Method for treating cancer and tumor patients by combining AKR1c3 enzyme-activated compound and immune checkpoint inhibitor

Abstract

A method for treating cancer or a tumor, in particular treating cancer by combining two drugs, relating to the field of cancer treatment. A method for treating cancer by combining a drug containing an AKR1C3 enzyme-activated compound and an immune checkpoint inhibitor / immune cell treatment drug. In particular, the immune checkpoint inhibitor is selected from an anti-PD -1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, etc., and the AKR1C3 enzyme-activated compound is selected from the compound of the following structure (I).

Description

A method of treating cancer and tumor patients by combining AKR1C3 enzyme-activated compounds with immune checkpoint inhibitors. Technical Field

[0001] This invention relates to treatment methods for cancer, particularly the combined use of two drugs to treat cancer, and belongs to the field of cancer treatment. Background Technology

[0002] The tumor microenvironment (TME) is a dynamic and complex micro-ecosystem composed of tumor cells and various immune cells. Inflammatory monocytes play an important role in the immune escape of cancer cells and also influence the tumor immune microenvironment through their interaction with T cells.

[0003] A recent study (Elewaut, A., Estivill, G., Bayerl, F. et al. Cancer cells impair monocyte-mediated T cell stimulation to evade immunity. Nature (2024). https: / / doi.org / 10.1038 / s41586-024-08257-4) found that inflammatory monocytes acquire antigens from tumor cells and activate T cells through a "cross-dressing" mechanism, thereby promoting anti-tumor immune responses. However, changes in signaling pathways in the tumor microenvironment, such as MAPK and PGE2, can inhibit this process, thereby weakening T cell activity and immune attack capabilities. Overactivation of the MAPK signaling pathway within tumor cells is the main reason for the suppression of this mechanism. Specifically, abnormal activation of MAPK signaling inhibits the production of type I interferon (IFN-I) (reducing it by 70%) and increases the secretion of prostaglandin E2 (PGE2) (increasing it by approximately 2.5 times). PGE2 not only directly inhibits the function of inflammatory monocytes but also blocks their effective interaction with T cells, thereby significantly weakening the reactivation capacity of T cells. Therefore, restoring the function of inflammatory monocytes is an effective way to treat cancer immunotherapy. For example, by increasing IFN-I levels and blocking PGE2 production, the function of inflammatory monocytes can be significantly restored, enhancing the tumor's immune attack mediated by T cells. By combining COX inhibitors (such as cyclooxygenase 2 inhibitors) and IFN-I agonists, the tumor microenvironment can be significantly improved, enhancing the effector function of T cells. By knocking out PGE2 synthesis-related genes using CRISPR / Cas9 technology, the production of PGE2 has been successfully reduced, restoring the activity of inflammatory monocytes.

[0004] The inventors of this application reported in patent PCT / CN2019 / 084604 and publication number WO2020010900A1 that DNA alkylating agents targeting the overexpression of aldehyde-ketone reductase AKR1C3 (compounds described in PCT / US2016 / 021581, PCT / US2016 / 025665, and PCT / US2016 / 062114) can inhibit the activity of AKR1C3 enzyme. Moreover, animal experiments have confirmed that DNA alkylating agents targeting the overexpression of aldehyde-ketone reductase AKR1C3 can reduce the content of prostaglandin E2 in animal blood. Interestingly, OBIPHARMA, INC. confirmed in patent PCT / US2021 / 029552 and publication number WO2022231580A1 that the combination of the AST-3424 (OBI-3424) compound described in PCT / US2016 / 062114 with immune checkpoint inhibitors (anti-PD-1 antibody (pembrolizumab) and anti-PD-L1 antibody (avelumab)) showed significant anti-tumor efficacy compared to AST-3424 monotherapy in a HepG2 tumor-humanized mouse model. Summary of the Invention

[0005] The compound (AST-3424) described in patents PCT / US2016 / 062114 and PCT / US2021 / 029552 is a DNA alkylating agent that targets the overexpression of aldehyde-ketone reductase AKR1C3, and AST-3424 can inhibit the activity of AKR1C3 enzyme and reduce the content of prostaglandin E2 in animal blood.

[0006] AST-3424 enters cancer cells and is activated by the AKR1C3 enzyme overexpressed by the cancer cells, releasing the metabolite AST-2660 (a DNA alkylating agent), thereby precisely killing cancer cells. Simultaneously, AST-3424 inhibits the activity of the AKR1C3 enzyme, reducing the level of prostaglandin E2. The combination immunotherapy of AST-3424 (OBI-3424) compound with immune checkpoint inhibitors (anti-PD-1 antibody (pembrolizumab)) and anti-PD-L1 antibody (avelumab)) described in patent PCT / US2021 / 029552 and publication number WO2022231580A1) may be because AST-3424 reduces the level of prostaglandin E2 by inhibiting the activity of the AKR1C3 enzyme, thereby restoring the activity of inflammatory monocytes. AST-3424 improves the tumor microenvironment, restores the activity of inflammatory monocytes, and enhances the effector function / reactivation ability of T cells, thereby achieving significant improvement in tumor treatment efficacy through combination immunotherapy of AST-3424 and PD-1 / PD-L1 inhibitors.

[0007] The discovery that cancer cells evade immune surveillance by inhibiting monocyte-mediated T cell reactivation reveals a complex and dynamic immune escape mechanism within the tumor microenvironment. By modulating two important signaling factors, IFN-I and PGE2, more effective combination immunotherapies can be developed, extending cancer treatment beyond traditional radiotherapy, chemotherapy, and surgery to achieve more durable anti-tumor effects by enhancing the body's immune response. Combination immunotherapy involves modulating key signaling factors to prevent cancer cells from escaping immune escape mechanisms and placing them back under T cell surveillance. For example, the combination of AST-3424 and PD-1 / PD-L1 inhibitors is an effective inhibitory therapy for hepatocellular carcinoma.

[0008] However, can all compounds that target the aldehyde-ketone reductase AKR1C3 inhibit the activity of AKR1C3 enzyme, reduce the content of prostaglandin E2, and restore the activity of inflammatory monocytes, thereby achieving significant combined immunotherapy effects?

[0009] Compounds that target the overexpression of aldehyde-ketone reductase AKR1C3 are prodrugs activated by the AKR1C3 enzyme. The prodrug itself is inactive, but under the action of the AKR1C3 enzyme, it is metabolized into drugs with tumor cell toxicity, such as DNA alkylating agents and AKR1C3 inhibitors.

[0010] The inventors of this application discovered through experimental research that other AKR1C3 enzyme-activated compounds also have combination immunotherapy effects.

[0011] In view of the above, this application proposes the following technical solution:

[0012] Treatment methods include using drugs containing AKR1C3 enzyme-activated compounds or their salts, esters, solvates, or isomers in combination with immune checkpoint inhibitors to treat cancer or tumor patients. The AKR1C3 enzyme-activated compounds do not include compounds with the following structures:

[0013] Treatment methods involve using drugs containing AKR1C3 enzyme-activated compounds or their salts, esters, solvates, or isotopic isomers in combination with immunotherapy drugs to treat cancer and tumor patients.

[0014] Immune checkpoint inhibitors (ICIs) are molecules that inhibit / block the inhibitory immune checkpoint system and have become an effective therapy for advanced neoplasmosis. Immunotherapy, represented by immune checkpoint inhibitors, works by activating the anti-tumor immune function of the patient's own T lymphocytes to kill tumor cells and has been widely used clinically. ICIs, as a series of antibodies developed targeting cytotoxic T-lymphocytocyte-associated protein 4 (CTLA-4) and programmed death 1 (PD-1) or their ligands PD-L1 / 2, have achieved good results in anti-tumor therapy.

[0015] Suppressive immune checkpoint antigens were selected from PD-1 / PD-L1 antigen, cytotoxic T-lymphocyte-Associated Protein 4 (CTLA-4), lymphocyte activation gene 3 (LAG-3), T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT), carcinoembryonic antigen-related cell adhesion molecule 1 (Cecam 1), leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1), T-cell immunoglobulin and mucin domain-3 (TIM-3), and V-domain Ig inhibitors for T-cell activation. Ig suppressor of T cell activation (VISTA), killer-cell immunoglobulin-like receptor (KIR), indoleamine-pyrrole 2,3-dioxygenase (IDO), B7-H3 (CD276), A2AR (adenosine A2A receptor), or CD47.

[0016] In some cases, immune checkpoint inhibitors are selected from anti-immune checkpoint antibodies that inhibit or block inhibitory immune checkpoint antigens.

[0017] In some cases, patients are cancer / tumor patients who are resistant to / insensitive to anti-immune checkpoint antibodies.

[0018] In some cases, anti-immune checkpoint antibodies are selected from one or more of the following: anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 (cytotoxic T lymphocyte-associated protein 4) antibody, anti-LAG-3 (lymphocyte activation gene 3) antibody, anti-TIGIT (inhibitory motif domain based on T cell immunoglobulin and immune receptor tyrosine) antibody, anti-Ceacam 1 (carcinoembryonic antigen-associated cell adhesion molecule 1) antibody, anti-LAIR-1 (leukocyte-associated immunoglobulin receptor-1) antibody, anti-TIM-3 (T cell immunoglobulin and mucin domain 3) antibody, anti-VISTA (Ig inhibitor of T cell activation V domain), anti-KIR (cytotoxic cell immunoglobulin receptor), anti-IDO (indoleamine-pyrrole 2,3-dioxygenase) antibody, anti-B7-H3 (anti-CD276) antibody, anti-A2AR (adenosine A2A receptor) antibody, and anti-CD47 antibody.

[0019] Preferably, the anti-PD-1 antibody is selected from pembrolizumab, toripalimab, sintilimab, camrelizumab, nivolumab, tislelizumab, pucotelimab, penpulimab, zimberelimab, serplulimab, cemiplimab, and dostarlimab.

[0020] The anti-PD-1 / CTLA-4 bispecific antibody was selected from catonilimab and ipalomlimab (a combination of ipalomlimab and tuvonralimab in a fixed ratio, QL1706, developed by Qilu Pharmaceutical).

[0021] The anti-PD-L1 antibodies were selected from durvalumab, atezolizumab, envafolimab, sugemalimab, avelumab, and adebrelimab.

[0022] The anti-CTLA-4 antibody was selected from ipilimumab and tremelimumab.

[0023] The anti-LAG-3 / PD-1 bispecific antibody was selected from OPDUALAG (a combination of nivolumab and relatlimab in a fixed ratio).

[0024] Clearly, these antibodies include monospecific antibodies, bispecific antibodies, and multispecific antibodies.

[0025] In some cases, for PD-1 or PD-L1, certain drugs require the detection of biomarkers and the detection results need to meet certain standards.

[0026] In some cases, for drugs containing AKR1C3 enzyme-activated compounds, certain drugs require the detection of biomarkers and the detection results need to meet certain standards.

[0027] In some embodiments, the cancer / tumor is hepatocellular carcinoma. The pathological paraffin blocks or sections of the ex vivo tumor tissue from the cancer / tumor patient are analyzed by immunohistochemical staining to detect the expression level of the AKR1C3 enzyme protein; the H-score score of the result is greater than or equal to 200.

[0028] or

[0029] The pathological paraffin blocks or sections of the ex vivo tumor tissue from the cancer / tumor patients were tested for AKR1C3 enzyme protein expression level by immunohistochemical staining. The results showed that the sum of the percentages of medium-intensity staining and high-intensity staining was greater than or equal to 70%.

[0030] Immunohistochemical staining (IHC) was used to detect the expression level of the AKR1C3 enzyme protein in pathological paraffin blocks or sections of isolated liver tumor tissue from patients. The specific IHC detection method is disclosed in patent application PCT / CN2021 / 114774, publication number WO2022048492A1, and the calculation method is explained as follows:

[0031] The percentage of cell staining in the focus area was evaluated on a semi-quantitative scale using AKR1C3 assays, which recorded the percentage of cytoplasmic and nuclear staining at four levels (0, 1+, 2+, and 3+).

[0032] (3) Tumor Sample Scoring Criteria

[0033] The H-score is used to assess the staining degree of tumor cells in the nuclear-cytoplasmic staining process (the total value from 0 to 3+ should not exceed 100), indicating the level of AKR1C3 enzyme expression.

[0034] 0 (Uncolored): Values ​​between 0 and 100

[0035] Tumor cell nucleus-cytoplasm 1+ (weak staining): values ​​between 0 and 100

[0036] Tumor cell nucleus-cytoplasm 2+ (moderate staining): values ​​between 0 and 100

[0037] Tumor cell nucleus-cytoplasm 3+ (high-grade staining, strong): values ​​between 0 and 100

[0038] Total percentage of positive staining in the nucleus and cytoplasm: values ​​between 0 and 100

[0039] The total H score will be calculated based on the proportion of tumors at each intensity. The final H score is calculated as follows: H-score = (%weak [1+] × 1) + (%medium [2+] × 2) + (%high [3+] × 3), with the final H score ranging from 0 to 300.

[0040] Obviously, the above-mentioned H-score of 200 is not theoretically equivalent to the sum of the percentages of moderate and high intensity staining being greater than or equal to 70%, as the latter does not take into account the case of weak staining. However, in the Phase II clinical trial conducted by the applicant, patients with advanced hepatocellular carcinoma whose test results showed a sum of the percentages of moderate and high intensity staining being greater than or equal to 70% all had corresponding H-scores greater than or equal to 200.

[0041] In some embodiments, the patient is a patient who tests negative for p53 gene mutation or whose p53 protein expression is normal.

[0042] In the Phase II clinical trial of AST-3424 in China, all enrolled patients were advanced, refractory hepatocellular carcinoma patients who had undergone multiple treatments, including:

[0043] Interventional therapies such as liver resection surgery, transarterial chemoembolization (TACE) and hepatic artery infusion chemotherapy, systemic single-agent chemotherapy and multi-agent combination chemotherapy, antibody-based immunotherapy (including monoclonal or bispecific antibodies such as PD-1 / PD-L1 / CTAL-4, and multi-antibodies), kinase inhibitor-based targeted therapy (i.e., molecular targeted therapy, such as tyrosine kinase inhibitors), combination therapy of antibody-based and kinase inhibitor-based drugs (targeted immunotherapy combination), radiotherapy, cell therapy, and traditional Chinese medicine treatment.

[0044] According to statistical classification, all 30 patients enrolled in the aforementioned Phase II clinical trial had received either antibody-based immunotherapy or molecular targeted therapy, or a combination of antibody-based immunotherapy and molecular targeted therapy (targeted immunotherapy combination). The preliminary results of the clinical trial indicate that AST-3424 monotherapy is still effective for patients with advanced refractory hepatocellular carcinoma who have progressed to disease progression (PD) after undergoing the above-mentioned treatments.

[0045] Further p53 gene mutation testing of the aforementioned 30 patients revealed that patients with negative p53 gene mutation results or positive p53 gene mutation results indicating a non-lethal mutation that does not affect normal p53 protein expression showed a significantly better response to AST-3424. In other words, compared to patients with positive p53 gene mutation results that affect normal p53 protein expression, these patients received AST-3424 treatment and achieved better therapeutic effects. Specifically, for patients whose AKR1C3 enzyme protein expression levels were measured by immunohistochemical staining, and whose H-score was greater than or equal to 200, and whose combined percentage of moderate and high-intensity staining was greater than or equal to 70%, if their p53 gene mutation results were negative or positive but indicated a non-lethal mutation that does not affect normal p53 protein expression, then these patients would benefit more from AST-3424 treatment. In other words, AST-3424 is more effective in combination immunotherapy for patients who test negative for p53 gene mutations or have normal p53 protein expression.

[0046] Fatal mutations, also known as pathogenic mutations, are called disease-causing mutations.

[0047] The p53 gene, also known as the tumor protein p53 gene or tp53 gene, is a tumor protein gene.

[0048] In this application, p53 (protein) and p53 (gene) are not distinguished and are used interchangeably.

[0049] In this application, p53 gene mutation negative includes no detected mutation and a situation where a mutation is detected but does not reach the defined positive level, i.e., a non-lethal mutation or a mutation that does not affect the normal expression of p53 protein.

[0050] Currently, relevant test kits have been approved for commercial use and can be purchased and used directly for testing, such as:

[0051] Produced by Xiamen AmoyDx Biotechnology Co., Ltd., China tp53 gene six mutation detection kit

[0052] The p53 gene amplification detection kit (fluorescence in situ hybridization) manufactured by Henan Sainote Biotechnology Co., Ltd. in China is a FISH detection kit for the p53 gene.

[0053] Manufactured by Integrated DNA Technologies, Inc., USA VariantPlex TM p53 kit for

[0054] p53 protein expression is considered normal, including normal expression as well as high expression (overexpression). Normal values ​​are artificially defined based on statistical analysis of clinical practice.

[0055] Normal expression of p53 protein can be determined by typical Western blotting (WB) or clinical infiltration chemoradiography (IHC). These methods directly measure the amount of p53 protein in the sample and then compare it with normal values ​​or a set threshold to determine whether it is normally or overexpressed.

[0056] Screening can also be performed through genetic testing. Generally, if a pathogenic gene mutation is detected, it can be determined that the p53 protein is neither normally expressed nor highly expressed. If no gene mutation is detected, or if a gene mutation is detected but it is not a pathogenic mutation, then it is highly likely that the p53 protein is expressed normally. Therefore, the normal / high expression of p53 protein can be determined by detecting whether a pathogenic gene mutation has occurred in the p53 gene.

[0057] The above-mentioned gene mutation or protein expression detection is generally obtained by detecting the patient's tumor or cancerous tissue, cells or other biological test samples.

[0058] Biological testing samples include peripheral blood samples, tumor tissue or suspected tumor tissue, thin-layer cytology samples, fine needle aspiration samples, bone marrow samples, lymph node samples, urine samples, ascites samples, irrigation samples, esophageal scrubbing samples, bladder or lung lavage samples, cerebrospinal fluid samples, cerebrospinal fluid samples, catheter aspiration samples, nipple discharge samples, pleural effusion samples, fresh frozen tissue samples, paraffin-embedded tissue samples, or extracts or processed samples derived from any of these samples. Venous whole blood or saliva are commonly used.

[0059] In some embodiments, immune cell therapy refers to adoptive immunotherapy.

[0060] Adoptive cell transfer therapy (ACT) refers to the isolation of immune-active cells (the main components of immunotherapy drugs) from cancer patients, their expansion and functional identification in vitro, and then the reinfusion of immunotherapy drugs into the patient. This aims to directly kill tumor cells or stimulate the body's immune response to kill tumor cells. Adoptive cell therapy mainly includes several categories such as TIL (Tumor Infiltrating Lymphocytes) cell therapy, LAK (lymphokine-activated killer cell therapy) cell therapy, CIK (cytokine-induced killer) cell therapy, DC (dendritic cell) cell therapy, NK (Natural Killer Cell) cell therapy, TCR-T (T-cell receptor engineered T cells) cell therapy, and CAR-T (Chimeric Antigen Receptor T-Cell) cell therapy.

[0061] In particular, the following CAR-T cell therapy products received marketing approval: Axicabtagene Ciloleucel (CD19), brexucabtagene autoleucel (CD19), Tisagenlecleucel (CD19), lisocabtagene maraleuce (CD19), idecabtagene vicleucel (BCMA), Relmacabtagene autoleucel (CD19), and Ciltacabtagene autoleucel (BCMA). CIK cell therapy products also received marketing approval. Green Cross Cell Corporation (BCMA) has received marketing approval for its DC cell therapy products: Sipuleucel-T and CreaVax-RCC.

[0062] In this application, drug resistance in cancer treatment can be broadly classified into primary (intrinsic) resistance and secondary (acquired) resistance. Primary resistance is characterized by a lack of objective clinical response after treatment, i.e., insensitivity. In a narrower sense, drug resistance refers to secondary (acquired) resistance, which means that there is a response upon initial administration of a certain drug, but the response decreases or ceases after a period of treatment.

[0063] In some embodiments, the AKR1C3 enzyme-activated compound is selected from the following structural formulas (4)-(15):

[0064] Formula (4) is a prodrug compound for AKR1C3 enzyme-activated alkylating agent, and more specifically, it is a prodrug compound for AKR1C3 enzyme-activated DNA alkylating agent.

[0065] Among them, X, Y, Z, R, T, A, and X 10 The definition is as described in the claims of patent application PCT / US2016 / 021581, publication number WO2016145092A1 (corresponding to Chinese application number 2016800150788, publication number CN107530556A), and the specific method for synthesizing and preparing the compound is also described in the above application. The entire text of the above application is incorporated herein by reference, and the specific definition is as follows:

[0066] X 10 It is O, S, SO or SO2;

[0067] A is C6-C 10 aryl, 5-15 quinone heteroaryl or -N=CR 1 R 2 ;

[0068] R 1 and R 2 Each can be independently hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, or C6-C 10 Aryl, 4-15 membered heterocycles, ethers, -CONR 13 R 14 or -NR 13 COR 14 ;

[0069] X, Y, and Z are each independently hydrogen, CN, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4-15 membered heterocycles, ethers, -CONR 13 R 14 or -NR 13 COR 14 ;

[0070] R represents hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4-15 membered heterocycles, ethers, -CONR 13 R 14 or -NR 13 COR 14 ;

[0071] R 13 and R 14 Each can be independently hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, or C6-C 10 Aryl, 4-15 membered heterocyclic or ether;

[0072] T contains an aminophosphate alkylating agent, said aminophosphate alkylating agent comprising one or more bonds to -OP(Z) 1 Z part 5 -X 5 -Y 5 Partial alkylating agent, of which Z 5 It is a heteroatom containing nitrogen, sulfur, or oxygen, X 5 It is a substituted or unsubstituted ethylened ethyl, Y 5 It is a halogen or another leaving group, or Z 5 -X 5 -Y 5 Together they form an aziridinyl (NCH2CH2) moiety and Z 1 It is O or S; and

[0073] These alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclic, heteroaryl, and etheryl groups may be substituted or unsubstituted.

[0074] Among them, X, Y, Z, R, D, L 1 A and X 10 The definition is as described in the claims of patent application PCT / US2016 / 025665, publication number WO2016161342A3 (corresponding to Chinese application number 2016800200132, publication number CN108136214A), and the specific method for synthesizing and preparing the compound is also described in the above application. The entire text of the above application is incorporated herein by reference, and the specific definition is as follows:

[0075] X 10 It can be O, S, SO or SO2;

[0076] A is C6-C 10 aryl, 5 to 15-membered heteroaryl, or -N=CR 1 R 2 ;

[0077] R1 and R 2 Each is independently hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4- to 15-membered heterocyclic, 5- to 15-membered heteroaryl, ether, -CONR 13 R 14 or -NR 13 COR 14 ;

[0078] 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-membered heterocyclic, 5- to 15-membered heteroaryl, ether, -CONR 13 R 14 or -NR 13 COR 14 ;

[0079] Each R is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4- to 15-membered heterocyclic, 5- to 15-membered heteroaryl, ether, -CONR 13 R 14 or -NR 13 COR 14 ;

[0080] R 13 and R 14 Each is independently hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4- to 15-membered heterocyclic, 5- to 15-membered heteroaryl or ether;

[0081] Where L 1 The definition of D is as follows:

[0082] L 1 Selected from:

[0083] R 40 and R 41 Independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4- to 15-membered heterocyclic or 5- to 15-membered heteroaryl;

[0084] R 42 C2-C3 alkyl or heteroalkyl groups substituted with 1 to 3 C1-C6 alkyl groups, depending on the case;

[0085] V(-) can be any anion, preferably a pharmaceutically acceptable anion;

[0086] D is the part that makes D-OH an anticancer drug, wherein OH is an aliphatic hydroxyl or phenolic hydroxyl, or an OH moiety attached to a phosphorus atom as provided herein; or

[0087] L 1 for:

[0088] R 40 As defined above, R 43 It is hydrogen or forms a heterocycle with D, and the extended phenyl moiety is substituted as appropriate, and

[0089] D is to make D-NR 43 H represents the portion of the anticancer drug; or

[0090] L 1 For the key, -OC(R) 40 R 41 )2-、-OC(R 40 R 41 )-NR 40 R 41 (+)-C(R 40 R 41 )-or

[0091] Where R 40 R 41 and V as defined above, and

[0092] D is an anticancer drug containing tertiary or secondary nitrogen atoms, wherein the tertiary or secondary nitrogen atoms are bonded to L. 1 ;

[0093] and

[0094] The alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclic, heteroaryl, and ether groups thereon may be substituted as appropriate.

[0095] Among them, R1, R2, R3, R4, R5, R8, R9, R 10 The definition is as described in the claims of patent application PCT / CN2020 / 089692, publication number WO2020228685A1, and the specific method for synthesizing and preparing the compound is also described in the above application. The entire text of the above application is incorporated herein by reference, and the specific definition is as follows:

[0096] R1 is C6-C 10 Aryl or Z-substituted aryl, 4-15 membered heterocycle or Z-substituted heterocycle, 5-15 membered heteroaryl or Z-substituted heteroaryl, 7-15 membered fused ring or Z-substituted fused ring;

[0097] R2 is hydrogen, a halogen atom, a cyano or isocyano group, a hydroxyl group, a mercapto group, an amino group, OTs, OMS, a C1-C6 alkyl or Z-substituted alkyl group, a C2-C6 alkenyl or Z-substituted alkenyl group, a C2-C6 ynyl or Z-substituted ynyl group, a C3-C8 cycloalkyl or Z-substituted cycloalkyl group, or a C6-C... 10 Aryl or Z-substituted aryl, 4-15 membered heterocycles or Z-substituted heterocycles, 5-15 membered heteroaryl or Z-substituted heteroaryl, ethers with 1-6 carbon atoms or Z-substituted alkoxy groups with 1-6 carbon atoms, -CONR 6 R 7 -SO2NR 6 R 7 -SO2R 6 -OCOO-R 6 -COOR 6 -NR 6 COR 7 -OCOR 6 -NR 6 SO2R 7 -NR 6 SO2NR 6 R 7 Alternatively, R2 and the atoms on the R1 group to which it is bonded can form a 7-15 member fused ring or a Z-substituted fused ring.

[0098] R3 is hydrogen, halogen, cyano or isocyano, hydroxyl, mercapto, amino, OTs, OMS, C1-C6 alkyl or Z-substituted alkyl, C2-C6 alkenyl or Z-substituted alkenyl, C2-C6 ynyl or Z-substituted ynyl, 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 -OCO-R 6 -OCOO-R 6 -COOR 6 -NR 6 COR 7 -OCOR 6 -NR 6 SO2R 7 ;

[0099] R4 and R5 are each independently hydrogen, halogen atom, cyano or isocyano, hydroxyl, mercapto, amino, OTs, OMS, C1-C6 alkyl or Z-substituted alkyl, C2-C6 alkenyl or Z-substituted alkenyl, C2-C6 ynyl or Z-substituted ynyl, 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 -NR 6 SO2R 7 Alternatively, R4, R5, and the atoms on the benzene ring to which they are bonded can form 7-15 fused rings or Z-substituted fused rings.

[0100] R 6 and R 7 Each of the following is independently hydrogen, cyano or isocyano, C1-C6 alkyl or Z-substituted alkyl, C2-C6 alkenyl or Z-substituted alkenyl, C2-C6 ynyl or Z-substituted ynyl, 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, or R 6 R 7 The group and the atoms it is bonded to together form a 5-7 membered heterocyclic group or a Z-substituted 5-7 membered heterocyclic group;

[0101] R8, R 10 Each of them 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 ynyl or Z-substituted ynyl, C3-C8 cycloalkyl or Z-substituted cycloalkyl, and at least one of them must be hydrogen or deuterium;

[0102] R9 is a substituted C6-C group having at least one fluorine atom or nitro group substitution. 10 Aryl, substituted 4-15-membered heterocyclic rings having at least one fluorine atom or nitro substitution, or substituted 5-15-membered heteroaryl rings having at least one fluorine atom or nitro substitution.

[0103] Z substituents are halogen atoms, cyano or isocyano, hydroxyl, mercapto, amino, OTs, OMS, C1-C3 alkyl or substituted alkyl, C1-C3 alkoxy or substituted alkoxy, C2-C3 alkenyl or substituted alkenyl, C2-C3 ynyl or substituted ynyl, C3-C8 cycloalkyl or substituted cycloalkyl, aromatic ring, heterocyclic, heteroaromatic ring and fused ring or substituted aromatic ring, heterocyclic, heteroaromatic ring and fused ring, and the substitution is monosubstituted or gemine disubstituted;

[0104] The substitution of C6-C in R9 10 The substituents of aryl, substituted 4-15-membered heterocyclic, and substituted 5-15-membered heteroaryl are halogen atoms, nitro, cyano or isocyano, hydroxyl, amino, C1-C3 alkyl or alkoxy, alkenyl, alkynyl, cycloalkyl or benzene ring, substituted benzene ring, C1-C3 alkoxy or halogen-substituted alkoxy.

[0105] Formula (8) is a prodrug compound for AKR1C3 enzyme activation alkylation, and more specifically, it is a prodrug compound for AKR1C3 enzyme activation DNA alkylation.

[0106] in:

[0107] A is a substituted or unsubstituted C6-C10 aryl, biaryl, or substituted biaryl, 5-15 membered heteroaryl, or -N=CR 1 R 2 The substituents used in the substitution process are selected from the following groups: halogens, -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 10 2 C1-C6 alkyl groups, C3-C10 heterocyclic groups;

[0108] Among them, R 100 R 101 and R 102 Each can be independently hydrogen, C1-C8 alkyl, or C6-C 12 Aryl; or R 101 and R 102 Together with the nitrogen atom to which it is attached, it forms a 5-7 membered heterocycle;

[0109] The alkyl and aryl groups are each substituted with 1-3 halogroups or 1-3 C1-C6 alkyl groups;

[0110] R 1 and R 2 Each can be either phenyl or methyl;

[0111] X, Y, and Z are each independently either hydrogen or a halide group;

[0112] R is hydrogen or a C1-C6 alkyl or a halogen-substituted alkyl.

[0113] The definition of Rw is as described in the claims of patent application PCT / CN2020 / 120281, publication number WO2021068952A1 (corresponding to Chinese application number 202080071652.8, publication number CN114555574A), and the specific method for synthesizing and preparing the compound is also described in the above application. The full text of the above application is incorporated herein by reference, and the specific definition is as follows:

[0114] Rw is

[0115] R1 represents H and C. 1-6 Alkyl, C 3-6 Cycloalkyl, 4-6-membered heterocycloalkyl, 5-6-membered heteroaryl or phenyl, wherein the C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-6-membered heterocycloalkyl, 5-6-membered heteroaryl, and phenyl groups are optionally surrounded by 1, 2, or 3 R groups. a Replaced;

[0116] Each R a Independently, it can be H, F, Cl, Br, I, -CN, -OH, or C. 1-3 Alkoxy or C 1-3 alkyl;

[0117] R2 is H or C 1-6 alkyl;

[0118] Alternatively, R1 and R2 can be linked together to form a 4-6 membered heterocyclic alkyl group with the attached N atom, wherein the 4-6 membered heterocyclic alkyl group is optionally surrounded by 1, 2, or 3 R atoms. b Replaced;

[0119] Each R b Independently, it can be H, F, Cl, Br, I, -CN, -OH, -NH2, -OCH3, -OCH2CH3, -CH3, or -CH2CH3;

[0120] R3 can be H, F, Cl, Br, I, -OH, -NH2, or C. 1-3 Alkoxy or C 1-3 alkyl;

[0121] Alternatively, R2 and R3 can be connected together to form a structural unit. for

[0122] T1 is -(CR) c R d ) m -or-(CR) c R d ) n -O-;

[0123] m is 1, 2, or 3;

[0124] n is 1 or 2;

[0125] T2 is either N or CH;

[0126] R c and R d Each is independently H, F, C 1-3 Alkyl or C 1-3 Alkoxy;

[0127] R4, R5, and R6 are each independently H, F, Cl, Br, I, and C, respectively. 1-3 Alkyl or C 1-3 Alkoxy;

[0128] T is either N or CH;

[0129] R7 and R8 are each independently H, F, Cl, Br or I;

[0130] R9 and R 10 Each can be independently H, F, Cl, Br, I, -CN or

[0131] The 4-6 membered heterocyclic alkyl and 5-6 membered heteroaryl groups each contain 1, 2, 3 or 4 heteroatoms independently selected from N, -O- and -S-.

[0132] The definitions of R1, R2, R3, R4, and T are as described in the claims of patent application PCT / CN2021 / 118597, publication number WO2022057838A1. The specific methods for synthesizing and preparing the compounds are also described in the aforementioned application, and are hereby incorporated in their entirety into this application, specifically defined as follows:

[0133] T is either N or CH;

[0134] R1 and R2 are each independently H, F, Cl, Br, I, or C. 1-3 Alkyl, wherein the C 1-3 Alkyl groups may be optionally surrounded by 1, 2, or 3 R's. a Replaced;

[0135] Each R a Independently, F, Cl, Br, I, -CN, -OH, or -NH2;

[0136] R3 and R4 are independently H, F, Cl, Br, I, CN, and C, respectively. 1-3 Alkyl, C 1-3 Alkoxy, Wherein, the C 1-3 Alkyl groups may be optionally surrounded by 1, 2, or 3 R's. e Replaced;

[0137] R b and R c Each can be independently H, -CH3, -CH2CH3, -(CH2)2CH3, or -CH(CH3)2;

[0138] R d It is -CH3, -CH2CH3, -(CH2)2CH3, -CH(CH3)2;

[0139] Each R e Independently F, Cl, Br, I, -CN, -OH or -NH2.

[0140] The definitions of A, E, G, X, and Y are as described in the claims of patent application PCT / NZ2019 / 050030, publication number WO2019190331A1 (corresponding to Chinese application number 2019800234236, publication number CN111918864A). The specific methods for synthesizing and preparing the compounds are also described in the aforementioned applications, and are hereby incorporated in their entirety into this application, specifically defined as follows:

[0141] A can be H, C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, CFH2, CF2H, CF3, F, Cl, Br, I, OCF3, COR, or CON(R)2;

[0142] E represents SO or SO2;

[0143] X is Cl, Br, I, or OSO2R;

[0144] Y is Cl, Br, I, or OSO2R;

[0145] Each R is independently H or a C1-C6 alkyl group;

[0146] G is a free radical selected from the group including formulas (B)-(AA):

[0147] in:

[0148] R1 is H, C1-C6 alkyl, CH2(CH2)nOH, CH2CH(OH)CH2OH, phenyl, pyridyl, benzyl, or pyridylmethyl, provided that when R1 is phenyl, pyridyl, benzyl, or pyridylmethyl, R1 is optionally substituted at any available position with C1-C6 alkyl, C1-C6 alkenyl, C1-C6 ynyl, OR6, N(R6)(R7), CFH2, CF2H, CF3, F, Cl, Br, I, OC, F3, COR6, CON(R6)(R7), SOR6, SON(R6)(R7), SO2R6, SO2N(R6)(R7), CN, or NO2;

[0149] R2 and R3 are each 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;

[0150] R4 can be N(R6)(R7), OH, OCH2(CH2)nN(R6)(R7) or CH2(CH2)nN(R6)(R7);

[0151] R5 is an H or C1-C6 alkyl group;

[0152] R6 and R7 are each independently H or C1-6 alkyl, or R6 and R7 together form a substituted or unsubstituted 5- or 6-membered heterocycle;

[0153] Z is CH or N;

[0154] W can be CH2, O, S, SO, or SO2;

[0155] n is between 0 and 6;

[0156] * indicates the connection point with equation (I).

[0157] 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 as described in the claims of patent application CN202210585771.6, publication number CN115403579A. The specific methods for synthesizing and preparing the compounds are also described in the aforementioned applications, and are hereby incorporated in their entirety into this application, specifically defined as follows:

[0158] G 1 G 2 G 3 or G4 The same or different, and each independently constitutes a CR 5 Or N atoms;

[0159] Each R 5 They may be the same or different, and each is independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, hydroxyl, cyano, amino, nitro, -NR a R b -C(O)NR a R b Cycloalkyl, heterocyclic, aryl, and heteroaryl, wherein each of the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups is independently and optionally substituted by one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, hydroxyl, oxo, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl, and heteroaryl groups;

[0160] Y is selected from -(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 -;

[0161] R y1Selected from hydrogen atoms, alkyl groups, haloalkyl groups, hydroxyalkyl groups, cycloalkyl groups, and heterocyclic groups;

[0162] R y2 and R y3 They may be the same or different, and each is independently selected from hydrogen atoms, halogens, alkyl groups, haloalkyl groups, hydroxyalkyl groups, cycloalkyl groups, and heterocyclic groups;

[0163] Or R y2 and R y3 Together they form = O;

[0164] Z is either O or OH;

[0165] For single or double bonds, when When it is a single bond, Z is OH. Z is 0 when it is a double bond;

[0166] E is selected from NH, O, and S atoms;

[0167] T is selected from -C(R) T1 R T2 )-、-NR T3 -or -O-;

[0168] R T1 and R T2 They may be the same or different, and each is independently selected from hydrogen atoms, deuterium atoms, halogens, alkyl groups, haloalkyl groups, hydroxyalkyl groups, cycloalkyl groups, and heterocyclic groups;

[0169] Or R T1 and R T2 Together with the carbon atom attached thereto, they form a cycloalkyl or heterocyclic group, each of which is independently and optionally replaced by one or more substituents selected from halogens, alkyl groups, and hydroxyl groups;

[0170] R T3 Selected from hydrogen atoms, alkyl groups, haloalkyl groups, hydroxyalkyl groups, cycloalkyl groups, and heterocyclic groups;

[0171] Ring A is a 6- to 10-membered aryl or a 5- to 10-membered heteroaryl;

[0172] Each R 1 They may be the same or different, and each is independently selected from hydrogen atom, deuterium atom, halogen, alkyl, alkenyl, alkoxy, hydroxyl, 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)Rc -S(O)2R c -B(OR) d 2. Nitro, cycloalkyl, heterocyclic, aryl, and heteroaryl, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups are each independently and optionally selected from 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. Substituted by one or more of the following substituents: nitro, cycloalkyl, heterocyclic, aryl, and heteroaryl;

[0173] R a and R b They may be the same or different, and each is independently selected from hydrogen atoms, alkyl groups, haloalkyl groups, hydroxyl groups, hydroxyalkyl groups, and -C(O)R groups. e , cycloalkyl and heterocyclic groups; or R a and R b Together with the nitrogen atom attached thereto, a cycloalkyl or heterocyclic group is formed, wherein the cycloalkyl or heterocyclic group is optionally substituted by one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, hydroxy, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl and heteroaryl.

[0174] R c The group is selected from alkyl, cycloalkyl, heterocyclic, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclic, aryl and heteroaryl groups are optionally substituted by one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, hydroxyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl and heteroaryl groups;

[0175] R d It is a hydrogen atom or a carbon atom. 1-6 alkyl;

[0176] R e The group is selected from alkyl, alkoxy, cycloalkyl, heterocyclic, aryl, and heteroaryl, wherein the alkyl, alkoxy, cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally substituted by one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, hydroxyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl, and heteroaryl groups;

[0177] Each R 2 They may be the same or different, and each is independently selected from hydrogen atom, deuterium atom, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, oxo, hydroxyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl and heteroaryl;

[0178] Each R 3 They may be the same or different, and each is independently selected from hydrogen atom, deuterium atom, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, oxo, hydroxyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl and heteroaryl;

[0179] R 4 Selected from hydrogen atoms, alkyl groups, haloalkyl groups, hydroxyl groups, and hydroxyalkyl groups;

[0180] n is 0, 1, 2, or 3;

[0181] v is 0, 1, or 2;

[0182] w is 0, 1, or 2;

[0183] f is 0, 1, or 2;

[0184] g is 0, 1, or 2;

[0185] h is 0, 1, or 2;

[0186] m can be 0, 1, 2, 3, 4, or 5;

[0187] s can be 0, 1, 2, 3, 4, 5, 6, 7, or 8;

[0188] t can be 0, 1, 2, 3, 4, 5, or 6;

[0189] The condition is that,

[0190] When Y is an -O- atom and E is an O atom, ring A is a phenyl or a 5- to 6-membered heteroaryl group, and G 3 For CR 5 Or N atom, R 5 Not a hydrogen atom;

[0191] When Y is an -O- atom and E is an S atom, ring A is a phenyl or a 5- to 6-membered heteroaryl group;

[0192] When G 1 G 2 G 3 and G 4 All are CR 5 Y is NR y1 When n, v, and w are all 1, and E is an O atom, 1) T is not CH2 or CD2, 2) there is at least one R.2 or R 3 For deuterium atoms, 3)R 4 Selected from alkyl, haloalkyl, hydroxy and hydroxyalkyl, 4) one of R 1 It is a 3- to 8-membered cycloalkyl or a 5- to 8-membered heterocyclic group, wherein the 3- to 8-membered cycloalkyl or 5- to 8-membered heterocyclic group is optionally substituted by one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, hydroxyl, oxo, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl and heteroaryl, 5) ring A is R d It is a hydrogen atom or a carbon atom. 1-6 alkyl.

[0193] Or its pharmaceutically acceptable salt.

[0194] Among them, R 1 R 2a R 2b R 3 R 4 R 5 The definitions of , n, and Z are as described in the claims of patent application PCT / IB2020 / 057285, publication number WO2021005586A1 (corresponding to Chinese application number CN202080053804.1, publication number CN114206870A). The specific methods for synthesizing and preparing the compounds are also described in the aforementioned applications, and are hereby incorporated in their entirety into this application, specifically defined as follows:

[0195] Is it a single bond or a double bond?

[0196] Z when When it is a single bond, it is OH; or when It is 0 when it is a double bond;

[0197] Each R 1 Independently selected from the group consisting of: (C1-C6)alkyl, (C1-C6)alkoxy, (C0-C4)alkylN(R) 8 2. and halogen groups;

[0198] R 2a and R 2b Each is independently selected from the group consisting of: H, (C1-C6)alkyl, and halogen;

[0199] Each R 3 Independently selected from the following groups: H, and halogen groups;

[0200] R 4The group consisting of: aryl groups, 5- to 6-membered heteroaryl groups comprising 1, 2, 3, or 4 heteroatoms independently selected from N, O, and S; and 9- to 10-membered fused bicyclic heteroaryl groups comprising 1, 2, 3, or 4 heteroatoms independently selected from N, O, and S; wherein any of the foregoing is optionally constituting one or more R 6 replace;

[0201] R 5 Choose from the following groups: H; (C1-C6)alkyl; (C2-C6)alkenyl; (C0-C4)alkyl OR 8 (C1-C4)alkyl (C3-C4)alkyl 10 )cycloalkyl; halo(C1-C6)alkyl; (C2-C3)ynyl; (C1-C4)alkylN(R) 10 )2;

[0202] Each R 6 Independently selected from the group consisting of: halogen; (C1-C6)alkyl; (C1-C6)alkoxy; halo(C1-C6)alkyl; OH; aryl; 3- to 6-membered heterocycle; 5- to 6-membered heteroaryl; (C0-C4)alkyl S(O) m (C1-C6)alkyl; halo(C1-C6)alkoxy; (C0-C4)alkylS(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 )cycloalkyl; wherein each R 6 When not a halogen group, OH, or OP(O)(OH)2, it is optionally subjected to one to three R groups. 9 Replacement; or two adjacent R 6 Together with the atoms to which they are attached, they form 5- to 7-membered heterocycles or (C5-C8) cycloalkyl groups;

[0203] Each R 7 and R 8 Independently select from the following groups: H or optionally selected by one to three Rs. 9 Substituted (C1-C6) alkyl groups;

[0204] Each R9 Independently selected from the group consisting of: halogen; -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- to 6-membered heterocycles optionally substituted with oxo (=O); (C0-C4)alkyl S(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 )2;

[0205] Each R 10 Independently selected from the group consisting of: H, (C1-C6)alkyl; or 3 to 6-membered heterocycles, wherein the 3 to 6-membered heterocycles are optionally substituted by one or more of: (C1-C6)alkyl; and oxo (=O);

[0206] Each R 11 Choose from the following groups: H; optionally selected by one to four Rs. 12 Replaced 4- to 6-membered heterocycles; optionally replaced by one to four R 12 Substituted (C3-C6)cycloalkyl; optionally substituted (C0-C3)alkyl, (C3-C6)cycloalkyl, (C1-C3)alkyl; optionally substituted with one to three R groups. 12 The substituted CH2-aryl; (C1-C6)alkyl; (C2-C6)alkenyl; or (C2-C6)ynyl, wherein each of the (C1-C6)alkyl; (C2-C6)alkenyl; and (C2-C6)ynyl is optionally substituted by one or more R 13 replace;

[0207] Each R 12 Independently selected from the group consisting of: OH, (C1-C3)alkoxy, NH2; or (C1-C3)alkyl groups optionally substituted with one or more OH groups;

[0208] Each R13 Independently selected from the group consisting of: halogen, OH, amino, (C1-C4)alkylamino, di(C1-C4)alkylamino, (C1-C3)alkoxy; and C(O)-(C3-C8)cycloalkyl;

[0209] m is 0, 1, or 2; and

[0210] n is 0, 1, or 2.

[0211] Or its pharmaceutically acceptable salt.

[0212] Among them, R w X, R4, R 10 R 13 R 14 The definition is as described in the claims of patent application PCT / CN2022 / 098082, publication number WO2022258043A1, and the specific method for synthesizing and preparing the compound is also described in the above application. The entire text of the above application is incorporated herein by reference, and the specific definition is as follows:

[0213] Each of the two X's is independently CR. 15 Or N;

[0214] R 13 R 14 Each of these groups is independently hydrogen, C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, C6-C 20 Aryl, a heterocyclic group of 5-20 membered rings, halogenated C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, halogenated C6-C 20 Aryl, or halogen-substituted 5-20 membered heterocyclic groups, and R 13 R 14 They are not both hydrogen;

[0215] R 10 It is hydrogen, C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, C6-C 20 Aryl, a heterocyclic group of 5-20 membered rings, halogenated C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, halogenated C6-C 20 Aryl, or halogen-substituted 5-20 membered heterocyclic groups;

[0216] Or R 10 With R 13 or R 14 It can meet the above R 10 R 13 R 14 Under the defined conditions, they are connected to form 5-9 elemental rings;

[0217] R4, R 15Each of these groups is independently hydrogen, halogen, C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, cyano, or a heterocyclic group with a 5-20 membered ring, C6-C 20 Aryl, or halogen-substituted C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, 5-20 membered heterocyclic groups, C6-C 20 Aryl;

[0218] Or R 10 With R 15 It can meet the above R 10 R 15 Under defined conditions, it forms 4-12 membered cyclic hydrocarbons or heterocycles;

[0219] R W yes

[0220] A is CR 16 Or N, where the position of A can vary on the ring;

[0221] R 16 Hydrogen, C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, C6-C 20 Aryl, a heterocyclic group of 5-20 membered rings, halogenated C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, halogenated C6-C 20 Aryl, or halogen-substituted 5-20 membered heterocyclic groups;

[0222] R6 and R7 satisfy the following conditions:

[0223] R6 and R7 are each independently hydrogen, halogen, cyano, hydroxyl, C1-C6 alkyl, cycloalkyl, alkenyl, alkoxy, heterocyclic group of 5-20 membered rings, or C6-C7. 20 Aryl, or halogen-substituted C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, 5-20 membered heterocyclic groups, C6-C 20 Aryl, or cyano-substituted C1-C6 alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, 5-20 membered heterocyclic groups, C6-C 20 Aryl or hydroxy-substituted C1-C6 alkyl, cycloalkyl, alkoxy, 5-20 membered heterocyclic groups, C6-C 20 Aryl, or -CONR 11 R 12 , or -CH2NR 11 R 12 ;

[0224] Or R6 and R7 are connected to form

[0225] A 5-8 member mono- or fused heterocyclic ring containing at least one N, S, or O, or two or three of N, S, and O simultaneously.

[0226] Alternatively, it may contain at least one N, S, or O, or two or three of N, S, and O, of a 5-8 member mono- or fused heterocyclic ring, wherein the mono- or fused heterocyclic ring is substituted with a C1-C6 alkyl group.

[0227] R6 can be used with CR 16 They connect to form 5-9 membered rings, heterocycles, or aromatic heterocycles;

[0228] R 11 R 12 The following conditions must be met:

[0229] R 11 R 12 Each is independently a C1-C6 alkyl group, a halogen-substituted C1-C6 alkyl group, or an R-substituted alkyl group. 11 R 12 Under the conditions defined above, -CONR 11 R 12 The N in it forms a 5-7 membered ring with -CH2NR. 11 R 12 The N in the ring forms a 5-7 member ring.

[0230] Or its pharmaceutically acceptable salt.

[0231] Among them, 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 patent application PCT / CN2023 / 123253, publication number WO2024078392A1. The specific methods for synthesizing and preparing the compounds are also described in the aforementioned application, and are hereby incorporated in their entirety into this application, specifically defined as follows:

[0232] in:

[0233] (II)R 1 R 2 Together with the inserted atoms, they connect to form arbitrarily substituted 4-8 membered carbon rings or heterocycles, R 4 and R 5 The definition is as described in (I); or

[0234] (III)R 1 R 5 Together with the inserted atoms, they connect to form arbitrarily substituted 4-8 membered carbon rings or heterocycles, R 2 and R 4The definition is as described in (I); or

[0235] (IV)R 4 R 5 Together with the inserted atoms, they connect to form arbitrarily substituted 4-8 membered carbon rings or heterocycles, R 1 and R 2 The definition is as described in (I) or (II); or

[0236] (I)R 1 It is hydrogen, deuterium, or any substituted C 1-4 Alkyl, any substituted C 2-4 alkenyl or any substituted C 2-4 alkynyl group;

[0237] R 2 R 4 R 5 Each can be independently hydrogen, halogen (e.g., F), or any substituted C. 1-4 Alkyl, any substituted C 2- 4-Alkenyl, any substituted C 2-4 Alkyne group, any substituted C 1-4 Alkoxy or optionally substituted 3- to 5-membered rings;

[0238] in:

[0239] X represents O, S, and NR. 10 Arbitrary substitution of C 1-4 Alkylene or any substituted C 1-4 Heteroalkylene, wherein R 10 It is hydrogen, and C with any substitution. 1-4 Alkyl groups, any substituted 3-6 membered rings, or nitrogen-protected groups;

[0240] R 3 It is hydrogen, and C with any substitution. 1-4 Alkyl or any substituted 3-10 membered rings;

[0241] R 6 It is hydrogen, deuterium, or any substituted C 1-4 Alkyl, any substituted C 2-4 alkenyl or any substituted C 2-4 alkynyl group;

[0242] Integers n1 and n2 are each independently 0, 1, 2, 3 or 4;

[0243] Each R a R b Each occurrence is independently an substituted C1-4 alkyl or optionally substituted C1-4 heteroalkylene; or two examples R a Or two instances R bTogether with the inserted atoms, they connect to form arbitrarily substituted 3-6 membered rings, and any remaining instances R a and / or R b As defined above.

[0244] Preferably, the AKR1C3 enzyme-activating compound of formula (4) is selected from compounds with the following structures:

[0245] The specific synthesis method and corresponding spectral data of the compound of formula (4) are disclosed in WO2016145092A1 (corresponding to Chinese public text CN107530556A), which are incorporated herein by reference in their entirety.

[0246] The AKR1C3 enzyme-activating compound of formula (5) is selected from compounds with the following structures:

[0247] The specific synthesis method and corresponding spectral data of the compound of formula (5) are disclosed in WO2016161342A1 (corresponding to Chinese public texts CN108136214A and CN112142692A), which are incorporated herein by reference in their entirety.

[0248] The AKR1C3 enzyme-activating compound of formula (6) is selected from compounds with the following structures:

[0249] The AKR1C3 enzyme-activating prodrug compound of formula (7) is selected from compounds with the following structures:

[0250] The specific synthesis methods and corresponding spectral data of the compounds in formulas (6) and (7) are disclosed in WO2020228685A1 (corresponding to Chinese publication CN113853379A), which are incorporated herein by reference in their entirety.

[0251] The AKR1C3 enzyme-activating compound of formula (8) is selected from compounds with the following structures:

[0252] as well as

[0253]

[0254] as well as

[0255] The specific synthesis method and corresponding spectral data of the compound of formula (8) are disclosed in WO2020228685A1 (corresponding to Chinese public text CN113853379A) ​​and WO2016145092A1 (corresponding to Chinese public text CN107530556A), which are incorporated herein by reference in their entirety.

[0256] The AKR1C3 enzyme-activating compound of formula (9) is selected from compounds with the following structures:

[0257] The specific synthesis method of the compound of formula (9) and the corresponding spectroscopic data are disclosed in WO2021068952A1 (corresponding to Chinese public text CN114555574A), which are incorporated herein by reference in their entirety.

[0258] The AKR1C3 enzyme-activating compound of formula (10) is selected from compounds with the following structures:

[0259] The specific synthesis method and corresponding spectral data of the compound of formula (10) are disclosed in WO2022057838A1, which are incorporated herein by reference in their entirety.

[0260] The AKR1C3 enzyme-activating compound of formula (11) is selected from compounds with the following structures:

[0261] The specific synthesis method of the compound of formula (11) and the corresponding spectroscopic data are disclosed in WO2019190331A1 (corresponding to Chinese public text CN111918864A), which are incorporated herein by reference in their entirety.

[0262] The AKR1C3 enzyme-activating compound (KARS inhibitor) of formula (12) is selected from compounds with the following structures:

[0263] The specific synthesis method of the compound of formula (12) and the corresponding spectroscopic data are disclosed in Chinese public text CN115403579A, which is incorporated herein by reference in its entirety.

[0264] The AKR1C3 enzyme-activating compound (KARS inhibitor) of formula (13) is selected from compounds with the following structures:

[0265] The specific synthesis method of the compound of formula (13) and the corresponding spectroscopic data are disclosed in WO2021005586A1 (corresponding to Chinese public text CN114206870A), which are incorporated herein by reference in their entirety.

[0266] The AKR1C3 enzyme-activating compound of formula (14) is selected from compounds with the following structures:

[0267] The specific synthesis method and corresponding spectral data of the compound of formula (14) are disclosed in WO2022258043A1, which are incorporated herein by reference in their entirety.

[0268] The AKR1C3 enzyme-activating compounds of formula (15) are selected from compounds 1-569 in Table 1-15 of patent application PCT / CN2023 / 123253, publication number WO2024078392A1. The structures of some compounds are listed below:

[0269] The specific synthesis method and corresponding spectral data of compound 1-569 of formula (15) are disclosed in WO2024078392A1 and are incorporated herein by reference in their entirety.

[0270] In some embodiments of this application, the AKR1C3 enzyme-activating compound is selected from the following structural compounds:

[0271] In some embodiments of this application, the drug containing an AKR1C3 enzyme-activated compound or its salt, ester, solvate, or isomer is selected from injectable formulations of compounds with the following structural formula, wherein the injectable formulation contains a compound with the following structural formula, N,N-dimethylacetamide, polyoxyethylene (35) castor oil, and ethanol.

[0272] In some embodiments of this application, the concentration of the compound with the following structure in the injectable formulation is 10-20 mg / ml.

[0273] In some embodiments of this application, the injectable formulation is a concentrated ethanol solution, labeled as 2 ml in volume, containing 20 mg of the following compound, 100 mg of N,N-dimethylacetamide, and 800 mg of polyoxyethylene (35) castor oil.

[0274] The drug described in this application refers to a pharmaceutical product or preparation, wherein the prepared pharmaceutical product contains an AKR1C3 enzyme-activated compound of the active ingredient formula (4)-(15) within a specific dosage range or its salt, ester, solvate, isomer, and / or the prepared pharmaceutical product is administered in a specific dosage form or by a specific method of administration.

[0275] In addition to containing AKR1C3 enzyme-activating compounds of formulas (4)-(15), the aforementioned drugs should also contain pharmaceutically acceptable excipients or excipients, depending on the specific drug, pharmaceutical preparation, or formulation. The drug can be any dosage form for clinical use, such as tablets, suppositories, dispersible tablets, enteric-coated tablets, chewable tablets, orally disintegrating tablets, capsules, sugar-coated tablets, granules, dry powders, oral solutions, small injection needles, lyophilized powder for injection, or large-volume infusions. Depending on the specific dosage form and administration method, the pharmaceutically acceptable excipients or excipients in the drug may include one or more of the following: diluents, solubilizers, disintegrants, suspending agents, lubricants, binders, fillers, flavoring agents, sweeteners, antioxidants, surfactants, preservatives, encapsulating agents, and pigments, etc.

[0276] The preparation of reagents from specific AKR1C3 enzyme-activating compounds includes the following implementations:

[0277] 1. An injectable preparation, a concentrated ethanol solution, labeled as 2 ml in volume, containing 20 mg of the following compound, 100 mg of N,N-dimethylacetamide, and 800 mg of polyoxyethylene (35) castor oil.

[0278] For specific preparation methods, please refer to PCT patent application PCT / CN2024 / 092639, publication number WO2024230831.

[0279] 2. Lyophilized formulation: Using TFX05-0110mg, sodium sulfobutyl betacyclodextrin (SBECD) 570-630mg, and sodium bicarbonate (NaHCO3) 0.1995-0.2205mg, the solution is dissolved in water for injection to a final volume of 5mL and then lyophilized to obtain the lyophilized formulation containing TFX05-0110mg, sodium sulfobutyl betacyclodextrin (SBECD) 570-630mg, and sodium bicarbonate (NaHCO3) 0.1995-0.2205mg. For specific preparation methods, please refer to patent CN118021737A and PCT / CN2020 / 120281.

[0280] 3. Lyophilized formulation: Using TFX05-0110.0mg, mannitol 142.5-157.5mg, disodium hydrogen phosphate dodecahydrate 62.7-69.3mg, sodium dihydrogen phosphate 1.425-1.575mg, and tert-butanol 769.5-850.5mg, the solution was diluted to 5mL with water for injection and then lyophilized to obtain the lyophilized formulation of TFX05-0110.0mg, mannitol 142.5-157.5mg, disodium hydrogen phosphate dodecahydrate 62.7-69.3mg, and sodium dihydrogen phosphate 1.425-1.575mg. For the specific preparation method, please refer to patent CN118021738A and PC T / CN2020 / 120281.

[0281] 4. Oral solid dosage form, the drug composition includes:

[0282] (i) an intraparticle blend, wherein the intraparticle blend comprises:

[0283] (a) Amorphous spray particles comprising: a compound having formula (I), wherein the compound is present in an amorphous form in an amount of 11.9 wt%; and about 26.6 wt% of hydroxypropyl methylcellulose succinate.

[0284] (b) Approximately 1.3 wt% of silicon dioxide;

[0285] (c) Approximately 30.3 wt% lactose;

[0286] (ii) Extragranular blends, wherein the extragranular blend comprises:

[0287] (d) Approximately 24.6 wt% lactose and microcrystalline cellulose;

[0288] (e) Approximately 2.8 wt% of croscarmellose sodium;

[0289] (f) approximately 1.0 wt% of silicon dioxide; and

[0290] (g) Approximately 1.5 wt% of sodium stearate fumarate.

[0291] For details on its preparation method, please refer to patent WO2024023659.

[0292] "Cancer" refers to leukemia, lymphoma, carcinoma, and other malignant tumors (including solid tumors) that can potentially grow without restriction, spreading locally through invasion and throughout the body through metastasis.

[0293] Examples of cancers that can be treated with AST-3424 or similar AKR1C3 enzyme-activated compounds include (but are not limited to) cancers of the adrenal glands, bone, brain, breast, bronchi, colon and / or rectum, gallbladder, head and neck, kidneys, larynx, liver, lungs, nervous tissue, pancreas, prostate, parathyroid glands, skin, stomach, and thyroid. Other examples of cancers include acute and chronic lymphocytic and granulocytic tumors, adenocarcinoma, adenoma, basal cell carcinoma, cervical dysdifferentiation and carcinoma in situ, Ewing's sarcoma, epidermoid carcinoma, giant cell tumor, glioblastoma multiforme, pilocytic tumor, enteroganglioma, proliferative corneal nerve tumor, islet cell carcinoma, Kaposi's sarcoma, leiomyoma, leukemia, lymphoma, malignant carcinoid tumors, malignant melanoma, malignant hypercalcemia, and Marfanoid body type. Tumors, medullary carcinoma, metastatic skin cancer, mucosal neuroma, myeloma, mycosis fungoides, neuroblastoma, osteosarcoma, osteogenic and other sarcomas, ovarian tumors, pheochromocytoma, polycythemia vera, primary brain tumors, small cell lung cancer, squamous cell carcinoma of both ulcerative and papillary types, hyperplasia, seminoma, soft tissue sarcoma, retinoblastoma, rhabdomyosarcoma, renal cell tumors, localized skin lesions, reticulum cell sarcoma, and Wilms' tumor.

[0294] Combination therapy, also known as combined drug therapy, refers to the use of two or more anticancer drugs simultaneously or sequentially during a single treatment course. Monotherapy refers to the use of only one anticancer drug in a single treatment course.

[0295] Generally speaking, combination therapy requires exploring different dosages and dosing cycles based on the characteristics of the disease and the types of drugs used in combination. Only by exploring combination therapy plans based on the above conditions can better therapeutic effects be achieved than single-drug therapy.

[0296] The dosage and dosing cycle of the combination therapy regimen need to be determined through clinical trials, referring to the dosage and dosing regimen of the alkylating prodrug compound and the combination drugs mentioned above. Attached Figure Description

[0297] Figure 1 shows the weight changes of tumor-bearing mice in the MH-22A vector#1 and MH-22A hAKR1C3#20 cell subcutaneous xenograft tumor models after administration of AST-3424, AST, and anti-PD-1Ab single drugs, as well as AST-3424, AST, and anti-PD-1Ab in combination. The data points represent the mean weight within the group, and the error bars represent the standard error (SEM).

[0298] Figure 2 shows the weight change (%) curves of MH-22A vector#1 and MH-22A hAKR1C3#20 xenograft tumor model mice after administration of AST-3424, AST, anti-PD-1Ab single drugs and AST-3424, AST and anti-PD-1Ab combination drugs respectively. The relative weight change was calculated based on the animal's weight at the beginning of administration. The data points represent the percentage change in average weight within the group, and the error bar represents the standard error (SEM).

[0299] Figure 3 shows the tumor growth curves of MH-22A vector#1 and MH-22A hAKR1C3#20 cell subcutaneous xenograft tumor models in tumor-bearing mice after administration of AST-3424, AST, and anti-PD-1Ab single drugs, as well as AST-3424, AST, and anti-PD-1Ab in combination. The data points represent the mean tumor volume within the group, and the error bars represent the standard error (SEM). Example

[0300] The chemical structural formula of AST-3424 is:

[0301] The synthesis method is described in patent application text PCT / US2016 / 062114, publication number WO2017087428A1 (corresponding to Chinese application number 2016800200132, publication number CN108136214A).

[0302] The chemical structure of AST is:

[0303] The synthesis method is described in patent application text PCT / CN2020 / 089692, publication number WO2020228685A1.

[0304] Example 1: Efficacy and safety evaluation of AST-3424 and AST alone, and in combination with mouse PD-1Ab, in the MH-22A hAKR1C3C3H syngeneic mouse model.

[0305] (1) Experimental materials and experimental plan

[0306] The information on the laboratory animals is as follows:

[0307] Species: Mouse

[0308] Strain: C3H / He mice

[0309] Age: 6-8 weeks

[0310] Sex: Female

[0311] Weight: 18–23 grams

[0312] Quantity: 120 (including any remaining mice from the group)

[0313] Supplier: Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0314] Animal Certificate Number: No. 110011210115205782

[0315] The key testing drug information is as follows:

[0316] AST-3424 injection, manufactured by a pharmaceutical company commissioned by Shenzhen Aixindawei Pharmaceutical Technology Co., Ltd., has a specification of 1mL:10mg, containing 0.75ml ethanol, 0.25ml propylene glycol and 10mg AST-3424.

[0317] AST injection, manufactured by a pharmaceutical company commissioned by Shenzhen Aixindawei Pharmaceutical Technology Co., Ltd., has a specification of 2mL: 20mg, containing 20mg of the following compound, 100mg of N,N-dimethylacetamide and 800mg of polyoxyethylene (35) castor oil.

[0318] Positive compounds

[0319] Name: Anti-PD-1Ab

[0320] Supplier: BioXCell

[0321] Brand code: BioXCell-BP0146

[0322] Batch number: 810421S1

[0323] Packaging: 55.4mg

[0324] Concentration: 9.90 mg / mL

[0325] Storage conditions: 4℃

[0326] Other biological and chemical reagents were purchased from commercial reagent companies and are not listed here.

[0327] This experiment evaluated the in vivo efficacy and safety of compounds AST-3424 and AST alone, as well as AST-3424 and AST in combination with mouse PD-1 antibody, using the MH-22A hAKR1C3 C3H syngeneic mouse model. The experimental animal groups and administration regimens are shown in Table 1.

[0328] Table 1: Grouping and Dosing Regimens of Animals in In Vivo Efficacy Experiments Note: Each group consists of 6 mice. The route of administration is iv (tail vein injection) and ip (intraperitoneal injection). The day of grouping is designated PG-D0, and administration begins at PG-D0. Vehicle 1 and Vehicle 2 are glucose injection D5W (pH = 7.4).

[0329] (2) Experimental methods and procedures

[0330] Cell culture

[0331] Mouse hepatocellular carcinoma cells MH-22A vector#1 and MH-22A hAKR1C3#20 (custom-made mouse hepatocellular carcinoma cells highly expressing human AKR1C3 enzyme protein from a CRO) were cultured in vitro as a monolayer. The culture conditions were: DMEM medium supplemented with 10% fetal bovine serum, 100 U / mL penicillin, 100 μg / mL streptomycin, 2 mM L-glutamine, and 1.5 μg / mL blisterdin, incubated at 37°C in a 5% CO2 incubator. Cells were passaged twice a week using trypsin-EDTA digestion. When the cell saturation of MH-22A vector#1 and MH-22A hAKR1C3#20 cells reached 80%-90% and the required cell count was achieved, the cells were harvested, counted, and seeded.

[0332] Tumor cell inoculation

[0333] 5 × 10⁵ cells were subcutaneously injected into the right nape of the neck of each mouse. 6 100 cells were inoculated, with 24 mice inoculated with MH-22A vector#1 cells and 96 mice inoculated with MH-22A hAKR1C3#20 cells. The inoculation volume was 0.1 mL, and the cell suspension was PBS. In vivo efficacy experiments were conducted on day 5 post-inoculation, when the average tumor volume reached 61 mm². 3 At that time, mice were randomly assigned to groups of 6 and started receiving medication. The specific experimental grouping and administration regimens are shown in Table 1.

[0334] Preparation of solvent and test drug

[0335] Table 2: Preparation methods of solvents and test drugs Note: Before administration, bring AST-3424 and AST solution to room temperature and mix thoroughly.

[0336] Daily observation of laboratory animals

[0337] The experimental protocol and any modifications thereof were assessed and approved by the Laboratory Animal Management and Use Committee (IACUC) of Nantong WuXi AppTec Pharmaceutical Technology Co., Ltd. The use and welfare of laboratory animals were conducted in accordance with the regulations of the International Committee for Evaluation and Accreditation of Laboratory Animals (AAALAC). Animal health and mortality were monitored daily. Routine examinations included observing the effects of tumor growth and drug treatment on daily behavior, such as activity levels, food and water intake (visual assessment only), weight changes (measured twice weekly), physical appearance, and any other abnormalities. The number of deaths and side effects within each group were recorded based on the number of animals in each group.

[0338] Tumor measurements and experimental indicators

[0339] The experimental endpoint is to examine whether tumor growth is inhibited, delayed, or cured. Tumor diameter is measured twice weekly using calipers. The formula for calculating tumor volume is: V = 0.5a × b 2 , where a and b represent the long and short diameters of the tumor, respectively.

[0340] The relative tumor-suppressive efficacy of the compounds was evaluated using TGI (%) or relative tumor proliferation rate T / C (%).

[0341] Tumor growth inhibition rate (TGI) (%): The calculation formula is as follows: TGI (%) = [1 - (Ti - T0) / (Ci - C0)] × 100%. Where Ti is the average tumor volume at a certain measurement after the drug administration group is grouped, T0 is the average tumor volume at the time of grouping of the drug administration group, Ci is the average tumor volume at a certain measurement after the solvent control group is grouped, and C0 is the average tumor volume at the time of grouping of the solvent control group.

[0342] Relative tumor proliferation rate (T / C%): The calculation formula is as follows: T / C% = TRTV / CRTV × 100% (TRTV: RTV of the treatment group; CRTV: RTV of the solvent control group). The relative tumor volume (RTV) is calculated based on the tumor measurement results. The calculation formula is RTV = Vi / V0, where V0 is the average tumor volume measured at the time of grouping (i.e., PG-D0), and Vi is the average tumor volume at a specific measurement. TRTV and CRTV are based on data from the same day.

[0343] Tumor clearance rate: The calculation formula is as follows: NTV = 0 / N × 100% (NTV = 0: Number of mice in a group with a tumor volume of 0 mm3. N: Total number of mice in a group.)

[0344] Sample collection

[0345] When the tumor volume of a single mouse exceeds 3,000 mmHg 3 Alternatively, on the 28th day after grouping, tumor samples are collected, weighed, and recorded. The tumors are then photographed and flash-frozen in liquid nitrogen. All samples are stored at -80°C before being sent to the customer.

[0346] Statistical analysis

[0347] Statistical analysis was performed, including the mean and standard error (SEM) of tumor volume at each time point for each group. Statistical analysis was conducted based on the data to assess differences between groups. Comparisons between two groups were performed using the T-test. Comparisons between three or more groups were performed using one-way ANOVA. If the variances were homogeneous (F-test p-value > 0.05), Dunnett's test was used; if the variances were unequal (F-test p-value < 0.05), Games-Howell's multiple comparison test was used. p > 0.05 was considered statistically insignificant, p < 0.05 was considered statistically significant, and p < 0.01 was considered highly significant.

[0348] (3) Experimental Results

[0349] Mouse weight change

[0350] The body weight of experimental animals is used as a reference indicator for indirectly determining drug toxicity. The effects of AST-3424 and AST alone, as well as AST-3424 and AST in combination with mouse PD-1Ab, on the body weight of MH-22A hAKR1C3 C3H syngeneic mouse model are shown in Figure 1 for the changes in body weight of animals in different groups and in Figure 2 for the changes in relative body weight (%) of animals in different groups.

[0351] Tumor volume

[0352] In the MH-22A vector#1 and MH-22A hAKR1C3#20 cell subcutaneous xenograft C3H syngeneic mouse models, the changes in tumor volume in each group after administration of AST-3424, AST, and anti-PD-1Ab as single drugs and in combination with anti-PD-1Ab are shown in Table 3.

[0353] Table 3: Tumor volume at different time points in each group (mm) 3 ) Note: Data are expressed as mean ± SEM, n = 6. Number of days after grouping.

[0354] Tumor change curve

[0355] The tumor growth curves of tumor-bearing mice after treatment with the test substance are shown in Figure 3.

[0356] Evaluation of antitumor drug efficacy

[0357] Starting from day 18 after grouping, some experimental animals in the MH-22A hAKR1C3#20 model developed tumors exceeding 3,000 mmHg. 3 The tumors were euthanized, and therefore the antitumor efficacy was evaluated based on the tumor volume on day 18 after grouping. The results of the antitumor efficacy evaluation are shown in Tables 4, 5, and 6.

[0358] On day 28 after grouping, tumors in the MH-22A hAKR1C3#20 model were completely cleared. Therefore, the tumor clearance rate was calculated based on the tumor volume on day 28 after grouping. The results of the tumor clearance rate are shown in Table 7.

[0359] Table 4: In vivo efficacy evaluation of AST-3424 on the MH-22Avector#1 model (based on tumor volume calculation on day 18 after grouping). Note: a. Average value ± SEM. b. Tumor growth inhibition rate TGI (%) = [1-(T 18 -T0) / (C 18 -C0)]×100%. c. Relative tumor proliferation rate T / C%=T RTV / C RTV ×100%, RTV=V 18 / V0. dp values ​​were analyzed using T-test based on tumor volume (compared to the solvent control group Vehicle 1). p>0.05 was considered as no significant difference, p<0.05 as a significant difference, and p<0.01 as a highly significant difference.

[0360] Table 5: In vivo efficacy evaluation of AST-3424 and AST alone, and AST-3424 and AST in combination with mouse PD-1Ab in the MH-22AhAKR1C3#20 model (based on tumor volume calculation on day 18 after grouping). Note: a. Average value ± SEM. b. Tumor growth inhibition rate. Tumor growth inhibition rate TGI (%) = [1-(T 18 -T0) / (C 18 -C0)]×100%. c. Relative tumor proliferation rate T / C%=T RTV / C RTV ×100%, RTV=V 18 / V0. dp values ​​were analyzed using one-way ANOVA based on tumor volume (compared to the solvent control group Vehicle 2). p>0.05 was considered as no significant difference, p<0.05 as a significant difference, and p<0.01 as a highly significant difference.

[0361] Table 6: In vivo efficacy evaluation of AST-3424 on MH-22Avector#1 and MH-22AhAKR1C3#20 models (based on tumor volume calculation on day 18 after grouping). Note: a. Mean ± SEM. bp values ​​were analyzed using a T-test based on tumor volume (compared to Group 2). p>0.05 was considered no significant difference, p<0.05 was considered significant, and p<0.01 was considered highly significant. sp values ​​were analyzed using a T-test based on tumor volume (compared to Group 4). p>0.05 was considered no significant difference, p<0.05 was considered significant, and p<0.01 was considered highly significant.

[0362] Table 7: Tumor clearance rates of AST-3424 and AST alone, and AST-3424 and AST in combination with mouse PD-1Ab in the MH-22AhAKR1C3 C3H syngeneic model (calculated based on tumor volume on day 28 after grouping). Note: a. Average value ± SEM. b. Tumor clearance rate (%) = N TV＝0 / N×100%(N) TV =0: Tumor volume in a certain group of mice = 0 mm 3 N: Number of mice in a group.

[0363] Experimental Results Analysis

[0364] The in vivo efficacy evaluation of AST-3424 against the MH-22A vector#1 and MH-22A hAKR1C3#20 models is shown in Table 6. In the MH-22A hAKR1C3#20 model, the tumor volume reached 8 mm in the treatment group (Group 7) with AST-3424, 1 mg / kg IV (5 days on, 2 days off, 2 weeks off, 5 days on). 3 Compared with Group 4, it showed better tumor suppression effect (p = 0.032).

[0365] This experiment compared three groups of single-drug and combination therapy, and the differences between the single-drug and combination therapy groups were detected by calculating the tumor clearance rate in mice. The first comparison experiment showed that in the Anti-PD-1Ab single-drug group (Group 5, 10 mg / kg, ip, (Day 2, 5, 9, 12)(MH-22AhAKR1C3#20)), one mouse's tumor was cleared, with a tumor clearance rate of 16.67%. Meanwhile, in the other single-drug group, the AST-3424 group (Group 4, AST-3424, 1 mg / kg, iv, (Day 0, 7)(MH-22AhAKR1C3#20), no mice had tumors cleared. Compared to the two single-drug groups, the combination therapy group (Group 6, AST-3424, 1 mg / kg, iv, (Day 0, 7) + Anti-PD-1Ab, 10 mg / kg, ip, (Day 1 ... In mice treated with AST-3424 (1 mg / kg iv, (Day 0, 7)) and Anti-PD-1Ab (10 mg / kg ip, (Day 2, 5, 9, 12)), tumors were completely cleared, achieving a clearance rate of 50%. These results indicate that the combined administration of AST-3424 (1 mg / kg iv, (Day 0, 7)) and Anti-PD-1Ab (10 mg / kg ip, (Day 2, 5, 9, 12)) significantly enhanced tumor clearance compared to the two single-drug groups.

[0366] The second comparative experiment of single-drug and combination therapy showed that in the Anti-PD-1Ab single-drug group (Group 5, 10 mg / kg, ip, (Day 2, 5, 9, 12)(MH-22AhAKR1C3#20)), one mouse had its tumor cleared, with a tumor clearance rate of 16.67%. Meanwhile, in the other single-drug group, the AST-3424 group (Group 7, 1 mg / kg iv, (5 days on, 2 days off, 2 weeks off, 5 days on)(MH-22AhAKR1C3#20)), two mice had their tumors cleared, with a tumor clearance rate of 33.33%. Compared to the two single-drug groups, the combination therapy group (Group 8, AST-3424, 1 mg / kg, iv, (5 days on, 2 days off, 2 weeks off, 5 days on) + Anti-PD-1Ab, 10 mg / kg, ip, (Day 2, 5, 9, 12)(MH-22AhAKR1C3#20)) In hAKR1C3#20), tumors were completely cleared in 4 mice, with a clearance rate of 66.67%. The results indicate that the combination of AST-3424 (1 mg / kg, IV, (5 days on, 2 days off, 2 weeks off, 5 days on)) and Anti-PD-1Ab (10 mg / kg, IP, (Day 2, 5, 9, 12)) also significantly improved tumor clearance compared to the two single-drug groups.

[0367] In the third comparative experiment, we found similar results. In the Anti-PD-1Ab monotherapy group (Group 5, 10 mg / kg, ip, (Day 2, 5, 9, 12)(MH-22AhAKR1C3#20)), one mouse had its tumor cleared, with a tumor clearance rate of 16.67%. Meanwhile, in the other monotherapy group, the AST group (Group 9, AST, 5 mg / kg iv, (Day 0, 7, 14)(MH-22AhAKR1C3#20)), no mice had their tumors cleared. Compared to the two monotherapy groups, in the combination therapy group (Group 10, AST, 5 mg / kg, iv, (Day 0, 7, 14) + Anti-PD-1Ab, 10 mg / kg, ip, (Day 2, 5, 9, 12, 16, 19)(MH-22AhAKR1C3#20)), three mice had their tumors completely cleared, with a clearance rate of 50%. The results showed that the combination of AST (5 mg / kg iv, (Day 0, 7, 14)) and Anti-PD-1Ab (10 mg / kg, ip, (Day 2, 5, 9, 12, 16, 19)) significantly enhanced the tumor clearance capacity in mice compared to the two single-drug groups.

[0368] Example 2: Experiment on changes in the levels of prostaglandins E2 and F2 in the blood of cynomolgus monkeys before and after administration of AST-3424.

[0369] Three cynomolgus monkeys were used in the experiment as shown in Table 8.

[0370] Table 8: Experiments with AST-3424 administration in cynomolgus monkeys

[0371] Four male cynomolgus monkeys were purchased from Guangxi Xiongsen Primate Development and Experimentation Co., Ltd. All animals were healthy and passed physical examinations without any abnormalities. Three were used for drug administration experiments, and the remaining animals were used to prepare blank plasma.

[0372] Before administration, and at 6, 24, 48, and 72 hours after administration. Collect 1 mL of blood via the femoral vein or other suitable vein and place it in an anticoagulant-free blood collection tube. After collection, place the blood sample on ice and centrifuge for 30–60 minutes to separate the serum (centrifugation conditions: 3500 rpm, 10 minutes, 2–8°C). Store the collected serum at –80°C before analysis.

[0373] Prostaglandins E2 and F2 in serum samples were analyzed using a conventional ELISA method.

[0374] The serum concentrations of prostaglandins E2 and F2 in cynomolgus monkeys after a single intravenous infusion are shown in Table 9 below.

[0375] Table 9: Changes in the levels of prostaglandins E2 and F2 in the blood of cynomolgus monkeys before and after administration of AST-3424 (the upper table shows the changes in prostaglandin F2 levels, and the lower table shows the changes in prostaglandin E2 levels).

[0376] After administration of AST-3424 to cynomolgus monkeys, both prostaglandin E2 and F2 decreased, but fluctuated and increased at 24 hours. This is presumably related to the timing characteristics of prostaglandin E2 and F2 secretion in the animals themselves, indicating that administration of AST-3424 can inhibit the secretion of prostaglandin E2 and F2 in cynomolgus monkeys.

[0377] AST-3424 reduces tumor-mediated suppression of immune responses by inhibiting the production of prostaglandin E2.

[0378] Example 3: In vitro AKR1C3 enzyme activity inhibition experiment

[0379] Experimental apparatus:

[0380] The Waters Acquity I Class UPLC ultra-high performance liquid chromatograph is equipped with a Xevo G2-XSQ TofHRMS quadrupole time-of-flight high-resolution mass spectrometer.

[0381] Buffer solution and materials:

[0382] 1. PBS phosphate buffered saline solution,

[0383] 2. PBS phosphate buffer solution with 20 mM NADPH

[0384] 3. 250 μg / mL AKR1C3 in PBS phosphate buffer solution

[0385] 4. 250 μM of 50% MeOH / H2O solution for the test compound.

[0386] 5. A 50% MeOH / H2O solution of 250 μM progesterone

[0387] 6. A 100% acetonitrile solution of 1 μg / mL propranolol

[0388] Experimental operation procedure

[0389] Step 1: Prepare Eppendorf tubes by dividing the reaction mixture into two portions (n=2) according to Table 10 below, and mix them gently.

[0390] Table 10: Reaction mixture processing

[0391] Step 2: Incubate the two portions of the mixture above at 37°C for 30 minutes.

[0392] Step 3: Add an additional 10 μL of 20 mM NADPH PBS phosphate buffer solution and 2 μL of 250 μM progesterone in 50% MeOH / H2O solution to each Eppendorf tube and mix gently.

[0393] Step 4: Immediately transfer the 50 μL mixture from the above steps to 100 μL of a 100% acetonitrile solution of 1 μg / mL propranolol (internal standard IS).

[0394] Step 5: Incubate the remaining sample at 37°C for 30 minutes and add 100 μL of 1 μg / mL propranolol (internal standard IS) in 100% acetonitrile solution.

[0395] Step 6: For all samples, add 100 μL of reagent water, vortex mix at 1100 rpm for 5 minutes, and centrifuge at 15000 rpm for 10 minutes at room temperature.

[0396] Step 7: Load all samples onto LC / MS to determine the content of reduced progesterone, i.e., 20α-dihydroprogesterone.

[0397] The testing conditions for the LC-MS instrument are as follows:

[0398] Liquid phase elution gradient:

[0399] Quadrupole time-of-flight mass spectrometry parameters:

[0400] Step 9, Calculation of reduced progesterone (20α-dihydroprogesterone): The peak areas of reduced progesterone (20α-dihydroprogesterone) and propranolol in each sample were determined by LC / MS. The peak area ratio of reduced progesterone to propranolol was calculated, and the ratio at time 0 was set to 0%.

[0401] AKR1C3 activity (%) = [(Amount of reduced progesterone after sample standardization)] 30min -(Reduced progesterone content after sample standardization) 0min [(Standardized reduced progesterone levels in the negative control group)] 30min -(Standardized reduced progesterone levels in the negative control group) 0min ]*100.

[0402] Based on the above calculations, the following table shows the relative activity % of AKR1C3 at a compound concentration of 5 μM / L.

[0403] Table 11: Relative Activity % of AKR1C3 at Compound Concentration of 5 μM / L

[0404] According to research literature (Samad TA, Moore KA, Sapirstein A, et al. Interleukin-1[beta]-mediated induction of Cox-2 in the CNS contributes to inflammatory pain hypersensitivity[J]. Nature, 2001, 410 (6827): 471-5.; Baojian Z, Yanbing Y, Gele A, et al. Tanshinone IIA Attenuates Diabetic Peripheral Neuropathic Pain in Experimental Rats via Inhibiting Inflammation[J]. Evidence-Based Complementary and Alternative Medicine, 2018, 2018: 1-8.; Lovering A, Ride J, Bunce C, et al. Crystal Structures of Prostaglandin D211-Ketoreductase(AKR1C3) in Complex with the Nonsteroidal Anti-Inflammatory Drugs Flufenamic Acid and Indomethacin[J].Cancer Research, 2004, 64(5): 1802-1810.;Matsuura K, Shiraishi H, Hara A, et al. Identification of a principal mRNA species for human 3alpha-hydroxysteroid dehydrogenase isoform (AKR1C3) that exhibits high prostaglandin D211-ketoreductase activity.[J]. Journal of Biochemistry, 1998, 124(5): 940-6.) It can be seen that aldehyde-ketoreductase AKR1C3 plays an important catalytic role in the biochemical pathway of prostaglandin H2 / D2 to prostaglandin E2 / F2:

[0405] Therefore, it can be concluded that AKR1C3 enzyme-activating compounds such as AST-3424 and AST are specific substrates for AKR1C3 enzyme. During the binding process with aldehyde-ketone reductase AKR1C3, they incidentally inhibit the activity of AKR1C3 enzyme, thereby incidentally reducing the prostaglandin E2 / F2 level.

[0406] It is further speculated that all AKR1C3 enzyme-activated compounds similar to AST-3424 and AST (selected from the following structural formulas (4)-(15)) are AKR1C3 enzyme-specific substrates. During their activation by binding with aldehyde reductase AKR1C3, they will incidentally inhibit the activity of AKR1C3 enzyme, thereby incidentally reducing the level of prostaglandin E2 / F2. Thus, like AST in Example 1 and AST-3424 in patent application PCT / US2021 / 029552 and publication number WO2022231580A1, they have a better tumor suppression effect when used in combination with immune checkpoint inhibitors and immune cell therapy drugs.

Claims

Treatment methods include using drugs containing AKR1C3 enzyme-activated compounds or their salts, esters, solvates, or isomers in combination with immune checkpoint inhibitors to treat cancer and tumor patients. The compounds activated by the AKR1C3 enzyme do not include the following structural compounds: Treatment methods involve using drugs containing AKR1C3 enzyme-activated compounds or their salts, esters, solvates, or isotopic isomers in combination with immunotherapy drugs to treat cancer and tumor patients. The treatment method according to claim 1, wherein, Immune checkpoint inhibitors are selected from anti-immune checkpoint antibodies that inhibit or block inhibitory immune checkpoint antigens; or the patient is a cancer / tumor patient who is resistant / insensitive to anti-immune checkpoint antibodies. The treatment method according to claim 3, wherein, The anti-immune checkpoint antibodies are selected from anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIGIT antibody, anti-Ceacam 1 antibody, anti-LAIR-1 antibody, anti-TIM-3 antibody, anti-VISTA antibody, anti-KIR antibody, anti-IDO antibody, anti-CD276 antibody, anti-A2AR antibody, or anti-CD47 antibody. The treatment method according to claim 4, wherein, The anti-PD-1 antibodies were selected from nivolumab, pembrolizumab, sintilimab, camrelizumab, tislelizumab, and toripalimab. The treatment method according to claim 4, wherein, The anti-PD-L1 antibodies were selected from avelumab, durvalumab, atezolizumab, sugemalimab, and atezolizumab. The treatment method according to claim 1 or 2, wherein, The cancer was selected from hepatocellular carcinoma. The treatment method according to claim 1 or 2, wherein, The patients were those who tested negative for p53 gene mutation or had normal p53 protein expression. The treatment method according to claim 1 or 2, wherein, Compounds activated by AKR1C3 enzyme are selected from the following structural formulas (4)-(15): Among them, X, Y, Z, R, T, A, and X 10 The definition is as set forth in the claims of patent application PCT / US2016 / 021581, publication number WO2016145092A1 (corresponding to Chinese application number 2016800150788, publication number CN107530556A); Among them, X, Y, Z, R, D, L 1 A and X 10 The definition is as set forth in the claims of patent application PCT / US2016 / 025665, publication number WO2016161342A3 (corresponding to Chinese application number 2016800200132, publication number CN108136214A); Among them, R1, R2, R3, R4, R5, R8, R9, R 10 The definition is as set forth in the claims of patent application PCT / CN2020 / 089692, publication number WO2020228685A9 (corresponding to Chinese application number 2020800358890, publication number CN113853379A); in: A is a substituted or unsubstituted C6-C10 aryl, biaryl, or substituted biaryl, 5-15 membered heteroaryl, or -N=CR 1 R 2 The substituents used in the substitution process are selected from the following groups: halogens, -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 10 2 C1-C6 alkyl groups, C3-C10 heterocyclic groups; Among them, R 100 R 101 and R 102 Each can be hydrogen, C1-C8 alkyl, or C6-C12 aryl; or R 101 and R 102 Together with the nitrogen atom to which it is attached, it forms a 5-7 membered heterocycle; The alkyl and aryl groups are each substituted with 1-3 halogroups or 1-3 C1-C6 alkyl groups; R 1 and R 2 Each can be either phenyl or methyl; X, Y, and Z are each independently either hydrogen or a halide group; R is hydrogen or a C1-C6 alkyl or a halogen-substituted alkyl; The definition of Rw is as described in the claims of patent application PCT / CN2020 / 120281, publication number WO2021068952A1 (corresponding to Chinese application number 202080071652.8, publication number CN114555574A); The definitions of R1, R2, R3, R4, and T are as described in the claims of patent application PCT / CN2021 / 118597, publication number WO2022057838A1; The definitions of A, E, G, X, and Y are as described in the claims of patent application PCT / NZ2019 / 050030, publication number WO2019190331A1 (corresponding to Chinese application number 2019800234236, publication number CN111918864A); 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 as described in the claims of patent application CN202210585771.6, publication number CN115403579A; Or its pharmaceutically acceptable salt. Among them, R 1 R 2a R 2b R 3 R 4 R 5 The definitions of , n, and Z are as described in the claims of patent application PCT / IB2020 / 057285, publication number WO2021005586A1 (corresponding to Chinese application number CN202080053804.1, publication number CN114206870A); Or its pharmaceutically acceptable salt. Among them, R w X, R4, R 10 R 13 R 14 The definition is as set forth in the claims of patent application PCT / CN2022 / 098082, publication number WO2022258043A1; Or its pharmaceutically acceptable salt. Among them, 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 patent application PCT / CN2023 / 123253, publication number WO2024078392A1. The treatment method according to claim 1, wherein, Compounds that activate AKR1C3 enzymes are selected from the following structural compounds: The treatment method according to claim 1 or 2, wherein, Drugs containing AKR1C3 enzyme-activated compounds or their salts, esters, solvates, or isomers are selected from injectable formulations of compounds with the following structural formula, wherein the injectable formulation contains a compound with the following structural formula, N,N-dimethylacetamide, polyoxyethylene (35) castor oil, and ethanol. The treatment method according to claim 11, wherein, In the aforementioned injectable formulation, the concentration of the compound with the following structure is 10-20 mg / ml. The treatment method according to claim 11, wherein, The injectable formulation is a concentrated ethanol solution, labeled as 2 ml in volume, containing 20 mg of the following compound, 100 mg of N,N-dimethylacetamide, and 800 mg of polyoxyethylene (35) castor oil.

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