WO2022152296A1 - 一种吡啶并[1,2-a]嘧啶酮类似物的应用 - Google Patents

一种吡啶并[1,2-a]嘧啶酮类似物的应用 Download PDF

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WO2022152296A1
WO2022152296A1 PCT/CN2022/072322 CN2022072322W WO2022152296A1 WO 2022152296 A1 WO2022152296 A1 WO 2022152296A1 CN 2022072322 W CN2022072322 W CN 2022072322W WO 2022152296 A1 WO2022152296 A1 WO 2022152296A1
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pik3ca
mutated
cancer
compound
time
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PCT/CN2022/072322
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English (en)
French (fr)
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李永国
隗维
宋艾云
叶未
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广州嘉越医药科技有限公司
上海嘉坦医药科技有限公司
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Priority to JP2023543006A priority Critical patent/JP2024502671A/ja
Priority to EP22739171.1A priority patent/EP4279489A1/en
Priority to US18/260,472 priority patent/US20240075038A1/en
Publication of WO2022152296A1 publication Critical patent/WO2022152296A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems

Definitions

  • the present invention relates to the technical field of biomedicine, in particular, the present invention relates to the application of pyrido[1,2-a]pyrimidinone analogs.
  • malignant tumor is a kind of disease that seriously threatens human life and health, and its morbidity and mortality are increasing year by year.
  • Human mortality due to cancer ranks second only to cardiovascular and cerebrovascular diseases.
  • the essence of carcinogenesis is that the molecular signals that regulate the physiological functions of cells are abnormal during the transduction process, resulting in the disorder of normal physiological functions of cells and the infinite proliferation.
  • Cell signal transduction is closely related to the occurrence, development, recurrence and metastasis of tumors.
  • Traditional cytotoxic drugs for the treatment of tumors generally have shortcomings such as low selectivity, strong toxic and side effects, and poor drug resistance, which promotes the transfer of anti-tumor drugs to the research direction of small molecule targeted drugs.
  • PI3K-AKT-mTOR is an important pathway in cell cycle regulation, which is crucial for cell growth, division, survival and reproduction, and its transitional activation is involved in the occurrence, development, survival and migration of various tumors.
  • PI3K phosphatidylinositol 3-kinase
  • AKT ammalian target of rapamycin
  • mTOR mimmalian target of rapamycin
  • PI3K or mTOR-specific inhibitors listed, and the dual inhibitors of these two molecules can theoretically have better anti-tumor efficacy.
  • PF-05212384 is a dual-target inhibitor of PI3K and mTOR developed by Pfizer and is currently in phase II clinical trials.
  • Everolimus is an oral mTOR single-target inhibitor developed by Novartis, trade name Afinitor, which was approved by the FDA in March 2009. Internationally, everolimus is approved for multiple indications: advanced renal cell carcinoma (RCC), tuberous sclerosis-associated subependymal giant cell astrocytoma (TSC-SEGA) and renal angiomyolipoma (TSC-AML), advanced pancreatic neuroendocrine tumors (pNET), postmenopausal estrogen receptor-positive/HER-2-negative advanced breast cancer (BC) and other tumors.
  • RCC advanced renal cell carcinoma
  • TSC-SEGA tuberous sclerosis-associated subependymal giant cell astrocytoma
  • TSC-AML renal angiomyolipoma
  • pNET pancreatic neuroendocrine tumors
  • BC postmenopausal estrogen receptor-positive
  • the object of the present invention is to provide the application of pyrido[1,2-a]pyrimidinone analogs, the compounds or pharmaceutically acceptable salts thereof have good antitumor activity against PIK3CA mutant cancers, such as PIK3CA mutant One or more of breast cancer, PIK3CA-mutated ovarian cancer, PIK3CA-mutated endometrial cancer, PIK3CA-mutated cervical cancer, and PIK3CA-mutated bladder cancer have good antitumor activity.
  • the present invention provides the use of compound I or a pharmaceutically acceptable salt thereof in the preparation of medicaments, and the structure of said compound I is as follows:
  • the medicament is for the treatment and/or prevention of PIK3CA-mutated cancer.
  • the PIK3CA-mutated cancer may be PIK3CA-mutated breast cancer.
  • the PIK3CA-mutated cancer may be a PIK3CA-mutated ovarian cancer.
  • the PIK3CA-mutated cancer may be a PIK3CA-mutated endometrial cancer.
  • the PIK3CA-mutated cancer may be a PIK3CA-mutated cervical cancer.
  • the PIK3CA-mutated cancer may be a PIK3CA-mutated bladder cancer.
  • the drug is presented in an oral dosage form.
  • the drug is presented as a tablet.
  • the PIK3CA-mutated ovarian cancer can be a PIK3CA-mutated ovarian clear cell carcinoma, such as a PIK3CA-mutated left ovarian clear cell carcinoma.
  • the PIK3CA-mutated ovarian cancer may be a PIK3CA-mutated ovarian cancer with metastases, and the metastases may be liver metastases.
  • the PIK3CA-mutated ovarian cancer may be ovarian cancer that is refractory to first-line treatment regimens or second-line treatment regimens.
  • the PIK3CA-mutated cervical cancer can be PIK3CA-mutated cervical squamous cell carcinoma.
  • the PIK3CA-mutated cervical cancer may be PIK3CA-mutated cervical cancer with metastases, and the metastases may be lymphatic and/or lung.
  • the lymph can be left supraclavicular and/or retroperitoneal lymph.
  • the PIK3CA-mutated cervical cancer may be ineffective for first-line, second- or third-line treatment.
  • the present invention also provides compound I or a pharmaceutically acceptable salt thereof for the treatment and/or prevention of PIK3CA-mutated cancer, the structure of said compound I is shown below:
  • the PIK3CA-mutated cancer is as described in any of the previous schemes.
  • the present invention provides methods of treating and/or preventing PIK3CA-mutated cancers by administering to a patient a therapeutically effective amount of said Compound I or a pharmaceutically acceptable salt thereof.
  • the PIK3CA mutated cancer is as described in any of the previous protocols.
  • the dosage of the compound I or a pharmaceutically acceptable salt thereof can be based on It is administered according to the body weight of the subject/patient, preferably, the administration dose of the compound I or its pharmaceutically acceptable salt is 0.1-2.0 mg/time, for example: 0.1 mg/time, 0.2 mg/time, 0.3 mg/time mg/time, 0.4mg/time, 0.5mg/time, 0.6mg/time, 0.7mg/time, 0.8mg/time, 0.9mg/time, 1.0mg/time, 1.1mg/time, 1.2mg/time, 1.3 mg/time, 1.4 mg/time, 1.5 mg/time, 1.6 mg/time, 1.7 mg/time, 1.8 mg/time, 1.9 mg/time or 2.0 mg/time.
  • the administration frequency of the compound I or a pharmaceutically acceptable salt thereof may be 1 time/day or 2 times/day.
  • the Compound I or a pharmaceutically acceptable salt thereof may be administered orally.
  • the compound I or a pharmaceutically acceptable salt thereof is orally administered, and the administration dose is 0.1-2.0 mg/time, such as 0.1 mg/time. times, 0.4 mg/time, 0.5 mg/time, 0.6 mg/time, 0.7 mg/time, 0.9 mg/time or 1.1 mg/time, and the administration frequency is 1 time/day or 2 times/day.
  • the application or the method for treating and/or preventing PIK3CA-mutated cancer may further include the step of detecting whether the patient carries a PIK3CA gene mutation.
  • the present invention also provides a combination medicine box, which includes medicine box A and medicine box B;
  • the kit A includes a reagent for detecting PIK3CA gene mutation; the kit B includes compound I or a pharmaceutically acceptable salt thereof.
  • the administration times of the kit A and the kit B are not sequential or the kit A is administered first in the kit.
  • the reagent for detecting PIK3CA gene mutation is used to detect whether a cancer patient carries a PIK3CA gene mutation; for example, the cancer patient is suffering from breast cancer, ovarian cancer, endometrial cancer, cervical cancer. and one or more of bladder cancer.
  • the content of the compound I or a pharmaceutically acceptable salt thereof is a therapeutically effective amount.
  • the kit B further includes pharmaceutically acceptable excipients.
  • the dosage and administration frequency of the compound I or a pharmaceutically acceptable salt thereof are as described in any of the previous schemes.
  • the combination kit is for the treatment and/or prevention of PIK3CA-mutated cancers as described in any of the preceding schemes.
  • the term "pharmaceutically acceptable” refers to those compounds, materials, compositions and/or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissue , without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • salts refers to salts of compounds of the present invention prepared with relatively non-toxic, pharmaceutically acceptable acids or bases.
  • base additions can be obtained by contacting neutral forms of such compounds with a sufficient amount of a pharmaceutically acceptable base in neat solution or in a suitable inert solvent.
  • Pharmaceutically acceptable base addition salts include, but are not limited to, lithium, sodium, potassium, calcium, aluminum, magnesium, zinc, bismuth, ammonium, diethanolamine.
  • acids additions can be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable acid in neat solution or in a suitable inert solvent.
  • a salt is not limited to, lithium, sodium, potassium, calcium, aluminum, magnesium, zinc, bismuth, ammonium, diethanolamine.
  • the pharmaceutically acceptable acids include inorganic acids, including but not limited to: hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, sulfuric acid, and the like.
  • Described pharmaceutically acceptable acid includes organic acid, described organic acid includes but is not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid , fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acid citric acid, oleic acid , tannic acid, pantothenic acid, hydrogen tartrate, ascorbic acid, gentisic acid, fumaric acid, gluc
  • treatment refers to therapeutic therapy.
  • treatment refers to: (1) ameliorating one or more biological manifestations of the disease or disorder, (2) interfering with (a) one or more points in the biological cascade leading to or causing the disorder or (b) ) one or more biological manifestations of the disorder, (3) amelioration of one or more symptoms, effects or side effects associated with the disorder, or one or more symptoms, effects or side effects associated with the disorder or its treatment, or (4) slowing the progression of the disorder or one or more biological manifestations of the disorder.
  • prevention refers to a reduced risk of acquiring or developing a disease or disorder.
  • terapéuticaally effective amount refers to an amount of a compound that, when administered to a patient, is sufficient to effectively treat the disease or disorder described herein.
  • a “therapeutically effective amount” will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, but can be adjusted as needed by those skilled in the art.
  • pharmaceutically acceptable excipients refers to the excipients and additives used in the manufacture of pharmaceuticals and formulation of prescriptions, and are all substances other than active ingredients that are included in pharmaceutical preparations. See Pharmacopoeia of the People's Republic of China (2020 Edition) Volume Four or Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009 Sixth Edition).
  • patient refers to any animal, preferably mammals, and most preferably humans, to which the compound is to be or has been administered according to embodiments of the present invention.
  • mammal includes any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., with humans being the most preferred.
  • the reagents and raw materials used in the present invention are all commercially available.
  • Compound I has the effect on one or more of PIK3CA-mutated breast cancer, PIK3CA-mutated ovarian cancer, PIK3CA-mutated cervical cancer, PIK3CA-mutated endometrial cancer and PIK3CA-mutated bladder cancer Good antitumor activity.
  • Figure 1 shows the in vivo efficacy results of compound I on human breast cancer BT-474 xenografts.
  • Compound I in the following examples refers to For pyrido [1,2-a] pyrimidinone analogs.
  • stop solution was added to stop the reaction.
  • the stop solution contains EDTA, biotinylated phosphatidylinositol-3,4,5-triphosphate.
  • Cell culture conditions are: 37°C, 5% CO 2 and 95% humidity;
  • IC 50 value the concentration of inhibitor when 50% inhibitory effect is achieved
  • BT-474 and T47D are PIK3CA mutated breast cancer cell lines, PIK3CA mutated BT-474 and T47D cells were purchased from ATCC, among which, Nos are HTB-20 TM and HTB-133 TM respectively.
  • FBS fetal bovine serum
  • Insulin purchased from Gibco, product number: EPX010-12003-901
  • Test article Compound I;
  • Positive control substance Cisplatin, molecular weight: 300.05; solvent: PBS (phosphate buffered saline); storage condition: 2-8° C.; supplier: Qilu Pharmaceutical.
  • the IC 50 of the compound cell proliferation was determined by CTG method.
  • Step 1 Harvest cells in exponential growth phase and perform a viable cell count using the Vi-Cell XR Cell Counter. Adjust the cell suspension to the appropriate concentration with the medium. Add 90 ⁇ L of cell suspension to each well in a 96-well cell culture plate, and the final cell concentration is 1500-6000 cells/well.
  • Step 2 Compound I was administered at an initial concentration of 3 ⁇ M, the control drug Cisplatin was administered at an initial concentration of 100 ⁇ M, three-fold serial dilution, a total of 9 concentration gradients and a DMSO control, the final concentration of DMSO in each well was 0.1%, set Incubate for 72 hours at 37°C, 5% CO 2 incubator.
  • Step 3 After 72 hours of drug treatment, add 50 ⁇ L (1/2 culture volume) of CTG solution pre-thawed and equilibrated to room temperature in each well according to the CTG operation instructions, mix with a microplate shaker for 2 minutes, and place at room temperature for 10 minutes. Fluorescence signal values were measured with an Envision 2104 plate reader after minutes.
  • the cell viability was calculated by the formula: V sample /V vehicle control ⁇ 100%.
  • V sample is the reading of the drug-treated group
  • V vehicle control is the average value of the solvent control group.
  • GraphPad Prism 5.0 software a nonlinear regression model was used to draw sigmoid dose-survival curves and calculate IC50 values.
  • Cell culture conditions are: 37°C, 5% CO 2 and 95% humidity.
  • ME-180, Ca Ski and C-33A are PIK3CA mutated cervical cancer cell lines, PIK3CA mutated ME-180, Ca Ski and C-33A cells were purchased from ATCC, wherein Nos are HTB-33 TM , CRL-1550 TM and HTB-31 TM , respectively.
  • FBS fetal bovine serum
  • Test article Compound I;
  • Cisplatin molecular weight: 300.05; solvent: PBS (phosphate buffered saline); storage condition: 2-8°C; supplier: Qilu Pharmaceutical.
  • Step 1 Harvest cells in exponential growth phase and perform a viable cell count using the Vi-Cell XR cytometer. Adjust the cell suspension to the appropriate concentration with the medium. Add 90 ⁇ L of cell suspension to each well in a 96-well cell culture plate, and the final cell concentration is 1500-6000 cells/well.
  • Step 2 Compound I and the control drug Alpelisib were administered at an initial concentration of 3 ⁇ M, and the control drug Cisplatin was administered at an initial concentration of 100 ⁇ M, three-fold serial dilution, a total of 9 concentration gradients and a DMSO control, the final concentration of DMSO in each well was 0.1% in a 37°C, 5% CO 2 incubator for 72 hours.
  • Step 3 After 72 hours of drug treatment, add 50 ⁇ L (1/2 culture volume) of CTG solution pre-thawed and equilibrated to room temperature in each well according to the CTG operation instructions, mix with a microplate shaker for 2 minutes, and place at room temperature for 10 minutes. Fluorescence signal values were measured with an Envision 2104 plate reader after minutes.
  • the cell viability was calculated by the formula: V sample /V vehicle control ⁇ 100%.
  • V sample is the reading of the drug-treated group
  • V vehicle control is the average value of the solvent control group.
  • GraphPad Prism 5.0 software a nonlinear regression model was used to draw sigmoid dose-survival curves and calculate IC50 values.
  • Cell culture conditions are: 37°C, 5% CO 2 and 95% humidity.
  • J82 and TCCSUP are PIK3CA-mutated bladder cancer cell lines
  • PIK3CA-mutated J82 and TCCSUP cells were purchased from ATCC, wherein, Nos are HTB-1 TM and HTB-5 TM , respectively.
  • FBS fetal bovine serum
  • Test article Compound I;
  • Cisplatin molecular weight: 300.05; solvent: PBS (phosphate buffered saline); storage condition: 2-8°C; supplier: Qilu Pharmaceutical.
  • Step 1 Harvest cells in exponential growth phase and perform a viable cell count using the Vi-Cell XR cytometer. Adjust the cell suspension to the appropriate concentration with the medium. Add 90 ⁇ L of cell suspension to each well in a 96-well cell culture plate, and the final cell concentration is 1500-6000 cells/well.
  • Step 2 Compound I and the control drug Erdafitinib were administered at an initial concentration of 3 ⁇ M, and the control drug Cisplatin was administered at an initial concentration of 100 ⁇ M, three-fold serial dilution, a total of 9 concentration gradients and a DMSO control, the final concentration of DMSO in each well was 0.1% in a 37°C, 5% CO 2 incubator for 72 hours.
  • Step 3 After 72 hours of drug treatment, add 50 ⁇ L (1/2 culture volume) of CTG solution pre-thawed and equilibrated to room temperature in each well according to the CTG operation instructions, mix with a microplate shaker for 2 minutes, and place at room temperature for 10 minutes. Fluorescence signal values were measured with an Envision 2104 plate reader after minutes.
  • the cell viability was calculated by the formula: V sample /V vehicle control ⁇ 100%.
  • V sample is the reading of the drug-treated group
  • V vehicle control is the average value of the solvent control group.
  • GraphPad Prism 5.0 software a nonlinear regression model was used to draw sigmoid dose-survival curves and IC50 values were calculated.
  • Cell culture conditions are: 37°C, 5% CO 2 and 95% humidity.
  • HEC-1-A and HEC-1-B are PIK3CA-mutated endometrial cancer cell lines, PIK3CA-mutated HEC-1-A and HEC-1-B cells were purchased from ATCC, wherein Nos are HTB- 112TM and HTB- 113TM , respectively.
  • FBS fetal bovine serum
  • Test article Compound I;
  • Positive control substance Alpelisib, molecular weight: 441.47; Solvent: DMSO; Storage condition after dissolution: -20°C; Supplier: Shanghai Taosu Biochemical Technology Co., Ltd., CAS No.: 1217486-61-7;
  • Step 1 Harvest cells in exponential growth phase and perform a viable cell count using the Vi-Cell XR cytometer. Adjust the cell suspension to the appropriate concentration with the medium. Add 90 ⁇ L of cell suspension to each well in a 96-well cell culture plate, and the final cell concentration is 1500-6000 cells/well.
  • Step 2 Compound I and control drug Alpelisib were administered at an initial concentration of 3 ⁇ M, three-fold serial dilution, a total of 9 concentration gradients and a DMSO control, the final concentration of DMSO in each well was 0.1%, placed at 37 ° C, 5% CO 2 in an incubator for 72 hours.
  • Step 3 After 72 hours of drug treatment, add 50 ⁇ L (1/2 culture volume) of CTG solution pre-thawed and equilibrated to room temperature in each well according to the CTG operation instructions, mix with a microplate shaker for 2 minutes, and place at room temperature for 10 minutes. Fluorescence signal values were measured with an Envision 2104 plate reader after minutes.
  • the cell viability was calculated by the formula: V sample /V vehicle control ⁇ 100%.
  • V sample is the reading of the drug-treated group
  • V vehicle control is the average value of the solvent control group.
  • GraphPad Prism 5.0 software a nonlinear regression model was used to draw sigmoid dose-survival curves and IC50 values were calculated.
  • Example 6 In vivo pharmacodynamic study of the test drug on human breast cancer BT-474 subcutaneous xenograft tumor BALB/c nude mouse model
  • OBJECTIVE To study the in vivo efficacy of the test drug on human breast cancer BT-474 subcutaneous xenograft tumor BALB/c nude mice model.
  • Cell culture Human breast cancer BT-474 cells were cultured in monolayer in vitro, and the culture conditions were Hybri-Care medium with 10% fetal bovine serum, 100U/ml penicillin (purchased from Gibco) and 100 ⁇ g/ml streptomycin (purchased from Gibco), cultured at 37°C with 5% CO 2 . Conventional digestion treatment and passage with trypsin-EDTA (purchased from Gibco, product number: 25200-072) were performed twice a week. When the cell saturation was 80%-90%, cells were harvested, counted, and seeded.
  • PF05212384 and everolimus were purchased from Shanghai Taosu Biochemical Technology Co., Ltd.
  • Tumor inoculation 0.2 ml (1 ⁇ 10 7 ) BT-474 cells (plus Matrigel, volume 1:1) were subcutaneously inoculated into the right back of each mouse, and the grouping started when the average tumor volume reached 122 mm 3 Dosing.
  • the experimental groupings and dosing schedules are shown in Table 10 below.
  • Vehicle 2 1% methylcellulose, the percentages are by volume.
  • the reference drug PF05212384 in the 20 mg/kg group had a significant antitumor effect compared with the vehicle control group.
  • Example 7 Efficacy data of compound I on cervical cancer patients with PIK3CA mutation in clinical trials.
  • Case data Subject 1, female, 50 years old, underwent "extensive total hysterectomy with double adnexectomy + pelvic lymph node dissection" on 2019-5-5; the postoperative pathological report showed cervical squamous cell carcinoma; postoperatively on 2019- Postoperative adjuvant therapy was performed from May to August, and recurrence was found on 2020-8-25; first-line treatment, disease progression on 2020-11-10; second-line treatment, on 2021-1-4 Disease progression; third-line treatment, until 2021-6, after disease progression, informed consent was obtained on 2021-6-18, and genetic testing was carried out by Yakangbo PIK3CA gene mutation detection kit.
  • Baseline target lesions are lymph (left supraclavicular) 26mm, right lower lobe 17mm, left lower lobe 14mm, lymph (retroperitoneal) 21mm, total diameter 78mm; 2021-08-25
  • the target lesions are Lymph (left supraclavicular) 13mm, right lower lobe 13mm, left lower lobe 8mm, lymph (retroperitoneal) 13mm, total diameter 47mm; compared with baseline, the tumor shrinkage was 39.7%, and the drug effect was evaluated as PR (partial response, target The sum of the largest diameter of the lesions is reduced by ⁇ 30% and maintained for at least 4 weeks); the second tumor assessment will be conducted on 21 October 2021, and the target lesions are lymphatic (left supraclavicular) 14mm, right lower lobe 14mm, left lower lobe NE (existing, unmeasurable, less than 5mm), lymphatic (retroperitoneal) 12mm, total diameter less than 45mm; shrinkage of tumor greater than 42.3% compared with baseline, eff
  • Compound I has shown curative effect on PIK3CA-mutated cervical cancer in clinical trials. Subject 1 was ineffective after third-line treatment such as radiotherapy and chemotherapy, chemotherapy plus angiogenesis inhibitor, and there was no treatment option available. After taking compound I, the curative effect is significant, and the tumor shrinks to PR (partial remission, the sum of the maximum diameter of the target lesions is reduced by ⁇ 30%, maintained for at least 4 weeks).
  • Example 8 Efficacy data of compound I on ovarian cancer patients with PIK3CA mutation in clinical trials.
  • Case data Subject 2, female, 44 years old, underwent "ovarian cancer tumor cytoreduction (whole uterus + double annex + omentum + appendectomy + right pelvic lymph node biopsy) on 2020-4-27) , postoperative pathology showed clear cell carcinoma of the left ovary; received first-line treatment, the last chemotherapy time was 2020-10-11; CT showed possible liver metastasis on 2020-10-27; performed "liver tumor resection + Abdominal lesion resection + partial diaphragm resection + diaphragm repair + intestinal adhesion release”; the postoperative pathology report was clear cell carcinoma metastasis in the liver; the patient received second-line treatment in January 2021, and the MRI showed on 2021-4-8 Tumor progression.
  • the baseline target lesion is the paracolic groove on the right side of the peritoneum (the longest diameter is 35 mm), the longest diameter of the peritoneal lesser omentum cavity is 47 mm, and the total diameter is 82 mm; 2021-07-06
  • the first time Tumor evaluation showed that the target lesions were (1) the right paracolic groove of the peritoneum (not measurable, calculated by the diameter of 5mm), the longest
  • Compound I showed efficacy in PIK3CA-mutated ovarian cancer in clinical trials. Subject 2 was resistant to second-line standard therapy, and no treatment options were available. After taking compound I, the curative effect was significant, and the tumor reached PR (partial remission, the sum of the maximum diameter of target lesions decreased by ⁇ 30%, maintained for at least 4 weeks) and continued to shrink (59.8% tumor shrinkage).

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Abstract

一种吡啶并[1,2-a]嘧啶酮类似物或其药学上可接受的盐在制备药物中的应用，所述药物用于治疗和/或预防PIK3CA突变的乳腺癌、PIK3CA突变的卵巢癌、PIK3CA突变的子宫内膜癌、PIK3CA突变的宫颈癌和PIK3CA突变的膀胱癌中的一种或多种。

Description

一种吡啶并[1,2-a]嘧啶酮类似物的应用
本申请要求申请日为2021/1/18的中国专利申请2021100617618的优先权。本申请引用上述中国专利申请的全文。
技术领域
本发明涉及生物医药技术领域，具体地，本发明涉及吡啶并[1,2-a]嘧啶酮类似物的应用。
背景技术
目前，恶性肿瘤是严重威胁人类生命健康的一类疾病，其发病率和死亡率呈逐年上升的趋势，人类因癌症引起的死亡率仅次于心脑血管疾病位列第二位。癌变的本质是调控细胞生理功能的分子信号在转导过程中出现异常，导致对细胞的正常生理功能失调而无限增生。细胞信号转导与肿瘤的发生、发展、复发和转移等密切相关。传统治疗肿瘤的细胞毒类药物普遍存在着选择性低、毒副作用强及耐药性差等缺点，这促进了抗肿瘤药物向小分子靶向药物研究方向转移。
PI3K-AKT-mTOR是细胞周期调控的一个重要通路，对细胞生长、分裂、生存和繁殖等至关重要，其过渡活化参与多种肿瘤的发生、发展、存活和迁移。PI3K(磷脂酰肌醇3-激酶，phosphatidylinositol 3-kinase)、AKT和mTOR(mammalian target of rapamycin)是此通路的关键分子，因此成为抗肿瘤治疗的靶标，已有PI3K或mTOR特异性的抑制剂上市，而此两个分子的双重抑制剂理论上可以有更好的抗肿瘤疗效。
PF-05212384(PKI-587)是辉瑞公司开发的PI3K和mTOR双靶点抑制剂，目前处于二期临床实验阶段。依维莫司(Everolimus)是诺华公司开发的口服mTOR单靶点抑制剂，商品名Afinitor，于2009年3月由FDA批准上市。 国际上依维莫司被批准用于多种适应症：晚期肾细胞癌(RCC)、结节性硬化症相关室管膜下巨细胞星型细胞瘤(TSC-SEGA)及肾血管平滑肌脂肪瘤(TSC-AML)、晚期胰腺神经内分泌肿瘤(pNET)、绝经后雌激素受体阳性/HER-2阴性的晚期乳腺癌(BC)等肿瘤。
发明内容
本发明的目的在于提供吡啶并[1,2-a]嘧啶酮类似物的应用，该类化合物或其药学上可接受的盐对PIK3CA突变的癌种具有良好的抗肿瘤活性，例如对PIK3CA突变的乳腺癌、PIK3CA突变的卵巢癌、PIK3CA突变的子宫内膜癌、PIK3CA突变的宫颈癌和PIK3CA突变的膀胱癌中的一种或多种具有良好的抗肿瘤活性。
本发明提供了化合物Ⅰ或其药学上可接受的盐在制备药物中的应用，所述化合物Ⅰ的结构如下所示：
Figure PCTCN2022072322-appb-000001
所述药物用于治疗和/或预防PIK3CA突变的癌症。
其中，所述PIK3CA突变的癌症可为PIK3CA突变的乳腺癌。
所述PIK3CA突变的癌症可为PIK3CA突变的卵巢癌。
所述PIK3CA突变的癌症可为PIK3CA突变的子宫内膜癌。
所述PIK3CA突变的癌症可为PIK3CA突变的宫颈癌。
所述PIK3CA突变的癌症可为PIK3CA突变的膀胱癌。
本发明中，所述药物以口服剂型呈现。
本发明中，所述药物以片剂呈现。
所述的PIK3CA突变的卵巢癌可为PIK3CA突变的卵巢透明细胞癌，例 如为PIK3CA突变的左卵巢透明细胞癌。
所述的PIK3CA突变的卵巢癌可为具有转移灶的PIK3CA突变的卵巢癌，所述转移灶可为肝转移灶。
所述PIK3CA突变的卵巢癌可为对一线治疗方案或二线治疗方案无效的卵巢癌。
所述的PIK3CA突变的宫颈癌可为PIK3CA突变的宫颈鳞癌。
所述的PIK3CA突变的宫颈癌可为具有转移灶的PIK3CA突变的宫颈癌，所述转移灶可为淋巴和/或肺。所述淋巴可为左锁骨上淋巴和/或腹膜后淋巴。
所述的PIK3CA突变的宫颈癌可为对一线治疗方案、二线治疗方案或三线治疗方案无效的宫颈癌。
本发明还提供了用于治疗和/或预防PIK3CA突变的癌症的化合物Ⅰ或其药学上可接受的盐，所述化合物Ⅰ的结构如下所示：
Figure PCTCN2022072322-appb-000002
其中，所述PIK3CA突变的癌症如前任一方案所述。
本发明提供了治疗和/或预防PIK3CA突变的癌症的方法，其通过向患者施用治疗有效量的所述化合物Ⅰ或其药学上可接受的盐。所述PIK3CA突变的癌症如前任一方案所述。
所述化合物Ⅰ或其药学上可接受的盐在制备药物中的应用或所述治疗和/或预防PIK3CA突变的癌症的方法中，所述化合物Ⅰ或其药学上可接受的盐的剂量可以根据受试者/患者的体重来施用，较佳地，所述化合物Ⅰ或其药学上可接受的盐的施用剂量为0.1-2.0mg/次，例如：0.1mg/次、0.2mg/次、0.3 mg/次、0.4mg/次、0.5mg/次、0.6mg/次、0.7mg/次、0.8mg/次、0.9mg/次、1.0mg/次、1.1mg/次、1.2mg/次、1.3mg/次、1.4mg/次、1.5mg/次、1.6mg/次、1.7mg/次、1.8mg/次、1.9mg/次或2.0mg/次。
所述应用或所述治疗和/或预防PIK3CA突变的癌症的方法中，所述化合物Ⅰ或其药学上可接受的盐的施用频率可为1次/日或2次/日。
所述应用或所述治疗和/或预防PIK3CA突变的癌症的方法中，所述化合物Ⅰ或其药学上可接受的盐可经口服施用。
较佳地，所述应用或所述治疗和/或预防癌症的方法中，所述化合物Ⅰ或其药学上可接受的盐经口服施用，施用剂量为0.1-2.0mg/次，例如0.1mg/次、0.4mg/次、0.5mg/次、0.6mg/次、0.7mg/次、0.9mg/次或1.1mg/次，施用频率为1次/日或2次/日。
所述应用或所述治疗和/或预防PIK3CA突变的癌症的方法中，还可包括检测患者是否携带PIK3CA基因突变的步骤。
本发明还提供了一种组合药盒，其包括药盒A和药盒B；
其中所述药盒A包括检测PIK3CA基因突变的试剂；所述药盒B包括化合物Ⅰ或其药学上可接受的盐。
较佳地，所述药盒A与药盒B的施用时间不分先后或者所述药盒中先施用所述药盒A。
较佳地，所述药盒A中，所述检测PIK3CA基因突变的试剂用于检测癌症患者是否携带PIK3CA基因突变；例如所述癌症患者为患有乳腺癌、卵巢癌、子宫内膜癌、宫颈癌和膀胱癌中的一种或多种的患者。
较佳地，所述药盒B中，所述化合物Ⅰ或其药学上可接受的盐的含量为治疗有效量。
较佳地，所述药盒B还包括药学上可接受的辅料。
较佳地，所述药盒B中，所述化合物Ⅰ或其药学上可接受的盐的剂量和施用频率如前任一方案所述。
较佳地，所述组合药盒用于治疗和/或预防PIK3CA突变的癌症，所述PIK3CA突变的癌症如前任一方案所述。
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。
这里所采用的术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。
术语“药学上可接受的盐”是指本发明化合物与相对无毒的、药学上可接受的酸或碱制备得到的盐。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的药学上可接受的碱与这类化合物的中性形式接触的方式获得碱加成盐。药学上可接受的碱加成盐包括但不限于：锂盐、钠盐、钾盐、钙盐、铝盐、镁盐、锌盐、铋盐、铵盐、二乙醇胺盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的药学上可接受的酸与这类化合物的中性形式接触的方式获得酸加成盐。所述的药学上可接受的酸包括无机酸，所述无机酸包括但不限于：盐酸、氢溴酸、氢碘酸、硝酸、碳酸、磷酸、亚磷酸、硫酸等。所述的药学上可接受的酸包括有机酸，所述有机酸包括但不限于：乙酸、丙酸、草酸、异丁酸、马来酸、丙二酸、苯甲酸、琥珀酸、辛二酸、反丁烯二酸、乳酸、扁桃酸、邻苯二甲酸、苯磺酸、对甲苯磺酸、柠檬酸、水杨酸、酒石酸、甲磺酸、异烟酸、酸式柠檬酸、油酸、单宁酸、泛酸、酒石酸氢、抗坏血酸、龙胆酸、富马酸、葡糖酸、糖酸、甲酸、乙磺酸、双羟萘酸(即4,4’-亚甲基-双(3-羟基-2-萘甲酸))、氨基酸(例如谷氨酸、精氨酸)等。当本发明的化合物中含有相对酸性和相对碱性的官能团时，可以 被转换成碱加成盐或酸加成盐。具体可参见Berge et al.,"Pharmaceutical Salts",Journal of Pharmaceutical Science 66:1-19(1977)、或、Handbook of Pharmaceutical Salts:Properties,Selection,and Use(P.Heinrich Stahl and Camille G.Wermuth,ed.,Wiley-VCH,2002)。
术语“治疗”指治疗性疗法。涉及具体病症时，治疗指：(1)缓解疾病或者病症的一种或多种生物学表现，(2)干扰(a)导致或引起病症的生物级联中的一个或多个点或(b)病症的一种或多种生物学表现，(3)改善与病症相关的一种或多种症状、影响或副作用，或者与病症或其治疗相关的一种或多种症状、影响或副作用，或(4)减缓病症或者病症的一种或多种生物学表现发展。
术语“预防”是指获得或发生疾病或障碍的风险降低。
术语“治疗有效量”是指在给予患者时足以有效治疗本文所述的疾病或病症的化合物的量。“治疗有效量”将根据化合物、病症及其严重度、以及欲治疗患者的年龄而变化，但可由本领域技术人员根据需要进行调整。
术语“药学上可接受的辅料”是指生产药品和调配处方时使用的赋形剂和附加剂，是除活性成分以外，包含在药物制剂中的所有物质。可参见中华人民共和国药典(2020年版)四部或Handbook of Pharmaceutical Excipients(Raymond C Rowe,2009 Sixth Edition)。
术语“患者”是指根据本发明的实施例，即将或已经接受了该化合物给药的任何动物，哺乳动物为优，人类最优。术语“哺乳动物”包括任何哺乳动物。哺乳动物的实例包括但不限于牛、马、羊、猪、猫、狗、小鼠、大鼠、家兔、豚鼠、猴、人等，以人类为最优。
在不违背本领域常识的基础上，上述各优选条件，可任意组合，即得本发明各较佳实例。
本发明所用试剂和原料均市售可得。
本发明的积极进步效果在于：化合物Ⅰ对PIK3CA突变的乳腺癌、PIK3CA突变的卵巢癌、PIK3CA突变的宫颈癌、PIK3CA突变的子宫内膜癌和 PIK3CA突变的膀胱癌中的一种或多种具有良好的抗肿瘤活性。
附图说明
图1为化合物Ⅰ对人源乳腺癌BT-474异种移植瘤的体内药效结果。
具体实施方式
下面通过实施例的方式进一步说明本发明，但并不因此将本发明限制在所述的实施例范围之中。下列实施例中未注明具体条件的实验方法，按照常规方法和条件，或按照商品说明书选择。
下述实施例中的化合物I是指
Figure PCTCN2022072322-appb-000003
为吡啶并[1,2-a]嘧啶酮类似物。
实施例1 测试化合物Ⅰ体外对PIK3CA突变体激酶活性的抑制作用的IC 50
1.实验材料和方法
主要试剂：人PI3K p110α/p85α(购于Promega，货号V1721)、人PI3K p110α(E542K)/p85α(购于Millipore，货号14-782)、人PI3K p110α(E545K)/p85α(购于Millipore，货号14-783)和人PI3K p110α(H1047R)/p85α(购于Millipore，货号14-792)
实验方法：
1)配置化合物，化合物Ⅰ的起始浓度为100nM，3倍递减稀释10个浓度并转移至测试板中。
2)将人PI3K p110α/p85α、人PI3K p110α(E542K)/p85α、人PI3K p110α(E545K)/p85α或人PI3K p110α(H1047R)/p85α与测定缓冲液 (10μM磷脂酰肌醇4,5-二磷酸和Mg/ATP)孵育。通过加入ATP溶液引发反应，孵育30分钟。
3)孵育30分钟后，加入终止液终止反应。终止液包含EDTA,生物素化磷脂酰肌醇-3,4,5-三磷酸。
4)添加检测缓冲液，其中含有铕标记的抗GST单克隆抗体、GST标记的GRP1PH结构域和链霉亲和素别藻蓝蛋白。
5)读板并根据公式HTRF＝10000x(Em665nm/Em620nm)，计算HTRF值。应用GraphPad Prism 5.0软件，使用非线性回归模型绘制S型剂量-存活率曲线并计算IC 50值。
2.实验结果
表1 化合物Ⅰ在检测的PI3Kα激酶以及其突变体上的IC 50数值
Figure PCTCN2022072322-appb-000004
从表1的实验结果可以看出，化合物Ⅰ不仅对PI3Kα激酶有着极强的抑制作用(IC 50＝1nM)，而且对其三种典型的突变体(E542K，H1047R和E545K)的抑制作用与野生型相当。
实施例2 用CTG方法测试化合物Ⅰ在PIK3CA突变的乳腺癌细胞系中的IC 50
1.实验材料和方法
(1)细胞系
表2 PIK3CA突变的乳腺癌细胞系
Figure PCTCN2022072322-appb-000005
备注：a.细胞培养条件为：37℃，5％CO 2和95％湿度；
b.IC 50值：达到50％抑制效果时抑制剂的浓度；
c.BT-474、T47D为PIK3CA突变的乳腺癌细胞系，PIK3CA突变的BT-474和T47D细胞购买于ATCC，其中，
Figure PCTCN2022072322-appb-000006
No分别为HTB-20 TM、HTB-133 TM。
d.表2中10％FBS中的百分数为体积百分数。
(2)试剂
1)FBS(胎牛血清)(购于ExCell，产品号：FND500)；
2)DMEM培养基(购于Gibco，产品号：C11995500BT)；
3)Insulin(购于Gibco，产品号：EPX010-12003-901)；
4)RPMI1640培养基(购于Hyclone，产品号：SH30809.01)；
5)MEM培养基(购于Hyclone，产品号：SH30024.01)。
(3)供试品和阳性对照品
供试品：化合物Ⅰ；
阳性对照品：Cisplatin，分子量：300.05；溶剂：PBS(磷酸盐缓冲液)；储存条件：2-8℃；供应商：齐鲁制药。
(4)CTG方法测定化合物细胞增殖IC 50。
步骤1：收集处于指数生长期的细胞并用Vi-Cell XR细胞计数仪进行活 细胞计数。用培养基将细胞悬液调整到适当浓度。每孔加90μL细胞悬液于96孔细胞培养板，最终细胞浓度为1500～6000细胞/孔。
步骤2：化合物Ⅰ给药起始浓度为3μM,对照药物Cisplatin给药起始浓度为100μM，三倍连续稀释，一共9个浓度梯度和一个DMSO的对照，每孔DMSO终浓度为0.1％，置于37℃，5％CO 2孵箱中培养72小时。
步骤3：药物处理72小时后，按照CTG操作说明，每孔加入50μL(1/2培养体积)预先融化并平衡到室温的CTG溶液，用微孔板震荡器混匀2分钟，于室温放置10分钟后用Envision2104读板仪测定荧光信号值。
(5)数据分析
细胞存活率用公式：V sample/V vehicle control×100％计算。其中V sample为药物处理组的读数，V vehicle control为溶剂对照组的平均值。应用GraphPad Prism 5.0软件，使用非线性回归模型绘制S型剂量-存活率曲线并计算IC 50值。
2.实验结果
表3 化合物Ⅰ和Cisplatin在检测的各细胞系上的IC 50数值
Figure PCTCN2022072322-appb-000007
从表3的实验结果可以看出，化合物Ⅰ对T47D和BT-474肿瘤细胞达到50％抑制效果时抑制剂的浓度都低于1μM。
实施例3用CTG方法测试化合物I在PIK3CA突变的宫颈癌细胞系中的IC 50
1.实验材料与方法
(1)细胞系
表4 PIK3CA突变的宫颈癌细胞系
Figure PCTCN2022072322-appb-000008
备注：a.细胞培养条件为：37℃，5％CO 2和95％湿度。
b.ME-180、Ca Ski和C-33A为PIK3CA突变的宫颈癌细胞系，PIK3CA突变的ME-180、Ca Ski和C-33A细胞购买于ATCC，其中
Figure PCTCN2022072322-appb-000009
No分别为HTB-33 TM、CRL-1550 TM和HTB-31 TM。
c.表4中10％FBS、1％MEM NEAA和1％丙酮酸钠中的百分数为体积百分数。
(2)试剂
1)FBS(胎牛血清)(购于ExCell，产品号：FND500)；
2)RPMI1640培养基(购于Hyclone，产品号：SH30809.01)；
3)McCoy's 5A培养基(购于Gibco,产品号：12330-031)；
4)MEM培养基(购于Hyclone，产品号：SH30024.01)；
5)MEM NEAA(购于Gibco，产品号：11140-050)；
6)丙酮酸钠(购于Gibco)。
(3)供试品和阳性对照品
供试品：化合物Ⅰ；
阳性对照品：
1)Alpelisib，分子量：441.47；溶剂：DMSO；溶解后储存条件：-20℃；供应商：上海陶素生化科技有限公司，CAS号：1217486-61-7；
2)Cisplatin，分子量：300.05；溶剂：PBS(磷酸盐缓冲液)；储存条件：2-8℃；供应商：齐鲁制药。
(4)CTG方法测定化合物细胞增殖IC 50
步骤1：收集处于指数生长期的细胞并用Vi-Cell XR细胞计数仪进行活细胞计数。用培养基将细胞悬液调整到适当浓度。每孔加90μL细胞悬液于96孔细胞培养板，最终细胞浓度为1500～6000细胞/孔。
步骤2：化合物Ⅰ和对照药物Alpelisib给药起始浓度为3μM,对照药物Cisplatin给药起始浓度为100μM，三倍连续稀释，一共9个浓度梯度和一个DMSO的对照，每孔DMSO终浓度为0.1％，置于37℃，5％CO 2孵箱中培养72小时。
步骤3：药物处理72小时后，按照CTG操作说明，每孔加入50μL(1/2培养体积)预先融化并平衡到室温的CTG溶液，用微孔板震荡器混匀2分钟，于室温放置10分钟后用Envision2104读板仪测定荧光信号值。
(5)数据分析
细胞存活率用公式：V sample/V vehicle control×100％计算。其中V sample为药物处理组的读数，V vehicle control为溶剂对照组的平均值。应用GraphPad Prism 5.0软件，使用非线性回归模型绘制S型剂量-存活率曲线并计算IC 50值。
2.实验结果
表5 化合物Ⅰ、Alpelisib和Cisplatin在检测的各细胞系上的IC 50数值
Figure PCTCN2022072322-appb-000010
结论：化合物I对于PIK3CA突变的宫颈癌细胞生长抑制明显，半数抑制浓度在0.015-0.180μM之间，而对照化合物Alpelisib基本无效，在两个细胞系中半数抑制浓度高于最大浓度3μM；另外，化合物Ⅰ对PIK3CA突变的 宫颈癌细胞生长抑制优于对照化合物Cisplatin。
实施例4 用CTG方法测试化合物I在PIK3CA突变的膀胱癌细胞系中的IC 50
1.实验材料与方法
(1)细胞系
表6 PIK3CA突变的膀胱癌细胞系
Figure PCTCN2022072322-appb-000011
备注：a.细胞培养条件为：37℃，5％CO 2和95％湿度。
b.J82和TCCSUP为PIK3CA突变的膀胱癌细胞系，PIK3CA突变的J82和TCCSUP细胞购买于ATCC，其中，
Figure PCTCN2022072322-appb-000012
No分别为HTB-1 TM和HTB-5 TM。
c.表6中10％FBS、1％MEM NEAA和1％丙酮酸钠中的百分数为体积百分数。
(2)试剂
1)FBS(胎牛血清)(购于ExCell，产品号：FND500)；
2)MEM培养基(购于Hyclone，产品号：SH30024.01)；
3)MEM NEAA(购于Gibco，产品号：11140-050)；
4)丙酮酸钠(购于Gibco)。
(3)供试品和阳性对照品
供试品：化合物Ⅰ；
阳性对照品：
1)Erdafitinib，分子量：446.55；溶剂：DMSO；溶解后储存条件：-8℃；供应商：上海陶素生化科技有限公司，CAS号：1346242-81-6；
2)Cisplatin，分子量：300.05；溶剂：PBS(磷酸盐缓冲液)；储存条件：2-8℃；供应商：齐鲁制药。
(4)CTG方法测定化合物细胞增殖IC 50
步骤1：收集处于指数生长期的细胞并用Vi-Cell XR细胞计数仪进行活细胞计数。用培养基将细胞悬液调整到适当浓度。每孔加90μL细胞悬液于96孔细胞培养板，最终细胞浓度为1500～6000细胞/孔。
步骤2：化合物Ⅰ和对照药物Erdafitinib给药起始浓度为3μM,对照药物Cisplatin给药起始浓度为100μM，三倍连续稀释，一共9个浓度梯度和一个DMSO的对照，每孔DMSO终浓度为0.1％，置于37℃，5％CO 2孵箱中培养72小时。
步骤3：药物处理72小时后，按照CTG操作说明，每孔加入50μL(1/2培养体积)预先融化并平衡到室温的CTG溶液，用微孔板震荡器混匀2分钟，于室温放置10分钟后用Envision2104读板仪测定荧光信号值。
(5)数据分析
细胞存活率用公式：V sample/V vehicle control×100％计算。其中V sample为药物处理组的读数，V vehicle control为溶剂对照组的平均值。应用GraphPad Prism 5.0软件，使用非线性回归模型绘制S型剂量-存活率曲线并计算IC 50值。
2.实验结果
表7 化合物Ⅰ、Erdafitinib和Cisplatin在检测的各细胞系上的IC 50数值
Figure PCTCN2022072322-appb-000013
Figure PCTCN2022072322-appb-000014
结论：化合物I对于PIK3CA突变的膀胱癌细胞生长抑制明显，半数抑制浓度在1μM以内，而对照化合物Erdafitinib基本无效，在两个细胞系中半数抑制浓度高于最大浓度3μM；另外，化合物Ⅰ对PIK3CA突变的膀胱癌细胞生长抑制优于对照化合物Cisplatin。
实施例5 用CTG方法测试化合物I在PIK3CA突变的子宫内膜癌细胞系中的IC 50
1.实验材料与方法
(1)细胞系
表8 PIK3CA突变的子宫内膜癌细胞系
Figure PCTCN2022072322-appb-000015
备注：a.细胞培养条件为：37℃，5％CO 2和95％湿度。
b.HEC-1-A和HEC-1-B为PIK3CA突变的子宫内膜癌细胞系，PIK3CA突变的HEC-1-A和HEC-1-B细胞购买于ATCC，其中
Figure PCTCN2022072322-appb-000016
No分别为HTB-112 TM和HTB-113 TM。
c.表8中10％FBS中的百分数为体积百分数。
(2)试剂
1)FBS(胎牛血清)(购于ExCell，产品号：FND500)；
2)McCoy's 5A培养基(购于Gibco,产品号：12330-031)；
3)MEM培养基(购于Hyclone，产品号：SH30024.01)。
(3)供试品和阳性对照品
供试品：化合物Ⅰ；
阳性对照品：Alpelisib，分子量：441.47；溶剂：DMSO；溶解后储存条件：-20℃；供应商：上海陶素生化科技有限公司，CAS号：1217486-61-7；
(4)CTG方法测定化合物细胞增殖IC 50
步骤1：收集处于指数生长期的细胞并用Vi-Cell XR细胞计数仪进行活细胞计数。用培养基将细胞悬液调整到适当浓度。每孔加90μL细胞悬液于96孔细胞培养板，最终细胞浓度为1500～6000细胞/孔。
步骤2：化合物Ⅰ和对照药物Alpelisib给药起始浓度为3μM，三倍连续稀释，一共9个浓度梯度和一个DMSO的对照，每孔DMSO终浓度为0.1％，置于37℃，5％CO 2孵箱中培养72小时。
步骤3：药物处理72小时后，按照CTG操作说明，每孔加入50μL(1/2培养体积)预先融化并平衡到室温的CTG溶液，用微孔板震荡器混匀2分钟，于室温放置10分钟后用Envision2104读板仪测定荧光信号值。
(5)数据分析
细胞存活率用公式：V sample/V vehicle control×100％计算。其中V sample为药物处理组的读数，V vehicle control为溶剂对照组的平均值。应用GraphPad Prism 5.0软件，使用非线性回归模型绘制S型剂量-存活率曲线并计算IC 50值。
2.实验结果
表9 化合物Ⅰ、Alpelisib在检测的各细胞系上的IC 50数值
Figure PCTCN2022072322-appb-000017
结论：化合物I对于PIK3CA突变的子宫内膜癌细胞生长抑制明显，半数抑制浓度在0.1μM以内，而对照化合物Alpelisib高于2uM。与对照药相 比，化合物I对于PIK3CA突变的子宫内膜癌疗效显著。
实施例6 研究受试药对人源乳腺癌BT-474皮下异种移植肿瘤BALB/c裸小鼠模型的体内药效学研究
实验目的：研究受试药对人源乳腺癌BT-474皮下异种移植肿瘤BALB/c裸小鼠模型具有体内药效。
实验设计
(1)细胞培养：人乳腺癌BT-474细胞体外单层培养，培养条件为Hybri-Care培养基中加10％胎牛血清，100U/ml青霉素(购于Gibco)和100μg/ml链霉素(购于Gibco)，37℃ 5％CO 2培养。一周两次用胰酶-EDTA(购于Gibco，产品号：25200-072)进行常规消化处理传代。当细胞饱和度为80％-90％时，收取细胞，计数，接种。
(2)动物：BALB/c裸小鼠，雌性，6周龄，体重16-18克。由上海西普尔-必凯实验动物有限公司提供。
(3)对照品
PF05212384和依维莫司都购于上海陶素生化科技有限公司。
(4)肿瘤接种：将0.2ml(1×10 7)BT-474细胞(加基质胶，体积1:1)皮下接种于每只小鼠的右后背，肿瘤平均体积达到122mm 3时开始分组给药。实验分组和给药方案见下表10。
表10 受试药对人乳腺癌BT-474细胞模型的实验分组和给药方案
Figure PCTCN2022072322-appb-000018
Figure PCTCN2022072322-appb-000019
备注：
溶媒1：丙二醇、Tween-80及5％葡萄糖溶液(v:v:v＝30:5:65)混合液。
溶媒2：1％甲基纤维素，所述百分数为体积百分数。
(4)体内药效结果：如图1和表11所示。
化合物Ⅰ在人源乳腺癌BT-474异种移植瘤模型中的体内药效。给药20天时，与溶媒对照组相比，化合物Ⅰ在0.05mg/kg、0.1mg/kg和0.3mg/kg的剂量下，T/C分别为：44％、23％和16％，TGI分别为77％、109％和118％，由此结果可知在人源乳腺癌BT-474异种移植瘤模型中，化合物Ⅰ在0.05，0.1和0.3mg/kg的剂量均具有显著抗肿瘤作用，且有剂量依赖的趋势。参照药PF05212384在20mg/kg组(T/C＝15％，TGI＝134％，p＝0.006)，与溶媒对照组相比具有显著的抗肿瘤作用。依维莫司(Everolimus)，5mg/kg(T/C＝19％，TGI＝113％，p<0.001)，与溶媒对照组相比具有显著的抗肿瘤作用，化合物Ⅰ在0.3mg/kg剂量具有与依维莫司(Everolimus)在5mg/kg，相当的抗肿瘤作用。
表11 化合物Ⅰ对BT-474异种移植瘤模型的抑瘤药效评价
(基于给药后第20天数据得出)
Figure PCTCN2022072322-appb-000020
Figure PCTCN2022072322-appb-000021
注：
a.平均值±SEM。
b.肿瘤生长抑制由T/C和TGI(TGI(％)＝[1-(T 20-T 0)/(V 20-V 0)]×100)计算。
结论：化合物I对于PIK3CA突变的乳腺癌肿瘤生长抑制明显，在极低的剂量组0.1mg/kg，0.3mg/kg可以引起肿瘤缩小，与对照化合物依维莫司(5mg/kg，高剂量)相当。
实施例7 临床试验中化合物I对于携带PIK3CA突变的宫颈癌患者的疗效数据。
病例资料：受试者1，女，50岁，于2019-5-5进行“广泛全子宫双附件切除术+盆腔淋巴清扫术”；术后病理报告显示为宫颈鳞癌；术后于2019-5月至8月期间进行术后辅助治疗，于2020-8-25影像学现复发；一线治疗，于2020-11-10影像学现疾病进展；二线治疗，于2021-1-4影像学现疾病进展；三线治疗，至2021-6疾病进展后于2021-6-18知情同意，通过雅康博人PIK3CA基因突变检测试剂盒进行基因检测，基因检测显示肿瘤携带PIK3CA突变，参加“化合物I治疗PIK3CA突变的妇科肿瘤的临床研究”：自2021-07-01日起至2021-10-21日第二次肿瘤评估期间，含化合物Ⅰ的片剂，剂量为1.1mg，单次给药，每日一次，1.1mg剂量含化合物Ⅰ为0.5mg的片剂两片和含化合物Ⅰ为0.1mg的片剂一片。基线靶病灶为淋巴(左锁骨上)26mm，右肺下叶17mm，左肺下叶14mm，淋巴(腹膜后)21mm，总径78mm；2021-08-25进行第一次肿瘤评估，靶病灶为淋巴(左锁骨上)13mm，右肺下叶13mm，左肺下叶8mm，淋巴(腹膜后)13mm，总径47mm；与基线相比缩瘤39.7％， 药效评估为PR(部分缓解，靶病灶最大径之和减少≥30％，至少维持4周)；2021-10-21进行第二次肿瘤评估，靶病灶为淋巴(左锁骨上)14mm，右肺下叶14mm，左肺下叶NE(存在，无法测量，小于5mm)，淋巴(腹膜后)12mm，总径小于45mm；与基线相比缩瘤大于42.3％，药效评估为PR(部分缓解，靶病灶最大径之和减少≥30％，至少维持4周)。
结论：化合物I在临床试验中对PIK3CA突变的宫颈癌显示出疗效，受试者1在经过放化疗，化疗加血管生成抑制剂等三线治疗无效，已无治疗方案可选。在服用化合物I后，疗效显著，肿瘤缩小至PR(部分缓解，靶病灶最大径之和减少≥30％，至少维持4周)。
实施例8 临床试验中化合物I对于携带PIK3CA突变的卵巢癌患者的疗效数据。
病例资料：受试者2，女，44岁，于2020-4-27进行“卵巢癌肿瘤细胞减灭术(全子宫+双附件+大网膜+阑尾切除术+右侧盆腔淋巴结活检术)，术后病理显示左卵巢透明细胞癌；接受一线治疗方案治疗，末次化疗时间为2020-10-11；2020-10-27CT显示肝转移可能；于2020-11-23进行“肝肿瘤切除术+腹腔病损切除术+膈肌部分切除术+膈肌修补术+肠粘连松解术”；术后病理报告肝部为透明细胞癌转移；2021年1月患者接受二线治疗，2021-4-8核磁显示肿瘤进展。2021-04-23获得知情同意，既往基于二代测序的基因检测报告显示肿瘤携带PIK3CA突变，参加“化合物I治疗PIK3CA突变的妇科肿瘤的临床研究”：自2021-05-11日起至2021-08-31日第二次肿瘤评估期间，含化合物Ⅰ的片剂，剂量为1.1mg，单次给药，每日一次，1.1mg剂量含化合物Ⅰ为0.5mg的片剂两片和含化合物Ⅰ为0.1mg的片剂一片；基线靶病灶为腹膜右侧结肠旁沟(最长直径35mm)，腹膜小网膜腔最长直径47mm，总径82mm；2021-07-06第一次肿瘤评估显示，靶病灶为(1)腹膜右侧结肠旁沟(不可测量，按直径5mm计算)，腹膜小网膜腔最长直径34mm；总径39mm；与基 线相比缩瘤52.4％；药效评估为PR(部分缓解，靶病灶最大径之和减少≥30％，至少维持4周)；2021-08-30：第二次肿瘤评估，靶病灶为(1)腹膜右侧结肠旁沟(不可测量，按直径5mm计算)，(2)腹膜小网膜腔最长直径28mm；总径33mm；与基线相比缩瘤59.8％；药效评估为PR(部分缓解，靶病灶最大径之和减少≥30％，至少维持4周)。
结论：化合物I在临床试验中对PIK3CA突变的卵巢癌显示出疗效，受试者2在经过二线的标准治疗后耐药，已无治疗方案可选。在服用化合物I后，疗效显著，肿瘤至PR(部分缓解，靶病灶最大径之和减少≥30％，至少维持4周)并持续缩小(缩瘤59.8％)。

Claims (13)

  1. 一种化合物Ⅰ或其药学上可接受的盐在制备药物中的应用，其特征在于，所述化合物Ⅰ的结构如下所示：
    Figure PCTCN2022072322-appb-100001
    所述药物用于治疗和/或预防PIK3CA突变的癌症。
  2. 如权利要求1所述的化合物Ⅰ或其药学上可接受的盐在制备药物中的应用，其特征在于，所述PIK3CA突变的癌症为PIK3CA突变的乳腺癌。
  3. 如权利要求1所述的化合物Ⅰ或其药学上可接受的盐在制备药物中的应用，其特征在于，所述PIK3CA突变的癌症为PIK3CA突变的卵巢癌。
  4. 如权利要求1所述的化合物Ⅰ或其药学上可接受的盐在制备药物中的应用，其特征在于，所述PIK3CA突变的癌症为PIK3CA突变的子宫内膜癌。
  5. 如权利要求1所述的化合物Ⅰ或其药学上可接受的盐在制备药物中的应用，其特征在于，所述PIK3CA突变的癌症为PIK3CA突变的宫颈癌。
  6. 如权利要求1所述的化合物Ⅰ或其药学上可接受的盐在制备药物中的应用，其特征在于，所述PIK3CA突变的癌症为PIK3CA突变的膀胱癌。
  7. 如权利要求3所述的化合物Ⅰ或其药学上可接受的盐在制备药物中的应用，其特征在于，所述PIK3CA突变的卵巢癌满足以下条件中的一种或多种：
    (1)所述的PIK3CA突变的卵巢癌为PIK3CA突变的卵巢透明细胞癌；
    (2)所述的PIK3CA突变的卵巢癌为具有转移灶的PIK3CA突变的卵巢癌；
    (3)所述PIK3CA突变的卵巢癌为对一线治疗方案或二线治疗方案无 效的卵巢癌；
    较佳地，所述PIK3CA突变的卵巢癌满足以下条件中的一种或多种：
    (I)所述的PIK3CA突变的卵巢癌为PIK3CA突变的左卵巢透明细胞癌；
    (II)所述的PIK3CA突变的卵巢癌为具有肝转移灶的PIK3CA突变的卵巢癌。
  8. 如权利要求5所述的化合物Ⅰ或其药学上可接受的盐在制备药物中的应用，其特征在于，所述的PIK3CA突变的宫颈癌满足以下条件中的一种或多种：
    (1)所述的PIK3CA突变的宫颈癌为PIK3CA突变的宫颈鳞癌；
    (2)所述的PIK3CA突变的宫颈癌为具有转移灶的PIK3CA突变的宫颈癌；
    (3)所述的PIK3CA突变的宫颈癌为对一线治疗方案、二线治疗方案或三线治疗方案无效的宫颈癌；
    较佳地，所述转移灶为淋巴和/或肺；更佳地，所述淋巴为左锁骨上淋巴和/或腹膜后淋巴。
  9. 如权利要求1-8至少一项所述的化合物Ⅰ或其药学上可接受的盐在制备药物中的应用，其特征在于，其满足以下条件中的一种或多种：
    (1)所述药物以口服剂型呈现；
    (2)所述药物以片剂呈现；
    (3)所述化合物Ⅰ或其药学上可接受的盐的施用剂量为0.1-2.0mg/次，例如：0.1mg/次、0.2mg/次、0.3mg/次、0.4mg/次、0.5mg/次、0.6mg/次、0.7mg/次、0.8mg/次、0.9mg/次、1.0mg/次、1.1mg/次、1.2mg/次、1.3mg/次、1.4mg/次、1.5mg/次、1.6mg/次、1.7mg/次、1.8mg/次、1.9mg/次或2.0mg/次；
    (4)所述化合物Ⅰ或其药学上可接受的盐的施用频率为1次/日或2次/ 日。
  10. 一种组合药盒，其包括药盒A和药盒B；
    其中所述药盒A包括检测PIK3CA基因突变的试剂；所述药盒B包括化合物Ⅰ或其药学上可接受的盐；
    较佳地，所述组合药盒满足以下条件中的一种或多种：
    (1)所述药盒A与药盒B的施用时间不分先后或者所述药盒中先施用所述药盒A；
    (2)所述药盒A中，所述检测PIK3CA基因突变的试剂用于检测癌症患者是否携带PIK3CA基因突变；例如所述癌症患者为患有乳腺癌、卵巢癌、子宫内膜癌、宫颈癌和膀胱癌中的一种或多种的患者；
    (3)所述药盒B中，所述化合物Ⅰ或其药学上可接受的盐的含量为治疗有效量；
    (4)所述药盒B还包括药学上可接受的辅料；
    (5)所述药盒B中，所述化合物Ⅰ或其药学上可接受的盐的剂量和施用频率如权利要求9所述；
    (6)所述组合药盒用于治疗和/或预防PIK3CA突变的癌症，所述PIK3CA突变的癌症如权利要求1-8至少一项所述。
  11. 一种用于治疗和/或预防PIK3CA突变的癌症的化合物Ⅰ或其药学上可接受的盐，其特征在于，所述化合物Ⅰ的结构如下所示：
    Figure PCTCN2022072322-appb-100002
    其中，所述PIK3CA突变的癌症如权利要求1-8至少一项所述。
  12. 一种治疗和/或预防PIK3CA突变的癌症的方法，其特征在于，其通过向患者施用治疗有效量的化合物Ⅰ或其药学上可接受的盐，
    Figure PCTCN2022072322-appb-100003
    其中，所述PIK3CA突变的癌症如权利要求1-8至少一项所述。
  13. 如权利要求12所述的治疗和/或预防PIK3CA突变的癌症的方法，其特征在于，所述方法还包括检测患者是否携带PIK3CA基因突变的步骤。
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015192761A1 (zh) * 2014-06-17 2015-12-23 辰欣药业股份有限公司 作为mTOR/PI3K抑制剂的吡啶并[1,2-a]嘧啶酮类似物
CN105461712A (zh) * 2014-06-17 2016-04-06 南京明德新药研发股份有限公司 作为mTOR/PI3K抑制剂的吡啶并[1,2-a]嘧啶酮类似物
WO2017101829A1 (zh) * 2015-12-16 2017-06-22 辰欣药业股份有限公司 吡啶并[1,2-a]嘧啶酮类似物的晶型及其制备方法和中间体

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015192761A1 (zh) * 2014-06-17 2015-12-23 辰欣药业股份有限公司 作为mTOR/PI3K抑制剂的吡啶并[1,2-a]嘧啶酮类似物
CN105461712A (zh) * 2014-06-17 2016-04-06 南京明德新药研发股份有限公司 作为mTOR/PI3K抑制剂的吡啶并[1,2-a]嘧啶酮类似物
WO2017101829A1 (zh) * 2015-12-16 2017-06-22 辰欣药业股份有限公司 吡啶并[1,2-a]嘧啶酮类似物的晶型及其制备方法和中间体

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
"Handbook of Pharmaceutical Salts: Properties, Selection, and Use", 2002, WILEY-VCH
BERGE ET AL.: "Pharmaceutical Salts", JOURNAL OF PHARMACEUTICAL SCIENCE, vol. 66, 1977, pages 1 - 19, XP002675560, DOI: 10.1002/jps.2600660104
CAS , no. 1346242-81-6
CAS, no. 1217486-61-7

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