Method for treating cancer with AST-001 and use thereof in preparing medicament for treating cancer

Abstract

Disclosed is AST-001 for treating cancer, in particular pharmaceutical use and a therapeutic method thereof. A medicament comprising AST-001 or a salt, an ester, a solvate, an isomer, or an isotopic variant thereof is used to treat a patient with a cancer or tumor. The dosage regimen is once on days 1, 8, and 15 of a 28-day treatment cycle. The patient with the cancer or tumor is selected from patients with pancreatic cancer, intrahepatic cholangiocarcinoma, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal adenocarcinoma, ampullary carcinoma, breast cancer, prostate cancer, and mixed hepatocellular-cholangiocarcinoma, or preferably selected from patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma. The dose at each administration is (25-55) mg / m2. The ex vivo tumor tissue pathological paraffin block or pathological section from the patients with cancers or tumors is tested for AKR1C3 enzyme protein expression level via immunohistochemical staining, with the test result, an H-score, being greater than or equal to 100.

Description

Methods for treating cancer with AST-001 and its use in the preparation of drugs for treating cancer. Technical Field

[0001] This invention relates to treatment methods for cancer, particularly to the treatment of specific types of cancer / tumors with AST-001, and belongs to the field of cancer treatment. Background Technology

[0002] The structure of AST-001, a DNA alkylating agent prodrug targeting overexpression of aldehyde-ketone reductase 1C3 (AKR1C3), is as follows:

[0003] It contains one chiral carbon atom (see "*" in the figure), is a racemic mixture, and is similar to AST-3424 (also known as OBI-3424, TH-3424). Both are activated by the AKR1C3 enzyme overexpressed by cancer cells after entering cancer cells, releasing the metabolite AST-2660 (also known as 2660):

[0004] AST-001 has completed Phase I clinical trials in China, and this invention is proposed based on the results of these Phase I clinical trials. Summary of the Invention

[0005] Based on existing technologies that disclose AST-001, similar to AST-3424, as a broad-spectrum anti-tumor DNA alkylating agent prodrug against multiple tumors (PCT application PCT / CN2021 / 129077, publication number WO2023077452) and concentrated solutions for injection (PCT / CN2024 / 092639, WO2024230831), and based on the overall data and relevant subgroup data from the completed Phase I clinical trial of AST-001 in China, those skilled in the art can determine that with a dosing regimen of 28 days per cycle, administered once on day 1, day 8, and day 15, AST-001 has good efficacy in treating patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma; the dosage during treatment is (25-55) mg / m². 2 It has good therapeutic effect. When the expression level of AKR1C3 enzyme protein is detected by immunohistochemical staining of pathological paraffin blocks or pathological sections of ex vivo tumor tissues of cancer and tumor patients, and the H-score of the detection result is greater than or equal to 100, it has good therapeutic effect.

[0006] In view of the above, this application proposes the following technical solution:

[0007] The treatment method involves treating cancer or tumor patients with drugs containing AST-001 or its salts, esters, solvates, isomers, or isotope variants. The dosing regimen is every 28 days, with one dose administered on day 1, day 8, and day 15. The cancer or tumor patients are selected from pancreatic cancer, intrahepatic cholangiocarcinoma, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal cancer, ampullary cancer, breast cancer, prostate cancer, and mixed-type liver cancer, preferably from pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma patients.

[0008] For pharmaceutical use, drugs for treating cancer and tumors are prepared using AST-001 or its salts, esters, solvates, isomers, or isotope variants. The dosing regimen is one cycle every 28 days, with one dose administered on day 1, day 8, and day 15. The cancer and tumor patients are selected from pancreatic cancer, intrahepatic cholangiocarcinoma, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal cancer, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, preferably from pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma patients.

[0009] Treatment method: Treatment of cancer and tumor patients with drugs containing AST-001 or its salts, esters, solvates, isomers, or isotopic variants. The dosing regimen is every 28 days, with administration on days 1, 8, and 15, at a dose of (25-55) mg / m² each time. 2 .

[0010] For pharmaceutical use, AST-001 or its salts, esters, solvates, isomers, or isotopic variants are used to prepare drugs for treating cancer and tumors. The dosing regimen is one cycle every 28 days, with administration on days 1, 8, and 15, and the dosage at each administration is (25-55) mg / m². 2 .

[0011] Treatment method: Cancer and tumor patients are treated with drugs containing AST-001 or its salts, esters, solvates, isomers, or isotope variants. The dosing regimen is one cycle every 28 days, with one dose each on day 1, day 8, and day 15. The expression level of AKR1C3 enzyme protein in the pathological paraffin blocks or pathological sections of the ex vivo tumor tissue of the cancer and tumor patients is detected by immunohistochemical staining. The H-score of the detection result is greater than or equal to 100.

[0012] For pharmaceutical use, drugs for treating cancer and tumors are prepared using AST-001 or its salts, esters, solvates, isomers, or isotope variants. The dosing regimen is one cycle every 28 days, with one dose administered on day 1, day 8, and day 15. The expression level of AKR1C3 enzyme protein in pathological paraffin blocks or sections of ex vivo tumor tissue from the cancer or tumor patients is detected by immunohistochemical staining, and the H-score of the detection result is greater than or equal to 100.

[0013] The treatment method involves treating cancer and tumor patients with drugs containing AST-001 or its salts, esters, solvates, isomers, or isotopic variants. The dosing regimen is a 28-day cycle, with administration once on day 1, day 8, and day 15. The method is characterized by:

[0014] The dosage for each administration is (25-55) mg / m². 2 The pathological paraffin blocks or pathological sections of the ex vivo tumor tissues of the cancer and tumor patients were tested for the expression level of AKR1C3 enzyme protein by immunohistochemical staining, and the H-score of the test results was greater than or equal to 100.

[0015] or

[0016] The dosage for each administration is (25-55) mg / m². 2 The cancer and tumor patients mentioned are selected from pancreatic cancer, intrahepatic bile duct cancer, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal cancer, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, preferably from pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma patients.

[0017] or

[0018] The pathological paraffin blocks or sections of the ex vivo tumor tissue from the cancer and tumor patients were tested for the expression level of AKR1C3 enzyme protein by immunohistochemical staining. The H-score of the test result was greater than or equal to 100. The cancer and tumor patients were selected from pancreatic cancer, intrahepatic bile duct cancer, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal adenocarcinoma, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, with pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma patients being preferred.

[0019] or

[0020] The dosage for each administration is (25-55) mg / m². 2The pathological paraffin blocks or sections of the ex vivo tumor tissue from the cancer or tumor patients were used to detect the expression level of AKR1C3 enzyme protein by immunohistochemical staining. The H-score of the detection result was greater than or equal to 100. The cancer or tumor patients were selected from pancreatic cancer, intrahepatic bile duct cancer, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal adenocarcinoma, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, preferably from pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma patients.

[0021] Pharmaceutical applications: The preparation of drugs for treating cancer and tumors using AST-001 or its salts, esters, solvates, isomers, or isotopic variants, with a dosing regimen of once every 28 days, administered on days 1, 8, and 15. The characteristic feature is:

[0022] The dosage for each administration is (25-55) mg / m². 2 The pathological paraffin blocks or pathological sections of the ex vivo tumor tissues of the cancer and tumor patients were tested for the expression level of AKR1C3 enzyme protein by immunohistochemical staining, and the H-score of the test results was greater than or equal to 100.

[0023] or

[0024] The dosage for each administration is (25-55) mg / m². 2 The cancer and tumor patients mentioned are selected from pancreatic cancer, intrahepatic bile duct cancer, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal cancer, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, preferably from pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma patients.

[0025] or

[0026] The pathological paraffin blocks or sections of the ex vivo tumor tissue from the cancer and tumor patients were tested for the expression level of AKR1C3 enzyme protein by immunohistochemical staining. The H-score of the test result was greater than or equal to 100. The cancer and tumor patients were selected from pancreatic cancer, intrahepatic bile duct cancer, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal adenocarcinoma, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, with pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma patients being preferred.

[0027] or

[0028] The dosage for each administration is (25-55) mg / m². 2The pathological paraffin blocks or sections of the ex vivo tumor tissue from the cancer or tumor patients were used to detect the expression level of AKR1C3 enzyme protein by immunohistochemical staining. The H-score of the detection result was greater than or equal to 100. The cancer or tumor patients were selected from pancreatic cancer, intrahepatic bile duct cancer or hepatocellular carcinoma patients.

[0029] Regarding the compounds described herein, chemical structures such as AST-001 contain organic amine structures and P=O double bonds. Therefore, these compounds may also be administered in the form of salts, i.e., the present invention provides pharmaceutically acceptable salts of these compounds. These salts can be basic salts, including salts formed by the compounds with inorganic bases (e.g., alkali metal hydroxides, alkaline earth metal hydroxides, etc.) or with organic bases (e.g., monoethanolamine, diethanolamine, or triethanolamine, etc.). Alternatively, the salts can be acidic salts, including salts formed by the compounds with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, perchloric acid, sulfuric acid, or phosphoric acid, etc.) or with organic acids (e.g., methanesulfonic acid, trifluoromethanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, fumaric acid, oxalic acid, maleic acid, citric acid, etc.). Similarly, they may also react with certain acids or alcohols to form esters, therefore, these compounds may also be administered in the form of esters.

[0030] For various reasons, these compounds may also form solvates with certain solvents, which may be hydrates or alcohols, and therefore the compounds may also be administered in solvate form. The selection and preparation of acceptable salts, esters, and solvates of the compounds are well known in the art.

[0031] The term "isotope variant" refers to a compound that contains one or more isotopes in a non-natural proportion at one or more of the atoms constituting such a compound. In some embodiments, the "isotope variant" of the compound contains one or more isotopes in a non-natural proportion, including (but not limited to) hydrogen. 1 H), deuterium ( 2 H), tritium ( 3 H), carbon-11 ( 11 C), Carbon-12 ( 12 C), Carbon-13 ( 13 C), Carbon-14 ( 14 C) Nitrogen-13 ( 13 N), nitrogen-14 ( 14 N), nitrogen-15 ( 15 N), Oxygen-14 ( 14 O), Oxygen-15 ( 15 O), Oxygen-16 ( 16 O), Oxygen-17 ( 17 O), Oxygen-18 ( 18 O), Fluorine-17 ( 17 F), Fluorine-18 (18 F), Phosphorus-31 ( 31 P), Phosphorus-32 ( 32 P), Phosphorus-33 ( 33 P), sulfur-32 ( 32 S), sulfur-33 ( 33 S), sulfur-34 ( 34 S), sulfur-35 ( 35 S), sulfur-36 ( 36 S), Chlorine-35 ( 35 Cl), Chlorine-36 ( 36 Cl), Chlorine-37 ( 37 Cl), Bromine-79 ( 79 Br), bromine-81 ( 81 Br), Iodine-123 ( 123 I), iodine-125( 125 I), iodine-127( 127 I), iodine-129( 129 I) and Iodine-131 131 I). In some embodiments, the "isotopic variant" of the compound is in a stable form, i.e., non-radioactive. In some embodiments, the "isotopic variant" of the compound contains one or more isotopes in non-natural proportions, including (but not limited to) hydrogen ( 1 H), deuterium ( 2 H), carbon-12 ( 12 C), Carbon-13 ( 13 C) Nitrogen-14 ( 14 N), nitrogen-15 ( 15 N), Oxygen-16 ( 16 O), Oxygen-17 ( 17 O), Oxygen-18 ( 18 O), Fluorine-17 ( 17 F), Phosphorus-31 ( 31 P), sulfur-32 ( 32 S), sulfur-33 ( 33 S), sulfur-34 ( 34 S), sulfur-36 ( 36 S), Chlorine-35 ( 35 Cl), Chlorine-37 ( 37 Cl), Bromine-79 ( 79 Br), bromine-81 ( 81 Br) and iodine-127 127 I). In some embodiments, the "isotopic variant" of the compound is in an unstable form, i.e., radioactive. In some embodiments, the "isotopic variant" of the compound contains one or more isotopes in non-natural proportions, including (but not limited to) tritium. 3 H), carbon-11 (11 C), Carbon-14 ( 14 C) Nitrogen-13 ( 13 N), Oxygen-14 ( 14 O), Oxygen-15 ( 15 O), Fluorine-18 ( 18 F), Phosphorus-32 ( 32 P), Phosphorus-33 ( 33 P), sulfur-35 ( 35 S), Chlorine-36 ( 36 Cl), Iodine-123 ( 123 I), iodine-125( 125 I), iodine-129( 129 I) and Iodine-131 131 I). It should be understood that in compounds as provided herein, any hydrogen may be, for example, where feasible to those skilled in the art. 2 H is D, or any carbon can be, for example 13 C, or any nitrogen, can be, for example... 15 N, and any oxygen can be 18 O. In some embodiments, the "isotopic variants" of the compound contain non-natural proportions of deuterium (D).

[0032] In this application, "isomer" refers to stereoisomer (R / S).

[0033] The term "drug" as used in this article refers to a pharmaceutical product or preparation, wherein the prepared pharmaceutical product contains AST-001 compound or its salt or solvate within a specific dosage range, and / or the prepared pharmaceutical product is administered in a specific dosage form or via a specific route of administration.

[0034] The resulting pharmaceutical products, drugs, and formulations may also contain pharmaceutically acceptable excipients or excipients. The drug may 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 colorants, etc.

[0035] Regarding the dosing regimen of AST-001: Each cycle is 28 days, with one dose administered on day 1, day 8, and day 15. This means that for a given patient, single-drug intravenous injection therapy is administered in a 28-day cycle, with one dose administered on day 1, day 8, and day 15, for a total of three doses per cycle.

[0036] In some embodiments, the body surface area is used to calculate the dosage per administration, with the dosage at each administration being (25-55) mg / m². 2 Based on the above dosage, for an average person (175cm tall, 75kg), the corresponding equivalent body surface area (BSA) is... 2 = ([height (cm) × weight (kg)] / 3600) 1 / 2 =1.90, then the corresponding dose is 47.5-104.5mg!

[0037] In some embodiments, immunohistochemical staining (IHC) is used to detect the expression level of AKR1C3 enzyme protein in pathological paraffin blocks or sections of excised liver tumor tissue from patients. The specific IHC detection method is disclosed in patent application PCT / CN2021 / 114774, publication number WO2022048492A1, and its calculation method is explained below:

[0038] 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+).

[0039] (3) Tumor Sample Scoring Criteria

[0040] 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.

[0041] 0 (Uncolored): Values ​​between 0 and 100

[0042] Tumor cell nucleus-cytoplasm 1+ (weak staining): values ​​between 0 and 100

[0043] Tumor cell nucleus-cytoplasm 2+ (moderate staining): values ​​between 0 and 100

[0044] Tumor cell nucleus-cytoplasm 3+ (high-grade staining, strong): values ​​between 0 and 100

[0045] Total percentage of positive staining in the nucleus and cytoplasm: values ​​between 0 and 100

[0046] 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.

[0047] In some embodiments, AST-001 is a concentrated solution for injection, with a specification of 10 mg AST-001 active pharmaceutical ingredient per 1 mL.

[0048] As a preferred embodiment, based on the physicochemical properties and stability of AST-001, the AST-001 is a concentrated solution for injection, with a specification of 10 mg AST-001 active pharmaceutical ingredient per 1 mL, and excipients including N,N-dimethylacetamide, polyoxyethylene (35) castor oil and anhydrous ethanol.

[0049] Further preferably, the AST-001 is a concentrated solution for injection, with a specification of 10 mg AST-001 raw material per 1 mL, and 1 mL of concentrated solution contains 10 mg AST-001 raw material, 50 mg N,N-dimethylacetamide, 400 mg polyoxyethylene (35) castor oil, and the remainder is anhydrous ethanol.

[0050] For the specific preparation process and method of the above-mentioned concentrated solution for injection of AST-001, please refer to patent application PCT / CN2024 / 092639, publication number WO2024230831A1.

[0051] The above-mentioned concentrated solution for injection cannot be administered directly via intravenous infusion and must be diluted with a diluent (glucose injection, normal saline injection) before use.

[0052] Before administering the medication, add 0.4 ml of 5% sodium bicarbonate injection to 100 ml of sterile 5% glucose injection in an intravenous infusion bag that does not contain di(2-ethylhexyl) phthalate to adjust the pH value.

[0053] Add the calculated amount of concentrated AST-001 injection solution, accurate to 0.01 ml, to the 5% glucose injection bag after pH adjustment to prepare AST-001 injection solution for intravenous infusion administration.

[0054] If the patient is not suitable for glucose injection, normal saline should be used instead:

[0055] Before administering the medication, add 0.4 ml of 5% sodium bicarbonate injection to 100 ml of sterile 0.9% saline for injection in an intravenous infusion bag that does not contain di(2-ethylhexyl) phthalate to adjust the pH value.

[0056] Add the calculated amount of concentrated AST-001 injection solution, accurate to 0.01 ml, to the pH-adjusted saline bag to prepare AST-001 injection solution for intravenous administration.

[0057] Considering the stability of the prepared injection solution, the prepared AST-001 intravenous injection solution should be injected within 6 hours, preferably within 25-35 minutes.

[0058] Considering that large-volume intravenous injections should preferably be prepared as isotonic solutions, the final concentration of AST-001 in the prepared AST-001 intravenous injection solution should be between 0.02 mg / ml and 1.00 mg / ml. Therefore, the amount of the above-mentioned concentrated AST-001 injection solution should be adjusted when preparing the AST-001 intravenous injection solution.

[0059] "Cancer" refers to leukemia, lymphoma, carcinoma, and other malignant tumors (including solid tumors) that can potentially grow without restriction, either by invasion and local spread or by metastasis and systemic spread.

[0060] Based on the anticancer mechanism of AST-001 as a DNA alkylating agent prodrug, AST-001 possesses broad-spectrum anticancer potential. Examples of cancers and tumors that AST-001 can treat 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.

[0061] Treatment can be monotherapy or combination therapy. Combination therapy refers to the use of only one anticancer drug in a single course of treatment. Combination therapy refers to the simultaneous or sequential use of two or more anticancer drugs in a single course of treatment.

[0062] 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.

[0063] 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. Example

[0064] The chemical structural formula of AST-001 is:

[0065] The synthesis method is described in the patent application text PCT / CN2020 / 089692, publication number WO2020228685A9 (corresponding to Chinese application number 202080035889.0, publication number CN113853379A).

[0066] Example: Phase I clinical trial of AST-001 conducted in China

[0067] The clinical trial registration number is CTR20220934.

[0068] The trial was approved by the ethics committees of the medical institutions participating in the clinical trial, conducted in accordance with the principles of the Declaration of Helsinki, and informed consent was obtained from all participants.

[0069] Inclusion criteria

[0070] 1. Participants must voluntarily participate in this study, fully understand the relevant risks, demonstrate good compliance, and sign an informed consent form.

[0071] 2. Male or female, aged 18-70.

[0072] 3. Pathological histology and / or cytology confirms a malignant solid tumor (including but not limited to pancreatic cancer, hepatocellular carcinoma, non-small cell lung cancer, breast cancer, gastric cancer, esophageal cancer, colon cancer, rectal cancer, renal cell carcinoma, glioma, prostate cancer), and it is a metastatic or unresectable advanced case, and standard treatment has failed, or there is no standard treatment, or standard treatment is not suitable at this stage.

[0073] 4. The Eastern Cooperative Oncology Group (ECOG) performance status score is 0 or 1.

[0074] 5. Life expectancy ≥ 12 weeks.

[0075] 6. Prior to the first dose, all toxicities of previous anticancer treatment (excluding hair loss, fatigue, or peripheral neuropathy) must have been reduced to Grade 1 or baseline levels (NCI CTCAE 5th edition).

[0076] 7. Laboratory tests must meet the following criteria, and the indicators cannot be corrected by blood transfusion or hematopoietic stimulating factors within 14 days prior to the screening period to meet the inclusion criteria: a) Hemoglobin ≥ 90 g / L; b) Platelet count ≥ 100 × 10⁻⁶.9 c) Absolute neutrophil count (ANC) ≥ 1.5 × 10⁹ / L; 9 / L; d) Total bilirubin ≤1.5×ULN; e) ALT and AST ≤3.0×ULN, or ALT and AST ≤5.0×ULN in the presence of liver tumors; f) Creatinine clearance >50 ml / min as determined by the Cockcroft-Gault equation.

[0077] 8. No history of alcoholism, drug abuse or substance abuse within the past year.

[0078] 9. Female patients of childbearing age should have a negative pregnancy test result within 5 days prior to the start of treatment and should not be breastfeeding (a positive urine pregnancy test result needs to be confirmed by a serum pregnancy test).

[0079] 10. Female and male participants of childbearing age must agree to use effective contraception with their partners from the start of the study (e.g., surgical sterilization or condom or diaphragm contraception combined with spermicide gel or intrauterine device [IUD], etc.) until 6 months after the last dose.

[0080] Exclusion criteria:

[0081] 1. History of other malignant tumors within 2 years prior to the first dose, excluding adequately treated basal cell carcinoma, carcinoma in situ at other sites, or natural disease history, and other tumors whose treatment would not interfere with the safety or efficacy assessment of the current study.

[0082] 2. Underwent major surgery (excluding diagnostic surgery) within 4 weeks prior to the first dose.

[0083] 3. Patients who have received anti-tumor treatments such as radiotherapy, chemotherapy, immunotherapy, biotherapy, targeted therapy, or endocrine therapy within 4 weeks prior to the first dose (nitrosourea or mitomycin C treatment requires a 6-week washout period; oral fluorouracil drugs require a 2-week washout period; small molecule targeted therapy requires a 2-week washout period).

[0084] 4. Participated in a study of the investigational drug (diagnostic or therapeutic) or device within 4 weeks prior to the first dose.

[0085] 5. Repaglinide, a medium-to-potency CYP2C8 / CYP2B6 / CYP2C9 inhibitor or inducer should be used concurrently during the study.

[0086] 6. Pleural effusion or ascites requiring drainage every week or more.

[0087] 7. Subjects with HBV infection and HBV-DNA ≥ 2,000 IU / mL (subjects with HBV infection and HBV-DNA < 2,000 IU / mL should receive antiviral treatment with entecavir, tenofovir disoproxil fumarate or tenofovir alafenamide fumarate according to the National Guidelines for the Prevention and Control of Hepatitis B after enrollment, and should maintain treatment during the study period for up to 6 months after the last dose).

[0088] 8. Uncontrolled active bacterial, viral, or fungal infections that require systemic treatment.

[0089] 9. Known infection with human immunodeficiency virus (HIV) or positive for syphilis.

[0090] 10. The patient's cardiac history meets any of the following conditions: a) New York Heart Association (NYHA) Class III or IV congestive heart failure; b) QTcF interval >450 ms for men and >470 ms for women; c) Myocardial infarction or bypass / stent surgery within 6 months prior to the first dose; d) Other cardiac diseases deemed unsuitable for enrollment by the investigator.

[0091] 11. Women who are pregnant, breastfeeding, or planning to become pregnant.

[0092] 12. Accompanying diseases or symptoms that may interfere with the study, or physical abnormalities that the researchers believe may pose an excessive risk to the patient, including but not limited to active peptic ulcers or gastritis, changes in mental state, or mental abnormalities that may interfere with the patient's understanding of the informed consent form.

[0093] 13. Previous allergy to ethanol, polyoxyethylene (35) castor oil, and N,N-dimethylacetamide.

[0094] 14. Subjects who are unwilling or unable to comply with the study protocol for any reason.

[0095] Test drug:

[0096] AST-001 concentrated solution for injection: manufactured by a pharmaceutical company commissioned by Shenzhen Aixindawei Pharmaceutical Technology Co., Ltd., specification 2mL: 20mg; contains 20mg AST-001 raw material, 100mg N,N-dimethylacetamide, 800mg polyoxyethylene (35) castor oil, and the remainder is anhydrous ethanol.

[0097] Dosage regimen:

[0098] Each cycle lasts 28 days, with the drug administered once on day 1, day 8, and day 15. A maximum of 26 cycles of treatment may be permitted.

[0099] AST-001 is administered via intravenous infusion according to the assigned dose level group. The dosage will be calculated based on baseline body surface area; if the patient's weight changes by more than 10% during treatment, the body surface area should be recalculated and the dosage adjusted accordingly.

[0100] The dose escalation cohort for the first human study was 5.0 mg / m². 2 10.0 mg / m 2 15.0 mg / m 2 20.0 mg / m 2 25.0 mg / m 2 30.0 mg / m 2 35.0 mg / m 2 and 40.0 mg / m 2 Wait until the MTD or a dose confirmed by the safety review committee.

[0101] Before administration, add 0.4 ml of 5% sodium bicarbonate injection to 100 ml of commercially available sterile 5% glucose water for injection (D5W) in an intravenous infusion bag free of DEHP (di(2-ethylhexyl) phthalate). Add the calculated required volume (accurate to 0.01 ml) of concentrated AST-001 injection solution to the pH-adjusted D5W bag to prepare the AST-001 injection solution for intravenous infusion. If the patient is not suitable for glucose injection, use normal saline instead.

[0102] Before administration, add 0.4 ml of 5% sodium bicarbonate injection to 100 ml of commercially available sterile 0.9% saline solution for injection in an intravenous infusion bag free of DEHP (di(2-ethylhexyl) phthalate). Add the calculated required volume (accurate to 0.01 ml) of concentrated AST-001 injection solution to the pH-adjusted saline bag to prepare the AST-001 injection solution for intravenous administration.

[0103] After pH adjustment, the pH of the intravenous injection solution was 7.4-8.1, and the final concentration of AST-001 was 0.02 mg / ml to 1.00 mg / ml.

[0104] The precise calculation method for the required volume of concentrated AST-001 solution for injection is as follows:

[0105] For a patient who is 175cm tall and weighs 75kg, the corresponding equivalent body surface area (BSA) is... 2 = ([height (cm) × weight (kg)] / 3600) 1 / 2 =1.90, then the corresponding dose is 1.90 × 40 = 76 mg (at 40 mg / m²). 2For example, if the above-mentioned AST-001 concentrated injection solution is drawn, the amount should be 76 ÷ 10 × 1 = 7.60 ml.

[0106] The prepared intravenous AST-001 solution should be administered within 6 hours. In practice, it can be administered via intravenous bolus injection using an infusion pump, completing the injection within 25-35 minutes.

[0107] Clinical evaluation

[0108] Effectiveness evaluation includes clinical efficacy assessment.

[0109] Clinical efficacy was assessed using RECIST 1.1, the efficacy evaluation criteria for solid tumors. MRI / CT was used to assess lesions, and the same assessment method was used for the same lesion throughout the study. Subjects must have measurable tumor lesions at baseline.

[0110] Efficacy evaluation indicators include complete response (CR), partial response (PR), stable disease (SD), and progressive disease (PD).

[0111] Complete remission (CR): All target lesions disappear and the short diameter of all pathological lymph nodes (including target nodules and non-target nodules) must be reduced to <10 mm.

[0112] Partial remission (PR): The sum of the diameters of the target lesions is reduced by at least 30% compared to the baseline level.

[0113] Disease progression (PD): The minimum sum of the diameters of all target lesions measured throughout the entire experimental study is used as a reference, with a relative increase of at least 20% in the sum of diameters (or the baseline value if the baseline measurement is the minimum); in addition, the absolute value of the sum of diameters must increase by at least 5 mm (the appearance of one or more new lesions is also considered disease progression).

[0114] Disease stability (SD): The degree of reduction in target lesions does not reach the PR level, nor does the degree of increase reach the PD level; it falls between the two. The minimum value of the sum of diameters can be used as a reference in studies.

[0115] Study endpoints

[0116] Based on the objective response rate (ORR), disease control rate (DCR), duration of response (DOR), and progression-free survival (PFS) of the subjects, the efficacy of AST-3424 monotherapy for HCC and other malignant tumors was initially evaluated.

[0117] Objective response rate (ORR): The percentage of cases that achieve complete remission (CR) or partial remission (PR) after treatment out of the total evaluable cases.

[0118] The Disease Control Rate (DCR) is the percentage of patients with confirmed complete remission (CR), partial remission (PR), and stable disease (SD) among those with evaluable response rates.

[0119] Test results

[0120] As of October 17, 2024, 36 subjects had been enrolled and received drug treatment. The specific clinical trial results are shown in Table 1 below.

[0121] Table 1: Clinical data on the efficacy of AST-001 in solid tumors as of October 17, 2024

[0122] For patients whose OS data is "greater than" a certain value, it means that as of October 17, 2024, the patient is still in the survival follow-up period.

[0123] The "*" indicates a patient whose OS data shows a certain value, meaning that the patient's survival follow-up had ended as of October 17, 2024 (the patient had passed away on or before October 17, 2024).

[0124] AKR1C3: H score. ICC, intrahepatic cholangiocarcinoma. HCC, hepatocellular carcinoma.

[0125] Progression-free survival (PFS) and overall survival (OS) are calculated in months by dividing the calculated number of calendar days by 30.4375.

[0126] Therapeutic effect analysis section

[0127] Of the 36 cases in the total population, 2 had partial response (PR), 15 had stable disease (SD), and 18 had progressive disease (PD), resulting in an overall response rate (ORR) of 5.6% (2 / 36). The disease control rate (DCR) was 47.2% (17 / 36). The median progression-free survival (PFS) was 1.41 months, the mean PFS was 2.72 months, the median overall survival (OS) was 5.71 months, and the mean OS was 7.20 months.

[0128] Subgroup analysis was performed:

[0129] Subgroup 1 analyzed data from patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma: 2 cases of HCC, 7 cases of ICC, and 10 cases of pancreatic cancer, totaling 19 patients. Of these, 2 achieved partial response (PR), 8 had stable disease (SD), and 9 had progressive disease (PD). In this subgroup, the disease control rate (DCR) was 52.63% (10 / 19), the objective response rate (ORR) was 10.53% (2 / 19), the median progression-free survival (PFS) was 1.51 months, and the mean PFS was 3.50 months; the median overall survival (OS) was 6.01 months, and the mean OS was 7.08 months.

[0130] Subgroup 2, analysis of 25-55 mg / m² 2 Dosage group data: A total of 26 patients were included, with 2 achieving partial response (PR), 13 experiencing stable disease (SD), 10 experiencing progressive disease (PD), and 1 patient not evaluated (at 25-55 mg / m²). 2 In the dose group, the ORR was 7.70% (2 / 26), the DCR was 57.70% (15 / 26), the median PFS was 2.12 months, the mean PFS was 3.15 months, the median OS was 5.49 months, and the mean OS was 6.78 months.

[0131] Subgroup 3 analyzed patients with an AKR1C3 H-score greater than or equal to 100, totaling 22 patients. Among them, 2 had PR, 7 had SD, and 13 had PD. In the subgroup of patients with an AKR1C3 H-score greater than or equal to 100, the ORR was 9.10% (2 / 22), the DCR was 40.10% (9 / 22), the median PFS was 1.41 months, the mean PFS was 2.91 months, the median OS was 5.01 months, and the mean OS was 7.10 months.

[0132] Based on the mechanism of action of AST-001, its efficacy is related to the expression level of the AKR1C3 enzyme: higher expression levels may result in better therapeutic effects. Furthermore, the therapeutic effect of chemotherapy drugs is related to the dosage: generally, under safe conditions, higher doses within a certain range will achieve better therapeutic effects. Therefore, further subgroup analysis of the two-group combination was conducted.

[0133] Subgroup 4 analyzed patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma, and whose blood glucose levels were 25-55 mg / m². 2 Data from patients in the dosage groups (intersection of subgroup 1 and subgroup 2): A total of 17 patients, 2 with partial response (PR), 7 with stable disease (SD), and 8 with progressive disease (PD), i.e., patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma and a dosage range of 25-55 mg / m². 2 In the dose subgroup, the DCR was 52.94% (9 / 17), the ORR was 11.76% (2 / 17), the median PFS was 2.73 months, the mean PFS was 3.75 months, the median OS was 6.01 months, and the mean OS was 7.27 months.

[0134] Subgroup 5 analyzed data from patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma with an AKR1C3 H-score greater than or equal to 100 (intersection of subgroups 1 and 3): a total of 12 patients, 2 with PR, 4 with SD, and 6 with PD. That is, in the subgroup of patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma and an AKR1C3 H-score greater than or equal to 100, the DCR was 50% (6 / 12), the ORR was 16.67% (2 / 12), the median PFS was 1.41 months, the mean PFS was 3.84 months, the median OS was 6.39 months, and the mean OS was 7.61 months.

[0135] Subgroup 6, whose AKR1C3 H-score was greater than or equal to 100 and who were administered 25-55 mg / m². 2 Dosage group patient data (intersection of subgroups 2 and 3): 14 patients in total, 2 with PR, 6 with SD, and 6 with PD, i.e., those with an AKR1C3 H-score greater than or equal to 100 and a dose of 25-55 mg / m². 2 In the dose subgroup, the disease control rate (DCR) was 57.14% (8 / 14), the objective response rate (ORR) was 14.29% (2 / 12), the median progression-free survival (PFS) was 2.08 months, the mean PFS was 3.73 months, the median overall survival (OS) was 5.19 months, and the mean OS was 7.86 months.

[0136] The comparison results of efficacy data (ORR, DCR) and mean and median PFS and OS between subgroups and double subgroups and the overall group are shown in Table 2 below.

[0137] Table 2: Comparison of mean and median efficacy data (ORR, DCR) and PFS, OS between subgroups, dual subgroups, and the overall group.

[0138] Based on the drug's mechanism of action and preliminary clinical efficacy data, subgroup analysis included the following factors: AKR1C3 expression (≥100 was considered intermediate to high expression), specific tumor type (pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma patients), and 25 mg / m². 2 Dosage levels (clinically observed in the PR dose group). Subgroup analysis revealed that 25 mg / m²... 2 At dose levels and above, in subjects with an AKR1C3 expression H-score ≥100, and in patients with specific tumor types (pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma), all efficacy endpoints (ORR, DCR, and mean PFS) were improved compared to the full analysis set. A specific examination of ORR in different subgroups revealed that the ORR values ​​in subgroups 1, 2, and 3 were significantly higher than the overall group; while the ORR values ​​in subgroups 4, 5, and 6 were generally 2-3 times higher than the overall group, and the average PFS quantified by the former was also generally more than 35% higher than the overall group. In other words, the comparison of efficacy data clearly leads to the following conclusions:

[0139] 1. AST-001 is more sensitive to patients with AKR1C3 expression (≥100 is considered intermediate to high expression), and these patients may have better efficacy when receiving AST-001 treatment;

[0140] 2. AST-001 is more sensitive to patients with certain types of tumors (pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma), and these patients may have better efficacy when receiving AST-001 treatment.

[0141] 3. Use (25-55) mg / m³ 2 AST-001 treatment at dose levels is more effective for cancer / tumor patients, or relatively speaking, the dose of AST-001 for cancer / tumor patients should not be lower than 25 mg / m². 2 ;

[0142] 4. The combination of the above three conditions will result in better treatment outcomes:

[0143] The dosage for each administration is (25-55) mg / m². 2 The pathological paraffin blocks or pathological sections of the ex vivo tumor tissues of the cancer and tumor patients were tested for the expression level of AKR1C3 enzyme protein by immunohistochemical staining, and the H-score of the test results was greater than or equal to 100.

[0144] or

[0145] The dosage for each administration is (25-55) mg / m². 2 The cancer or tumor patients mentioned are selected from patients with pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma;

[0146] or

[0147] The pathological paraffin blocks or sections of the ex vivo tumor tissues of the cancer and tumor patients were tested for the expression level of AKR1C3 enzyme protein by immunohistochemical staining. The H-score of the test result was greater than or equal to 100. The cancer and tumor patients were selected from pancreatic cancer, intrahepatic bile duct cancer or hepatocellular carcinoma patients.

[0148] or

[0149] The dosage for each administration is (25-55) mg / m². 2 The pathological paraffin blocks or sections of the ex vivo tumor tissue from the cancer or tumor patients were used to detect the expression level of AKR1C3 enzyme protein by immunohistochemical staining. The H-score of the detection result was greater than or equal to 100. The cancer or tumor patients were selected from pancreatic cancer, intrahepatic bile duct cancer or hepatocellular carcinoma patients.

[0150] Safety assessment

[0151] Adverse events (AEs) refer to all adverse medical events that occur after a subject receives the study drug. These events can manifest as symptoms, signs, illnesses, or abnormal laboratory test results, but are not necessarily causally related to the study drug. Therefore, adverse events can be the exacerbation of pre-existing symptoms, signs, or laboratory abnormalities, or newly diagnosed diseases, or clinically significant abnormal laboratory values, regardless of whether they are considered to be related to the study drug.

[0152] The severity of adverse events should be rated according to the NCI Common Terminology Standard for Adverse Events (CTCAE) (version 5.0) grading system, whenever possible. Adverse events not covered by CTCAE should be graded on a five-point scale (mild, moderate, severe, life-threatening, and fatal).

[0153] Serious adverse events (SAEs) are adverse medical events that occur in a subject after receiving the study drug, including death, life-threatening events, permanent or serious disability or loss of function, hospitalization or prolonged hospitalization, congenital abnormalities or birth defects.

[0154] Suspected and unexpected serious adverse reaction (SUSAR) refers to a suspected and unexpected serious adverse reaction whose clinical manifestations exceed the information available in existing data such as the investigator's manual for investigational drugs.

[0155] Treatment-related adverse events (TEAEs) refer to any adverse medical event that occurs in a subject after receiving the investigational drug. Treatment-related adverse events (TRAEs) are defined as TEAEs whose relevance to the investigational drug is judged to be "certainly relevant," "probably relevant," or "possibly relevant."

[0156] Dose-limiting toxicity (DLT) is defined as the occurrence of any of the following during the first treatment cycle, which the investigator considers at least

[0157] Events possibly related to AST-001:

[0158] • Grade 4 neutropenia lasting ≥7 days.

[0159] • Febrile neutropenia is defined as an absolute neutrophil count (ANC) <1000 / mm3 accompanied by a single body temperature >38.3℃ or a sustained body temperature ≥38℃ for more than 1 hour.

[0160] • Grade 3 thrombocytopenia, accompanied by grade 2 or higher bleeding or requiring platelet transfusion.

[0161] • Grade 4 thrombocytopenia, regardless of duration.

[0162] Grade 4 anemia, regardless of duration.

[0163] • Grade ≥3 non-hematologic toxicity, except for the following:

[0164] I. With appropriate supportive care, nausea and / or vomiting and diarrhea resolve to grade 1 or baseline within 72 hours;

[0165] II can relieve grade 1 or baseline level grade 3 fever (without neutropenia) or fatigue within 72 hours;

[0166] Grade III asymptomatic laboratory abnormalities that can recover to Grade 1 or baseline levels within 72 hours or have no clinical significance;

[0167] IV can alleviate asymptomatic liver enzyme elevation to grade 1 or baseline levels within 7 days;

[0168] Researchers determined that aspartate aminotransferase (AST) or alanine aminotransferase (ALT) levels at least potentially associated with AST-001 were >3 × the upper limit of normal (ULN), and that bilirubin levels were >2 × ULN.

[0169] • Any other significant toxicity that the SRC considers to be dose-limiting.

[0170] The safety dataset included all 36 participants who received the investigational drug. During the trial, the vast majority of participants maintained stable laboratory parameters, vital signs, physical examination results, electrocardiograms, and weight relative to baseline. DLT was not reported. A safety overview is provided below:

[0171] • 485 TEAEs occurred in 36 (100%) subjects. Compared to other dose groups, 25 mg / m² 2 30mg / m 2 55mg / m 2 The dosage group had a higher incidence of TEAEs. Very common TEAEs (incidence ≥10%) included anemia (77.8%), decreased white blood cell count (38.9%), vomiting (38.9%), weakness (36.1%), decreased platelet count (33.3%), nausea (30.6%), decreased neutrophil count (27.8%), hypoalbuminemia (27.8%), hyponatremia (25.0%), decreased lymphocyte count (22.2%), diarrhea (19.4%), decreased appetite (19.4%), weight loss (16.7%), fever (13.9%), hiccups (13.9%), elevated AST (11.1%), constipation (11.1%), sinus tachycardia (11.1%), proteinuria (11.1%), and insomnia (11.1%).

[0172] • 34 patients (94.4%) experienced 313 episodes of TRAE. Compared to other dosage groups, 25 mg / m²... 230mg / m 2 55mg / m 2 The dose group had a higher incidence of TRAEs. Very common TRAEs (incidence ≥10%) included anemia (72.2%), decreased white blood cell count (38.9%), decreased platelet count (33.3%), vomiting (33.3%), nausea (27.8%), asthenia (25.0%), decreased neutrophil count (22.2%), diarrhea (19.4%), decreased appetite (13.9%), hiccups (13.9%), decreased lymphocyte count (11.1%), and increased AST (11.1%).

[0173] • 15 cases (41.7%) had 59 grade 3-5 TEAEs, of which the grade 3-5 TEAEs with an incidence of ≥10% were decreased platelet count (11.1%) and anemia (13.9%).

[0174] • 11 cases (30.6%) had 40 episodes of grade 3-5 TRAEs, including anemia (13.9%), decreased platelet count (11.1%), decreased white blood cell count (8.3%), decreased neutrophil count (8.3%), diarrhea (2.8%), and small bowel obstruction (2.8%), all of which were grade 3. No grade 4-5 TRAEs were reported.

[0175] • 25 subjects (69.4%) died. No subjects died due to TRAE. The causes of death were: disease progression in 16 cases (44.4%), adverse events in 1 case (2.8%) (grade 5 acute respiratory failure in the 20 mg / m2 dose group, unrelated to the investigational drug), other causes in 7 cases (19.4%) (cardiac arrest, rectal cancer, respiratory failure, liver cancer, tumor recurrence and failure death, bowel cancer, shock), and unknown causes in 1 case (2.8%).

[0176] • 10 subjects (27.8%) experienced 16 serious adverse events (SAEs).

[0177] • Three subjects (8.3%) experienced three serious adverse events (SARs), including one at 20 mg / m². 2 One patient (3.3%) in the dosage group experienced small bowel obstruction; 35 mg / m² 2 In the 2-dose group, 1 patient (3.3%) experienced a decrease in platelet count; 45 mg / m² 2 One patient (3.3%) in the dosage group had anemia. All cases had a SAR grade of 3.

[0178] • Three subjects (8.3%) experienced four dose-reduction TEAEs.

[0179] • Three subjects (8.3%) experienced four dose-dependent adverse events (TRAEs) leading to a decrease in platelet count, including three instances of decreased platelet count at 25 mg / m².2 and 50mg / m 2 One patient in each dosage group (both 2.8%); one case of anemia, 45 mg / m². 2 One case (2.8%) was in the dosage group.

[0180] • 14 subjects (38.9%) experienced 43 instances of TEAE leading to discontinuation of medication.

[0181] • 10 subjects (27.8%) experienced 33 instances of transarterial epidemiological events (TRAEs) leading to drug discontinuation, including 12 instances of decreased platelet counts at 25, 30, 35, and 50 mg / m². 2 One patient (2.8%) in each dosage group received 55 mg / m². 2 Three patients (8.3%) in the dosage group experienced decreased white blood cell counts in six instances, at doses of 20, 25, 35, and 55 mg / m². 2 One case (2.8%) was observed in each dosage group; three cases of anemia occurred at doses of 25, 35, and 45 mg / m². 2 One case (2.8%) in each dosage group; 7 cases of decreased neutrophil count, 25, 55 mg / m². 2 One patient (2.8%) in each dosage group; two patients experienced weakness, 35 mg / m². 2 Two cases (5.6%) were in the dosage group; others included one case of decreased lymphocyte count (35 mg / m²). 2 One case in the dosage group and one case of diarrhea (25 mg / m²). 2 One case in the dosage group and one case of small bowel obstruction (20 mg / m²). 2 (1 case in the dosage group).

[0182] The TEAE and TRAE that lead to permanent discontinuation are both 55 mg / m². 2 One case of grade 3 anemia was reported in one subject (2.8%) in the dose group.

[0183] Safety data showed that the laboratory indicators, vital signs, physical examination, electrocardiogram, and weight of the vast majority of subjects remained stable relative to baseline during the study. Common adverse events leading to adverse reactions (TRAEs) (incidence ≥10%) included anemia, decreased white blood cell count, decreased platelet count, decreased neutrophil count, decreased lymphocyte count, asthenia, decreased appetite, hiccups, gastrointestinal adverse reactions (vomiting, nausea, diarrhea), and elevated AST. TRAE severity was mostly grade 1-2; grade 3 TRAEs included anemia, decreased platelet count, decreased white blood cell count, decreased neutrophil count, diarrhea, and small bowel obstruction. No grade 4-5 TRAEs were reported, and no fatal TRAEs occurred. TRAEs leading to dose reduction were decreased platelet count and anemia. TRAEs leading to discontinuation of medication were mainly decreased platelet count, decreased neutrophil count, decreased white blood cell count, and anemia. One case was treated at 55 mg / m².2 Subjects in the dosage group permanently discontinued the medication due to grade 3 anemia.

[0184] Based on the resolution of the Safety Review Committee, and after a comprehensive assessment of the risks and benefits for the subjects, the dose was increased to 55 mg / m². 2 Terminated; MTD confirmation will no longer be performed.

[0185] A Phase I clinical trial of AST-001 monotherapy in previously treated advanced solid tumor patients demonstrated good safety.

[0186] Survival follow-up data updated.

[0187] As of February 24, 2026, 36 enrolled subjects had received the drug treatment. The specific clinical trial results are shown in Table 1 below.

[0188] Table 3: Clinical data on the efficacy of AST-001 in solid tumors as of February 24, 2026

[0189] For patients whose OS data is "greater than" a certain value, it means that as of February 24, 2026, the patient is still in the survival follow-up period.

[0190] "*" indicates a patient whose OS data shows a certain value, meaning that the patient's survival follow-up had ended as of February 24, 2026 (the patient had passed away on or before February 24, 2026).

[0191] The efficacy analysis section has been updated.

[0192] Of the 36 cases, 4 had partial response (PR), 13 had stable disease (SD), and 18 had progressive disease (PD), resulting in an overall response rate (ORR) of 11.1% (4 / 36). The disease control rate (DCR) was 47.2% (17 / 36). The median progression-free survival (PFS) was 1.41 months, the mean PFS was 3.15 months, the median overall survival (OS) was 6.38 months, and the mean OS was 10.26 months. When calculating the mean and median OS, values ​​greater than or equal to were treated as equal to the mean.

[0193] Subgroup analysis was performed:

[0194] Subgroup 1 analyzed data from patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma: 2 cases of HCC, 7 cases of ICC, and 10 cases of pancreatic cancer, totaling 19 cases. 3 patients achieved partial response (PR), 7 had stable disease (SD), and 9 had progressive disease (PD). In this subgroup, the disease control rate (DCR) was 52.63% (10 / 19), the objective response rate (ORR) was 15.79% (3 / 19), the median progression-free survival (PFS) was 1.43 months, and the mean PFS was 3.52 months; the median overall survival (OS) was 6.21 months, and the mean OS was 10.63 months.

[0195] Subgroup 2, analysis of 25-55 mg / m² 2 Dosage group data: A total of 26 patients were included, with 4 achieving partial response (PR), 11 experiencing stable disease (SD), 10 experiencing progressive disease (PD), and 1 patient not evaluated (at 25-55 mg / m²). 2 In the dose group, the ORR was 15.40% (4 / 26), the DCR was 57.70% (15 / 26), the median PFS was 2.14 months, the mean PFS was 3.75 months, the median OS was 6.38 months, and the mean OS was 11.02 months.

[0196] Subgroup 3 analyzed patients with an AKR1C3 H-score greater than or equal to 100, totaling 22 patients. Among them, 4 had PR, 5 had SD, and 13 had PD. In the subgroup of patients with an AKR1C3 H-score greater than or equal to 100, the ORR was 18.20% (4 / 22), the DCR was 40.10% (9 / 22), the median PFS was 1.41 months, the mean PFS was 3.55 months, the median OS was 6.34 months, and the mean OS was 9.82 months.

[0197] Based on the mechanism of action of AST-001, its efficacy is related to the expression level of the AKR1C3 enzyme: higher expression levels may result in better therapeutic effects. Furthermore, the therapeutic effect of chemotherapy drugs is related to the dosage: generally, under safe conditions, higher doses within a certain range will achieve better therapeutic effects. Therefore, further subgroup analysis of the two-group combination was conducted.

[0198] Subgroup 4 analyzed patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma, and whose blood glucose levels were 25-55 mg / m². 2 Data from patients in the dosage groups (intersection of subgroup 1 and subgroup 2): A total of 17 patients, 3 with partial response (PR), 7 with stable disease (SD), and 7 with progressive disease (PD), i.e., patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma and a dosage range of 25-55 mg / m². 2 In the dose subgroup, the DCR was 58.82% (10 / 17), the ORR was 17.64% (3 / 17), the median PFS was 2.15 months, the mean PFS was 3.78 months, the median OS was 6.21 months, and the mean OS was 11.24 months.

[0199] Subgroup 5 analyzed data from patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma with an AKR1C3 H-score greater than or equal to 100 (intersection of subgroups 1 and 3): a total of 12 patients, 3 with PR, 3 with SD, and 6 with PD. That is, in the subgroup of patients with pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma with an AKR1C3 H-score greater than or equal to 100, the DCR was 50% (6 / 12), the ORR was 25% (3 / 12), the median PFS was 1.43 months, the mean PFS was 4.01 months, the median OS was 6.21 months, and the mean OS was 10.23 months.

[0200] Subgroup 6, whose AKR1C3 H-score was greater than or equal to 100 and who were administered 25-55 mg / m². 2 Dosage group patient data (intersection of subgroups 2 and 3): 14 patients in total, 4 with partial response (PR), 4 with stable disease (SD), and 6 with progressive disease (PD), i.e., those with an AKR1C3 H-score greater than or equal to 100 and a dose of 25-55 mg / m². 2 In the dose subgroup, the DCR was 57.14% (8 / 14), the ORR was 28.57% (4 / 14), the median PFS was 2.76 months, the mean PFS was 4.73 months, the median OS was 6.34 months, and the mean OS was 12.14 months.

[0201] The results of comparing the efficacy data (ORR, DCR) of subgroups and double subgroups with the overall group, as well as the mean and median data of PFS and OS, are shown in Table 4 below.

[0202] Table 4: Comparison of mean and median efficacy data (ORR, DCR) and PFS, OS between subgroups and combination subgroups and the overall group

[0203] Based on the drug's mechanism of action and preliminary clinical efficacy data, subgroup analysis included the following factors: AKR1C3 expression (≥100 was considered intermediate to high expression), specific tumor type (pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma patients), and 25 mg / m². 2 Dosage levels (clinically observed in the PR dose group). Subgroup analysis revealed that 25 mg / m²... 2 At dose levels and above, in subjects with an AKR1C3 expression H-score ≥100, and in patients with specific tumor types (pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma), all efficacy endpoints (ORR, DCR, and mean PFS) were improved compared to the overall analysis set (general population). A specific examination of ORR in different subgroups revealed that the ORR values ​​in subgroups 1, 2, and 3 were significantly higher than those in the general population; while the ORR values ​​in subgroups 5 and 6 were generally 2-3 times higher than those in the general population, and the average PFS quantified by the subgroups was also generally higher in the subgroups than in the general population.

[0204] In particular, the mean OS of subgroups 2, 4, and 6 was significantly higher than that of the overall group, indicating that the dose and AKR1C3 target enzyme had a significant impact on the efficacy of AST-001 in cancer treatment.

Claims

1. Treatment method: treating cancer or tumor patients with drugs containing AST-001 or its salts, esters, solvates, isomers, or isotope variants, with a dosing regimen of once every 28 days, administered on day 1, day 8, and day 15. The cancer or tumor patients are selected from pancreatic cancer, intrahepatic cholangiocarcinoma, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal adenocarcinoma, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, preferably from pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma patients.

2. Pharmaceutical use: To prepare a drug for treating cancer and tumors using AST-001 or its salts, esters, solvates, isomers, or isotope variants. The dosing regimen is one cycle of 28 days, with one dose administered on day 1, day 8, and day 15. The cancer or tumor patients are selected from pancreatic cancer, intrahepatic cholangiocarcinoma, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, squamous cell carcinoma of the lung, endometrial cancer, cervical cancer, appendiceal adenocarcinoma, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, preferably from pancreatic cancer, intrahepatic cholangiocarcinoma, or hepatocellular carcinoma patients.

3. Treatment methods: Cancer and tumor patients are treated with drugs containing AST-001 or its salts, esters, solvates, isomers, or isotopic variants. The dosing regimen is one cycle every 28 days, with administration on days 1, 8, and 15, at a dose of (25-55) mg / m² each time. 2 .

4. Pharmaceutical Use: This drug is prepared using AST-001 or its salts, esters, solvates, isomers, or isotope variants to treat cancer and tumors. The dosing regimen is a 28-day cycle, with administration on days 1, 8, and 15, at a dose of (25-55) mg / m². 2 .

5. Treatment method: Cancer and tumor patients are treated with drugs containing AST-001 or its salts, esters, solvates, isomers, or isotope variants. The dosing regimen is one cycle every 28 days, with one dose each on day 1, day 8, and day 15. The expression level of AKR1C3 enzyme protein in the pathological paraffin blocks or pathological sections of the ex vivo tumor tissue of the cancer and tumor patients is detected by immunohistochemical staining. The H-score of the detection result is greater than or equal to 100.

6. Pharmaceutical use: To prepare drugs for treating cancer and tumors using AST-001 or its salts, esters, solvates, isomers, or isotope variants. The dosing regimen is one cycle every 28 days, with one dose administered on day 1, day 8, and day 15. The expression level of AKR1C3 enzyme protein in pathological paraffin blocks or pathological sections of ex vivo tumor tissue from the cancer or tumor patients is detected by immunohistochemical staining, and the H-score of the detection result is greater than or equal to 100.

7. A treatment method involving the treatment of cancer or tumor patients with a drug containing AST-001 or its salts, esters, solvates, isomers, or isotopic variants, wherein the dosing regimen is a 28-day cycle, with administration once on day 1, day 8, and day 15, characterized in that: The dosage for each administration is (25-55) mg / m². 2 The pathological paraffin blocks or pathological sections of the ex vivo tumor tissues of the cancer and tumor patients were tested for the expression level of AKR1C3 enzyme protein by immunohistochemical staining, and the H-score of the test results was greater than or equal to 100. or The dosage for each administration is (25-55) mg / m². 2 The cancer and tumor patients mentioned are selected from pancreatic cancer, intrahepatic bile duct cancer, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal cancer, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, preferably from pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma patients. or The pathological paraffin blocks or sections of the ex vivo tumor tissue from the cancer and tumor patients were tested for the expression level of AKR1C3 enzyme protein by immunohistochemical staining. The H-score of the test result was greater than or equal to 100. The cancer and tumor patients were selected from pancreatic cancer, intrahepatic bile duct cancer, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal adenocarcinoma, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, with pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma patients being preferred. or The dosage for each administration is (25-55) mg / m². 2 The pathological paraffin blocks or sections of the ex vivo tumor tissue from the cancer or tumor patients were used to detect the expression level of AKR1C3 enzyme protein by immunohistochemical staining. The H-score of the detection result was greater than or equal to 100. The cancer or tumor patients were selected from pancreatic cancer, intrahepatic bile duct cancer, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal adenocarcinoma, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, preferably from pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma patients.

8. Pharmaceutical use: A drug for treating cancer and tumors is prepared using AST-001 or its salts, esters, solvates, isomers, or isotopic variants. The dosing regimen is a 28-day cycle, with administration once on day 1, day 8, and day 15. The characteristic feature is that: The dosage for each administration is (25-55) mg / m². 2 The pathological paraffin blocks or pathological sections of the ex vivo tumor tissues of the cancer and tumor patients were tested for the expression level of AKR1C3 enzyme protein by immunohistochemical staining, and the H-score of the test results was greater than or equal to 100. or The dosage for each administration is (25-55) mg / m². 2 The cancer and tumor patients mentioned are selected from pancreatic cancer, intrahepatic bile duct cancer, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal cancer, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, preferably from pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma patients. or The pathological paraffin blocks or sections of the ex vivo tumor tissue from the cancer and tumor patients were tested for the expression level of AKR1C3 enzyme protein by immunohistochemical staining. The H-score of the test result was greater than or equal to 100. The cancer and tumor patients were selected from pancreatic cancer, intrahepatic bile duct cancer, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal adenocarcinoma, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, with pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma patients being preferred. or The dosage for each administration is (25-55) mg / m². 2 The pathological paraffin blocks or sections of the ex vivo tumor tissue from the cancer or tumor patients were used to detect the expression level of AKR1C3 enzyme protein by immunohistochemical staining. The H-score of the detection result was greater than or equal to 100. The cancer or tumor patients were selected from pancreatic cancer, intrahepatic bile duct cancer, rectal cancer, lung adenocarcinoma, hepatocellular carcinoma, colon cancer, lung squamous cell carcinoma, endometrial cancer, cervical cancer, appendiceal adenocarcinoma, ampullary cancer, breast cancer, prostate cancer, and mixed liver cancer, preferably from pancreatic cancer, intrahepatic bile duct cancer, or hepatocellular carcinoma patients.

9. The method or use according to any one of claims 1-8, wherein the AST-001 is a concentrated solution for injection, and its specification is that each 1 mL contains 10 mg of AST-001 active pharmaceutical ingredient.

10. The use or method according to claim 9, wherein the AST-001 is a concentrated solution for injection, and its specification is that each 1 mL contains 10 mg of AST-001 active pharmaceutical ingredient, and the excipients include N,N-dimethylacetamide, polyoxyethylene (35) castor oil and anhydrous ethanol.

11. The use or method according to claim 9, wherein the AST-001 is a concentrated solution for injection, the specification of which is 10 mg AST-001 raw material per 1 mL, 1 mL of concentrated solution contains 10 mg AST-001 raw material, 50 mg N,N-dimethylacetamide, 400 mg polyoxyethylene (35) castor oil, and the balance is anhydrous ethanol.

12. The use or method according to claim 11, characterized in that, Before administering the medication, add 0.4 ml of 5% sodium bicarbonate injection to 100 ml of sterile 5% glucose injection in an intravenous infusion bag that does not contain di(2-ethylhexyl) phthalate to adjust the pH value. Add the calculated amount of concentrated AST-001 injection solution, accurate to 0.01 ml, to the 5% glucose injection bag after pH adjustment to prepare AST-001 injection solution for intravenous infusion administration. or Before administering the medication, add 0.4 ml of 5% sodium bicarbonate injection to 100 ml of sterile 0.9% saline for injection in an intravenous infusion bag that does not contain di(2-ethylhexyl) phthalate to adjust the pH value. Add the calculated amount of concentrated AST-001 injection solution, accurate to 0.01 ml, to the pH-adjusted saline bag to prepare AST-3424 injection solution for intravenous administration.

13. The use or method according to claim 12, characterized in that, The prepared AST-001 intravenous injection solution should be administered within 6 hours, preferably within 25-35 minutes.

14. The use or method according to claim 12, characterized in that, The final concentration of AST-001 in the prepared AST-001 intravenous injection solution is 0.02 mg / ml to 1.00 mg / ml.

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