WO2023036282A1 - 嘧啶并杂环类化合物的晶型及其制备方法 - Google Patents

嘧啶并杂环类化合物的晶型及其制备方法 Download PDF

Info

Publication number
WO2023036282A1
WO2023036282A1 PCT/CN2022/118023 CN2022118023W WO2023036282A1 WO 2023036282 A1 WO2023036282 A1 WO 2023036282A1 CN 2022118023 W CN2022118023 W CN 2022118023W WO 2023036282 A1 WO2023036282 A1 WO 2023036282A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
crystal form
add
formula
test
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2022/118023
Other languages
English (en)
French (fr)
Inventor
徐洋洋
孙继奎
伍文韬
张杨
陈曙辉
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
D3 Bio Wuxi Co Ltd
Original Assignee
D3 Bio Wuxi Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to IL311340A priority Critical patent/IL311340A/en
Priority to AU2022342220A priority patent/AU2022342220A1/en
Priority to JP2024515669A priority patent/JP7688447B2/ja
Priority to MX2024003027A priority patent/MX2024003027A/es
Priority to US18/690,417 priority patent/US20250129102A1/en
Priority to CN202280060673.9A priority patent/CN117957220A/zh
Application filed by D3 Bio Wuxi Co Ltd filed Critical D3 Bio Wuxi Co Ltd
Priority to CA3231329A priority patent/CA3231329A1/en
Priority to KR1020247011544A priority patent/KR20240056582A/ko
Priority to EP22866743.2A priority patent/EP4385986A4/en
Publication of WO2023036282A1 publication Critical patent/WO2023036282A1/zh
Anticipated expiration legal-status Critical
Priority to JP2025082474A priority patent/JP2025124698A/ja
Ceased legal-status Critical Current

Links

Images

Classifications

    • 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
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C07D491/052—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being six-membered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • 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
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D419/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen, oxygen, and sulfur atoms as the only ring hetero atoms
    • C07D419/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen, oxygen, and sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D419/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen, oxygen, and sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/13—Crystalline forms, e.g. polymorphs

Definitions

  • the present invention relates to a crystal form of a class of pyrimidoheterocyclic compounds and a preparation method thereof, in particular to a preparation method and application of a compound represented by formula (II) and a crystal form thereof.
  • RAS oncogene mutations are the most common activating mutations in human cancers, occurring in 30% of human tumors.
  • the RAS gene family includes three subtypes (KRAS, HRAS, and NRAS), and 85% of RAS-driven cancers are caused by mutations in KRAS subtypes.
  • KRAS mutations are commonly found in solid tumors, such as lung adenocarcinoma, pancreatic ductal carcinoma, and colorectal cancer. In KRAS-mutant tumors, 80% of oncogenic mutations occurred at codon 12, and the most common mutations included: p.G12D (41%), p.G12V (28%), and p.G12C (14%).
  • KRAS plays a pivotal role in the signal regulation of cell growth.
  • the upstream cell surface receptors such as EGFR (ErbB1), HER2 (ErbB2), ErbB3, and ErbB4, after receiving external signals, pass the RAS protein to transmit the signal passed downstream.
  • EGFR EGFR
  • HER2 ErbB2
  • ErbB3 ErbB4
  • the KRAS protein is not activated, it is tightly bound to GDP (guanine nucleotide diphosphate).
  • guanine nucleotide exchange factors such as SOS1
  • GTP guanine nucleotide triphosphate
  • KRAS gene After being activated by guanine nucleotide exchange factors such as SOS1, it binds to GTP (guanine nucleotide triphosphate) and becomes a kinase active state.
  • GTP guanine nucleotide triphosphate
  • the KRAS gene After being activated by guanine nucleotide exchange factors such as SOS1, it binds to GTP (guanine nucleotide triphosphate) and becomes a kinase active state.
  • the KRAS gene After the KRAS gene is mutated, it can independently transmit growth and proliferation signals to the downstream pathway independently of the upstream growth factor receptor signal, resulting in uncontrolled cell growth and tumor progression. Important indicators.
  • KRAS was the first oncogene discovered, it was long considered an undruggable target.
  • Amgen and Mirati Therapeutics successively announced the clinical research results of the KRAS small molecule inhibitors AMG510 and MRTX849, which for the first time confirmed the clinical effectiveness of KRAS inhibitors in the clinical treatment of tumors.
  • Both AMG 510 and MRTX849 are irreversible sub-inhibitors, which inhibit KRAS activity by forming an irreversible covalent bond with the cysteine residue of the KRAS G12C mutein.
  • KRAS plays a pivotal role in the signal regulation of cell growth.
  • the upstream cell surface receptors such as EGFR (ErbB1), HER2 (ErbB2), ErbB3, and ErbB4, after receiving external signals, pass the RAS protein to transmit the signal passed downstream.
  • EGFR EGFR
  • HER2 ErbB2
  • ErbB3 ErbB4
  • GTP GTP
  • KRAS is an important member of RAS protein. After KRAS gene mutation, it can independently transmit growth and proliferation signals to downstream pathways independently of upstream growth factor receptor signals, resulting in uncontrolled cell growth and tumor progression. At the same time, KRAS Gene mutation is also an important indicator of tumor prognosis.
  • KRAS (G12C) mutation the replacement of glycine at position 12 by cysteine provides a good direction for the development of covalent inhibitors.
  • the present invention provides a compound of formula (II),
  • n is selected from 0-3.
  • the n is selected from 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8 , 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, and 3.0.
  • said n is selected from 0.5, 1, 1.5, 2, 2.5 and 3.
  • said n is selected from 2.
  • the present invention also provides a compound of formula (II),
  • n is selected from 0-2.
  • the n is selected from 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8 , 1.9 and 2.
  • said n is selected from 0.5, 1, 1.5 and 2.
  • said n is selected from 2.
  • the present invention also provides crystal form A of the compound of formula (II), which is characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2 ⁇ angles: 8.514 ⁇ 0.200°, 14.689 ⁇ 0.200°, 18.122 ⁇ 0.200°.
  • the X-ray powder diffraction pattern of the above crystal form A has characteristic diffraction peaks at the following 2 ⁇ angles: 6.218 ⁇ 0.200°, 8.514 ⁇ 0.200°, 12.299 ⁇ 0.200°, 14.689 ⁇ 0.200°, 16.903 ⁇ 0.200°, 18.122 ⁇ 0.200°, 18.927 ⁇ 0.200°, 25.580 ⁇ 0.200°.
  • the X-ray powder diffraction pattern of the above crystal form A has characteristic diffraction peaks at the following 2 ⁇ angles: 6.218 ⁇ 0.200°, 8.514 ⁇ 0.200°, 11.663 ⁇ 0.200°, 12.299 ⁇ 0.200°, 14.689 ⁇ 0.200°, 16.903 ⁇ 0.200°, 18.122 ⁇ 0.200°, 18.927 ⁇ 0.200°, 19.364 ⁇ 0.200°, 20.386 ⁇ 0.200°, 21.914 ⁇ 0.200°, 25.580 ⁇ 0.200°.
  • the X-ray powder diffraction pattern of the above crystal form A has characteristic diffraction peaks at the following 2 ⁇ angles: 6.218 ⁇ 0.200°, 8.514 ⁇ 0.200°, 11.663 ⁇ 0.200°, 12.299 ⁇ 0.200°, 14.689 ⁇ 0.200°, 16.903 ⁇ 0.200°, 18.122 ⁇ 0.200°, 18.927 ⁇ 0.200°, 19.364 ⁇ 0.200°, 20.386 ⁇ 0.200°, 21.914 ⁇ 0.200°, 22.640 ⁇ 0.200°, 25.580 ⁇ 0.7, 4 ⁇ 0.200°, 25.9088 0.200°, 27.715 ⁇ 0.200°.
  • the X-ray powder diffraction pattern of the above crystal form A has characteristic diffraction peaks at the following 2 ⁇ angles: 6.218°, 8.514°, 11.663°, 12.299°, 14.689°, 16.903°, 18.122°, 18.554 °, 18.927°, 19.364°, 20.386°, 21.914°, 22.640°, 23.867°, 24.553°, 24.806°, 25.580°, 25.988°, 27.147°, 27.715°, 29.135°, 31.799°.
  • the X-ray powder diffraction pattern of the above crystal form A has characteristic diffraction peaks at the following 2 ⁇ angles: 8.514 ⁇ 0.200°, 14.689 ⁇ 0.200°, and may also be at 6.218 ⁇ 0.200°, and/or 11.663 ⁇ 0.200°, and/or 12.299 ⁇ 0.200°, and/or 16.903 ⁇ 0.200°, and/or 18.122 ⁇ 0.200°, and/or 18.554 ⁇ 0.200°, and/or 18.927 ⁇ 0.200°, and/or 19.364 ⁇ 0.200 °, and/or 20.386 ⁇ 0.200°, and/or 21.914 ⁇ 0.200°, and/or 22.64 ⁇ 0.200°, and/or 23.867 ⁇ 0.200°, and/or 24.553 ⁇ 0.200°, and/or 24.806 ⁇ 0.200°, and/or 25.58 ⁇ 0.200°, and/or 25.988 ⁇ 0.200°, and/or 27.147 ⁇ 0.200°, and/or 27.715 ⁇
  • the XRPD spectrum of the above-mentioned crystal form A of the A crystal form is shown in FIG. 1 .
  • the differential scanning calorimetry curve of the above crystal form A has an endothermic peak at 115.37°C ⁇ 3°C.
  • the DSC spectrum of the above crystal form A is shown in FIG. 2 .
  • the weight loss of the thermogravimetric analysis curve of the above crystal form A reaches 5.379% at 150.0°C ⁇ 3°C.
  • the TGA spectrum of the above crystal form A is shown in FIG. 3 .
  • the present invention also provides a preparation method of formula (II) compound A crystal form, comprising:
  • the present invention also provides the application of the above-mentioned compound and crystal form A in the preparation of medicaments for treating solid tumors.
  • the aforementioned solid tumors are lung cancer and rectal cancer.
  • the compound of the present invention has good cell proliferation inhibitory activity on MIA-PA-CA-2 cell line and NCI-H358 cells with KRASG12C mutation, good stability of liver microsomes, liver cells, plasma and whole blood, and good PK properties, and has a significant antitumor effect. Crystal form A is stable, less affected by light and heat, and has good PK properties.
  • the intermediate compound of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by its combination with other chemical synthesis methods, and the methods described by those skilled in the art. Known equivalents, preferred embodiments include, but are not limited to, the examples of the present invention.
  • the structure of the compounds of the present invention can be confirmed by conventional methods known to those skilled in the art. If the present invention involves the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, in single crystal X-ray diffraction (SXRD), the cultured single crystal is collected with a Bruker D8 venture diffractometer to collect diffraction intensity data, the light source is CuK ⁇ radiation, and the scanning method is: After scanning and collecting relevant data, the absolute configuration can be confirmed by further analyzing the crystal structure by direct method (Shelxs97).
  • SXRD single crystal X-ray diffraction
  • Test method About 10-20 mg of sample is used for XRPD detection.
  • Phototube voltage 30kV
  • phototube current 10mA
  • Anti-scatter slit 0mm
  • Step size 0.2 seconds
  • the present invention 's differential thermal analysis (Differential Scanning Calorimeter, DSC) method
  • Test method Take a sample ( ⁇ 4.02mg) and place it in a DSC aluminum pot for testing. Under the condition of 50mL/min N 2 , heat the sample from 30°C (room temperature) to 400°C at a heating rate of 10°C/min.
  • Thermogravimetric Analysis (Thermal Gravimetric Analyzer, TGA) method of the present invention
  • Test method Take a sample (2 ⁇ 5mg) and place it in a TGA platinum pot for testing. Under the condition of 25mL/min N 2 , at a heating rate of 10°C/min, heat the sample from room temperature to 350°C or lose 20% of its weight.
  • Fig. 1 is the XRPD spectrogram of the Cu-K ⁇ radiation of formula (II) compound A crystal form;
  • Fig. 2 is the DSC spectrogram of formula (II) compound A crystal form
  • Fig. 3 is the TGA spectrogram of formula (II) compound A crystal form
  • Figure 4 is the change of tumor volume over time with different doses of the compound of formula (I);
  • Fig. 5 is the animal body weight change of formula (I) compound different dosage prolongs with time
  • Fig. 6 is a single crystal X-ray diffraction three-dimensional structure ellipsoid diagram of the compound of formula (III).
  • Embodiment 1 the preparation of formula (I) compound
  • reaction solution Slowly add the reaction solution to 3.5L of 1M hydrochloric acid solution, add 2L of water for liquid extraction, continue to extract the aqueous phase with 2L of ethyl acetate, combine the organic phase and continue, continue to add 1L of ethyl acetate, and use 5L of saturated Wash with salt water, dry and filter over anhydrous sodium sulfate, concentrate, divide the crude product into 4 batches, add 200 mL of methyl tert-butyl ether, stir for 10 min, slowly add 200 mL of petroleum ether, and stir for 0.5-1 hour to precipitate a solid.
  • Compound 1-11 was resolved by SFC, chromatographic column: DAICEL CHIRALPAK AD (250mm*50mm, 10 ⁇ m); mobile phase: [0.1% NH 3 . H 2 O EtOH]; EtOH %: 45%-45%, 6.3 min.
  • Compound 1-12B peak elution time: 1.665min
  • compound 1-12A peak elution time: 2.446min
  • the reaction solution was poured into 500 mL of ice water, stirred until clear, and separated.
  • the organic phase was extracted with water (300 mL x 3).
  • the two batches of aqueous phase were combined, then back extracted with dichloromethane (500 mL x 4), and the organic phase was discarded.
  • Cool the water phase to 10°C add 500mL 2-methyltetrahydrofuran (exothermic) to the water phase, then adjust the pH to 9 with sodium carbonate, extract with 2-methyltetrahydrofuran (400mL x 2), combine the organic phases, Wash with water (300mL x 4), wash with saturated brine (300mL), dry over anhydrous sodium sulfate, filter and concentrate to obtain compound 1-20 as a gray solid.
  • LCMS m/z 608.27 [M+H] + .
  • Embodiment 2 Single crystal X-ray diffraction detection analysis of the compound of formula (III)
  • Embodiment 3 Preparation of formula (II) compound A crystal form
  • Method 1 Add the compound of formula (I) (0.2g) into ethanol (2.4mL), heat to 50°C, dissolve and slowly add water (0.48mL) to the reaction solution, and stir at 50°C for 72 hours , filtered, and the solid was collected to obtain the compound A crystal form of formula (II).
  • Method 2 Add the compound of formula (I) (5.0g) into ethanol (4V) and stir until clarification, slowly add water (1V) into the system, add seed crystals (0.5g) at 20-30°C, and then Stir at high temperature for 15 hours, then slowly add water (3V) dropwise, and continue stirring for 1 to 3 hours. The filter cake was collected by filtration to obtain the compound A crystal form of formula (II).
  • Embodiment 4 the solid stability test of formula (II) compound A crystal form
  • each of the pharmaceutical low-density polyethylene bags is tied tightly with cable ties, and then put into a single-layer aluminum foil bag and heat-sealed. Finally, the samples were stored in plastic buckets in a stable constant temperature and humidity box.
  • the packaging used in the stability test simulates the material storage package. The material is divided into 1.5g/package stability samples under red light. Each sample is packaged in the same way and the stability sample label is attached. Under different conditions The placed samples were sampled and tested (XRPD) at the 3rd and 6th months, and the test results were compared with the initial test results on day 0. The test results are shown in Table 3 below.
  • Illumination control *1 The light control sample should be placed at the same time, and the light control sample should be sealed with a screw cap, and then completely wrapped with tinfoil.
  • the main reagents used in this study include CellTiter-Glo (Promega, catalog number: G7573).
  • the main instrument used in this study is the PerkinElmer EnVision multifunctional microplate reader.
  • F Con is the fluorescence reading value of the Con group after 72 hours of culture.
  • F Cpd is the fluorescence reading of each compound well after 72 hours of incubation.
  • the compound of the present invention is to the test result of the MIA-PA-CA-2 cell proliferation inhibition of KRASG12C mutation
  • Test article IC 50 (nM) Compound of formula (I) 0.44
  • the main reagents used in this study included RPMI-1640 medium, penicillin/streptomycin antibiotics were purchased from Vicente, and fetal bovine serum was purchased from Biosera.
  • CellTiter-Glo (Cell Viability Chemiluminescence Detection Reagent) reagent was purchased from Promega.
  • the NCI-H358 cell line was purchased from the Cell Bank of the Chinese Academy of Sciences.
  • the main instrument used in this study is the Nivo multi-label analyzer (PerkinElmer).
  • Example-Min Use the equation (Sample-Min)/(Max-Min)*100% to convert the original data into an inhibition rate, and the value of IC50 can be obtained by curve fitting with four parameters ("log(inhibitor) vs. response--Variable slope" mode).
  • Table 5 provides the inhibitory activity of the compounds of the present invention on the proliferation of NCI-H358 cells.
  • Test article NCI-H358IC 50 (nM) Compound of formula (I) 0.6
  • the incubation plate into the shaker in the incubator, and start the timer to start the reaction.
  • Two replicate samples were prepared for each time point for each compound.
  • the incubation conditions were 37° C., saturated humidity, and 5% CO 2 .
  • the final concentration of the test substance is 1 ⁇ M
  • the final concentration of the control substance is 3 ⁇ M
  • the final concentration of the liver cells is 0.5 ⁇ 106 cells/mL
  • the final concentration of the total organic solvent is 0.96%
  • the final concentration of DMSO is 0.1 %.
  • the incubation plate was taken out, and 25 ⁇ L of the mixture of compound and control compound and cells was taken out and added to the sample plate containing 125 ⁇ L of stop solution (acetonitrile solution containing 200 ng/mL tolbutamide and Rabenol).
  • stop solution acetonitrile solution containing 200 ng/mL tolbutamide and Rabenol.
  • For Blank sample plates directly add 25 ⁇ L of incubation medium without hepatocytes. After all the sample plates were sealed, they were shaken at 600 rpm for 10 minutes on a plate shaker, and then centrifuged at 3220 ⁇ g for 20 minutes. The supernatants of the test product and the reference product were diluted with ultrapure water at a ratio of 1:3. All samples were mixed and analyzed by LC/MS/MS.
  • T60 incubation plate and NCF60 incubation plate were prepared two 96-well incubation plates, named T60 incubation plate and NCF60 incubation plate respectively. Add 445 ⁇ L microsomal working solution (hepatic microsomal protein concentration is 0.56 mg/mL) to the T60 incubation plate and NCF60 incubation plate, and then place the above incubation plate in a 37°C water bath for pre-incubation for about 10 minutes.
  • microsomal working solution hepatic microsomal protein concentration is 0.56 mg/mL
  • the final concentration of the compound, testosterone, diclofenac and propafenone in the reaction is 1 ⁇ M
  • the concentration of liver microsomes is 0.5 mg/mL
  • the final concentration of DMSO and acetonitrile in the reaction system 0.01% (v/v) and 0.99% (v/v), respectively.
  • stop solution containing 200 ng/mL tolbutamide and 200 ng/mL labetalol
  • acetonitrile solution remove 60 ⁇ L sample from the T60 incubation plate to terminate the reaction. All sample plates were shaken and centrifuged at 3220 ⁇ g for 20 minutes, and then 80 ⁇ L of supernatant was diluted into 240 ⁇ L of pure water for liquid chromatography-tandem mass spectrometry analysis, and all samples were injected for analysis by liquid chromatography-tandem mass spectrometry.
  • the final incubation concentration of compounds and control compounds bisacodyl, enalapril maleate, procaine and propensine was 2 ⁇ M, and the final organic phase content was 2.0%.
  • the corresponding incubation plate was taken out, and 400 ⁇ L of acetonitrile solution containing 200 ng/mL tolbutamide and Rabenol was added to each corresponding sample well to precipitate proteins.
  • the anticoagulant EDTA-K2 was used to collect fresh whole blood of CD-1 mice, SD rats, beagle dogs, and cynomolgus monkeys. Before the experiment started, the whole blood was mixed with PBS 1:1 (v:v), and placed in a 37°C water bath to preheat for 10-20 minutes. Prepare 96-well incubation plates and name them T0, T30, T60, and T240 respectively.
  • incubation plates including T0, T30, T60, and T240 incubation plates, mix 2 ⁇ L of the compound or control compound working solution with 98 ⁇ L of mouse, rat, dog, monkey, and human blank whole blood, and each sample is prepared Two parallel holes. All samples were incubated in a 37°C water bath. Compounds were incubated at a final concentration of 5 ⁇ M and control compounds were incubated at a final concentration of 2 ⁇ M.
  • Plasma samples with a compound concentration of 2 ⁇ M were respectively prepared from the plasma of the above five species, placed in a 96-well equilibrium dialysis device, and dialyzed with phosphate buffered saline solution for 4 hours at 37 ⁇ 1°C.
  • warfarin was used as a control compound.
  • concentrations of analytes in plasma and dialysis buffer were determined by LC-MS/MS.
  • test compound was mixed with 5% dimethyl sulfoxide/95% (10% hydroxypropyl- ⁇ -cyclodextrin) solution, vortexed and sonicated to prepare a 1 mg/mL clear solution, which was filtered through a microporous membrane for use .
  • Male SD rats aged 7 to 10 weeks were selected, and the candidate compound solution was administered intravenously or orally.
  • Whole blood was collected for a certain period of time to prepare plasma, and the drug concentration was analyzed by LC-MS/MS method, and the pharmacokinetic parameters were calculated by Phoenix WinNonlin software (Pharsight, USA).
  • Table 11 The experimental results are shown in Table 11:
  • test compound was mixed with 5% dimethyl sulfoxide/95% (10% hydroxypropyl- ⁇ -cyclodextrin) solution, vortexed and sonicated to prepare a 1 mg/mL clear solution, which was filtered through a microporous membrane for use .
  • Male CD mice aged 7 to 10 weeks were selected, and the candidate compound solution was administered intravenously or orally.
  • Whole blood was collected for a certain period of time to prepare plasma, and the drug concentration was analyzed by LC-MS/MS method, and the pharmacokinetic parameters were calculated by Phoenix WinNonlin software (Pharsight, USA).
  • Table 13 The experimental results are shown in Table 13:
  • Cell culture In vitro monolayer culture of human pancreatic cancer Mia PaCa-2 cells (ATCC-CRL-1420), the culture conditions are DMEM medium plus 10% fetal bovine serum, 2.5% horse serum, 37 °C 5% carbon dioxide incubator culture . Routine digestion with trypsin-EDTA was performed twice a week for passaging. When the cell saturation is 80%-90% and the number reaches the requirement, the cells are collected, counted, resuspended in an appropriate amount of PBS, and added Matrigel at 1:1 to obtain a cell suspension with a cell density of 25x 106 cells/mL.
  • PO oral administration
  • QD once a day
  • Tumor diameters were measured twice a week with vernier calipers.
  • TGI percent or relative tumor proliferation rate T/C (%).
  • Relative tumor proliferation rate T/C (%) TRTV/CRTV ⁇ 100% (TRTV: RTV of the treatment group; CRTV: RTV of the negative control group).
  • TGI (%) reflects tumor growth inhibition rate.
  • TGI (%) [(1-(Average tumor volume at the end of administration of a certain treatment group-Average tumor volume at the beginning of administration of this treatment group))/(Average tumor volume at the end of treatment of the solvent control group-Start of treatment of the solvent control group Time-average tumor volume)] ⁇ 100%.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Epidemiology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Plural Heterocyclic Compounds (AREA)
  • Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)

Abstract

一类嘧啶并杂环类化合物的晶型及其制备方法，具体公开了式(II)化合物及其晶型的制备方法和应用。

Description

嘧啶并杂环类化合物的晶型及其制备方法
本发明主张如下优先权：
CN202111062619.1，申请日：2021年09月10日；
CN202211034826.0，申请日：2022年08月26日。
技术领域
本发明涉及一类嘧啶并杂环类化合物的晶型及其制备方法，具体涉及式(II)所示化合物及其晶型的制备方法和应用。
背景技术
RAS癌基因突变是人类癌症中最常见的激活突变，发生在30％的人类肿瘤中。RAS基因家族包括三个亚型(KRAS、HRAS和NRAS)，其中85％的RAS驱动的癌症是由KRAS亚型突变引起的。KRAS突变常见于实体肿瘤中，如：肺腺癌、胰腺导管癌和结直肠癌等。在KRAS突变肿瘤中，80％的致癌突变发生在密码子12上，最常见的突变包括：p.G12D(41％)、p.G12V(28％)和p.G12C(14％)。
KRAS基因的全名是Kirsten rat sarcoma viraloncogene homolog(Kristen大鼠肉瘤病毒癌基因同源物)。KRAS在细胞生长的信号调控中起着一个枢纽的作用，上游的EGFR(ErbB1)、HER2(ErbB2)、ErbB3和ErbB4等细胞表面受体，在接受了外界信号之后，会通过RAS蛋白，把信号传递到下游。KRAS蛋白没有被激活的时候，与GDP(鸟嘌呤核苷酸二磷酸)紧密结合。在被SOS1等鸟嘌呤核苷酸交换因子激活后，与GTP(鸟嘌呤核苷酸三磷酸)结合，变成激酶活性的状态。KRAS基因突变后，可以不依赖于上游生长因子受体信号，独立向下游通路传输生长和增殖的信号，造成不受控制的细胞生长和肿瘤进展，同时KRAS基因是否有突变，也是肿瘤预后的一个重要指标。
尽管KRAS是第一个被发现的癌基因，但长期以来被认为是不可成药靶点。直到2019年，Amgen和Mirati Therapeutics先后公布了的KRAS小分子抑制剂AMG510和MRTX849的临床研究结果，首次在临床上证实了KRAS抑制剂在临床上治疗肿瘤的有效性。AMG 510和MRTX849都属于不可逆小分抑制剂，通过与KRAS G12C突变蛋白的半胱氨酸残基形成不可逆共价键，从而抑制KRAS活性。
KRAS在细胞生长的信号调控中起着一个枢纽的作用，上游的EGFR(ErbB1)、HER2(ErbB2)、ErbB3和ErbB4等细胞表面受体，在接受了外界信号之后，会通过RAS蛋白，把信号传递到下游。KRAS蛋白在没有被激活的时候，是与GDP(鸟嘌呤核苷酸二磷酸)紧密结合的，在被SOS1等鸟嘌呤核苷酸交换因子激活后，转到与GTP(鸟嘌呤核苷酸三磷酸)结合，变成激酶活性的状态。KRAS是RAS蛋白中的重要一员，KRAS基因突变后，可以不依赖于上游生长因子受体信号，独立向下游通路传输生长和增殖的信号，造成不受控制的细胞生长和肿瘤进展，同时KRAS基因是否有突变，也是肿瘤预后的一个重要指标。
统计结果显示，肺腺癌有12-36％病人KRAS突变驱动的；27-56％结肠癌病人由KRAS驱动，此外90％的胰腺癌、21％的子宫内膜癌和12-36％的肺腺癌等都是由KRAS驱动的，病人群体巨大；在KRAS的基因突变中，97％是发生在第12位或者第13位的氨基酸残基发生了突变，其中G12D，G12V和G13D这三种突变，但是这三种突变的成药较差，KRAS(G12C)突变：12位的甘氨酸被半胱氨酸替换后为共价抑制剂的开发提供了一个很好的方向。
发明内容
本发明提供了式(II)化合物，
Figure PCTCN2022118023-appb-000001
其中，n选自0～3。
在本发明的一些方案中，所述n选自0、0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1.0、1.1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9、2、2.1、2.2、2.3、2.4、2.5、2.6、2.7、2.8、2.9和3.0。
在本发明的一些方案中，所述n选自0.5、1、1.5、2、2.5和3。
在本发明的一些方案中，所述n选自2。
本发明还提供了式(II)化合物，
Figure PCTCN2022118023-appb-000002
其中，n选自0～2。
在本发明的一些方案中，所述n选自0、0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1.0、1.1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9和2。
在本发明的一些方案中，所述n选自0.5、1、1.5和2。
在本发明的一些方案中，所述n选自2。
本发明还提供了式(II)化合物的A晶型，其特征在于其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.514±0.200°、14.689±0.200°、18.122±0.200°。
在本发明的一些方案中，上述A晶型的X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：6.218±0.200°、8.514±0.200°、12.299±0.200°、14.689±0.200°、16.903±0.200°、18.122±0.200°、18.927±0.200°、25.580±0.200°。
在本发明的一些方案中，上述A晶型的X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：6.218±0.200°、8.514±0.200°、11.663±0.200°、12.299±0.200°、14.689±0.200°、16.903±0.200°、18.122±0.200°、18.927±0.200°、19.364±0.200°、20.386±0.200°、21.914±0.200°、25.580±0.200°。
在本发明的一些方案中，上述A晶型的X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：6.218±0.200°、8.514±0.200°、11.663±0.200°、12.299±0.200°、14.689±0.200°、16.903±0.200°、18.122±0.200°、18.927±0.200°、19.364±0.200°、20.386±0.200°、21.914±0.200°、22.640±0.200°、25.580±0.200°、25.988±0.200°、27.147±0.200°、27.715±0.200°。
在本发明的一些方案中，上述A晶型的X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：6.218°、8.514°、11.663°、12.299°、14.689°、16.903°、18.122°、18.554°、18.927°、19.364°、20.386°、21.914°、22.640°、23.867°、24.553°、24.806°、25.580°、25.988°、27.147°、27.715°、29.135°、31.799°。
在本发明的一些方案中，上述A晶型的X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.514±0.200°、14.689±0.200°，还可以在6.218±0.200°，和/或11.663±0.200°，和/或12.299±0.200°，和/或16.903±0.200°，和/或18.122±0.200°，和/或18.554±0.200°，和/或18.927±0.200°，和/或19.364±0.200°，和/或20.386±0.200°，和/或21.914±0.200°，和/或22.64±0.200°，和/或23.867±0.200°，和/或24.553±0.200°，和/或24.806±0.200°，和/或25.58±0.200°，和/或25.988±0.200°，和/或27.147±0.200°，和/或27.715±0.200°，和/或29.135±0.200°，和/或31.799±0.200°处有特征衍射峰。
在本发明的一些方案中，上述A晶型的A晶型，其XRPD图谱如图1所示。
本发明的一些方案中，上述A晶型的XRPD图谱解析数据如表1所示：
表1
Figure PCTCN2022118023-appb-000003
Figure PCTCN2022118023-appb-000004
在本发明的一些方案中，上述A晶型的差示扫描量热曲线在115.37℃±3℃处具有吸热峰的峰值。
在本发明的一些方案中，上述A晶型的DSC图谱如图2所示。
在本发明的一些方案中，上述A晶型的热重分析曲线在150.0℃±3℃时失重达5.379％。
在本发明的一些方案中，上述A晶型的TGA图谱如图3所示。
本发明还提供式(II)化合物A晶型的制备方法，包括：
Figure PCTCN2022118023-appb-000005
(a)将式(I)化合物加入乙醇中搅拌至澄清；
(b)在搅拌下将水缓慢加入到体系中，在20～30℃加入晶种；
(c)20～30℃下搅拌15小时；
(d)20～30℃缓慢将水滴加到反应体系中，继续搅拌1～3小时。
(e)过滤，收集固体。
本发明还提供了上述化合物和A晶型在制备治疗实体瘤药物中的应用。
在本发明的一些方案中，上述实体瘤为肺癌和直肠癌。
技术效果
本发明化合物对KRASG12C突变的MIA-PA-CA-2细胞系、NCI-H358细胞具有良好的细胞增殖抑制活性，有良好的肝微粒体、肝细胞、血浆、全血稳定性，有良好的PK性质，并且有显著的抑瘤作用。A晶型稳定、受光热影响小，有良好的PK性质。
定义和说明
除非另有说明，本文所用的下列术语和短语旨在含有下列含义。一个特定的短语或术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。当本文出现商品名时，旨在指代其对应的商品或其活性成分。
本发明的中间体化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。
本发明具体实施方式的化学反应是在合适的溶剂中完成的，所述的溶剂须适合于本发明的化学变化及其所需的试剂和物料。为了获得本发明的化合物，有时需要本领域技术人员在已有实施方式的基础上对合 成步骤或者反应流程进行修改或选择。
下面会通过实施例具体描述本发明，这些实施例并不意味着对本发明的任何限制。
本发明所使用的所有溶剂是市售的，无需进一步纯化即可使用。
本发明的化合物可以通过本领域技术人员所熟知的常规方法来确认结构，如果本发明涉及化合物的绝对构型，则该绝对构型可以通过本领域常规技术手段予以确证。例如单晶X射线衍射法(SXRD)，把培养出的单晶用Bruker D8 venture衍射仪收集衍射强度数据，光源为CuKα辐射，扫描方式：
Figure PCTCN2022118023-appb-000006
扫描，收集相关数据后，进一步采用直接法(Shelxs97)解析晶体结构，便可以确证绝对构型。
化合物依据本领域常规命名原则或者使用
Figure PCTCN2022118023-appb-000007
软件命名，市售化合物采用供应商目录名称。
本发明粉末X-射线衍射(X-ray powder diffractometer,XRPD)方法
仪器型号：布鲁克Bruker D2PhaserX-射线衍射仪
测试方法：大约10～20mg样品用于XRPD检测。
详细的XRPD参数如下：
光管：Cu,kα,
Figure PCTCN2022118023-appb-000008
光管电压：30kV，光管电流：10mA
发散狭缝：0.60mm
探测器狭缝：5.827mm
防散射狭缝：0mm
扫描范围：3-40deg
步径：0.02deg
步长：0.2秒
本发明差热分析(Differential Scanning Calorimeter,DSC)方法
仪器型号：NETZSCH DSC 214DSC21400A-0958-L
测试方法：取样品(～4.02mg)置于DSC铝锅内进行测试，在50mL/min N 2条件下，以10℃/min的升温速率，加热样品从30℃(室温)到400℃。
本发明热重分析(Thermal Gravimetric Analyzer,TGA)方法
仪器型号：TA Discovery TGA 5500热重分析仪
测试方法：取样品(2～5mg)置于TGA铂金锅内进行测试，在25mL/min N 2条件下，以10℃/min的升温速率，加热样品从室温到350℃或失重20％。
本发明X-射线单晶衍射方法
仪器型号：Bruker D8 VENTURE CMOS Photon II diffractometer with helios mx multilayer monochrmator.
测试方法：取0.0133g式(II)化合物A晶型在室温条件下溶解于2mL乙腈中。将样品溶液置于4mL半密封样品瓶中，在室温下缓慢挥发。十天后得到无色块状晶体。衍射实验温度T＝173(2)K。
仪器参数：
Bruker D8 VENTURE CMOS Photon II diffractometer with helios mx multilayer monochrmator.
Cryogenic system:Oxford Cryostream 800
Cu:
Figure PCTCN2022118023-appb-000009
2.5kW,
Distance from the crystal to detector:d＝45mm
Tube Voltage:50kV
Tube Current:50mA
附图说明
图1为式(II)化合物A晶型的Cu-Kα辐射的XRPD谱图；
图2为式(II)化合物A晶型的DSC谱图；
图3为式(II)化合物A晶型的TGA谱图；
图4为式(I)化合物不同给药量随着时间延长的肿瘤体积变化；
图5为式(I)化合物不同给药量随着时间延长的动物体重变化；
图6为式(III)化合物的单晶X射线衍射立体结构椭球图。
具体实施方式
为了更好的理解本发明的内容，下面结合具体实施例来做进一步的说明，但具体的实施方式并不是对本发明的内容所做的限制。
实施例1：式(I)化合物的制备
Figure PCTCN2022118023-appb-000010
Figure PCTCN2022118023-appb-000011
步骤1：化合物1-2的合成
准备好5升三口瓶，将化合物1-1(250g,2.00mol,1eq)和无水碳酸钾(690.26g,4.99mol,2.5eq)，碘化钾(331.62g,2.00mol,1eq)加入到N-甲基吡咯烷酮(2.5L)中，然后滴加对甲氧基苯甲基氯(641.36g,4.10mol,557.71mL,2.05eq)，反应液成黄色浑浊，在氮气保护下置于120℃油浴中搅拌反应5小时。合并6批反应液(250×6)，倒入20升水中，再加入10升甲基叔丁基醚搅拌，分液后收集有机相，水相用甲基叔丁基醚萃取(5L x 1)，合并有机相，饱和食盐水洗涤(10L x 2)，无水硫酸钠干燥，减压浓缩得到化合物1-2品。向粗品中加入3升石油醚，打浆过夜。打浆体系成乳白色浑浊，通过布氏漏斗过滤。滤饼用石油醚(500mL x 3)淋洗，得到的滤饼，得到化合物1-2。 1H NMR(400MHz,CDCl 3)δ＝7.23-7.18(m,4H),6.91-6.87(m,1H),6.82-6.76(m,4H),6.65-6.59(m,2H),4.20(s,4H),3.79(s,6H),2.19(s,3H)。LCMS:MS m/z＝366.1[M+H] +。
步骤2：化合物1-3的合成
将2,2,6,6-四甲基哌啶(2.44kg,17.29mol,2.94L,4eq)加入到无水四氢呋喃(15L)中，降温至-5-0℃，氮气置换三次，氮气保护下滴加正丁基锂(2.5M,6.92L,4eq)，-5-0℃反应15min，降温至-60℃，滴加化合物1-2(1.58kg,4.32mol,1eq)的四氢呋喃(1.5L)溶液，滴加完毕后-65～-60℃反应0.5小时，快速加入N,N-二甲基甲酰胺(3.16kg,43.24mol,3.33L,10eq)，-60℃反应10min，向反应液加入20L饱和氯化铵，用5L的甲基叔丁基醚萃取分液，有机相继续用20L饱和氯化铵洗涤，然后水相用10L的甲基叔丁基醚继续萃取分液，合并有机相，有机相用饱和食盐水洗涤(12L x 2)，无水硫酸钠干燥过滤后浓缩，粗品用石油醚/甲基叔丁基醚(3/1,3L)打浆5小时，过滤，收集滤饼得到化合物1-3。 1H NMR(400MHz,CDCl 3)δ＝10.43-10.35(m,1H),7.21-7.18(m,5H),6.92-6.81(m,5H),4.25(s,4H),3.80(s,6H),2.23(s,3H)。LCMS:MS m/z＝394.2[M+H] +。
步骤3：化合物1-4的合成
将化合物1-3(1.17kg,2.83mol,95％纯度,1eq)加入到N,N-二甲基甲酰胺(5.7L)中，5℃下分批加入溴代丁二酰亚胺(603.35g,3.39mol,1.2eq)，5-15℃反应1小时，向反应液中，缓慢加入5.7L水，有固体析出，搅拌20min后，继续缓慢加入11.4L水，继续搅拌40min，过滤，将滤饼用水(2L x 2)的淋洗。粗品用石油醚：甲基叔丁基醚＝10:1的混合溶剂7.7L打浆12小时，过滤，滤饼用石油醚：甲基叔丁基醚 ＝10:1的混合溶剂500mL淋洗后进行真空浓缩后，用氮气吹12小时，得到化合物1-4。 1H NMR(400MHz,CDCl 3)δ＝10.39(s,1H),7.17(d,J＝8.8Hz,4H),6.89(d,J＝8.8Hz,1H),6.85-6.82(m,4H),4.22(s,4H),3.79(s,6H),2.28(s,3H)。LCMS:MS m/z＝472.1[M+H] +，474.1[M+H] +。
步骤4：化合物1-6的合成
将化合物1-4(130g,275.22mmol,1eq)，碘化亚酮(104.83g,550.44mmol,2eq)，化合物1-5(264.37g,1.38mol,175.08mL,5eq)溶于DMF(1.3L)中，氮气保护，在100℃下搅拌4小时。降温，体系过滤，倒入水(1.3L)中淬灭，甲基叔丁基醚(400mL x 2)萃取，饱和食盐水洗涤(300mL)洗涤，无水硫酸钠干燥，过滤，收集母液浓缩得到粗品。粗品打浆(石油醚/甲基叔丁基醚＝8/1,300mL)得到化合物1-6，收集母液，母液浓缩柱层析(石油醚/乙酸乙酯＝100/1～5/1)得到粗品，粗品打浆(石油醚/甲基叔丁基醚＝8/1,100mL)得到化合物1-6。两批次固体用石油醚打浆混合，过滤，收集固体，得到化合物1-6。 1H NMR(400MHz,CDCl 3)δ＝10.37(q,J＝4.0Hz,1H),7.18-7.11(m,4H),6.89-6.82(m,4H),6.73(d,J＝8.8Hz,1H),4.36(s,4H),3.81(s,6H),2.37-2.29(m,3H)。LCMS:MS m/z＝484.0[M+Na] +。
步骤5：化合物1-8的合成
将钠氢(70.21g,1.76mol,60％纯度,1.8eq)加入到无水四氢呋喃(4.5L)中，降温至-5℃，置换氮气三次，氮气保护下滴加化合物1-7(203.82g,1.76mol,188.72mL,1.8eq)，-5～0℃反应10min后，滴加正丁基锂(2.5M,702.14mL,1.8eq)，继续在氮气保护下-5-0℃反应10min，降温至-10℃，滴加化合物1-6(450g,975.19mmol,1eq)的四氢呋喃(450mL)溶液，-10℃下反应10min，将反应液缓慢加入到5L饱和氯化铵中，分液萃取，有机相用4L的饱和食盐水洗涤，无水硫酸钠干燥过滤后浓缩，向浓缩后的粗品中加入900mL x 4的石油醚醚，晃荡洗涤后，将石油醚上清液倾倒掉，将粗品进行真空浓缩得到化合物1-8。 1H NMR(400MHz,CDCl 3)δ＝7.18-7.15(m,4H),6.90-6.78(m,4H),6.61(d,J＝8.8Hz,1H),5.72-5.57(m,1H),4.31(m,4H),3.81(s,6H)，3.76(s,3H),3.56(s,2H),3.50-3.38(m,1H),2.98-2.93(m,1H),2.38-2.26(m,3H)。LCMS:MS m/z＝578.1[M+H] +。
步骤6：化合物1-9的合成
将化合物1-8(1.15kg,1.77mol,89％纯度,1eq)加入到二氯甲烷(5.7L)中，加入N,N-二甲基甲酰胺二甲基缩醛(337.86g,2.84mol,376.66mL,1.6eq)，25℃反应1小时，降温至0℃，0-5℃下滴加入三氟化硼乙醚(377.27g,2.66mol,328.06mL,1.5eq)，反应10min。LCMS表明原料消失，出现产物Ms信号。将反应液缓慢加入到10L的半饱和碳酸氢钠溶液中，分液萃取，有机相用5L的饱和食盐水洗涤，无水硫酸钠干燥过滤后浓缩将粗品平均分成六份，每份加入0.83L的甲基叔丁基醚，搅拌20min后析出固体，继续加入石油醚：甲基叔丁基醚＝1:1的混合溶剂0.5L后，打浆过夜16小时，六份一起过滤，将滤饼用石油醚：甲基叔丁基醚＝1:1的混合溶剂800mL x 2淋洗后，旋干，母液经柱层析石油醚：乙酸乙酯＝100:0-0:1)得到化合物1-9。 1H NMR(400MHz,CDCl 3)δ＝8.43(d,J＝0.8Hz,1H),7.21-7.10(m,4H),6.91-6.81(m,4H),6.70(d,J＝8.8Hz,1H),5.93(dd,J＝3.2,14.8Hz,1H),4.35(s,4H),3.8(s,3H)，3.81(s,6H),3.38-3.29(m,1H),2.68(dd,J＝3.6,16.8Hz,1H),2.39-2.24(m,3H)。LCMS:MS m/z＝588.2[M+H] +。
步骤7：化合物1-10的合成
将化合物1-9(775g,1.32mol,1eq)加入到四氢呋喃(4L)中，降温至-60℃，置换氮气三次，氮气保护下滴加三仲丁基硼氢化理(1M,1.45L,1.1eq)，-60℃下反应10min。将反应液缓慢加入到3.5L的1M 盐酸溶液中，加入2L的水分液萃取，水相继续用2L的乙酸乙酯萃取，合并有机相中并继，续加入1L的乙酸乙酯，用5L饱和食盐水洗涤，无水硫酸钠干燥过滤后浓缩，将粗品分为4批，分别加入200mL的甲基叔丁基醚，搅拌10min，缓慢加入200mL的石油醚，搅拌0.5-1小时后析出固体，继续分批次加入1.6L的石油醚，打浆搅拌12小时，过滤，将滤饼用石油醚：甲基叔丁基醚＝10:1的溶剂300mL x 3淋洗，收集固体得到化合物1-10(1.15kg)。 1H NMR(400MHz,CDCl 3)δ＝7.167-7.14(m,4H),6.87-6.83(m，4H),6.63(d,J＝8.8Hz,1H),5.05-5.00(m,1H),4.61-4.58(m,1H),4.42-4.24(m,5H),3.85-3.73(m,10H)，3.13-3.05(m,1H),2.47-2.38(m,1H),2.35-2.31(m,3H)。LCMS:MS m/z＝590.3.[M+H] +。
步骤8：化合物1-11的合成
将化合物1-10(310g,525.80mmol,1eq)加入到乙醇(1.55L)中，加入S-甲基异硫脲硫酸盐(439.11g,1.58mol,3eq)和碳酸钠(111.46g,1.05mol,2eq)，45～50℃(内温)氮气保护下反应16小时。将大部分乙醇浓缩除去，向粗品中加入500毫升水和400毫升乙酸乙酯，搅拌，用500mL 1M盐酸调节pH＝3～4后有灰白色固体析出，再加入600mL石油醚，搅拌，体系中有大量灰白色固体析出。将体系经布氏漏斗过滤，滤饼用乙酸乙酯淋洗(200mL x 2)，得到的滤饼即为产品。将滤饼用2L二氯甲烷溶解，分液，有机相用无水硫酸钠干燥，浓缩得到化合物1-11。 1H NMR(400MHz,CDCl 3)δ＝7.22-7.14(m,4H),6.91-6.82(m,4H),6.65(dd,J＝8.4Hz 1H),5.12-5.08(m,1H),4.97-4.91(m,1H),4.67-4.57(m,1H),4.45-4.22(m,4H),3.88-3.74(m,6H),3.43-3.35(m,1H),2.77-2.72(m,1H),2.59(m,3H),2.40-2.31(m,3H)。LCMS:MS m/z＝630.2[M+H] +。
步骤9：化合物1-12B的合成
化合物1-11进行SFC拆分，色谱柱:DAICEL CHIRALPAK AD(250mm*50mm,10μm)；流动相:[0.1％NH 3。H 2O EtOH]；EtOH％:45％-45％,6.3min。得化合物1-12B(出峰时间:1.665min)，化合物1-12A(出峰时间:2.446min)。
步骤10：化合物1-13的合成
将化合物1-12B(2g,3.18mmol,1eq)溶于二氯甲烷(20mL)中，加入N,N-二异丙基乙胺(1.23g,9.53mmol,1.66mL,3eq)，降温至0～10℃，将三氟甲磺酸酐(1.34g,4.76mmol,786.11μL,1.5eq)缓慢加入到体系中，在此温度下反应15min。倒入饱和氯化铵水溶液(15mL)，分液，水相用二氯甲烷(15mL x 2)萃取，合并有机相，无水硫酸钠干燥，过滤，浓缩得到粗品。粗品经柱层析(石油醚/乙酸乙酯＝100/1～0/1)分离得到化合物1-13。LCMS m/z＝762.2[M+H] +。 1H NMR(400MHz,CDCl 3)δ＝7.21-7.11(m,4H),6.90-6.80(m,4H),6.66(d,J＝8.4Hz,1H),5.19-5.15(m,1H),5.04-4.93(m,1H),4.77-4.72(m,1H),4.41-4.19(m,4H),3.80(s,6H),3.62-3.54(m,1H),3.11-2.97(m,1H),2.56(s,3H),2.42-2.31(m,3H)。LCMS:MS m/z＝762.2[M+H] +。
步骤11：化合物1-15的合成
将化合物1-13(147g,186.74mmol,96.767％纯度,1eq)溶于N,N-二甲基甲酰胺(1.5L)中，加入N,N-二异丙基乙胺(72.40g,560.23mmol,97.58mL,3eq)，再加入化合物1-14(42.54g,214.76mmol,1.15eq,2HCl)，升温至50℃搅拌0.5小时。反应液直接用于下一步，得到化合物1-15的N,N-二甲基甲酰胺溶液。
步骤12：化合物1-16的合成
将化合物1-15(137.8g,187.02mmol,1eq)的N,N-二甲基甲酰胺溶液(1.5L)溶液加入到搅拌器中，加入三乙胺(18.92g,187.02mmol,26.03mL,1eq)，将二碳酸二叔丁酯(48.98g,224.42mmol,51.56mL,1.2eq)加到反应液中，18℃搅拌10小时。倒入水中(1.5L)，加入乙酸乙酯(400mL x 3),饱和氯化铵水溶液(400mL x 4)，无水硫酸钠干燥，过滤，浓缩得到化合物1-16。 1H NMR(400MHz,CDCl 3)δ＝7.16(d,J＝8.4Hz,4H),6.85(d,J＝8.6Hz,4H),6.64(d,J＝8.0Hz,1H),5.22(d,J＝7.2Hz,1H),4.90-4.68(m,2H),4.61(s,1H),4.41-4.21(m,4H),4.04(s,1H),3.80(s,6H),3.71(s,1H),3.50(d,J＝11.0Hz,2H),3.30(s,1H),3.24-3.02(m,2H),2.90(d,J＝2.0Hz,1H),2.78-2.58(m,2H),2.55(s,3H),2.34(d,J＝4.0Hz,3H),1.51(s,9H)。LCMS m/z＝837.2[M+H] +。
步骤13：化合物1-17的合成
将化合物1-16(245g,278.10mmol,95％纯度,1eq)溶于无水二氯甲烷(2500mL)，降温至0～10℃，然后将间氯过氧苯甲酸(56.46g,278.10mmol,85％纯度,1eq)分批加入，10℃搅拌0.5小时。补加间氯过氧苯甲酸(8.47g,41.71mmol,85％纯度,0.15eq)，10℃继续搅拌0.5小时。将该反应与(化合物1-16 10g批次)合并处理。反应液用饱和碳酸氢钠洗(1500mL)，5％硫代硫酸钠洗(1500mL)(湿润淀粉碘化钾试纸检测合格后)，半饱和食盐水洗(1500mL)，无水硫酸钠干燥过滤浓缩。柱层析纯化乙酸乙酯:石油醚＝10％-15％-20％-30％。得到化合物1-17。 1HNMR(400MHz,CDCl 3)δppm 7.15(d,J＝8.00Hz,4H),6.85(d,J＝8.80Hz,4H),6.65(d,J＝8.80Hz,1H),5.27(m,1H),4.78-4.91(m,2H),4.61(s,1H),4.24-4.38(m,4H),3.90-4.18(m,2H),3.78-3.82(m,6H),3.42-3.70(m,3H),3.33(br s,1H),3.06-3.28(m,2H),2.90(s,3H),2.66(m,2H),2.29-2.41(s,3H),1.51(s,9H)。LCMS m/z＝853.2[M+H] +。
步骤14：化合物1-19的合成
将化合物1-18(18.24g,114.59mmol,1.2eq)溶于无水四氢呋喃(900mL)，降温至-20℃，然后加入叔丁醇钠(11.01g,114.59mmol,1.2eq)，搅拌15min。然后加入化合物1-17(90.5g,95.49mmol,90％纯度,1eq)的无水四氢呋喃(180mL)，继续搅拌0.5小时。该反应与(化合物1-18 50g批次)合并处理。向反应液中加入1000mL饱和氯化铵淬灭反应，分液，水相用乙酸乙酯萃取(1000mL)分液，无水硫酸钠干燥过滤，浓缩。柱层析纯化(乙酸乙酯：石油醚＝20％-50％-100％)得白色固体化合物1-19。LCMS m/z＝948.4[M+H] +。
步骤15：化合物1-20的合成
将化合物1-19(97.00g,97.20mmol,95％纯度,1eq)溶于2-甲基四氢呋喃(500mL)，降温至0℃，然后将盐酸(8M,614.80mL,50.6eq)滴加到反应液中，升至25℃搅拌2小时。加入正庚烷(200mL)分液，收集水相，加入2-甲基四氢呋喃(300mL)，用碳酸钠调节pH＝8～9，搅拌澄清，分液，收集有机相，水相用2-甲基四氢呋喃(300mL x 2),收集有机相，饱和食盐水洗(200mL)，无水硫酸镁干燥，过滤，收集有机相，浓缩得到粗品，用二氯甲烷(1000mL)溶解，将三氟乙酸(283.73g,2.49mol,184.24mL,25.6eq)缓慢加入到二氯甲烷中控制温度在10℃以下，25℃搅拌3小时。将反应液倒入500mL冰水，搅拌至澄清，分液。有机相用水(300mL x 3)萃取。合并两个批次水相，然后用二氯甲烷(500mL x 4)反萃，弃去有机相。将水相降温至10℃，向水相在加入500mL 2-甲基四氢呋喃(会放热)，然后用碳酸钠调pH＝9，2-甲基四氢呋喃(400mL x 2)萃取，合并有机相，水洗(300mL x 4)，饱和食盐水洗(300mL)，无水硫酸钠干燥过滤浓缩得灰色固体化合物1-20。LCMS m/z＝608.27[M+H] +。
步骤16：式(Ⅰ)化合物的合成
将化合物1-20(58g,84.00mmol,88％纯度,1eq)，N,N-二异丙基乙胺(21.71g,168.00mmol,29.26mL,2eq)、2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(38.33g,100.80mmol,1.2eq)溶于DMF(600mL)，然后降温至0℃，分批加入2-氟丙烯酸(6.81g,75.60mmol,0.9eq)，0℃搅拌10min。将反应液倒入900mL水中，然后乙酸乙酯萃取(200mL x 3)，合并有机相，饱和氯化铵洗(200mL x 3)，饱和碳酸钠洗(200mL x 3)，饱和食盐水洗(200mL)，无水硫酸钠干燥过滤浓缩。然后用乙腈：水(100mL：150mL)打浆16小时，过滤得到式(Ⅰ)化合物。SFC分析方法(柱子：Chiralcel OD-3,50×4.6mm I.D.,3μm；流动相：A(CO2)和B(甲醇，含0.05％二异丙胺胺)；梯度：B％＝5～50％，3min；流速:3.4mL/min；波长:220nm；压力:1800psi，光学纯度：99.21％，出峰时间：1.840min。 1H NMR(400MHz,CD 3OD)δ＝6.80-6.68(m,1H),5.73-5.51(m,1H),5.46-5.19(m,3H),5.05-4.90(m,3H),4.74-4.58(m,2H),4.37-4.26(m,1H),4.20-4.06(m,2H),4.05-3.84(m,3H),3.79-3.59(m,2H),3.54-3.43(m,1H),3.42-3.35(m,1H),3.31-3.24(m,1H),3.13-2.89(m,3H),2.82-2.52(m,2H),2.50-2.42(m,1H),2.41-2.30(m,5H),2.29-2.18(m,1H)。
实施例2：式(III)化合物的单晶X-射线衍射检测分析
Figure PCTCN2022118023-appb-000012
取0.0133g式(II)化合物A晶型在室温条件下溶解于2mL乙腈中。将样品溶液置于4mL半密封样品瓶中，在室温下缓慢挥发。十天后得到无色块状晶体。收集晶体，用单晶X射线衍射仪(D8-VENTURE)收集衍射强度数据。式(III)化合物的晶体结构数据见表2，式(III)化合物立体结构椭球图见附图6。
表2.式(III)化合物的晶体数据
Figure PCTCN2022118023-appb-000013
Figure PCTCN2022118023-appb-000014
实施例3：式(II)化合物A晶型的制备
方法一：将式(Ⅰ)化合物(0.2g)加入到乙醇(2.4mL)中，加热至50℃，溶清后将水(0.48mL)缓慢加入到反应液中，在50℃下搅拌72小时，过滤，收集固体得到式(II)化合物A晶型。
方法二：将式(Ⅰ)化合物(5.0g)加入到乙醇(4V)搅拌至澄清，将水(1V)缓慢加入到体系中，在20～30℃下加入晶种(0.5g)，在此温度下搅拌15小时，再缓慢滴加水(3V)，继续搅拌1～3小时。过滤收集滤饼，得到式(II)化合物A晶型。
实施例4：式(II)化合物A晶型的固体稳定性试验
依据《原料药与制剂稳定性试验指导原则》(中国药典2015版四部通则9001)，考察式(II)化合物A晶型在高温(60℃，敞口)，高湿(室温/相对湿度92.5％，敞口)及强光照(5000lx，敞口)条件下的稳定性，以及长期实验(25±2℃/60±5％RH)，加速实验(40±2℃/75±5％RH)条件下的晶型稳定性。
称取式(II)化合物A晶型约20mg，置于玻璃样品瓶的底部，摊成薄薄一层。高温及高湿条件下放置的样品用铝箔纸封瓶口，并在铝箔纸上扎些小孔，保证样品能与环境空气充分接触；强光照条件下放置的样品用铝箔纸封瓶口，并在铝箔纸上扎些小孔。不同条件下放置的样品于第5天，10天取样检测(XRPD)，检测结果与0天的初始检测结果进行比较，试验结果见下表3所示。
将式(II)化合物A晶型放入双层药用低密度聚乙烯袋中，每份药用低密度聚乙烯袋分别用扎带扎紧，再放入单层铝箔袋中并热封，最后将样品储存在塑料桶中，储存在稳定性恒温恒湿箱中。稳定性试验所用的包装模拟物料存储包装，在红光下将物料分装为1.5g/包的稳定性样品，每份样都以相同的方式包装并贴好稳定性样品标签，在不同条件下放置的样品于第3个月，第6个月取样检测(XRPD)，检测结果与0天的初始检测结果进行比较，试验结果见下表3所示。
表3式(II)化合物A晶型的固体稳定性试验结果
Figure PCTCN2022118023-appb-000015
Figure PCTCN2022118023-appb-000016
光照对照 *1：需同时放置光照对照样品，光照对照样品采用螺纹瓶盖密封，再用锡箔纸完全包裹。
结论：式(II)化合物A晶型在高温、高湿、强光照条件下，以及长期实验和加速实验的条件下具有良好的稳定性。
生物测试数据：
实验例1：化合物对KRAS G12C突变的MIA-PA-CA-2细胞增殖抑制作用的测试
1.1实验目的
测试化合物对KRAS G12C突变的MIA-PA-CA-2细胞增殖抑制的IC 50。
1.2试剂
本研究使用的主要试剂包括CellTiter-Glo(Promega，货号：G7573)。
1.3仪器
本研究所使用主要仪器为PerkinElmer EnVision多功能酶标仪。
1.4实验方法
1)贴壁细胞经胰酶消化处理成细胞悬液，并对细胞悬液进行计数备用。
2)取适量细胞至离心管并用细胞培养液补足至需要体积，铺至96孔板，终密度为2000细胞/孔，100μL培养液。
3)培养24小时后，将化合物用DMSO配置成10mM，并用DPBS(杜比可磷酸缓冲盐)以3倍梯度稀释9个点，每孔加入10μL，两复孔。实验对照孔(Con)每孔加入10μL DPBS。
4)同天，取一块未加药处理的细胞培养板，加入50μL CellTiter Glo，利用EnVision进行荧光读值，标记为Day0读值。
5)加药处理后的细胞培养72h后，去除培养板，向细胞板中加入50μL CellTiter Glo，利用EnVision进行荧光读值。
6)数据分析：按下列公式计算各孔对细胞的抑制率：
Figure PCTCN2022118023-appb-000017
*F Day0为未加药处理的原始细胞数目测试孔的读值；
F Con为培养72小时后，Con组的荧光读值。
F Cpd为培养72小时后，各化合物孔的荧光读值。
7)使用GraphPad Prism软件对化合物的抑制率数据(抑制率％)进行log(agonist)vs.response--Variable slope非线性拟合分析，得到化合物的IC 50值，拟合公式为：
Y＝Bottom+(Top-Bottom)/(1+10^((LogIC 50-X)*HillSlope))
1.5实验结果
表4.本发明化合物对KRASG12C突变的MIA-PA-CA-2细胞增殖抑制的测试结果
受试品 IC 50(nM)
式(I)化合物 0.44
实验结果显示式(I)化合物对KRASG12C突变的MIA-PA-CA-2细胞系具有良好的细胞增殖抑制活性。
实验例2：化合物对KRAS G12C突变的H358细胞增殖抑制作用的测试
2.1实验目的
测试化合物对KRAS G12C突变的H358细胞增殖抑制的IC 50。
2.2试剂
本研究使用的主要试剂包括RPMI-1640培养基，盘尼西林/链霉素抗生素购自维森特，胎牛血清购自Biosera。CellTiter-Glo(细胞活率化学发光检测试剂)试剂购自Promega。NCI-H358细胞系购自中国科学院细胞库。
2.3仪器
本研究所使用主要仪器为Nivo多标记分析仪(PerkinElmer)。
2.4实验方法：
1)将NCI-H358细胞种于白色96孔板中，80μL细胞悬液每孔，其中包含4000个NCI-H358细胞。细胞板置于二氧化碳培养箱中过夜培养。
2)将待测化合物用排枪进行5倍稀释至第9个浓度，即从2mM稀释至5.12nM，设置双复孔实验。向中间板中加入78μL培养基，再按照对应位置，转移2μL每孔的梯度稀释化合物至中间板，混匀后转移20μL每孔到细胞板中。转移到细胞板中的化合物浓度范围是10μM至0.0256nM。细胞板置于二氧化碳培养箱中培养5天。另准备一块细胞板，在加药当天读取信号值作为最大值(下面方程式中Max值)参与数据分析。向此细胞板每孔加入25μL细胞活率化学发光检测试剂，室温孵育10分钟使发光信号稳定。采用多标记分析仪读数。
3)向细胞板中加入每孔25μL的细胞活率化学发光检测试剂，室温孵育10分钟使发光信号稳定。采用多标记分析仪读数。
数据分析：
利用方程式(Sample-Min)/(Max-Min)*100％将原始数据换算成抑制率，IC 50的值即可通过四参数进行曲线拟合得出(GraphPad Prism中"log(inhibitor)vs.response--Variable slope"模式得出)。表5提供了本发明的化合物对NCI-H358细胞增殖的抑制活性。
表5.本发明化合物对KRASG12C突变的H358细胞增殖抑制的测试结果
受试品 NCI-H358IC 50(nM)
式(I)化合物 0.6
结论：式(I)化合物对NCI-H358细胞增殖展现出较好的抑制活性。
实验例3：肝细胞代谢稳定性
实验目的：评定受试化合物分别在CD-1小鼠、SD大鼠、比格犬、食蟹猴、人肝细胞中的代谢稳定性
实验操作：准备若干96孔样品沉淀板，分别命名为T0、T15、T30、T60、T90、T120、T0-MC、T120-MC和空白基质。提前取出复苏培养液和孵育培养液，放置在37℃水浴锅中预热。从液氮罐中取出冻存的肝细胞，立即浸没到37℃水浴中(约90秒)。待冻存部分溶化松动后，分别倒入含有40mL复苏培养液的离心管中，轻柔的颠倒让细胞在复苏培养液中重悬。室温条件下，100×g离心5分钟，移除上清液，用适当体积的孵育培养液重悬肝细胞，用台盼蓝染色法计算细胞活率。将198μL的肝细胞混悬液(0.51×106cells/mL)加入到已预热的孵育板中，培养液对照组加入198μL不含肝细胞的孵育培养液至T0-MC和T120-MC孵育板中，所有孵育板在37℃培养箱中预孵育10分钟。然后加入2μL供试品和对照化合物工作液，混匀，立即将孵育板放入培养箱内的摇板机中，启动计时器开始反应。每个化合物的每个时间点准备2个重复样本。孵育条件为37℃、饱和湿度、含5％CO 2。测试体系中，供试品的终浓度为1μM，对照品的终浓度为3μM，肝细胞的终浓度为0.5×106cells/mL，总有机溶剂的终浓度为0.96％，其中DMSO的终浓度为0.1％。相应时间点孵育结束时，取出孵育板，取出25μL化合物和对照化合物与细胞的混合液加入到含有125μL终止液(含有200ng/mL甲苯磺丁脲和拉贝诺尔的乙腈溶液)的样品板中。对于Blank样品板，直接加入25μL不含肝细胞的孵育培养液。所有样品板封膜后在摇板机上以600rpm摇10分钟后，3220×g离心20分钟。供试品和对照品上清液用超纯水以1:3的比例稀释。所有样品混匀后用LC/MS/MS的方法进行分析。
实验结果：实验结果如表6所示。
表6.受试化合物CD-1小鼠、SD大鼠、比格犬、食蟹猴、人肝细胞中的代谢稳定性
Figure PCTCN2022118023-appb-000018
实验结论：多种属肝细胞代谢实验显示式(I)化合物具有良好的代谢稳定性。
实验例4:体外肝微粒体稳定性研究
实验目的：评定受试化合物分别在CD-1小鼠、SD大鼠、比格犬、食蟹猴、人肝微粒体中的代谢稳定性
实验操作：准备2块96孔孵育板，分别命名为T60孵育板和NCF60孵育板。在T60孵育板和NCF60孵育板上 分别加入445μL微粒体工作液(肝微粒体蛋白浓度为0.56mg/mL)，然后将上述孵育板放置于37℃水浴锅中预孵育大约10分钟。
预孵育结束后，在T60孵育板和NCF60孵育板上分别加入5μL供试品或对照化合物工作液，混匀。在NCF60孵育板上每孔添加50μL磷酸钾盐缓冲液启动反应；在T0终止板中加入180μL的终止液(含200ng/mL甲苯磺丁脲和200ng/mL拉贝洛尔的乙腈溶液)和6uL的NADPH再生体系工作液，从T60孵育板中取出54μL样品至T0终止板(T0样品产生)。在T60孵育板上每孔添加44μLNADPH再生体系工作液启动反应。在Blank板中只添加54μL微粒体工作液、6uL的NADPH再生体系工作液和180μL的终止液。因此，在供试品或对照化合物的样品中，化合物、睾酮、双氯芬酸和普罗帕酮的反应终浓度为1μM，肝微粒体的浓度为0.5mg/mL，DMSO和乙腈在反应体系中的终浓度分别为0.01％(v/v)和0.99％(v/v)。孵育适当时间(如5、15、30、45和60分钟)后，分别在每个终止板的样品孔中加入180μL的终止液(含200ng/mL甲苯磺丁脲和200ng/mL拉贝洛尔的乙腈溶液)，之后从T60孵育板中取出60μL样品以终止反应。所有样品板摇匀并在3220×g离心20分钟，然后每孔取80μL上清液稀释到240μL纯水中用于液相色谱串联质谱分析，液相色谱串联质谱分析所有样品进样分析。
实验结果：实验结果如表7所示。
表7.受试化合物CD-1小鼠、SD大鼠、比格犬、食蟹猴、人肝微粒体的代谢稳定性
Figure PCTCN2022118023-appb-000019
实验结论：肝微粒体代谢稳定性研究显示式(I)化合物具有良好的代谢稳定性
实验例5:血浆稳定研究
实验目的：评定受试化合物分别在CD-1小鼠、人的血浆稳定性
实验操作：将冻存的血浆解冻10～20分钟，待血浆完全解冻后，将其置于离心机中以3220×g离心5分钟，去除其中存在的悬浮物和沉淀物。准备96孔孵育板，分别命名为T0、T10、T30、T60、T120。将孵育板中对应的加入98μL的小鼠、大鼠、犬、猴和人空白血浆，然后加入2μL化合物或对照化合物的工作液加入到对应的孵育板中，每个样品准备两个平行孔。所有的样品在37℃水浴锅中进行孵育。化合物和对照化合物比沙可啶、马来酸依那普利、普鲁卡因和普鲁本辛的最终孵育浓度为2μM，最终有机相含量为2.0％。每一个孵育时间点结束时，取出相应的孵育板，向每个对应的样品孔中加入400μL含有200ng/mL甲苯磺丁脲和拉贝诺尔的乙腈溶液沉淀蛋白。所有样品板封膜并摇匀后，3220×g离心20分钟。取50μL上清液加入100μL超纯水稀释，所有样品混匀后用LC/MS/MS的方法分析。
实验结果：实验结果如表8所示。
表8.受试化合物CD-1小鼠、人的血浆稳定性
Figure PCTCN2022118023-appb-000020
实验结论：式(I)化合物在人和小鼠血浆中的稳定性良好。
实验例6:全血稳定研究
实验目的：评定受试化合物分别在评定受试化合物分别在CD-1小鼠、SD大鼠、比格犬、食蟹猴的全血稳定性
实验操作：在实验当天或实验的前一天，采用抗凝剂EDTA-K2采集新鲜的CD-1小鼠、SD大鼠、比格犬、食蟹猴全血。在实验开始前，将全血与PBS进行1：1(v：v)混合，放置于37℃水浴锅中预热10～20分钟。准备96孔孵育板，分别命名为T0、T30、T60、T240。在对应的孵育板中，包括T0、T30、T60和T240孵育板，将2μL化合物或对照化合物的工作液与98μL的小鼠、大鼠、犬、猴和人空白全血混合，每个样品准备两个平行孔。所有的样品在37℃水浴锅中进行孵育。化合物的最终孵育浓度为5μM，对照化合物的最终孵育浓度为2μM。每一个时间点孵育结束时，取出相应的孵育板，立即向对应的样品孔中加入100μL超纯水，混匀，然后加入800μL含有200ng/mL甲苯磺丁脲和拉贝诺尔的乙腈溶液沉淀蛋白。样品板封膜并摇匀后，3220×g离心20分钟。取150μL上清液用LC/MS/MS的方法分析。
实验结果：实验结果如表9所示。
表9.受试化合物CD-1小鼠、SD大鼠、比格犬、食蟹猴的全血稳定性
Figure PCTCN2022118023-appb-000021
实验结论：多种属全血稳定性研究显示式(I)化合物在全血中的稳定性良好。
实验例7:蛋白结合率研究
实验目的：采用平衡透析法测定受试化合物在CD-1小鼠、SD大鼠、比格犬、食蟹猴和人血浆中的蛋白结合率。
实验操作：采用上述五个物种的血浆分别配制化合物浓度为2μM的血浆样品，置于96孔平衡透析装置中，在37±1℃下用磷酸盐缓冲溶液透析4h。本实验采用华法林作为对照化合物。血浆和透析缓冲液中待测物的浓度用LC-MS/MS法进行测定。
实验结果：实验结果如表10所示。
表10.受试化合物CD-1小鼠、SD大鼠、比格犬、食蟹猴、人的蛋白结合率
Figure PCTCN2022118023-appb-000022
实验结论：多种属血浆结合率研究显示式(I)化合物在血浆中具有更高的蛋白未结合率。
实验例8:体内药代动力学研究
1)SD大鼠口服及静脉注射受试化合物的药代动力学研究
受试化合物与5％二甲基亚砜/95％(10％羟丙基-β-环糊精)溶液混合，涡旋并超声，制备得到1mg/mL澄清溶液，微孔滤膜过滤后备用。选取7至10周龄的雄性SD大鼠，静脉注射、口服给予候选化合物溶液。收集一定时间的全血，制备得到血浆，以LC-MS/MS方法分析药物浓度，并用Phoenix WinNonlin软件(美国Pharsight公司)计算药代参数。实验结果如表11所示：
表11.受试化合物的药代动力学结果
Figure PCTCN2022118023-appb-000023
注：Vd ss,u为血浆蛋白未结合下的表观容积分布(Vd ss,u＝Vd ss/PPB(Unbond％))；C max,u，AUC 0-last，u，都是血浆蛋白未结合下的相应数值(C max,u＝C max x PPB(Unbond％)；AUC 0-last，u＝AUC 0-lastx PPB(Unbond％))
实验结论：PK研究显示，式(I)化合物在大鼠中具有较高的非结合血浆暴露量和良好的口服生物利用度。
2)SD大鼠口服式(II)化合物的A晶型的药代动力学研究
精确称量109.72mg的受试化合物到玻璃瓶中，加入774uL 0.5％甲基纤维素(400粘度)水溶液，搅拌5分钟，加入10mL的0.5％甲基纤维素(400粘度)水溶液，搅拌5分钟，得均匀的不透明的混悬液。选取7至10周龄的雄性SD大鼠，口服给予候选化合物溶液。收集一定时间的全血，制备得到血浆，以LC-MS/MS方法分析药物浓度，并用Phoenix WinNonlin软件(美国Pharsight公司)计算药代参数。实验结果如表12所示：
表12.受试化合物的药代动力学结果
Figure PCTCN2022118023-appb-000024
实验结论：PK研究显示，式(II)化合物A晶型在大鼠中具有高的暴露量
3)CD小鼠口服及静脉注射受试化合物的药代动力学研究
受试化合物与5％二甲基亚砜/95％(10％羟丙基-β-环糊精)溶液混合，涡旋并超声，制备得到1mg/mL澄清溶液，微孔滤膜过滤后备用。选取7至10周龄的雄性CD小鼠，静脉注射、口服给予候选化合物溶液。收集一定时间的全血，制备得到血浆，以LC-MS/MS方法分析药物浓度，并用Phoenix WinNonlin软件(美国Pharsight公司)计算药代参数。实验结果如表13所示：
表13.受试化合物的药代动力学结果
Figure PCTCN2022118023-appb-000025
注：Vd ss,u为血浆蛋白未结合下的表观容积分布(Vd ss,u＝Vd ss/PPB(Unbond％))；C max,u，AUC 0-last，u，都是血 浆蛋白未结合下的相应数值(C max,u＝C max x PPB(Unbond％)；AUC 0-last，u＝AUC 0-lastx PPB(Unbond％))
实验结论：PK研究显示，式(I)化合物在小鼠中具有较高的非结合血浆暴露量和良好的口服生物利用度。
4)比格犬口服受试化合物的药代动力学研究
精确称量680.397mg受试化合物的粉末，加入50mL的0.5％甲基纤维素(400粘度)水溶液，搅拌10min，超声10min。加入50mL0.5％甲基纤维素(400粘度)水溶液，使用匀浆机均质化10min。加入11mL，超声5min，搅拌10min，加入358μL 0.5％甲基纤维素(400粘度)水溶液，搅拌2min。选取六个月以上雄性比格犬，口服给予候选化合物溶液。收集一定时间的全血，制备得到血浆，以LC-MS/MS方法分析药物浓度，并用Phoenix WinNonlin软件(美国Pharsight公司)计算药代参数。实验结果如表14所示：收集一定时间的全血，制备得到血浆，以LC-MS/MS方法分析药物浓度，并用Phoenix WinNonlin软件(美国Pharsight公司)计算药代参数。实验结果如表14所示：
表14.受试化合物的药代动力学结果
Figure PCTCN2022118023-appb-000026
实验结论：PK研究显示，式(II)化合物的A晶型在犬中具有高的暴露量。
实验例9：体内药效学研究
人胰腺癌Mia PaCa-2细胞裸小鼠皮下移植肿瘤Balb/c Nude小鼠模型的体内药效学研究
1.细胞培养和肿瘤组织准备
细胞培养：人胰腺癌Mia PaCa-2细胞(ATCC-CRL-1420)体外单层培养，培养条件为DMEM培养基中加10％胎牛血清，2.5％马血清，37℃5％二氧化碳孵箱培养。一周两次用胰酶-EDTA进行常规消化处理传代。当细胞饱和度为80％-90％，数量到达要求时，收取细胞，计数，重悬于适量PBS中，1:1加入基质胶，获取细胞密度为25x 106cells/mL的细胞悬液。
细胞接种：将0.2mL(5×106cells/mouse个)Mia PaCa-2细胞(加基质胶，体积比为1:1)皮下接种于每只小鼠的右后背，肿瘤平均体积达到190mm 3时，根据肿瘤体积进行随机分组，按照表15中的方案开始给药。
表15.实验动物分组及给药方案
Figure PCTCN2022118023-appb-000027
Figure PCTCN2022118023-appb-000028
注：PO代表口服；QD代表每日一次。
2.肿瘤测量和实验指标
每周两次用游标卡尺测量肿瘤直径。肿瘤体积的计算公式为：V＝0.5a×b 2，a和b分别表示肿瘤的长径和短径。
化合物的抑瘤疗效用TGI(％)或相对肿瘤增殖率T/C(％)评价。相对肿瘤增殖率T/C(％)＝TRTV/CRTV×100％(TRTV：治疗组RTV；CRTV：阴性对照组RTV)。根据肿瘤测量的结果计算出相对肿瘤体积(relative tumor volume，RTV)，计算公式为RTV＝Vt/V0，其中V0是分组给药时(即D0)测量所得平均肿瘤体积，Vt为某一次测量时的平均肿瘤体积，TRTV与CRTV取同一天数据。
TGI(％)，反映肿瘤生长抑制率。TGI(％)＝[(1-(某处理组给药结束时平均瘤体积－该处理组开始给药时平均瘤体积))/(溶剂对照组治疗结束时平均瘤体积－溶剂对照组开始治疗时平均瘤体积)]×100％。
3.实验结果
实验结果如图4、5所示。
给药22天时结果如表16所示
表16.给药第22天下的T/C及TGI
化合物 给药量 肿瘤体积均值 T/C TGI
溶媒 N/A 2016.29mm 3 N/A N/A
式(I)化合物 10mg/kg 249.87mm 3 12.39％ 93.06％
式(I)化合物 30mg/kg 124.14mm 3 6.16％ 99.64％
实验结论：式(I)化合物具有显著的抑瘤作用，且小鼠各个剂量组的体重平稳，无明显不耐受现象。

Claims (14)

  1. 式(II)化合物，
    Figure PCTCN2022118023-appb-100001
    其中，n选自0～3。
  2. 根据权利要求1所述的化合物，其中n选自0、0.5、1、1.5、2、2.5和3。
  3. 根据权利要求2所述的化合物，其中n选自2。
  4. 根据权利要求1～3任意一项所述的式(II)化合物的A晶型，
    Figure PCTCN2022118023-appb-100002
    其特征在于其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.514±0.200°、14.689±0.200°、18.122±0.200°。
  5. 根据权利要求4所述的A晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：6.218±0.200°、8.514±0.200°、12.299±0.200°、14.689±0.200°、16.903±0.200°、18.122±0.200°、18.927±0.200°、25.580±0.200°。
  6. 根据权利要求5所述的A晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：6.218±0.200°、8.514±0.200°、11.663±0.200°、12.299±0.200°、14.689±0.200°、16.903±0.200°、18.122±0.200°、18.927±0.200°、19.364±0.200°、20.386±0.200°、21.914±0.200°、25.580±0.200°。
  7. 根据权利要求6所述的A晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：6.218°、8.514°、11.663°、12.299°、14.689°、16.903°、18.122°、18.554°、18.927°、19.364°、20.386°、21.914°、22.640°、23.867°、24.553°、24.806°、25.580°、25.988°、27.147°、27.715°、29.135°、31.799°。
  8. 根据权利要求7所述的A晶型，其XRPD图谱如图1所示。
  9. 根据权利要求4～8任意一项所述的A晶型，其差示扫描量热曲线在115.37℃±3℃处具有吸热峰的峰值。
  10. 根据权利要求9所述的A晶型，其DSC图谱如图2所示。
  11. 根据权利要求4～8任意一项所述的A晶型，其热重分析曲线在150.0℃±3℃时失重达5.379％。
  12. 根据权利要求11所述的A晶型，其TGA图谱如图3所示。
  13. 根据权利要求1～3任意一项所述的化合物或4～12任意一项所述的A晶型在制备治疗实体瘤药物中的应用。
  14. 根据权利要求13所述的应用，其中，实体瘤为肺癌和直肠癌。
PCT/CN2022/118023 2021-09-10 2022-09-09 嘧啶并杂环类化合物的晶型及其制备方法 Ceased WO2023036282A1 (zh)

Priority Applications (10)

Application Number Priority Date Filing Date Title
CA3231329A CA3231329A1 (en) 2021-09-10 2022-09-09 Crystal form of pyrimidine heterocyclic compound and preparation method therefor
AU2022342220A AU2022342220A1 (en) 2021-09-10 2022-09-09 Crystal form of pyrimidine heterocyclic compound and preparation method therefor
JP2024515669A JP7688447B2 (ja) 2021-09-10 2022-09-09 ピリミジン複素環化合物の結晶形及びその調製方法
MX2024003027A MX2024003027A (es) 2021-09-10 2022-09-09 Forma cristalina del compuesto heterociclico de pirimidina y metodo de preparacion del mismo.
US18/690,417 US20250129102A1 (en) 2021-09-10 2022-09-09 Crystal form of pyrimidine heterocyclic compound and preparation method therefor
IL311340A IL311340A (en) 2021-09-10 2022-09-09 A crystalline form of a heterocyclic pyrimidine compound and a method for its preparation
EP22866743.2A EP4385986A4 (en) 2021-09-10 2022-09-09 CRYSTALLINE FORM OF A PYRIMIDIC HETEROCYCLIC COMPOUND AND PROCESS FOR ITS PREPARATION
CN202280060673.9A CN117957220A (zh) 2021-09-10 2022-09-09 嘧啶并杂环类化合物的晶型及其制备方法
KR1020247011544A KR20240056582A (ko) 2021-09-10 2022-09-09 피리미딘 헤테로사이클릭 화합물의 결정형 및 이의 제조 방법
JP2025082474A JP2025124698A (ja) 2021-09-10 2025-05-16 ピリミジン複素環化合物の結晶形及びその調製方法

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
CN202111062619.1 2021-09-10
CN202111062619 2021-09-10
CN202211034826.0 2022-08-26
CN202211034826 2022-08-26

Publications (1)

Publication Number Publication Date
WO2023036282A1 true WO2023036282A1 (zh) 2023-03-16

Family

ID=85507187

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2022/118023 Ceased WO2023036282A1 (zh) 2021-09-10 2022-09-09 嘧啶并杂环类化合物的晶型及其制备方法

Country Status (11)

Country Link
US (1) US20250129102A1 (zh)
EP (1) EP4385986A4 (zh)
JP (2) JP7688447B2 (zh)
KR (1) KR20240056582A (zh)
CN (1) CN117957220A (zh)
AU (1) AU2022342220A1 (zh)
CA (1) CA3231329A1 (zh)
IL (1) IL311340A (zh)
MX (1) MX2024003027A (zh)
TW (1) TWI820901B (zh)
WO (1) WO2023036282A1 (zh)

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023240263A1 (en) 2022-06-10 2023-12-14 Revolution Medicines, Inc. Macrocyclic ras inhibitors
WO2024206858A1 (en) 2023-03-30 2024-10-03 Revolution Medicines, Inc. Compositions for inducing ras gtp hydrolysis and uses thereof
WO2024229406A1 (en) 2023-05-04 2024-11-07 Revolution Medicines, Inc. Combination therapy for a ras related disease or disorder
WO2025034702A1 (en) 2023-08-07 2025-02-13 Revolution Medicines, Inc. Rmc-6291 for use in the treatment of ras protein-related disease or disorder
WO2025067459A2 (en) 2023-09-29 2025-04-03 D3 Bio (Wuxi) Co., Ltd. Therapies for the treatment of cancer
WO2025080946A2 (en) 2023-10-12 2025-04-17 Revolution Medicines, Inc. Ras inhibitors
WO2025137519A1 (en) * 2023-12-22 2025-06-26 Erasca, Inc. Kras inhibitors
WO2025171296A1 (en) 2024-02-09 2025-08-14 Revolution Medicines, Inc. Ras inhibitors
WO2025218730A1 (en) * 2024-04-18 2025-10-23 D3 Bio (Wuxi) Co., Ltd. Solid form of kras inhibitor
WO2025232765A1 (en) * 2024-05-07 2025-11-13 D3 Bio (Wuxi) Co., Ltd. Treatment of cancer with pyrimidoheterocyclic compounds
WO2025240847A1 (en) 2024-05-17 2025-11-20 Revolution Medicines, Inc. Ras inhibitors
WO2025255438A1 (en) 2024-06-07 2025-12-11 Revolution Medicines, Inc. Methods of treating a ras protein-related disease or disorder
WO2025265060A1 (en) 2024-06-21 2025-12-26 Revolution Medicines, Inc. Therapeutic compositions and methods for managing treatment-related effects
WO2026006747A1 (en) 2024-06-28 2026-01-02 Revolution Medicines, Inc. Ras inhibitors
WO2026015796A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026015801A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026015790A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026015825A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Use of ras inhibitor for treating pancreatic cancer

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112119075A (zh) * 2018-08-16 2020-12-22 豪夫迈·罗氏有限公司 稠环化合物
CN112390788A (zh) * 2019-08-13 2021-02-23 苏州闻天医药科技有限公司 一种用于抑制krasg12c突变蛋白的化合物及其制备方法和用途
CN112979664A (zh) * 2019-12-02 2021-06-18 上海璎黎药业有限公司 一种含氧杂环化合物、其制备方法及应用
WO2021180181A1 (zh) * 2020-03-12 2021-09-16 南京明德新药研发有限公司 嘧啶并杂环类化合物及其应用
WO2022081655A1 (en) * 2020-10-14 2022-04-21 Accutar Biotechnology, Inc. Substituted dihydropyranopyrimidine compounds as kras inhibitors

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI811656B (zh) * 2020-04-22 2023-08-11 大陸商德昇濟醫藥（無錫）有限公司 嘧啶并雜環類化合物及其應用

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112119075A (zh) * 2018-08-16 2020-12-22 豪夫迈·罗氏有限公司 稠环化合物
CN112390788A (zh) * 2019-08-13 2021-02-23 苏州闻天医药科技有限公司 一种用于抑制krasg12c突变蛋白的化合物及其制备方法和用途
CN112979664A (zh) * 2019-12-02 2021-06-18 上海璎黎药业有限公司 一种含氧杂环化合物、其制备方法及应用
WO2021180181A1 (zh) * 2020-03-12 2021-09-16 南京明德新药研发有限公司 嘧啶并杂环类化合物及其应用
WO2022081655A1 (en) * 2020-10-14 2022-04-21 Accutar Biotechnology, Inc. Substituted dihydropyranopyrimidine compounds as kras inhibitors

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
"Chinese Pharmacopoeia", 2015, article "Guiding Principles for Stability Testing of Raw Materials and Preparations"
See also references of EP4385986A4

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023240263A1 (en) 2022-06-10 2023-12-14 Revolution Medicines, Inc. Macrocyclic ras inhibitors
WO2024206858A1 (en) 2023-03-30 2024-10-03 Revolution Medicines, Inc. Compositions for inducing ras gtp hydrolysis and uses thereof
WO2024229406A1 (en) 2023-05-04 2024-11-07 Revolution Medicines, Inc. Combination therapy for a ras related disease or disorder
WO2025034702A1 (en) 2023-08-07 2025-02-13 Revolution Medicines, Inc. Rmc-6291 for use in the treatment of ras protein-related disease or disorder
WO2025067459A2 (en) 2023-09-29 2025-04-03 D3 Bio (Wuxi) Co., Ltd. Therapies for the treatment of cancer
WO2025080946A2 (en) 2023-10-12 2025-04-17 Revolution Medicines, Inc. Ras inhibitors
WO2025137519A1 (en) * 2023-12-22 2025-06-26 Erasca, Inc. Kras inhibitors
WO2025171296A1 (en) 2024-02-09 2025-08-14 Revolution Medicines, Inc. Ras inhibitors
WO2025218730A1 (en) * 2024-04-18 2025-10-23 D3 Bio (Wuxi) Co., Ltd. Solid form of kras inhibitor
WO2025232765A1 (en) * 2024-05-07 2025-11-13 D3 Bio (Wuxi) Co., Ltd. Treatment of cancer with pyrimidoheterocyclic compounds
WO2025240847A1 (en) 2024-05-17 2025-11-20 Revolution Medicines, Inc. Ras inhibitors
WO2025255438A1 (en) 2024-06-07 2025-12-11 Revolution Medicines, Inc. Methods of treating a ras protein-related disease or disorder
WO2025265060A1 (en) 2024-06-21 2025-12-26 Revolution Medicines, Inc. Therapeutic compositions and methods for managing treatment-related effects
WO2026006747A1 (en) 2024-06-28 2026-01-02 Revolution Medicines, Inc. Ras inhibitors
WO2026015796A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026015801A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026015790A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Methods of treating a ras related disease or disorder
WO2026015825A1 (en) 2024-07-12 2026-01-15 Revolution Medicines, Inc. Use of ras inhibitor for treating pancreatic cancer

Also Published As

Publication number Publication date
MX2024003027A (es) 2024-03-26
JP2025124698A (ja) 2025-08-26
CN117957220A (zh) 2024-04-30
IL311340A (en) 2024-05-01
TW202313627A (zh) 2023-04-01
TWI820901B (zh) 2023-11-01
US20250129102A1 (en) 2025-04-24
JP7688447B2 (ja) 2025-06-04
AU2022342220A1 (en) 2024-03-28
KR20240056582A (ko) 2024-04-30
EP4385986A1 (en) 2024-06-19
CA3231329A1 (en) 2023-03-16
EP4385986A4 (en) 2024-12-18
JP2024535792A (ja) 2024-10-02

Similar Documents

Publication Publication Date Title
TWI820901B (zh) 嘧啶並雜環類化合物的晶型及其製備方法
BR112022018140B1 (pt) Compostos pirimidoheterocíclicos e uso dos mesmos
US11760756B2 (en) Crystalline form of a PD-1/PD-L1 inhibitor
CN104109151B (zh) N‑(2‑{2‑二甲氨基乙基‑甲氨基}‑4‑甲氧基‑5‑{[4‑(1‑甲基吲哚‑3‑基)嘧啶‑2‑基]氨基}苯基)丙‑2‑烯酰胺的甲磺酸盐的多晶型
JP7573019B2 (ja) Ａｔｒ阻害剤の結晶形及びその使用
WO2021110168A1 (zh) 作为erk抑制剂的螺环类化合物及其应用
WO2020221358A1 (zh) Wee1抑制剂化合物的晶型及其应用
JP2021073246A (ja) Ｂ−ｒａｆキナーゼのマレイン酸塩、結晶形、調整方法、及びその使用
TW200409754A (en) 3-z-[1-(4-(N-((4-methyl-piperazin-1-yl)-methylcarbonyl)-N-methyl-amino)-anilino)-1-phenyl-methylene]-6-methoxycarbonyl-2-indolinone-monoethanesulphonate and the use thereof as a pharmaceutical composition
US9834528B2 (en) Alpha-amino esters of hydroxypropylthiazolidine carboxamide derivative and salt form, crystal polymorph thereof
TW201534601A (zh) 替匹拉希（Ｔｉｐｉｒａｃｉｌ）鹽酸鹽之穩定形結晶及其結晶化方法
CN113956269A (zh) 用于抑制蛋白激酶活性的二(杂)芳基大环化合物
CN109593102A (zh) 一种氘代二苯氨基嘧啶类化合物的制备方法及其晶型
CN103204843B (zh) 结晶型艾普拉唑钠乙醇化物及其制备方法
TWI811656B (zh) 嘧啶并雜環類化合物及其應用
EA049584B1 (ru) Кристаллическая форма пиримидинового гетероциклического соединения и способ ее получения
CN114057700B (zh) 成纤维细胞生长因子受体抑制剂的晶型及其制备方法
RU2832707C2 (ru) Кристаллическая форма ингибитора atr и ее применение
BR122023026304B1 (pt) Compostos pirimidoheterocíclicos e uso dos mesmos para a fabricação de um medicamento para tratar doenças relacionadas
HK40098899A (zh) 嘧啶并杂环类化合物及其应用
HK40078182B (zh) 嘧啶并杂环类化合物及其应用
BRPI0616374A2 (pt) formas de sal sólido para um pirrol substituìdo 2-indolinona, composição farmacêutica compreendendo as mesmas, método para modulação de atividade catalìtica de proteìna quinase, proteìna quinase, métodos pra preparação de cristais e de polimorfos e uso
TW202404973A (zh) Ｎ—甲基—Ｎ—（（１Ｓ，３Ｓ）—３—甲基—３—（（６—（１—甲基—１Ｈ—吡唑—４—基）吡唑并［１，５—ａ］吡嗪—４—基）氧基）環丁基）丙烯醯胺之結晶多形體
WO2021037156A1 (zh) 一种致癌性融合激酶抑制剂的晶型及其应用
BR122025019554A2 (pt) Forma sólida de 3-(5-flúor-benzofuran-3-il)-4-(5-metil-5h-[1,3]dioxolo[4,5- f]indol-7-il)pirrol-2,5-diona, seus usos, composição farmacêutica e processo para preparar a mesma

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 22866743

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 202280060673.9

Country of ref document: CN

Ref document number: 311340

Country of ref document: IL

WWE Wipo information: entry into national phase

Ref document number: 12024550617

Country of ref document: PH

Ref document number: 3231329

Country of ref document: CA

ENP Entry into the national phase

Ref document number: 2024515669

Country of ref document: JP

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 202427017399

Country of ref document: IN

WWE Wipo information: entry into national phase

Ref document number: 2022342220

Country of ref document: AU

Ref document number: AU2022342220

Country of ref document: AU

ENP Entry into the national phase

Ref document number: 2022866743

Country of ref document: EP

Effective date: 20240311

REG Reference to national code

Ref country code: BR

Ref legal event code: B01A

Ref document number: 112024004708

Country of ref document: BR

ENP Entry into the national phase

Ref document number: 2022342220

Country of ref document: AU

Date of ref document: 20220909

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 20247011544

Country of ref document: KR

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 202490694

Country of ref document: EA

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 11202401609V

Country of ref document: SG

ENP Entry into the national phase

Ref document number: 112024004708

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20240308

WWP Wipo information: published in national office

Ref document number: 18690417

Country of ref document: US

WWG Wipo information: grant in national office

Ref document number: 202490694

Country of ref document: EA