Guise et al., Cancer Res, 2010(70):1573), where "weak" corresponds to "Focal," "medium" to "Moderate," and "strong" to "Diffuse."

[0218] 13. In vivo efficacy experiment of compound S in gastric cancer GA6201 using the BRCA pathogenic mutation model HuPrime®. A subcutaneous xenograft tumor model of human gastric cancer was established by subcutaneous inoculation of BALB / c nude mice with HuPrime® model GA6201 tumor block. The experiment was divided into three groups: a group receiving the test drug ifosfamide 60 mg / kg, a group receiving the test drug compound S 2.5 mg / kg, and a group using a physiological saline (pH 7.0-7.6) vehicle control. Five mice were used in each group. The physiological saline (pH 7.0-7.6) vehicle control group and the group receiving the test drug compound S 2.5 mg / kg were administered once a week via tail vein injection for three weeks and observed for four weeks. The ifosfamide 60 mg / kg group was administered once daily via intraperitoneal injection for five consecutive days, followed by a two-day discontinuation, for a total of two weeks of administration and five weeks of observation. Specific administration regimens and TGIs are shown in Table 11.

[0219] [Table 13]

[0220] The corresponding curves of tumor volume and mouse body weight percentage over time are shown in Figures 17 and 18.

[0221] The experimental results showed that in the HuPrime® gastric cancer GA6201 model with BRCA pathogenic mutations, the group treated with the test drug ifosfamide (60 mg / kg) showed a specific tumor inhibitory effect on day 38 (3 days after completion of all administration cycles), with no statistically significant difference compared to the control group (p=0.857), and a relative tumor growth inhibition rate (TGI) of 58.83%. The group treated with test drug compound S (2.5 mg / kg) showed a significant tumor inhibitory effect on day 38 (3 days after completion of all administration cycles), with a statistically significant difference compared to the control group (p=0.000135), and a relative tumor growth inhibition rate (TGI) of 96.85%. The mice treated with test drug compound S (2.5 mg / kg) did not experience a decrease in body weight and did not exhibit obvious drug toxicity or good tolerance during treatment.

[0222] 14. In vivo efficacy experiment of compound S in the non-BRCA pathogenic mutation model HuPrime® lung cancer LU5173. To establish a subcutaneous xenograft model for human lung cancer, female NOD / SCID mice were subcutaneously inoculated with HuPrime® model LU5173 tumor block. The experiment was divided into three groups: a positive control drug ifosfamide 60 mg / kg group, a test drug compound S 4 mg / kg group, and a 7.5% anhydrous ethanol + 7.5% polyoxyethylene(35) castor oil + 85% glucose injection D5W vehicle control group, with 6 mice in each group. Of these, the 7.5% anhydrous ethanol + 7.5% polyoxyethylene(35) castor oil + 85% glucose injection D5W vehicle control group and the test drug compound S group were administered once a week via tail vein injection for 3 consecutive weeks. The positive control drug ifosfamide 60 mg / kg group was administered once daily via intraperitoneal injection for 5 consecutive days, followed by a 2-day interruption, and then administration for 2 consecutive weeks. Table 12 shows the specific administration regimens and TGIs.

[0223] [Table 14]

[0224] The corresponding curves of tumor volume and mouse body weight percentage over time are shown in Figures 19 and 20.

[0225] The experimental results showed that in a non-BRCA pathogenic variant HuPrime® lung cancer LU5173 model, the group treated with the test drug ifosfamide (60 mg / kg) showed a certain tumor inhibitory effect 32 days after the first dose, with a relative tumor growth inhibition rate (TGI) of 44.45%, which was not statistically significant compared to the control group (p>0.05). The group treated with test drug compound S (4 mg / kg) showed a significant tumor inhibitory effect 32 days after the first dose, with a relative tumor growth inhibition rate (TGI) of 52.58%, which was statistically significant compared to the control group (p<0.05). Mice treated with test drug compound S (4 mg / kg) did not show significant weight loss and were well tolerated during treatment.

[0226] Summary of the experiment From the experimental examples 12-14 above, the inventors found a significant difference in the in vivo efficacy of compound S between two models with similar AKR1C3 expression levels. In the BRCA pathogenic mutation HuPrime® gastric cancer GA6201 model, the group treated with the test drug compound S 2.5 mg / kg (administered once a week for 3 weeks) showed a significant tumor inhibitory effect, statistically significant compared to the control group (p=0.000135), with a relative tumor growth inhibition rate (TGI) of 96.85%. In the non-BRCA pathogenic mutation HuPrime® lung cancer LU5173 model, the group treated with a higher dose of the test drug compound S 4 mg / kg showed a significant tumor inhibitory effect 32 days after the first administration, with a relative tumor growth inhibition rate (TGI) of 52.58%, which was significantly lower than that of the HuPrime® gastric cancer GA6201 model. The results above show that even when compound S was used at a higher dose of 4 mg / kg in the non-BRCA pathogenic mutation model, the tumor inhibitory effect of compound S was lower than that of the low-dose 2.5 mg / kg group in the BRCA pathogenic mutation model. This also demonstrates that BRCA pathogenic mutations enhance the sensitivity of the HuPrime® gastric cancer GA6201 model to compound S and improve the killing effect of compound S on tumor tissue.

[0227] 15. Clinical detection of KRAS mutations and AKR1C3 enzyme protein expression levels in China In clinical trials approved by the Chinese regulatory authorities (CTR20220934), clinical detection of KRAS mutations and AKR1C3 enzyme protein expression levels was performed in enrolled Chinese clinical trial patients.

[0228] The trials were approved by the ethics committees of the participating medical institutions and were conducted in accordance with the principles of the Declaration of Helsinki and with the informed consent of all participants.

[0229] The detection and results of AKR1C3 enzyme protein expression levels were performed according to the method described in International Publication No. 2022048492. The H-score system was used to characterize the expression levels, with an H-score of ≥135 defined as high expression.

[0230] Among the 10 KRAS-positive patients, 7 had high AKR1C3 expression. The specific detection results are shown in Table 13 below.

[0231] [Table 15]

[0232] Similar to the toxic structures of AST-3424 and compound S, compounds with general formulas (1), (3)-(8), and (12) are all AKR1C3-activated anticancer prodrugs. These are lysed and, after activation by the AKR1C3 enzyme, release DNA alkylating agents with similar toxic structures. Therefore, combined with the experimental results for AST-3424 and compound S, it can be estimated that AKR1C3-activated DNA alkylating agent prodrugs with similar toxic structures (general formulas (1), (3)-(8), (12)) have a more significant therapeutic effect on cancer and tumor patients with homology-dependent recombination repair deficiency (HRD). Thus, for therapies utilizing homology-dependent recombination repair deficiency (HRD) positivity, it is appropriate to further select patients with high AKR1C3 expression (accompanied by KRAS gene mutations).

[0233] Furthermore, all of the AKR1C3-activated DNA alkylating agent toxins described above are alkylating agents that act on DNA double strands to induce crosslinking. These compounds should exhibit superior therapeutic effects in tumor models with mutations in homologous DNA double-strand break repair genes (HRRm) compared to non-mutated tumor models. In other words, AKR1C3-activated DNA alkylating agent prodrugs with similar toxin structures should have a more significant therapeutic effect on patients with cancer and tumors with homologous DNA double-strand break repair gene mutations when used alone or in combination with other therapeutic agents.

Claims

1. A method for administering cancer, tumors, or disorders or cell proliferation disorders resulting from cancer or tumors, comprising administering an AKR1C3 enzyme-activating prodrug to a patient if, without detecting the AKR1C3 protein expression level or AKR1C3 RNA content in the patient's tumor or cancer tissue, it is detected that the patient's tumor or cancer tissue has a KRAS gene mutation, or if it is detected that the patient has a KRAS gene mutation.

2. A method for treating cancer, tumors, or disorders or proliferative diseases caused by cancer or tumors, comprising the steps of administering a drug or formulation containing an AKR1C3 enzyme-activating prodrug, and detecting a KRAS gene mutation, but not comprising the step of detecting the amount of AKR1C3 protein expression or AKR1C3 RNA content in the patient's tumor or cancer tissue. A method comprising administering a drug or preparation containing an AKR1C3 enzyme-activating prodrug to the patient if it is detected that the patient's tumor or cancerous tissue has a KRAS gene mutation, or if it is detected that the patient has a KRAS gene mutation.

3. A pharmaceutically acceptable use of an AKR1C3 enzyme-activated prodrug in the manufacture of a drug for the treatment of cancer, tumors, or disorders or proliferative disorders resulting from cancer or tumors, characterized in that the drug is formulated for administration to a patient in whom a tumor or cancerous tissue has been detected to have a KRAS gene mutation, or to a patient in whom a KRAS gene mutation has been detected, and the detection of AKR1C3 protein expression levels or AKR1C3 RNA content in the patient's tumor or cancerous tissue is not required.

4. The method or use according to any one of claims 1, 2, or 3, wherein the KRAS gene mutation is selected from KRAS-G12D mutation, KRAS-G12V mutation, KRAS-G12C mutation, KRAS-G12A, and KRAS-G13D.

5. The AKR1C3 enzyme-activating prodrug is selected from the compounds of formulas (1) to (12) or their pharmaceutically acceptable salts, solvates, or isotopic isomers. 【Chemistry 1】 X, Y, Z, R, T, A and X １０ The definition is described in the claims of patent application PCT / US2016 / 021581, publication number WO2016145092A1 (corresponding to Chinese application No. 2016800150788, publication number CN107530556A), 【Chemistry 2】 X, Y, Z, R, D, L １ A and X １０ The definition is described in the claims of patent application PCT / US2016 / 025665, publication number WO2016 / 161342A1 (corresponding to Chinese application No. 2016800200132, publication number CN108136214A), 【Transformation 3】 R １ 、R ２ 、R ３ 、R ４ 、R ５ 、R ８ 、R ９ and R １０ are defined in the claims of the patent application PCT / CN2020 / 089692 with publication number WO2020228685A1 (corresponding to Chinese application No. 2020800358890 with publication number CN113853379A), 【Chemistry 4】 A is either substituted or non-substituted C ６ ~C １０ Aryl, biaryl or substituted biaryl, 5-15 member heteroaryl, or -N=CR １ R ２ Here, the substituents are halogen, -CN, and -NO. ２ , -O-(CH ２ )-O-,-CO ２ H and its salts, -OR １００ , -CO ２ R １００ , -CONR １０１ R １０２ , -NR １０１ R １０２ , -NR １００ SO ２ R １００ , -SO ２ R １００ , -SO ２ NR １０１ R １０２ , C １ ~C ６ Alkyl and C ３ ~C １０ Selected from a group consisting of heterocyclines, R １００ , R １０１ and R １０２ These are, independently, hydrogen and C １ ~C ８ Alkyl or C ６ ~C １２ It is either aryl or R １０１ and R １０２ These, together with the nitrogen atoms to which they are bonded, form a 5-7 membered heterocycle. Here, the alkyl group and aryl group are each composed of 1 to 3 halogen groups or 1 to 3 carbon atoms. １ ~C ６ It is replaced with an alkyl group, R １ and R ２ These are, independently, phenyl or methyl, X, Y, and Z are each independently hydrogen or halogen, R is hydrogen or C １ ~C ６ Alkyl or halogen-substituted alkyl, 【Transformation 5】 The definition of Rw is described in the claims of patent application PCT / CN2020 / 120281, publication number WO2021068952A1 (corresponding to Chinese application No. 202080071652.8, publication number CN114555574A), 【Transformation 6】 R １ , R ２ , R ３ , R ４ The definitions of and T are described in the claims of patent application PCT / CN2021 / 118597, publication number WO2022057838A1. 【Transformation 7】 The definitions of A, E, G, X, and Y are described in the claims of patent application PCT / NZ2019 / 050030, publication number WO2019190331A1 (corresponding to Chinese patent application No. 2019800234236, publication number CN111918864A). 【Transformation 8】 or a pharmaceutically acceptable salt thereof R ｗ X, R ４ , R １０ , R １３ , and R １４ The definition is described in the claims of patent application PCT / CN2022 / 098082, publication number WO2022258043A1. 【Chemistry 9】 R １ , R ２ , R ３ , R ４ G １ G ２ G ３ G ４ The definitions of E, T, Y, Z, m, n, s, t, v, w and ring A are described in the claims of patent application CN202210585771.6, publication number CN115403579A. 【Chemistry 10】 or a pharmaceutically acceptable salt thereof R １ , R ２ａ , R ２ｂ , R ３ , R ４ , R ５ The definitions of n and Z are described in the claims of patent application PCT / IB2020 / 057285, publication number WO2021005586A1 (corresponding to Chinese patent application CN202080053804.1, publication number CN114206870A), 【Chemistry 11】 or a pharmaceutically acceptable salt thereof R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ａ , R ｂ The definitions of n1 and n2 are as described in the claims of any one of claims 1, 2, or 3 of the patent application PCT / CN2023 / 123253, publication number WO2024078392A1.

6. When the AKR1C3 enzyme-activating prodrug is selected from the compounds of formulas (1), (3) to (8), (12) or their pharmaceutically acceptable salts, solvates, or isotopic isomers, The method or use of claim 5, wherein the tumor or cancerous tissue of the patient is detected to have a homology-dependent recombination repair deficiency (HDR), or the patient is detected to have a homology-dependent recombination repair deficiency (HDR).

7. Having a homology-dependent recombination repair defect is possible under the following conditions: A mutation occurs in one or more genes involved in the homology-dependent recombination repair (HRR) process. The genomic scarring (GS) score reaches a predetermined value. The method or use of claim 6, which must satisfy one or both of the following conditions.

8. Genes associated with homology-dependent recombination repair processes were selected from ATM, ATR, BARD1, BLM, BRCA1, BRCA2, BRIP1, CDK12, CHEK1, CHEK2, FAAP20, FAN1, FANCE, FANCL, FANCM, MRE11A, NBN, PALB2, POLQ, PPP2R2A, RAD51B, RAD51C, RAD51D, RAD54L, RBBP8, SLX4, and XRCC2. The method or use of claim 7, wherein the genomic scar is selected from loss of heterozygosity (LOH), telomere allele imbalance (TAI), and large-scale state transition (LST).

9. A drug formulation unit package comprising a separate packaging container for holding a drug formulation, an outer packaging component for housing the separate packaging container, and a drug usage instruction manual, wherein the drug formulation comprises an AKR1C3 enzyme-activated prodrug, and the drug usage instruction manual is The aforementioned drug is used to treat patients with cancer, tumors, or disorders or proliferative diseases resulting from cancer or tumors. Without detecting the AKR1C3 protein expression level or AKR1C3 RNA content in the tumor or cancerous tissue, KRAS gene mutation detection is performed on the patient. A drug unit package that records that if the tumor or cancerous tissue of the patient is detected to have a KRAS gene mutation, or if the patient is detected to have a KRAS gene mutation, a drug or formulation containing an AKR1C3 enzyme-activated prodrug will be administered to the patient.

10. KRAS gene mutations were selected from KRAS-G12D, KRAS-G12V, KRAS-G12C, KRAS-G12A, and KRAS-G13D. The AKR1C3 enzyme-activating prodrug is selected from the compounds of formulas (1) to (12) or their pharmaceutically acceptable salts, solvates, or isotopic isomers. 【Chemistry 12】 X, Y, Z, R, T, A and X １０ The definition is described in the claims of patent application PCT / US2016 / 021581, publication number WO2016145092A1 (corresponding to Chinese application No. 2016800150788, publication number CN107530556A), 【Chemistry 13】 X, Y, Z, R, D, L １ A and X １０ The definition is described in the claims of patent application PCT / US2016 / 025665, publication number WO2016 / 161342A1 (corresponding to Chinese application No. 2016800200132, publication number CN108136214A), 【Chemistry 14】 R １ 、R ２ 、R ３ 、R ４ 、R ５ 、R ８ 、R ９ and R １０ are defined in the claims of patent application PCT / CN2020 / 089692 (corresponding to Chinese application No. 2020800358890, published as CN113853379A) with publication number WO2020228685A1, 【Chemistry 15】 A is a substituted or unsubstituted C ６ ~C １０ aryl, biaryl or substituted biaryl, 5- to 15-membered heteroaryl, or -N=CR １ R ２ where the substituent is halogeno, -CN, -NO ２ , -O-(CH ２ )-O-, -CO ２ H and its salts, -OR １００ , -CO ２ R １００ , -CONR １０１ R １０２ , -NR １０１ R １０２ , -NR １００ SO ２ R １００ , -SO ２ R １００ , -SO ２ NR １０１ R １０２ , C １ ~C ６ alkyl and C ３ ~C １０ heterocyclyl, and is selected from the group consisting of, R １００ , R １０１ and R １０２ These are, independently, hydrogen and C １ ~C ８ Alkyl or C ６ ~C １２ It is either aryl or R １０１ and R １０２ These, together with the nitrogen atoms to which they are bonded, form a 5-7 membered heterocycle. Here, the alkyl group and aryl group are each composed of 1 to 3 halogen groups or 1 to 3 carbon atoms. １ ~C ６ It is replaced with an alkyl group, R １ and R ２ These are, independently, phenyl or methyl, X, Y, and Z are each independently hydrogen or halogen, R is hydrogen or C １ ~C ６ Alkyl or halogen-substituted alkyl, 【Chemistry 16】 The definition of Rw is described in the claims of patent application PCT / CN2020 / 120281, publication number WO2021068952A1 (corresponding to Chinese application No. 202080071652.8, publication number CN114555574A), 【Chemistry 17】 R １ , R ２ , R ３ , R ４ The definitions of and T are described in the claims of patent application PCT / CN2021 / 118597, publication number WO2022057838A1. [Chemistry 18] The definitions of A, E, G, X, and Y are described in the claims of patent application PCT / NZ2019 / 050030, publication number WO2019190331A1 (corresponding to Chinese patent application No. 2019800234236, publication number CN111918864A). 【Chemistry 19】 or a pharmaceutically acceptable salt thereof R ｗ X, R ４ , R １０ , R １３ , and R １４ The definition is described in the claims of patent application PCT / CN2022 / 098082, publication number WO2022258043A1. The AKR1C3 enzyme-activating prodrug is selected from the compounds of formulas (10) to (11) or their pharmaceutically acceptable salts, solvates, or isotopic isomers. 【Chemistry 20】 R １ , R ２ , R ３ , R ４ G １ G ２ G ３ G ４ The definitions of E, T, Y, Z, m, n, s, t, v, w and ring A are described in the claims of patent application CN202210585771.6, publication number CN115403579A. 【Chemistry 21】 or a pharmaceutically acceptable salt thereof R １ , R ２ａ , R ２ｂ , R ３ , R ４ , R ５ The definitions of n and Z are described in the claims of patent application PCT / IB2020 / 057285, publication number WO2021005586A1 (corresponding to Chinese patent application CN202080053804.1, publication number CN114206870A), 【Chemistry 22】 or a pharmaceutically acceptable salt thereof R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ａ , R ｂ The definitions of n1 and n2 are described in the claims of patent application PCT / CN2023 / 123253, publication number WO2024078392A1, for the formulation part packaging according to claim 9.

11. If the AKR1C3 enzyme-activating prodrug is selected from the compounds of formulas (1) to (9), (12) or their pharmaceutically acceptable salts, solvates, or isotopic isomers, then the tumor or cancerous tissue of the patient is detected to have a homology-dependent recombination repair deficiency (HDR), or the patient is detected to have a homology-dependent recombination repair deficiency (HDR). Preferably, Having a homology-dependent recombination repair deficiency means that one or both of the following conditions must be met: a mutation in one or more genes involved in the homology-dependent recombination repair (HRR) process, or a genomic scarring (GS) score reaching a predetermined value. Preferably, Genes related to homology-dependent recombination repair processes were selected from ATM, ATR, BARD1, BLM, BRCA1, BRCA2, BRIP1, CDK12, CHEK1, CHEK2, FAAP20, FAN1, FANCE, FANCL, FANCM, MRE11A, NBN, PALB2, POLQ, PPP2R2A, RAD51B, RAD51C, RAD51D, RAD54L, RBBP8, SLX4, and XRCC2. The formulation packaging according to claim 10, wherein the genomic scarring is selected from loss of heterozygosity (LOH), telomere allele mismatch (TAI), and large-scale state transition (LST).

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