WO2023246839A1 - Salt of thieno[2,3-d]imidazole compound, crystal form thereof and pharmaceutical use thereof - Google Patents

Salt of thieno[2,3-d]imidazole compound, crystal form thereof and pharmaceutical use thereof Download PDF

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WO2023246839A1
WO2023246839A1 PCT/CN2023/101590 CN2023101590W WO2023246839A1 WO 2023246839 A1 WO2023246839 A1 WO 2023246839A1 CN 2023101590 W CN2023101590 W CN 2023101590W WO 2023246839 A1 WO2023246839 A1 WO 2023246839A1
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crystal form
ray powder
radiation
diffraction pattern
powder diffraction
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PCT/CN2023/101590
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French (fr)
Chinese (zh)
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宫正
朱凤飞
范江
刘振平
窦赢
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西藏海思科制药有限公司
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Publication of WO2023246839A1 publication Critical patent/WO2023246839A1/en

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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
    • A61K31/4965—Non-condensed pyrazines
    • A61K31/497—Non-condensed pyrazines containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems

Definitions

  • the present invention relates to the field of medicine. Specifically, it relates to a salt crystal form of a thiophene[2,3-d]imidazole compound and its use in medicine.
  • Diabetes is a group of metabolic diseases characterized by high blood sugar. Hyperglycemia is caused by defective insulin secretion, impaired biological action, or both. The long-term high blood sugar in diabetes leads to chronic damage and dysfunction of various tissues, especially the eyes, kidneys, heart, blood vessels, and nerves. It is mainly divided into two types. Type 1 diabetes: Destruction of pancreatic B cells leads to absolute insulin deficiency. Type 2 diabetes: Insulin resistance is mainly associated with relative insulin deficiency or impaired insulin secretion is mainly associated with insulin resistance.
  • Drugs for type 2 diabetes can be divided into six major categories (insulins, insulin secretagogues, biguanides, glucosidase inhibitors, thiazolidinediones, SGLT2 inhibitors), each of which works through a different primary mechanism. .
  • these drugs have limited efficacy and do not address the most important issue, which is reduced cell function and associated obesity.
  • GLP-1 is a 30-amino acid long incretin hormone secreted by L cells in the intestine. GLP-1 stimulates insulin secretion, reduces glucagon secretion, inhibits gastric emptying, reduces appetite, and stimulates ⁇ -cell proliferation in a physiological and glucose-dependent manner. In non-clinical experiments, GLP-1 promotes ⁇ -cell persistence by stimulating the transcription of genes important for glucose-dependent insulin secretion and promoting ⁇ -cell regeneration. In healthy humans, GLP-1 plays an important role in regulating postprandial blood, leading to increased peripheral glucose absorption by stimulating glucose-dependent insulin secretion from the pancreas. GLP-1 also inhibits glucagon secretion, resulting in reduced hepatic glucose output. In addition, GLP-1 delays gastric emptying, slows small intestinal motility, and delays food absorption.
  • the object of the present invention is to provide a pharmaceutically acceptable salt (such as trihydroxymethylaminomethane salt or triethanolamine salt) of the compound represented by formula (I) and its solvate, crystal form and preparation method, its pharmaceutical composition and Its use in the preparation of medicines for diabetes or diabetes-related diseases.
  • a pharmaceutically acceptable salt such as trihydroxymethylaminomethane salt or triethanolamine salt
  • the crystal of the invention is easy to process, crystallize and handle; has good stability, is easy to take orally, and has good solubility and bioavailability.
  • the pharmaceutically acceptable salt is selected from tris(hydroxymethylaminomethane) salts or triethanolamine salts;
  • the molar ratio of compound represented by formula (I): pharmaceutically acceptable salt is 1:0.9 ⁇ 1:1.1;
  • the solvate is methanol
  • the present invention provides the trishydroxymethylaminomethane salt crystal form A of the compound represented by formula (I), using Cu-K ⁇ radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2 ⁇ positions: 8.97° ⁇ 0.2°, 9.32° ⁇ 0.2°, 9.92° ⁇ 0.2°, 10.68° ⁇ 0.2°, 17.50° ⁇ 0.2°, 20.17° ⁇ 0.2°;
  • the aforementioned trishydroxymethylaminomethane salt form A of the compound represented by formula (I) uses Cu-K ⁇ radiation, and its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2 ⁇ position: 6.02 ° ⁇ 0.2°, 8.97° ⁇ 0.2°, 9.32° ⁇ 0.2°, 9.92° ⁇ 0.2°, 10.68° ⁇ 0.2°, 12.03 ⁇ 0.2°, 13.77° ⁇ 0.2°, 15.07° ⁇ 0.2°, 16.18° ⁇ 0.2 °, 17.50° ⁇ 0.2°, 17.71° ⁇ 0.2°, 18.27° ⁇ 0.2°, 18.70° ⁇ 0.2°, 19.15° ⁇ 0.2°, 19.95° ⁇ 0.2°, 20.17° ⁇ 0.2°, 20.90° ⁇ 0.2°, 21.89° ⁇ 0.2°, 22.35° ⁇ 0.2°, 24.05° ⁇ 0.2°, 24.27° ⁇ 0.2°, 24.65° ⁇ 0.2°, 25.03° ⁇ 0.2°, 25.54° ⁇ 0.2°, 26.81° ⁇ 0.2°, 28.03°
  • the differential scanning calorimetry analysis curve and the thermogravimetric analysis curve of the trishydroxymethylaminomethane salt crystal form A of the compound represented by formula (I) are shown in Figure 2.
  • the present invention provides the trishydroxymethylaminomethane salt crystal form C of the compound represented by formula (I), using Cu-K ⁇ radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2 ⁇ positions: 10.55° ⁇ 0.2°, 13.69° ⁇ 0.2°, 19.41° ⁇ 0.2°, 20.70 ⁇ 0.2°;
  • the crystalline form C of the trishydroxymethylaminomethane salt of the compound represented by formula (I) uses Cu-K ⁇ radiation, and its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2 ⁇ position: 10.55 ° ⁇ 0.2°, 10.92° ⁇ 0.2°, 13.69° ⁇ 0.2°, 17.98° ⁇ 0.2°, 19.41° ⁇ 0.2°, 20.70 ⁇ 0.2°, 21.97 ⁇ 0.2°, 22.31 ⁇ 0.2°, 26.44 ⁇ 0.2°;
  • the crystalline form C of the trishydroxymethylaminomethane salt of the compound represented by formula (I) uses Cu-K ⁇ radiation, and its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2 ⁇ position: 5.44 ° ⁇ 0.2°, 8.12° ⁇ 0.2°, 10.17° ⁇ 0.2°, 10.55° ⁇ 0.2°, 10.92° ⁇ 0.2°, 11.56° ⁇ 0.2°, 13.08° ⁇ 0.2°, 13.69° ⁇ 0.2°, 14.49° ⁇ 0.2°, 15.26° ⁇ 0.2°, 17.06° ⁇ 0.2°, 17.78° ⁇ 0.2°, 17.98° ⁇ 0.2°, 18.41° ⁇ 0.2°, 19.41° ⁇ 0.2°, 20.70 ⁇ 0.2°, 21.25 ⁇ 0.2°, 21.97 ⁇ 0.2°, 22.31 ⁇ 0.2°, 23.27 ⁇ 0.2°, 24.23 ⁇ 0.2°, 25.24 ⁇ 0.2°, 25.70 ⁇ 0.2°, 26.44 ⁇ 0.2°, 27.49 ⁇ 0.2°, 28.71 ⁇ 0.2°;
  • the differential scanning calorimetry analysis curve and the thermogravimetric analysis curve of the trishydroxymethylaminomethane salt crystal Form C of the compound represented by formula (I) are shown in Figure 4.
  • the present invention provides the trishydroxymethylaminomethane salt crystal form D of the compound represented by formula (I), using Cu-K ⁇ radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2 ⁇ positions: 5.55° ⁇ 0.2°, 10.46° ⁇ 0.2°, 11.15° ⁇ 0.2°, 13.12° ⁇ 0.2°, 13.98° ⁇ 0.2°, 19.66° ⁇ 0.2°, 22.47° ⁇ 0.2°;
  • the trishydroxymethylaminomethane salt crystal form D of the compound represented by formula (I), using Cu-K ⁇ radiation is 5.55° ⁇ 0.2°, 8.34° ⁇ 0.2°, 10.46° ⁇ 0.2°, 11.15° ⁇ 0.2°, 11.50 ⁇ 0.2°, 13.12° ⁇ 0.2°, 13.98° ⁇ 0.2°, 19.66° ⁇ 0.2°, 20.80 ⁇ 0.2°, 22.08 ⁇ 0.2°, 22.47° ⁇ 0.2°, 23.26 ⁇ 0.2°, 26.73 ⁇ 0.2°;
  • the crystalline form D of the trishydroxymethylaminomethane salt of the compound represented by formula (I) uses Cu-K ⁇ radiation, and its X-ray powder diffraction pattern is as shown in Figure 5;
  • the present invention provides the triethanolamine salt crystal form A of the compound represented by formula (I).
  • its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2 ⁇ positions: 10.42° ⁇ 0.2°, 12.68° ⁇ 0.2 °, 14.80° ⁇ 0.2°, 16.37° ⁇ 0.2°, 16.94° ⁇ 0.2°, 23.39° ⁇ 0.2°;
  • the triethanolamine salt crystal form A of the compound represented by formula (I) uses Cu-K ⁇ radiation, and its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2 ⁇ position: 8.42° ⁇ 0.2°. , 10.42° ⁇ 0.2°, 12.68° ⁇ 0.2°, 13.28° ⁇ 0.2°, 14.80° ⁇ 0.2°, 16.37° ⁇ 0.2°, 16.94° ⁇ 0.2°, 18.04° ⁇ 0.2°, 22.26° ⁇ 0.2°, 23.39 ° ⁇ 0.2°, 24.98° ⁇ 0.2°, 25.55° ⁇ 0.2°;
  • the triethanolamine salt crystal form A of the compound represented by formula (I) uses Cu-K ⁇ radiation, and its X-ray powder diffraction pattern is as shown in Figure 7;
  • the present invention also provides a pharmaceutical composition, wherein the pharmaceutical composition contains a therapeutically effective amount of a pharmaceutically acceptable salt of the compound represented by formula (I) and its solvate or formula (I)
  • a pharmaceutically acceptable salt of the compound represented by formula (I) and its solvate or formula (I) The trishydroxymethylaminomethane salt crystal form A of the compound shown, the trishydroxymethylaminomethane salt crystal form C of the compound represented by formula (I), the trishydroxymethylaminomethane salt crystal form C of the compound represented by formula (I) Trishydroxymethylaminomethane salt crystal form D, triethanolamine salt crystal form A of the compound represented by formula (I) and pharmaceutically acceptable carriers or excipients.
  • the X-ray powder diffraction, DSC diagram, and TGA diagram disclosed in the present invention which are substantially the same, also belong to the scope of the present invention.
  • ester solvent refers to a combination of a lower organic acid containing 1 to 4 carbon atoms and a lower alcohol containing 1 to 6 carbon atoms. Specific examples include but are not limited to: acetic acid Ethyl ester, isopropyl acetate or butyl acetate.
  • the crystal structure of the present invention can be analyzed using various analytical techniques known to those of ordinary skill in the art, including but not limited to, X-ray powder diffraction (XRD), differential scanning calorimetry (DSC) and/or thermogravimetric analysis (Thermogravimetric Analysis (TGA), also called thermogravimetry (TG).
  • XRD X-ray powder diffraction
  • DSC differential scanning calorimetry
  • TGA thermogravimetric Analysis
  • TG thermogravimetry
  • the "2 ⁇ or 2 ⁇ angle" mentioned in the present invention refers to the diffraction angle, ⁇ is the Bragg angle, and the unit is ° or degree.
  • the error of 2 ⁇ The difference range can be ⁇ 0.3, ⁇ 0.2 or ⁇ 0.1.
  • crystal form of the present invention is not limited to the characteristic patterns that are exactly the same as those described in the drawings disclosed in the present invention, such as XRD, DSC, TGA, which patterns are basically the same as those described in the drawings or Any crystalline form with essentially the same characteristic pattern falls within the scope of the invention.
  • the melting peak height of the DSC curve depends on many factors related to sample preparation and instrument geometry, while the peak position is relatively insensitive to experimental details.
  • the crystalline compounds of the invention are characterized by a DSC pattern having characteristic peak positions with substantially the same properties as the DSC patterns provided in the drawings of the invention with an error tolerance of ⁇ 3°C.
  • the crystal form disclosed in the present invention can be prepared by the following common methods for preparing crystal forms:
  • the volatilization experiment is to evaporate the clarified solution of the sample at different temperatures until the solvent is dry.
  • the crystal slurry experiment is to stir the supersaturated solution of the sample (with insoluble solids present) at a certain temperature in different solvent systems.
  • the anti-solvent test is to take the sample and dissolve it in a good solvent, add the anti-solvent, stir the precipitated solid for a short time and then filter it immediately.
  • the cooling crystallization experiment is to dissolve a certain amount of sample into the corresponding solvent at high temperature, and then stir and crystallize directly at room temperature or low temperature.
  • the polymer template experiment is to add different types of polymer materials to the sample clarification solution and leave it open at room temperature to evaporate until the solvent dries.
  • the thermal method experiment is to treat the sample according to certain thermal method crystallization conditions and cool it to room temperature.
  • the water vapor diffusion experiment is to place the sample in a certain humidity environment at room temperature.
  • Figure 1 is an X-ray powder diffraction pattern of the trishydroxymethylaminomethane salt form A of compound I.
  • Figure 2 is a differential scanning calorimetry analysis curve and a thermogravimetric analysis chart of the trishydroxymethylaminomethane salt crystal form A of Compound I.
  • Figure 3 is an X-ray powder diffraction pattern of Form C of the trishydroxymethylaminomethane salt of Compound I.
  • Figure 4 is a differential scanning calorimetry analysis curve and a thermogravimetric analysis chart of the trishydroxymethylaminomethane salt crystal Form C of Compound I.
  • Figure 5 is an X-ray powder diffraction pattern of Form D of the trishydroxymethylaminomethane salt of Compound I.
  • Figure 6 is a differential scanning calorimetry analysis curve and a thermogravimetric analysis diagram of the trishydroxymethylaminomethane salt crystal form D of Compound I. Spectrum.
  • Figure 7 is the X-ray powder diffraction pattern of the triethanolamine salt crystal form A of compound I.
  • Figure 8 is a differential scanning calorimetry analysis curve and a thermogravimetric analysis chart of the triethanolamine salt crystal form A of Compound I.
  • HPLC measurement used LC-20AT (Shimadzu) high-pressure liquid chromatograph (Kromasil 100-5-C18, 4.6mm ⁇ 250mm).
  • XRD X-ray powder diffractometer Bruker D8Advance Diffractometer.
  • the 2 ⁇ scanning angle ranged from 3° to 45°, the scanning step was 0.02°, and the exposure time was 0.08 seconds.
  • the light tube voltage and current are 40kV and 40mA respectively, and the sample disk is a zero-background sample disk.
  • thermogravimetric analyzer is TA Discovery 550 (TA, US). Place 1-10mg sample into a balanced sample pan and automatically weigh in the TGA heating furnace. The sample was heated to the final temperature at a rate of 10°C/min, the nitrogen purge rate at the sample was 60 mL/min, and the nitrogen purge rate at the balance was 40 mL/min.
  • the known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from Titan Technology, Anaiji Chemical, Shanghai Demer, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, and Bailingwei Technology. Waiting for the company.
  • the room temperature is 20°C to 30°C.
  • Step 5 (S)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2-fluorobenzyl)-1-(oxy Alk-2-ylmethyl)-1H-thieno[2,3-d]imidazole-5-carboxylic acid methyl ester (1h)
  • Step 6 (S)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2-fluorobenzyl)-1-(oxy Alk-2-ylmethyl)-1H-thieno[2,3-d]imidazole-5-carboxylic acid (Compound I)
  • the compounds of the present invention were subjected to X-ray powder diffraction testing according to the following method.
  • DSC spectra were collected on TA Instruments discovery DSC 2500 and DSC 250 differential scanning calorimeters.
  • the test parameters of the trishydroxymethylaminomethane salt of compound I are shown in Table 2-1.
  • the test parameters of the triethanolamine salt of compound I As shown in Table 2-2.
  • TGA spectra were collected on TA Instruments discovery TGA 55 and TGA 550 thermogravimetric analyzers.
  • the test parameters of the trishydroxymethylaminomethane salt of compound I are shown in Table 3-1, and the test parameters of the triethanolamine salt of compound I are shown in the table. As shown in 3-2.
  • Freezing solution 90% FBS, 10% (V/V) DMSO
  • Compound I has a good agonistic effect on GLP-1 receptor, and its EC 50 value is less than 10 nM.
  • Cell line Chinese hamster ovary (CHO) cells stably expressing hERG potassium channel
  • CHO (Chinese Hamster Ovary) cells stably expressing hERG potassium channels were tested at room temperature using whole-cell patch clamp technology to record hERG potassium channel currents.
  • the glass microelectrode is drawn from a glass electrode blank (BF150-86-10, Sutter) by a drawing instrument.
  • the tip resistance after infusion of the electrode liquid is about 2-5M ⁇ .
  • the glass microelectrode can be connected by inserting it into the amplifier probe. to the patch clamp amplifier. Clamp voltage and data recording by pClamp 10
  • the software controls and records through the computer, with a sampling frequency of 10kHz and a filtering frequency of 2kHz.
  • the cells were clamped at -80mV, and the step voltage of induced hERG potassium current (I hERG ) was given a 2s depolarization voltage from -80mV to +20mV, and then repolarization to -50mV for 1s. then returns to -80mV.
  • This voltage stimulation was given every 10 s, and the administration process was started after confirming that the hERG potassium current was stable (at least 1 minute).
  • Compounds were administered for at least 1 minute at each concentration tested, and at least 2 cells (n ⁇ 2) were tested at each concentration.
  • Inhibition% represents the inhibition percentage of the hERG potassium current by the compound
  • I and Io represent the amplitude of the hERG potassium current after and before the addition of the drug, respectively.
  • X is the Log value of the detected concentration of the test product
  • Y is the inhibition percentage at the corresponding concentration
  • Bottom and Top are the minimum and maximum inhibition percentages respectively.
  • the compound of the present invention has good stability in human liver microsomes. For example, compared with the control compound, the clearance rate of compound 1 in human liver microsomes was significantly slower and the half-life was significantly longer.
  • the T 1/2 ratio of compound 1 to compound DZ-6 is greater than 3.
  • the compounds of the present invention have good oral properties.
  • the bioavailability of compound 1 in C57 mice is >25%.
  • mice Male cynomolgus monkeys (raised in Suzhou Leo Biotechnology Co., Ltd.), weighing 3 to 5 kg, were fasted overnight (14 to 18 hours). On the day of the experiment, 6 male cynomolgus monkeys were divided into 2 groups, with 3 monkeys in each group. The test compounds were administered intravenously and intragastrically at doses of 2 mg/kg and 10 mg/kg respectively, before and after administration. 5,15, 1 ml of blood was taken from the veins of the limbs at 30 minutes, 1, 2, 4, 6, 8, 10, 12 and 24 hours, placed in EDTAK2 centrifuge tubes, centrifuged at 6000 rpm and 4°C for 5 minutes, and then the plasma was collected.
  • the compounds of the present invention have good metabolic properties.
  • Compound I has a low clearance rate and a long half-life in monkeys, which has the advantages of longer drug action time and lower administration frequency.
  • each CYP isoenzyme is reflected by the production rate of each metabolite.
  • the compounds of the present invention have weak inhibitory activity against cytochrome P450 isoenzymes.
  • the inhibitory activity of compound I against CYP2C19, CYP1A2, CYP2D6 and CYP3A4-M is IC 50 >50 ⁇ M.
  • the assay uses a monolayer of Caco-2 cells incubated in triplicate in a 96-well Transwell plate.
  • Compounds containing compounds of the invention (2 ⁇ M) or control compounds: compound DZ-6, digoxin (10 ⁇ M), nadolol (2 ⁇ M) and metoprolol A transport buffer solution (HBSS, 10mM HEPES, pH 7.4 ⁇ 0.05) of lorol (2 ⁇ M) was added to the administration port hole on the apical or basal side.
  • Add DMSO-containing transport buffer solution to the corresponding receiving port hole. After incubating for 2 hours at 37 ⁇ 1°C, remove the cell plate and take appropriate amounts of samples from the top and bottom ends into a new 96-well plate.
  • Acetonitrile containing internal standard was then added to precipitate the protein.
  • Samples were analyzed using LC MS/MS and the concentrations of compounds of the invention and control compounds were determined. Concentration data were used to calculate apparent permeability coefficients for transport from the apical side to the basal side, and from the basal side to the apical side of the cell monolayer, and thus the efflux rate. The integrity of the cell monolayer after 2 hours of incubation was assessed by leakage of Lucifer Yellow.

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Abstract

The present invention provides a salt of a compound with GLP-1R agonist activity, as represented by formula (I), and/or a crystal form, and preparation and pharmaceutical use, wherein a pharmaceutically acceptable salt is selected from a tris(hydroxymethyl)aminomethane salt or a triethanolamine salt:

Description

一种噻吩[2,3-d]咪唑类化合物的盐及其晶型和在医药上的用途A salt of thiophene[2,3-d]imidazole compound, its crystal form and its use in medicine 技术领域Technical field
本发明涉及医药领域，具体的说，是涉及一种噻吩[2,3-d]咪唑类化合物的盐晶型和在医药上的用途。The present invention relates to the field of medicine. Specifically, it relates to a salt crystal form of a thiophene[2,3-d]imidazole compound and its use in medicine.
背景技术Background technique
糖尿病是一组以高血糖为特征的代谢性疾病。高血糖则是由于胰岛素分泌缺陷或其生物作用受损，或两者兼有引起。糖尿病时长期存在的高血糖，导致各种组织，特别是眼、肾、心脏、血管、神经的慢性损害、功能障碍，主要分为两种类型。1型糖尿病：胰岛B细胞破坏导致胰岛素绝对缺乏。2型糖尿病：胰岛素抵抗为主伴胰岛素相对性缺乏或胰岛素分泌受损为主伴胰岛素抵抗。Diabetes is a group of metabolic diseases characterized by high blood sugar. Hyperglycemia is caused by defective insulin secretion, impaired biological action, or both. The long-term high blood sugar in diabetes leads to chronic damage and dysfunction of various tissues, especially the eyes, kidneys, heart, blood vessels, and nerves. It is mainly divided into two types. Type 1 diabetes: Destruction of pancreatic B cells leads to absolute insulin deficiency. Type 2 diabetes: Insulin resistance is mainly associated with relative insulin deficiency or impaired insulin secretion is mainly associated with insulin resistance.
针对2型糖尿病的药物可分为六大类(胰岛素，促胰岛素分泌类，双胍类，葡萄糖苷酶抑制剂，噻唑烷二酮类，SGLT2抑制剂)，每一类通过不同的主要机制起作用。然而，除了GLP-1受体激动剂和SGLT2抑制剂外，这些药物的疗效有限，不能解决最重要的问题，即细胞功能下降和相关的肥胖。Drugs for type 2 diabetes can be divided into six major categories (insulins, insulin secretagogues, biguanides, glucosidase inhibitors, thiazolidinediones, SGLT2 inhibitors), each of which works through a different primary mechanism. . However, with the exception of GLP-1 receptor agonists and SGLT2 inhibitors, these drugs have limited efficacy and do not address the most important issue, which is reduced cell function and associated obesity.
GLP-1是一种30氨基酸的长肠促胰岛素激素，由肠内的L细胞分泌。GLP-1以生理和葡萄糖依赖的方式刺激胰岛素分泌，减少胰高血糖素分泌，抑制胃排空，减少食欲，刺激β细胞增殖。在非临床实验中，GLP-1通过刺激葡萄糖依赖性胰岛素分泌重要基因的转录和促进β细胞新生来促进β细胞的持续能力。在健康人中，GLP-1在调节餐后血液中起着重要作用，通过刺激胰腺的葡萄糖依赖性胰岛素分泌而导致周围葡萄糖吸收增加。GLP-1也抑制胰高血糖素的分泌，导致肝葡萄糖输出减少。此外，GLP-1延缓胃排空，减缓小肠运动，延缓食物吸收。GLP-1 is a 30-amino acid long incretin hormone secreted by L cells in the intestine. GLP-1 stimulates insulin secretion, reduces glucagon secretion, inhibits gastric emptying, reduces appetite, and stimulates β-cell proliferation in a physiological and glucose-dependent manner. In non-clinical experiments, GLP-1 promotes β-cell persistence by stimulating the transcription of genes important for glucose-dependent insulin secretion and promoting β-cell regeneration. In healthy humans, GLP-1 plays an important role in regulating postprandial blood, leading to increased peripheral glucose absorption by stimulating glucose-dependent insulin secretion from the pancreas. GLP-1 also inhibits glucagon secretion, resulting in reduced hepatic glucose output. In addition, GLP-1 delays gastric emptying, slows small intestinal motility, and delays food absorption.
GLP-1受体激动剂，如GLP-1、利拉鲁肽和exendin-4，都为多肽类药物，多用于注射。小分子GLP-1受体激动剂由于其具有口服生物利用度较高的潜力，成为近年来药物开发的热点。GLP-1 receptor agonists, such as GLP-1, liraglutide and exendin-4, are peptide drugs and are mostly used for injection. Small molecule GLP-1 receptor agonists have become a hot topic in drug development in recent years due to their potential for high oral bioavailability.
发明内容Contents of the invention
本发明的目的是提供一种式(I)所示化合物的可药用盐(如三羟甲基氨基甲烷盐或三乙醇胺盐)及其溶剂化物、晶型和制备方法，其药物组合物以及其在制备糖尿病或糖尿病相关的疾病的药物中的用途。 The object of the present invention is to provide a pharmaceutically acceptable salt (such as trihydroxymethylaminomethane salt or triethanolamine salt) of the compound represented by formula (I) and its solvate, crystal form and preparation method, its pharmaceutical composition and Its use in the preparation of medicines for diabetes or diabetes-related diseases.
本发明的晶体易于加工和结晶、处理；稳定性好、便于口服、具有较好的溶解度和生物利用度。The crystal of the invention is easy to process, crystallize and handle; has good stability, is easy to take orally, and has good solubility and bioavailability.
本发明提供一种式(I)所示化合物的可药用盐及其溶剂化物，
The present invention provides a pharmaceutically acceptable salt of the compound represented by formula (I) and its solvate,
在一些实施方案中，可药用盐选自三羟甲基氨基甲烷盐或三乙醇胺盐；In some embodiments, the pharmaceutically acceptable salt is selected from tris(hydroxymethylaminomethane) salts or triethanolamine salts;
在一些实施方案中，式(I)所示化合物：可药用盐的摩尔比为1:0.9～1:1.1；In some embodiments, the molar ratio of compound represented by formula (I): pharmaceutically acceptable salt is 1:0.9～1:1.1;
在一些实施方案中，式(I)所示化合物：可药用盐的摩尔比为1:1；In some embodiments, the molar ratio of compound represented by formula (I): pharmaceutically acceptable salt is 1:1;
在一些实施方案中，溶剂化物为甲醇；In some embodiments, the solvate is methanol;
本发明提供式(I)所示化合物的三羟甲基氨基甲烷盐晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：8.97°±0.2°、9.32°±0.2°、9.92°±0.2°、10.68°±0.2°、17.50°±0.2°、20.17°±0.2°；The present invention provides the trishydroxymethylaminomethane salt crystal form A of the compound represented by formula (I), using Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 8.97°±0.2°, 9.32°±0.2°, 9.92°±0.2°, 10.68°±0.2°, 17.50°±0.2°, 20.17°±0.2°;
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：8.97°±0.2°、9.32°±0.2°、9.92°±0.2°、10.68°±0.2°、15.07°±0.2°、17.50°±0.2°、17.71°±0.2°、19.95°±0.2°、20.17°±0.2°、24.27°±0.2°、26.81°±0.2°；In some embodiments, the aforementioned trishydroxymethylaminomethane salt form A of the compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2θ position: 8.97 °±0.2°, 9.32°±0.2°, 9.92°±0.2°, 10.68°±0.2°, 15.07°±0.2°, 17.50°±0.2°, 17.71°±0.2°, 19.95°±0.2°, 20.17°± 0.2°, 24.27°±0.2°, 26.81°±0.2°;
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：6.02°±0.2°、8.97°±0.2°、9.32°±0.2°、9.92°±0.2°、10.68°±0.2°、12.03±0.2°、13.77°±0.2°、15.07°±0.2°、16.18°±0.2°、17.50°±0.2°、17.71°±0.2°、18.27°±0.2°、18.70°±0.2°、19.15°±0.2°、19.95°±0.2°、20.17°±0.2°、20.90°±0.2°、21.89°±0.2°、22.35°±0.2°、24.05°±0.2°、24.27°±0.2°、24.65°±0.2°、25.03°±0.2°、25.54°±0.2°、26.81°±0.2°、28.03°±0.2°；In some embodiments, the aforementioned trishydroxymethylaminomethane salt form A of the compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2θ position: 6.02 °±0.2°, 8.97°±0.2°, 9.32°±0.2°, 9.92°±0.2°, 10.68°±0.2°, 12.03±0.2°, 13.77°±0.2°, 15.07°±0.2°, 16.18°±0.2 °, 17.50°±0.2°, 17.71°±0.2°, 18.27°±0.2°, 18.70°±0.2°, 19.15°±0.2°, 19.95°±0.2°, 20.17°±0.2°, 20.90°±0.2°, 21.89°±0.2°, 22.35°±0.2°, 24.05°±0.2°, 24.27°±0.2°, 24.65°±0.2°, 25.03°±0.2°, 25.54°±0.2°, 26.81°±0.2°, 28.03° ±0.2°;
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱如图1所示；In some embodiments, the trishydroxymethylaminomethane salt crystal form A of the compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern is as shown in Figure 1;
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型A，其差示扫描量热分析曲线(DSC)显示一条吸热曲线，其中T初始＝159.5℃，T峰值＝164.6℃，△H＝84.9J/g，其热重分析曲线(TGA)显示在120℃之前失重1.6％。In some embodiments, the differential scanning calorimetry curve (DSC) of the trishydroxymethylaminomethane salt crystal form A of the compound represented by formula (I) shows an endothermic curve, where T initial = 159.5°C , T peak = 164.6°C, ΔH = 84.9J/g, and its thermogravimetric analysis curve (TGA) shows a weight loss of 1.6% before 120°C.
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型A，其差示扫描量热分析曲线、热重分析曲线如图2所示。 In some embodiments, the differential scanning calorimetry analysis curve and the thermogravimetric analysis curve of the trishydroxymethylaminomethane salt crystal form A of the compound represented by formula (I) are shown in Figure 2.
本发明提供式(I)所示化合物的三羟甲基氨基甲烷盐晶型C，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：10.55°±0.2°、13.69°±0.2°、19.41°±0.2°、20.70±0.2°；The present invention provides the trishydroxymethylaminomethane salt crystal form C of the compound represented by formula (I), using Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 10.55°±0.2°, 13.69°±0.2°, 19.41°±0.2°, 20.70±0.2°;
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型C，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：10.55°±0.2°、10.92°±0.2°、13.69°±0.2°、17.98°±0.2°、19.41°±0.2°、20.70±0.2°、21.97±0.2°、22.31±0.2°、26.44±0.2°；In some embodiments, the crystalline form C of the trishydroxymethylaminomethane salt of the compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2θ position: 10.55 °±0.2°, 10.92°±0.2°, 13.69°±0.2°, 17.98°±0.2°, 19.41°±0.2°, 20.70±0.2°, 21.97±0.2°, 22.31±0.2°, 26.44±0.2°;
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型C，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：5.44°±0.2°、8.12°±0.2°、10.17°±0.2°、10.55°±0.2°、10.92°±0.2°、11.56°±0.2°、13.08°±0.2°、13.69°±0.2°、14.49°±0.2°、15.26°±0.2°、17.06°±0.2°、17.78°±0.2°、17.98°±0.2°、18.41°±0.2°、19.41°±0.2°、20.70±0.2°、21.25±0.2°、21.97±0.2°、22.31±0.2°、23.27±0.2°、24.23±0.2°、25.24±0.2°、25.70±0.2°、26.44±0.2°、27.49±0.2°、28.71±0.2°；In some embodiments, the crystalline form C of the trishydroxymethylaminomethane salt of the compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2θ position: 5.44 °±0.2°, 8.12°±0.2°, 10.17°±0.2°, 10.55°±0.2°, 10.92°±0.2°, 11.56°±0.2°, 13.08°±0.2°, 13.69°±0.2°, 14.49°± 0.2°, 15.26°±0.2°, 17.06°±0.2°, 17.78°±0.2°, 17.98°±0.2°, 18.41°±0.2°, 19.41°±0.2°, 20.70±0.2°, 21.25±0.2°, 21.97 ±0.2°, 22.31±0.2°, 23.27±0.2°, 24.23±0.2°, 25.24±0.2°, 25.70±0.2°, 26.44±0.2°, 27.49±0.2°, 28.71±0.2°;
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型C，使用Cu-Kα辐射，其X-射线粉末衍射图谱如图3所示；In some embodiments, the crystal form C of the trishydroxymethylaminomethane salt of the compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern is as shown in Figure 3;
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型C，其差示扫描量热分析曲线(DSC)显示三个吸热峰，其中T峰值＝49.6℃、T峰值＝112.8℃和T峰值＝161.4℃，其热重分析曲线(TGA)显示在130℃之前失重4.6％。In some embodiments, the differential scanning calorimetry curve (DSC) of the trishydroxymethylaminomethane salt crystal form C of the compound represented by formula (I) shows three endothermic peaks, wherein T peak value = 49.6 ℃, T peak = 112.8° C. and T peak = 161.4° C., and its thermogravimetric analysis curve (TGA) shows a weight loss of 4.6% before 130° C.
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型C，其差示扫描量热分析曲线、热重分析曲线如图4所示。In some embodiments, the differential scanning calorimetry analysis curve and the thermogravimetric analysis curve of the trishydroxymethylaminomethane salt crystal Form C of the compound represented by formula (I) are shown in Figure 4.
本发明提供式(I)所示化合物的三羟甲基氨基甲烷盐晶型D，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：5.55°±0.2°、10.46°±0.2°、11.15°±0.2°、13.12°±0.2°、13.98°±0.2°、19.66°±0.2°、22.47°±0.2°；The present invention provides the trishydroxymethylaminomethane salt crystal form D of the compound represented by formula (I), using Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 5.55°±0.2°, 10.46°±0.2°, 11.15°±0.2°, 13.12°±0.2°, 13.98°±0.2°, 19.66°±0.2°, 22.47°±0.2°;
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型D，使用Cu-Kα辐射，5.55°±0.2°、8.34±0.2°、10.46°±0.2°、11.15°±0.2°、11.50±0.2°、13.12°±0.2°、13.98°±0.2°、19.66°±0.2°、20.80±0.2°、22.08±0.2°、22.47°±0.2°、23.26±0.2°、26.73±0.2°；In some embodiments, the trishydroxymethylaminomethane salt crystal form D of the compound represented by formula (I), using Cu-Kα radiation, is 5.55°±0.2°, 8.34°±0.2°, 10.46°±0.2°, 11.15°±0.2°, 11.50±0.2°, 13.12°±0.2°, 13.98°±0.2°, 19.66°±0.2°, 20.80±0.2°, 22.08±0.2°, 22.47°±0.2°, 23.26±0.2°, 26.73±0.2°;
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型D，使用Cu-Kα辐射，其X-射线粉末衍射图谱如图5所示；In some embodiments, the crystalline form D of the trishydroxymethylaminomethane salt of the compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern is as shown in Figure 5;
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型D，其差示扫描量热分析曲线(DSC)显示两个吸热曲线，其中T峰值＝119.2℃，T峰值＝133.9℃，其热重分析曲线(TGA)显示在140℃之前失重3.7％。 In some embodiments, the differential scanning calorimetry curve (DSC) of the trishydroxymethylaminomethane salt crystal form D of the compound represented by formula (I) shows two endothermic curves, in which T peak value = 119.2 ℃, T peak = 133.9 ℃, and its thermogravimetric analysis curve (TGA) shows a weight loss of 3.7% before 140 ℃.
在一些实施方案中，前述的式(I)所示化合物的三羟甲基氨基甲烷盐晶型D，其差示扫描量热分析曲线、热重分析曲线如图6所示。In some embodiments, the differential scanning calorimetry analysis curve and the thermogravimetric analysis curve of the trishydroxymethylaminomethane salt crystal form D of the compound represented by formula (I) are shown in Figure 6.
本发明提供式(I)所示化合物的三乙醇胺盐晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：10.42°±0.2°、12.68°±0.2°、14.80°±0.2°、16.37°±0.2°、16.94°±0.2°、23.39°±0.2°；The present invention provides the triethanolamine salt crystal form A of the compound represented by formula (I). Using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 10.42°±0.2°, 12.68°±0.2 °, 14.80°±0.2°, 16.37°±0.2°, 16.94°±0.2°, 23.39°±0.2°;
在一些实施方案中，前述的式(I)所示化合物的三乙醇胺盐晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：8.42°±0.2°、10.42°±0.2°、12.68°±0.2°、13.28°±0.2°、14.80°±0.2°、16.37°±0.2°、16.94°±0.2°、18.04°±0.2°、22.26°±0.2°、23.39°±0.2°、24.98°±0.2°、25.55°±0.2°；In some embodiments, the triethanolamine salt crystal form A of the compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2θ position: 8.42°±0.2°. , 10.42°±0.2°, 12.68°±0.2°, 13.28°±0.2°, 14.80°±0.2°, 16.37°±0.2°, 16.94°±0.2°, 18.04°±0.2°, 22.26°±0.2°, 23.39 °±0.2°, 24.98°±0.2°, 25.55°±0.2°;
在一些实施方案中，前述的式(I)所示化合物的三乙醇胺盐晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：4.21±0.2°、6.30°±0.2°、8.42°±0.2°、10.42°±0.2°、12.68°±0.2°、13.28°±0.2°、14.35°±0.2°、14.80°±0.2°、15.11°±0.2°、16.10°±0.2°、16.37°±0.2°、16.94°±0.2°、17.32°±0.2°、18.04°±0.2°、18.72°±0.2°、19.83°±0.2°、20.96°±0.2°、21.23°±0.2°、21.39°±0.2°、21.70°±0.2°、21.85°±0.2°、22.26°±0.2°、22.61°±0.2°、23.39°±0.2°、24.22°±0.2°、24.55°±0.2°、24.98°±0.2°、25.55°±0.2°、26.48°±0.2°、28.07°±0.2°、29.90°±0.2°In some embodiments, the triethanolamine salt crystal form A of the compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 4.21±0.2°, 6.30°±0.2°, 8.42°±0.2°, 10.42°±0.2°, 12.68°±0.2°, 13.28°±0.2°, 14.35°±0.2°, 14.80°±0.2°, 15.11°±0.2°, 16.10° ±0.2°, 16.37°±0.2°, 16.94°±0.2°, 17.32°±0.2°, 18.04°±0.2°, 18.72°±0.2°, 19.83°±0.2°, 20.96°±0.2°, 21.23°±0.2 °, 21.39°±0.2°, 21.70°±0.2°, 21.85°±0.2°, 22.26°±0.2°, 22.61°±0.2°, 23.39°±0.2°, 24.22°±0.2°, 24.55°±0.2°, 24.98°±0.2°, 25.55°±0.2°, 26.48°±0.2°, 28.07°±0.2°, 29.90°±0.2°
在一些实施方案中，前述的式(I)所示化合物的三乙醇胺盐晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱如图7所示；In some embodiments, the triethanolamine salt crystal form A of the compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern is as shown in Figure 7;
在一些实施方案中，前述的式(I)所示化合物的三乙醇胺盐晶型A，其差示扫描量热分析曲线(DSC)显示一个吸热曲线，其中T初始＝146.2℃，T峰值＝149.9℃，△H＝73.6J/g，其热重分析曲线(TGA)显示在120℃之前失重1％。In some embodiments, the differential scanning calorimetry curve (DSC) of the triethanolamine salt crystal form A of the compound represented by formula (I) shows an endothermic curve, in which T initial = 146.2°C and T peak = 149.9°C, ΔH=73.6J/g, and its thermogravimetric analysis curve (TGA) shows a weight loss of 1% before 120°C.
在一些实施方案中，前述的式(I)所示化合物的三乙醇胺盐晶型A，其差示扫描量热分析曲线、热重分析曲线如图8所示。In some embodiments, the differential scanning calorimetry analysis curve and the thermogravimetric analysis curve of the triethanolamine salt crystal form A of the compound represented by formula (I) are shown in Figure 8.
本发明还提供一种式(I)所示化合物的可药用盐的制备方法，其中，所述方法包括：以式(I)所示化合物和酸成盐的步骤，所述的酸优选三羟甲基氨基甲烷盐或三乙醇胺盐。The present invention also provides a method for preparing a pharmaceutically acceptable salt of the compound represented by formula (I), wherein the method includes: the step of forming a salt from the compound represented by formula (I) and an acid, and the acid is preferably three Hydroxymethylaminomethane salt or triethanolamine salt.
在一些实施方案中，前述制备方法中所使用的溶剂选自C1-6卤代烷烃类溶剂、C2-6酯类溶剂、C2-6醚类溶剂、C1-6醇类溶剂或水中的一种或多种；In some embodiments, the solvent used in the aforementioned preparation method is selected from C 1-6 halogenated alkane solvents, C 2-6 ester solvents, C 2-6 ether solvents, C 1-6 alcohol solvents or water one or more;
在一些实施方案中，前述制备方法中所使用的溶剂选自二氯甲烷、1,2-二氯乙烷、乙酸乙酯、甲醇、乙醇、异丙醇、丙醇、***、四氢呋喃和水中的一种或多种。In some embodiments, the solvent used in the aforementioned preparation method is selected from the group consisting of dichloromethane, 1,2-dichloroethane, ethyl acetate, methanol, ethanol, isopropyl alcohol, propanol, diethyl ether, tetrahydrofuran and water. one or more.
又一方面，本发明还提供了一种药物组合物，其中，所述药物组合物含有治疗有效量的前述的式(I)所示化合物的可药用盐及其溶剂化物或者式(I)所示化合物的三羟甲基氨基甲烷盐晶型A、式(I)所示化合物的三羟甲基氨基甲烷盐晶型C、式(I)所示化合物的 三羟甲基氨基甲烷盐晶型D、式(I)所示化合物的三乙醇胺盐晶型A及药学上可接受的载体或赋形剂。In another aspect, the present invention also provides a pharmaceutical composition, wherein the pharmaceutical composition contains a therapeutically effective amount of a pharmaceutically acceptable salt of the compound represented by formula (I) and its solvate or formula (I) The trishydroxymethylaminomethane salt crystal form A of the compound shown, the trishydroxymethylaminomethane salt crystal form C of the compound represented by formula (I), the trishydroxymethylaminomethane salt crystal form C of the compound represented by formula (I) Trishydroxymethylaminomethane salt crystal form D, triethanolamine salt crystal form A of the compound represented by formula (I) and pharmaceutically acceptable carriers or excipients.
本申请中所述“有效量”或“治疗有效量”是指给予足够量的本申请公开的化合物，其将在某种程度上缓解所治疗的疾病或病症(例如糖尿病)的一种或多种症状。As used herein, an "effective amount" or a "therapeutically effective amount" refers to administration of a sufficient amount of a compound disclosed herein that will alleviate to some extent one or more of the diseases or conditions being treated (e.g., diabetes). symptoms.
在一些实施方案中，结果是减少和/或缓和疾病的体征、症状或原因，或生物***的任何其它希望改变。In some embodiments, the result is reduction and/or alleviation of signs, symptoms, or causes of disease, or any other desired change in a biological system.
例如，针对治疗用途的“有效量”是提供临床上显著的疾病症状降低所需的包含本申请公开的化合物的组合物的量。治疗有效量的实例(以游离碱形式计算)包括但不限于1-600mg、2-600mg、3-600mg、4-600mg、5-600mg、6-600mg、10-600mg、20-600mg、25-600mg、30-600mg、40-600mg、50-600mg、60-600mg、70-600mg、75-600mg、80-600mg、90-600mg、100-600mg、200-600mg、1-500mg、2-500mg、3-500mg、4-500mg、5-500mg、6-500mg、10-500mg、20-500mg、25-500mg、30-500mg、40-500mg、50-500mg、60-500mg、70-500mg、75-500mg、80-500mg、90-500mg、100-500mg、125-500mg、150-500mg、200-500mg、250-500mg、300-500mg、400-500mg、5-400mg、10-400mg、20-400mg、25-400mg、30-400mg、40-400mg、50-400mg、60-400mg、70-400mg、75-400mg、80-400mg、90-400mg、100-400mg、125-400mg、150-400mg、200-400mg、250-400mg、300-400mg、1-300mg、2-300mg、5-300mg、10-300mg、20-300mg、25-300mg、30-300mg、40-300mg、50-300mg、60-300mg、70-300mg、75-300mg、80-300mg、90-300mg、100-300mg、125-300mg、150-300mg、200-300mg、250-300mg、1-200mg、2-200mg、5-200mg、10-200mg、20-200mg、25-200mg、30-200mg、40-200mg、50-200mg、60-200mg、70-200mg、75-200mg、80-200mg、90-200mg、100-200mg、125-200mg、150-200mg、1-100mg、2-100mg、5-100mg、10-100mg、15-100mg、20-100mg、25-100mg、30-100mg、40-100mg、50-100mg、60-100mg、70-100mg、75-100mg、80-100mg、90-100mg；在一些实施方案中，治疗有效量的实例包括但不限于1mg、5mg、10mg、15mg、20mg、25mg、30mg、35mg、40mg、45mg、50mg、55mg、65mg、70mg、75mg、80mg、85mg、90mg、95mg、100mg、110mg、120mg、125mg、130mg、140mg、150mg、160mg、170mg、180mg、190mg、200mg、210mg、220mg、230mg、240mg、250mg、300mg。For example, an "effective amount" for therapeutic use is the amount of a composition containing a compound disclosed herein that is required to provide a clinically significant reduction in disease symptoms. Examples of therapeutically effective amounts (calculated as free base) include, but are not limited to, 1-600 mg, 2-600 mg, 3-600 mg, 4-600 mg, 5-600 mg, 6-600 mg, 10-600 mg, 20-600 mg, 25- 600mg, 30-600mg, 40-600mg, 50-600mg, 60-600mg, 70-600mg, 75-600mg, 80-600mg, 90-600mg, 100-600mg, 200-600mg, 1-500mg, 2-500mg, 3-500mg, 4-500mg, 5-500mg, 6-500mg, 10-500mg, 20-500mg, 25-500mg, 30-500mg, 40-500mg, 50-500mg, 60-500mg, 70-500mg, 75- 500mg, 80-500mg, 90-500mg, 100-500mg, 125-500mg, 150-500mg, 200-500mg, 250-500mg, 300-500mg, 400-500mg, 5-400mg, 10-400mg, 20-400mg, 25-400mg, 30-400mg, 40-400mg, 50-400mg, 60-400mg, 70-400mg, 75-400mg, 80-400mg, 90-400mg, 100-400mg, 125-400mg, 150-400mg, 200- 400mg, 250-400mg, 300-400mg, 1-300mg, 2-300mg, 5-300mg, 10-300mg, 20-300mg, 25-300mg, 30-300mg, 40-300mg, 50-300mg, 60-300mg, 70-300mg, 75-300mg, 80-300mg, 90-300mg, 100-300mg, 125-300mg, 150-300mg, 200-300mg, 250-300mg, 1-200mg, 2-200mg, 5-200mg, 10- 200mg, 20-200mg, 25-200mg, 30-200mg, 40-200mg, 50-200mg, 60-200mg, 70-200mg, 75-200mg, 80-200mg, 90-200mg, 100-200mg, 125-200mg, 150-200mg, 1-100mg, 2-100mg, 5-100mg, 10-100mg, 15-100mg, 20-100mg, 25-100mg, 30-100mg, 40-100mg, 50-100mg, 60-100mg, 70- 100 mg, 75-100 mg, 80-100 mg, 90-100 mg; in some embodiments, examples of therapeutically effective amounts include, but are not limited to, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg , 55mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 110mg, 120mg, 125mg, 130mg, 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 210mg, 220mg, 230mg, 240mg, 250mg , 300mg.
再一方面，本发明还提供了前述的式(I)所示化合物的可药用盐及其溶剂化物或者式(I)所示化合物的三羟甲基氨基甲烷盐晶型A、式(I)所示化合物的三羟甲基氨基甲烷盐晶型C、式(I)所示化合物的三羟甲基氨基甲烷盐晶型D、式(I)所示化合物的三乙醇胺盐晶型A、或者前述药物组合物在制备用于治疗糖尿病或或糖尿病相关的疾病药物中的应用。On the other hand, the present invention also provides pharmaceutically acceptable salts and solvates thereof of the compound represented by formula (I) or trishydroxymethylaminomethane salt crystal form A and formula (I) of the compound represented by formula (I). ) trishydroxymethylaminomethane salt crystal form C of the compound represented by formula (I), trishydroxymethylaminomethane salt crystal form D of the compound represented by formula (I), triethanolamine salt crystal form A of the compound represented by formula (I), Or the application of the aforementioned pharmaceutical composition in the preparation of drugs for treating diabetes or diabetes-related diseases.
其中可以理解的是，本发明所述的“优选地，……，其X-射线粉末衍射图谱进一步在 以下2θ位置具有特征衍射峰”，或者“更优选地，……，其X-射线粉末衍射图谱进一步在以下2θ位置具有特征衍射峰”等等诸如此类的表达，是指在前面所述2θ位置具有特征衍射峰的基础上，进一步还在所述的“以下2θ位置”具有特征衍射峰。It can be understood that the "preferably,..." of the present invention, its X-ray powder diffraction pattern is further in Expressions such as "having a characteristic diffraction peak at the following 2θ position", or "more preferably, ..., whose X-ray powder diffraction pattern further has a characteristic diffraction peak at the following 2θ position" and the like refer to having a characteristic diffraction peak at the aforementioned 2θ position. In addition to the characteristic diffraction peaks, it further has a characteristic diffraction peak at the "lower 2θ position".
本发明公开的X-射线粉末衍射或DSC图、TGA图，与其实质上相同的也属于本发明的范围。The X-ray powder diffraction, DSC diagram, and TGA diagram disclosed in the present invention, which are substantially the same, also belong to the scope of the present invention.
除非有相反的陈述，在说明书和权利要求书中使用的术语具有下述含义。Unless stated to the contrary, the terms used in the specification and claims have the following meanings.
“IC50”指半数抑制浓度，指达到最大抑制效果一半时的浓度。“IC 50 ” refers to the half inhibitory concentration, which is the concentration at which half of the maximum inhibitory effect is achieved.
本发明所述的“醚类溶剂”是指含有醚键-O-且碳原子数为1至10个的链状化合物或环状化合物，具体实例包括但不限于：四氢呋喃、***、丙二醇甲醚、甲基叔丁基醚、异丙醚或1,4-二氧六环。The "ether solvent" mentioned in the present invention refers to a chain compound or a cyclic compound containing an ether bond -O- and having 1 to 10 carbon atoms. Specific examples include but are not limited to: tetrahydrofuran, ether, and propylene glycol methyl ether. , methyl tert-butyl ether, isopropyl ether or 1,4-dioxane.
本发明所述的“醇类溶剂”是指一个或多个“羟基”取代“C1-6烷基”上的一个或多个氢原子所衍生的基团，所述“羟基”和“C1-6烷基”如前文所定义，具体实例包括但不限于：甲醇、乙醇、异丙醇、正丙醇、异戊醇或三氟乙醇。The "alcoholic solvent" mentioned in the present invention refers to a group derived from one or more "hydroxyl groups" replacing one or more hydrogen atoms on the "C 1-6 alkyl group". The "hydroxyl group" and "C "1-6 alkyl" is as defined above. Specific examples include but are not limited to: methanol, ethanol, isopropanol, n-propanol, isoamyl alcohol or trifluoroethanol.
本发明所述的“酯类溶剂”是指含碳原子数为1至4个的低级有机酸与含碳原子数为1至6个的低级醇的结合物，具体实例包括但不限于：乙酸乙酯、乙酸异丙酯或乙酸丁酯。The "ester solvent" mentioned in the present invention refers to a combination of a lower organic acid containing 1 to 4 carbon atoms and a lower alcohol containing 1 to 6 carbon atoms. Specific examples include but are not limited to: acetic acid Ethyl ester, isopropyl acetate or butyl acetate.
本发明所述的“酮类溶剂”是指羰基(-C(O)-)与两个烃基相连的化合物，根据分子中烃基的不同，酮可分为脂肪酮、脂环酮、芳香酮、饱和酮和不饱和酮，具体实例包括但不限于：丙酮、苯乙酮、4-甲基-2-戊酮。The "ketone solvent" mentioned in the present invention refers to a compound in which a carbonyl group (-C(O)-) is connected to two hydrocarbon groups. According to the different hydrocarbon groups in the molecule, ketones can be divided into aliphatic ketones, alicyclic ketones, aromatic ketones, Specific examples of saturated ketones and unsaturated ketones include, but are not limited to: acetone, acetophenone, and 4-methyl-2-pentanone.
本发明所述的“腈类溶剂”是指一个或多个“氰基”取代“C1-6烷基”上的一个或多个氢原子所衍生的基团，所述“氰基”和“C1-6烷基”如前文所定义，具体实例包括但不限于：乙腈或丙腈。The "nitrile solvent" mentioned in the present invention refers to a group derived from one or more "cyano groups" replacing one or more hydrogen atoms on the "C 1-6 alkyl group", and the "cyano group" and "C 1-6 alkyl" is as defined above, and specific examples include but are not limited to: acetonitrile or propionitrile.
本发明所述的“卤代烃类溶剂”是指一个或多个“卤素原子”取代“C1-6烷基”上的一个或多个氢原子所衍生的基团，所述“卤素原子”和“C1-6烷基”如前文所定义，具体实例包括但不限于：二氯甲烷、1,2-二氯乙烷、氯仿或四氯化碳。The "halogenated hydrocarbon solvent" mentioned in the present invention refers to a group derived from one or more "halogen atoms" replacing one or more hydrogen atoms on the "C 1-6 alkyl group". The "halogen atoms"" and "C 1-6 alkyl" are as defined above. Specific examples include but are not limited to: methylene chloride, 1,2-dichloroethane, chloroform or carbon tetrachloride.
如本发明所用，“本发明的晶体”、“本发明的晶型”、“本发明的多晶型物”等可互换使用。As used herein, "crystals of the present invention", "crystalline forms of the present invention", "polymorphs of the present invention", etc. are used interchangeably.
本发明晶型结构可以使用本领域普通技术人员已知的各种分析技术分析，包括但不限于，X-射线粉末衍射(XRD)、示差扫描热法(DSC)和/或热重分析(Thermogravimetric Analysis，TGA)，又叫热重法(Thermogravimetry，TG)。The crystal structure of the present invention can be analyzed using various analytical techniques known to those of ordinary skill in the art, including but not limited to, X-ray powder diffraction (XRD), differential scanning calorimetry (DSC) and/or thermogravimetric analysis (Thermogravimetric Analysis (TGA), also called thermogravimetry (TG).
本发明所述的“2θ或2θ角度”是指衍射角，θ为布拉格角，单位为°或度，所述2θ的误 差范围可以是±0.3、±0.2或±0.1。The "2θ or 2θ angle" mentioned in the present invention refers to the diffraction angle, θ is the Bragg angle, and the unit is ° or degree. The error of 2θ The difference range can be ±0.3, ±0.2 or ±0.1.
可以理解的是，本发明描述的和保护的数值为近似值。数值内的变化可能归因于设备的校准、设备误差、晶体的纯度、晶体大小、样本大小以及其他因素。It is understood that the numerical values described and claimed herein are approximations. Variations within values may be attributed to calibration of the equipment, equipment errors, purity of the crystals, crystal size, sample size, and other factors.
可以理解的是，本发明的晶型不限于与本发明公开的附图中描述的特征图谱完全相同的特征图谱，比如XRD、DSC、TGA，具有与附图中描述的哪些图谱基本上相同或本质上相同的特征图谱的任何晶型均落入本发明的范围内。It can be understood that the crystal form of the present invention is not limited to the characteristic patterns that are exactly the same as those described in the drawings disclosed in the present invention, such as XRD, DSC, TGA, which patterns are basically the same as those described in the drawings or Any crystalline form with essentially the same characteristic pattern falls within the scope of the invention.
可以理解的是，差示扫描量热(DSC)领域中所熟知的，DSC曲线的熔融峰高取决于与样品制备和仪器几何形状有关的许多因素，而峰位置对实验细节相对不敏感。因此，在一些实施方案中，本发明的结晶化合物的特征在于具有特征峰位置的DSC图，具有与本发明附图中提供的DSC图实质上相同的性质，误差容限为±3℃。It can be understood that, as is well known in the field of differential scanning calorimetry (DSC), the melting peak height of the DSC curve depends on many factors related to sample preparation and instrument geometry, while the peak position is relatively insensitive to experimental details. Thus, in some embodiments, the crystalline compounds of the invention are characterized by a DSC pattern having characteristic peak positions with substantially the same properties as the DSC patterns provided in the drawings of the invention with an error tolerance of ±3°C.
本发明公开的晶型可以经如下的常见的制备晶型的方法制备：The crystal form disclosed in the present invention can be prepared by the following common methods for preparing crystal forms:
1、挥发实验是将样品澄清溶液在不同温度下敞口挥发至溶剂干。1. The volatilization experiment is to evaporate the clarified solution of the sample at different temperatures until the solvent is dry.
2、晶浆实验是将样品的过饱和溶液(有不溶固体存在)在不同溶剂体系中某个温度下进行搅拌。2. The crystal slurry experiment is to stir the supersaturated solution of the sample (with insoluble solids present) at a certain temperature in different solvent systems.
3、抗溶剂实验是取样品溶解在良溶剂中，加入抗溶剂，析出固体短时搅拌后立即过滤处理。3. The anti-solvent test is to take the sample and dissolve it in a good solvent, add the anti-solvent, stir the precipitated solid for a short time and then filter it immediately.
4、冷却结晶实验是在高温下将一定量的样品溶解到相应溶剂中，然后直接在室温或低温搅拌析晶。4. The cooling crystallization experiment is to dissolve a certain amount of sample into the corresponding solvent at high temperature, and then stir and crystallize directly at room temperature or low temperature.
5、高分子模板实验是在样品澄清溶液中加入不同种类的高分子材料，置于室温下敞口挥发至溶剂干。5. The polymer template experiment is to add different types of polymer materials to the sample clarification solution and leave it open at room temperature to evaporate until the solvent dries.
6、热方法实验是将样品按一定热方法结晶条件处理并冷却至室温。6. The thermal method experiment is to treat the sample according to certain thermal method crystallization conditions and cool it to room temperature.
7、水汽扩散实验是将样品在室温下一定湿度环境中放置。7. The water vapor diffusion experiment is to place the sample in a certain humidity environment at room temperature.
附图说明Description of the drawings
图1为化合物I的三羟甲基氨基甲烷盐晶型A的X-射线粉末衍射图谱。Figure 1 is an X-ray powder diffraction pattern of the trishydroxymethylaminomethane salt form A of compound I.
图2为化合物I的三羟甲基氨基甲烷盐晶型A的差示扫描量热分析曲线及热重分析图谱。Figure 2 is a differential scanning calorimetry analysis curve and a thermogravimetric analysis chart of the trishydroxymethylaminomethane salt crystal form A of Compound I.
图3为化合物I的三羟甲基氨基甲烷盐晶型C的X-射线粉末衍射图谱。Figure 3 is an X-ray powder diffraction pattern of Form C of the trishydroxymethylaminomethane salt of Compound I.
图4为化合物I的三羟甲基氨基甲烷盐晶型C的差示扫描量热分析曲线及热重分析图谱。Figure 4 is a differential scanning calorimetry analysis curve and a thermogravimetric analysis chart of the trishydroxymethylaminomethane salt crystal Form C of Compound I.
图5为化合物I的三羟甲基氨基甲烷盐晶型D的X-射线粉末衍射图谱。Figure 5 is an X-ray powder diffraction pattern of Form D of the trishydroxymethylaminomethane salt of Compound I.
图6为化合物I的三羟甲基氨基甲烷盐晶型D的差示扫描量热分析曲线及热重分析图 谱。Figure 6 is a differential scanning calorimetry analysis curve and a thermogravimetric analysis diagram of the trishydroxymethylaminomethane salt crystal form D of Compound I. Spectrum.
图7为化合物I的三乙醇胺盐晶型A的X-射线粉末衍射图谱Figure 7 is the X-ray powder diffraction pattern of the triethanolamine salt crystal form A of compound I.
图8为化合物I的三乙醇胺盐晶型A的差示扫描量热分析曲线及热重分析图谱。Figure 8 is a differential scanning calorimetry analysis curve and a thermogravimetric analysis chart of the triethanolamine salt crystal form A of Compound I.
具体实施方式Detailed ways
以下结合附图及实施例详细说明本发明的技术方案，但本发明的保护范围包括但是不限于此。The technical solution of the present invention will be described in detail below with reference to the accompanying drawings and examples, but the protection scope of the present invention includes but is not limited thereto.
化合物的结构是通过核磁共振(NMR)或(和)质谱(MS)来确定的。NMR位移(δ)以10-6(ppm)的单位给出。NMR的测定是用(Bruker Avance III 400和Bruker Avance 300)核磁仪，测定溶剂为氘代二甲基亚砜(DMSO-d6)，氘代氯仿(CDCl3)，氘代甲醇(CD3OD)，内标为四甲基硅烷(TMS)。The structure of the compound is determined by nuclear magnetic resonance (NMR) or/and mass spectrometry (MS). NMR shifts (δ) are given in units of 10 -6 (ppm). NMR was measured using (Bruker Avance III 400 and Bruker Avance 300) nuclear magnetic instruments, and the measurement solvents were deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated chloroform (CDCl 3 ), and deuterated methanol (CD 3 OD ), the internal standard is tetramethylsilane (TMS).
MS的测定用(Agilent 6120B(ESI)和Agilent 6120B(APCI))。For MS measurement (Agilent 6120B (ESI) and Agilent 6120B (APCI)).
HPLC的测定使用LC-20AT(岛津)高压液相色谱仪(Kromasil 100-5-C18,4.6mm×250mm)。HPLC measurement used LC-20AT (Shimadzu) high-pressure liquid chromatograph (Kromasil 100-5-C18, 4.6mm×250mm).
XRD的测定使用X射线粉末衍射仪Bruker D8Advance Diffractometer进行分析。2θ扫描角度从3°到45°，扫描步长为0.02°，曝光时间为0.08秒。测试样品时光管电压和电流分别为40kV和40mA，样品盘为零背景样品盘。XRD was measured using an X-ray powder diffractometer Bruker D8Advance Diffractometer. The 2θ scanning angle ranged from 3° to 45°, the scanning step was 0.02°, and the exposure time was 0.08 seconds. When testing the sample, the light tube voltage and current are 40kV and 40mA respectively, and the sample disk is a zero-background sample disk.
TGA测试条件：热重分析仪的型号为TA Discovery 550(TA，US)。将1-10mg样品置于已平衡的样品盘中，在TGA加热炉内自动称量。样品以10℃/min的速率加热至最终温度，样品处氮气吹扫速度为60mL/min，天平处氮气吹扫速度为40mL/min。TGA test conditions: The model of thermogravimetric analyzer is TA Discovery 550 (TA, US). Place 1-10mg sample into a balanced sample pan and automatically weigh in the TGA heating furnace. The sample was heated to the final temperature at a rate of 10°C/min, the nitrogen purge rate at the sample was 60 mL/min, and the nitrogen purge rate at the balance was 40 mL/min.
DSC测试条件：差示扫描量热分析仪的型号为TA Discovery 250(TA，US)。1-2mg样品经精确称重后置于标准盘或扎孔的DSC Tzero样品盘中，以10℃/min的速率加热至最终温度，炉内氮气吹扫速度为50mL/min。DSC test conditions: The model of differential scanning calorimetry analyzer is TA Discovery 250 (TA, US). The 1-2mg sample is accurately weighed and placed in a standard plate or a perforated DSC Tzero sample plate, heated to the final temperature at a rate of 10°C/min, and the nitrogen purge rate in the furnace is 50mL/min.
本发明的己知的起始原料可以采用或按照本领域已知的方法来合成，或可购买于泰坦科技、安耐吉化学、上海德默、成都科龙化工、韶远化学科技、百灵威科技等公司。The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from Titan Technology, Anaiji Chemical, Shanghai Demer, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, and Bailingwei Technology. Waiting for the company.
实施例中无特殊说明，溶液是指水溶液。There is no special explanation in the examples, and the solution refers to an aqueous solution.
实施例中无特殊说明，室温为20℃～30℃。There are no special instructions in the examples, and the room temperature is 20°C to 30°C.
以下通过具体实施例详细说明本发明的实施过程和产生的有益效果，旨在帮助阅读者更好地理解本发明的实质和特点，不作为对本案可实施范围的限定。The implementation process and beneficial effects of the present invention are described in detail below through specific examples, which are intended to help readers better understand the essence and characteristics of the present invention, and are not intended to limit the implementable scope of the present invention.
实施例1：化合物I的制备
Example 1: Preparation of Compound I
第一步：2-(4-溴-2-氟苯基)乙酰氨基甲酸乙酯盐酸盐(1b)Step 1: 2-(4-Bromo-2-fluorophenyl)acetylcarbamate ethyl ester hydrochloride (1b)
ethyl 2-(4-bromo-2-fluorophenyl)acetimidate hydrochlorideethyl 2-(4-bromo-2-fluorophenyl)acetimidate hydrochloride
将1a(5.0g，26.04mmol)加入到无水乙醇(3.0mL)中，将反应置于0℃充分搅拌的同时，向反应中通入干燥的氯化氢气体直到饱和，反应继续在该温度下搅拌10小时。然后向反应加入***(40mL)，过滤，得到固体继续用***洗涤(100mL×2)，将固体干燥后得到1b(5.0g，产率72％)。Add 1a (5.0g, 26.04mmol) to absolute ethanol (3.0mL), place the reaction at 0°C and while stirring thoroughly, pass dry hydrogen chloride gas into the reaction until saturated, and the reaction continues to stir at this temperature. 10 hours. Then, diethyl ether (40 mL) was added to the reaction, filtered, and the solid obtained was washed with diethyl ether (100 mL × 2). The solid was dried to obtain 1b (5.0 g, yield 72%).
1H NMR(400MHz,DMSO-d6)δ12.04(s,2H),7.61(d,1H),7.51–7.43(m,2H),4.46(q,2H),4.14(s,2H),1.26(t,3H)。 1 H NMR (400MHz, DMSO-d 6 ) δ12.04(s,2H),7.61(d,1H),7.51–7.43(m,2H),4.46(q,2H),4.14(s,2H), 1.26(t,3H).
19F NMR(400MHz,DMSO-d6)δ111.30。 19 F NMR (400MHz, DMSO-d 6 ) δ 111.30.
第二步：4-溴-2-氟-1-(2,2,2-三乙氧基乙基)苯(1c)Step 2: 4-bromo-2-fluoro-1-(2,2,2-triethoxyethyl)benzene (1c)
4-bromo-2-fluoro-1-(2,2,2-triethoxyethyl)benzene4-bromo-2-fluoro-1-(2,2,2-triethoxyethyl)benzene
将1b(5.0g,19.30mmol)加入到无水乙醇(40mL)中，置于室温(15℃)搅拌48小时。再将反应液直接过滤得到滤液，将滤液旋干后再用石油醚和乙酸乙酯混合溶剂(100mL)(石油醚/乙酸乙酯(v/v)＝10:1)打浆，过滤，将滤液旋干后得到1c(4.0g，产率66％)。Add 1b (5.0 g, 19.30 mmol) to absolute ethanol (40 mL), and stir at room temperature (15°C) for 48 hours. Then directly filter the reaction solution to obtain a filtrate, spin the filtrate to dryness, then use a mixed solvent of petroleum ether and ethyl acetate (100 mL) (petroleum ether/ethyl acetate (v/v) = 10:1) to make a slurry, filter, and slurry the filtrate. After spin drying, 1c (4.0 g, yield 66%) was obtained.
1H NMR(400MHz,CDCl3)7.45(t,1H),7.23–7.14(m,2H),3.50(q,6H),3.07(s,2H),1.16(t,9H)。1H NMR (400MHz, CDCl 3 )7.45(t,1H),7.23–7.14(m,2H),3.50(q,6H),3.07(s,2H),1.16(t,9H).
第三步：(S)-2-(4-溴-2-氟苄基)-1-(恶烷-2-基甲基)-1H-噻吩并[2,3-d]咪唑-5-羧酸甲酯(1e)Step 3: (S)-2-(4-bromo-2-fluorobenzyl)-1-(oxan-2-ylmethyl)-1H-thieno[2,3-d]imidazole-5- Methyl carboxylate (1e)
methyl(S)-2-(4-bromo-2-fluorobenzyl)-1-(oxetan-2-ylmethyl)-1H-thieno[2,3-d]imidazole-5-carboxylatemethyl(S)-2-(4-bromo-2-fluorobenzyl)-1-(oxetan-2-ylmethyl)-1H-thieno[2,3-d]imidazole-5-carboxylate
将1d(500mg,2.06mmol)和1c(1.0g,3.19mmol)加入到冰乙酸(20mL)中，置于60℃搅拌10小时。将反应体系冷却至室温后缓慢加入到碳酸氢钠的饱和水溶液(200mL)中淬灭，再用乙酸乙酯萃取(50mL×3)，合并有机相，用无水硫酸钠干燥后过滤，滤液旋干， 经过硅胶柱色谱分离纯化(石油醚/乙酸乙酯(v/v)＝1:1)，得到1e(430mg，产率47％)。Add 1d (500 mg, 2.06 mmol) and 1c (1.0 g, 3.19 mmol) to glacial acetic acid (20 mL), and stir at 60°C for 10 hours. After the reaction system was cooled to room temperature, it was slowly added to a saturated aqueous solution of sodium bicarbonate (200 mL) to quench it, and then extracted with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was vortexed Dry, After separation and purification by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:1), 1e (430 mg, yield 47%) was obtained.
LCMS m/z＝439.0[M+1]+。LCMS m/z=439.0[M+1] + .
第四步：(S)-2-(2-氟-4-(4,4,5,5-四甲基-1,3,2-二氧代苯并呋喃-2-基)苄基)-1-(氧代烷-2-基甲基)-1H-噻吩并[2,3-d]咪唑-5-羧酸甲酯(1f)Step 4: (S)-2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxobenzofuran-2-yl)benzyl) -1-(Oxoalk-2-ylmethyl)-1H-thieno[2,3-d]imidazole-5-carboxylic acid methyl ester (1f)
methyl(S)-2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)-1-(oxetan-2-ylmethyl)-1H-thieno[2,3-d]imidazole-5-carboxylatemethyl(S)-2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)-1-(oxetan-2-ylmethyl)- 1H-thieno[2,3-d]imidazole-5-carboxylate
将1e(400mg,0.98mmol)和联硼酸频哪醇酯(262mg,1.0mmol)，Pd(dppf)Cl2·DCM(81mg,0.1mmol),乙酸钾(196mg,2.0mmol)加入到1,4-二氧六环中(20mL)，将反应置换为氮气保护后，置于100℃搅拌5小时。将反应液冷却至室温后加入水(100mL)，再用乙酸乙酯萃取(50mL×3)，合并有机相，用无水硫酸钠干燥后过滤，滤液旋干，经过硅胶柱色谱分离纯化(石油醚/乙酸乙酯(v/v)＝1:1)，得到1f(340mg，产率71％)。Add 1e (400mg, 0.98mmol) and pinacol diborate (262mg, 1.0mmol), Pd(dppf)Cl 2 ·DCM (81mg, 0.1mmol), potassium acetate (196mg, 2.0mmol) to 1,4 - dioxane (20 mL), replace the reaction with nitrogen protection, and stir at 100°C for 5 hours. After the reaction solution was cooled to room temperature, water (100 mL) was added, and then extracted with ethyl acetate (50 mL Ether/ethyl acetate (v/v)=1:1), 1f (340 mg, yield 71%) was obtained.
LCMS m/z＝487.2[M+1]+。LCMS m/z=487.2[M+1] + .
第五步：(S)-2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2-氟苄基)-1-(氧代烷-2-基甲基)-1H-噻吩并[2,3-d]咪唑-5-羧酸甲酯(1h)Step 5: (S)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2-fluorobenzyl)-1-(oxy Alk-2-ylmethyl)-1H-thieno[2,3-d]imidazole-5-carboxylic acid methyl ester (1h)
methyl(S)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2-fluorobenzyl)-1-(oxetan-2-ylmethyl)-1H-thieno[2,3-d]imidazole-5-carboxylatemethyl(S)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2-fluorobenzyl)-1-(oxetan-2-ylmethyl)-1H-thieno [2,3-d]imidazole-5-carboxylate
将1f(340mg,0.70mmol)和6g(220mg,0.84mmol)，Pd(dppf)Cl2·DCM(83mg,0.1mmol),碳酸钾(290mg,2.1mmol)加入到1,4-二氧六环(10mL)和水(2mL)的混合溶液中，将反应置换为氮气保护后，置于100℃搅拌5小时。将反应液冷却至室温后加入水(50mL)，再用乙酸乙酯萃取(30mL×3)，合并有机相，用无水硫酸钠干燥后过滤，滤液旋干，经过硅胶柱色谱分离纯化(石油醚/乙酸乙酯(v/v)＝1:1)，得到1h(200mg，产率48％)。Add 1f (340mg, 0.70mmol) and 6g (220mg, 0.84mmol), Pd(dppf)Cl 2 ·DCM (83mg, 0.1mmol), potassium carbonate (290mg, 2.1mmol) to 1,4-dioxane (10 mL) and water (2 mL), the reaction was replaced with nitrogen protection, and then placed at 100°C and stirred for 5 hours. After the reaction solution was cooled to room temperature, water (50 mL) was added, and then extracted with ethyl acetate (30 mL Ether/ethyl acetate (v/v)=1:1), 1h (200 mg, yield 48%) was obtained.
LCMS m/z＝587.1[M+1]+。LCMS m/z=587.1[M+1] + .
第六步：(S)-2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2-氟苄基)-1-(氧代烷-2-基甲基)-1H-噻吩并[2,3-d]咪唑-5-羧酸(化合物I)Step 6: (S)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2-fluorobenzyl)-1-(oxy Alk-2-ylmethyl)-1H-thieno[2,3-d]imidazole-5-carboxylic acid (Compound I)
(S)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2-fluorobenzyl)-1-(oxetan-2-ylmethyl)-1H-thieno[2,3-d]imidazole-5-carboxylic acid(S)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2-fluorobenzyl)-1-(oxetan-2-ylmethyl)-1H-thieno[ 2,3-d]imidazole-5-carboxylic acid
将1h(100mg,0.17mmol)和1i(120mg,0.85mmol)加入到乙腈(5mL)和水(1mL)的混合溶液中，置于12℃搅拌10小时。再将反应液用1M的盐酸调pH＝6，继续搅拌至白色固体析出，过滤，用水洗涤固体，在将固体用二氯甲烷溶解后，经过无水硫酸钠干燥，过滤，有机相减压浓缩后HPLC制备分离，得到化合物式I的三氟乙酸盐，再经过硅胶柱纯化得到化合物I(40mg，产率41％)。 1h (100 mg, 0.17 mmol) and 1i (120 mg, 0.85 mmol) were added to a mixed solution of acetonitrile (5 mL) and water (1 mL), and stirred at 12°C for 10 hours. Then adjust the pH of the reaction solution to 6 with 1M hydrochloric acid, continue stirring until a white solid precipitates, filter, wash the solid with water, dissolve the solid with dichloromethane, dry over anhydrous sodium sulfate, filter, and concentrate the organic phase under reduced pressure. After HPLC preparative separation, the trifluoroacetate salt of compound formula I was obtained, and then purified through silica gel column to obtain compound I (40 mg, yield 41%).
LCMS m/z＝573.1[M+1]+。LCMS m/z=573.1[M+1] + .
1H NMR(400MHz,CD3OD)δ12.90(br,1H),7.91(d,1H),7.88–7.80(m,4H),7.79–7.68(m,2H)7.64(d,1H),7.39(t,1H),6.92(d,1H),5.61(s,2H),5.06–4.98(m,1H),4.60(dd,1H),4.54–4.44(m,2H),4.43–4.28(m,3H),2.73–2.61(m,1H),2.40–2.27(m,1H)。 1 H NMR (400MHz, CD 3 OD) δ12.90(br,1H),7.91(d,1H),7.88–7.80(m,4H),7.79–7.68(m,2H)7.64(d,1H), 7.39(t,1H),6.92(d,1H),5.61(s,2H),5.06–4.98(m,1H),4.60(dd,1H),4.54–4.44(m,2H),4.43–4.28( m,3H),2.73–2.61(m,1H),2.40–2.27(m,1H).
实施例2：化合物I的三羟甲基氨基甲烷盐晶型A的制备Example 2: Preparation of Trishydroxymethylaminomethane Salt Form A of Compound I
称取化合物式I(2.920g)，加入76.8mL正丙醇，室温(约25℃)悬浮约10min；称取三羟甲基氨基甲烷(0.615g)，加入7.68mL水，室温(约25℃)悬浮并超声至溶液澄清；将三羟甲基氨基甲烷的澄清水溶液滴加至游离态的正丙醇悬浊液中，室温悬浮约16h，离心分离后(10000rpm,4min)将固体部分50℃真空干燥约17h。Weigh compound formula I (2.920g), add 76.8mL n-propanol, and suspend at room temperature (about 25°C) for about 10 minutes; weigh trishydroxymethylaminomethane (0.615g), add 7.68mL water, and suspend at room temperature (about 25°C). ) and sonicate until the solution is clear; add the clear aqueous solution of trishydroxymethylaminomethane dropwise into the free n-propanol suspension, suspend at room temperature for about 16 hours, and then centrifuge (10000 rpm, 4 min) and vacuum the solid part at 50°C Dry for about 17 hours.
1H NMR(400MHz,DMSO-d6)δ7.94-7.88(m,1H),7.88-7.80(m,3H),7.79-7.69(m,2H),7.66-7.60(m,1H),7.47(s,1H),7.40-7.32(m,1H),6.95-6.87(m,1H),5.61(s,2H),5.02-4.93(m,1H),4.57-4.21(m,6H),3.50-3.36(m,6H),2.71-2.60(m,1H),2.38–2.27(m,1H)。 1 H NMR (400MHz, DMSO-d6) δ7.94-7.88(m,1H),7.88-7.80(m,3H),7.79-7.69(m,2H),7.66-7.60(m,1H),7.47( s,1H),7.40-7.32(m,1H),6.95-6.87(m,1H),5.61(s,2H),5.02-4.93(m,1H),4.57-4.21(m,6H),3.50- 3.36(m,6H),2.71-2.60(m,1H),2.38-2.27(m,1H).
LCMS m/z＝573.1[M+1]+。LCMS m/z=573.1[M+1] + .
实施例3：化合物I的三羟甲基氨基甲烷盐晶型A的制备Example 3: Preparation of Trishydroxymethylaminomethane Salt Form A of Compound I
称取220.0mg三羟甲基氨基甲烷盐晶型C，加入0.5mL二甲基亚砜，在室温(约25℃)搅拌直至溶清；取60μL澄清液，于室温(约25℃)下滴加至0.6mL 4-甲基-2-戊酮中，搅拌约1h，离心分离后(10000rpm,4min)将固体部分室温真空干燥17h。Weigh 220.0 mg of trishydroxymethylaminomethane salt crystal form C, add 0.5 mL of dimethyl sulfoxide, and stir at room temperature (about 25°C) until the solution is clear; take 60 μL of the clear solution and drop it at room temperature (about 25°C). Add to 0.6 mL of 4-methyl-2-pentanone, stir for about 1 hour, centrifuge (10000 rpm, 4 minutes) and vacuum dry the solid part at room temperature for 17 hours.
实施例4：化合物I的三羟甲基氨基甲烷盐晶型C的制备Example 4: Preparation of Trishydroxymethylaminomethane Salt Form C of Compound I
称取220.2mg三羟甲基氨基甲烷盐晶型A，加入0.8mL N-甲基吡咯烷酮，在室温(约25℃)搅拌直至溶清；取70μL澄清液，于室温(约25℃)向澄清液中滴加二氯甲烷3.0mL，搅拌约1h，离心分离后(10000rpm,4min)将固体部分室温真空干燥17h。Weigh 220.2 mg of trishydroxymethylaminomethane salt crystal form A, add 0.8 mL of N-methylpyrrolidone, and stir at room temperature (about 25°C) until the solution is clear; take 70 μL of clarified liquid, and add 0.8 mL of N-methylpyrrolidone to the clarified solution at room temperature (about 25°C). Add 3.0 mL of methylene chloride dropwise to the solution, stir for about 1 hour, centrifuge (10000 rpm, 4 minutes) and dry the solid part under vacuum at room temperature for 17 hours.
实施例5：化合物I的三羟甲基氨基甲烷盐晶型D的制备Example 5: Preparation of Trishydroxymethylaminomethane Salt Form D of Compound I
称取220.0mg三羟甲基氨基甲烷盐晶型A，加入14.0mL甲醇，室温(约25℃)悬浮约16h，离心分离后(10000rpm,4min)将固体部分室温真空干燥约17h。Weigh 220.0 mg of trishydroxymethylaminomethane salt crystal form A, add 14.0 mL of methanol, and suspend at room temperature (about 25°C) for about 16 hours. After centrifugation (10000 rpm, 4 minutes), the solid part is vacuum dried at room temperature for about 17 hours.
实施例6：化合物I的三乙醇胺盐晶型A的制备Example 6: Preparation of Triethanolamine Salt Form A of Compound I
称取20.5mg三乙醇胺盐原料，加入6.5mL氯仿，在室温搅拌直至溶清；将澄清液体置于室温(约25℃)敞口挥发8天至有固体析出。Weigh 20.5 mg of the triethanolamine salt raw material, add 6.5 mL of chloroform, and stir at room temperature until the solution is clear; place the clear liquid at room temperature (about 25°C) to evaporate in the open for 8 days until solid precipitates.
1H NMR(400MHz,DMSO-d6)δppm 2.33(m,1H),2.64(m,7H),3.45(t,7H),4.40(m,5H),4.59(m,1H),5.00(br dd,1H),5.61(s,2H),6.92(d,1H),7.39(t,1H),7.65(d,1H),7.75(m,3H),7.85(m,3H),7.93(d,1H)。1H NMR(400MHz,DMSO-d6)δppm 2.33(m,1H),2.64(m,7H),3.45(t,7H),4.40(m,5H),4.59(m,1H),5.00(br dd, 1H),5.61(s,2H),6.92(d,1H),7.39(t,1H),7.65(d,1H),7.75(m,3H),7.85(m,3H),7.93(d,1H ).
晶型测试例Crystal form test example
1.化合物的XRD测试 1. XRD test of compounds
将本发明化合物按照如下方法进行X-射线粉末衍射测试。The compounds of the present invention were subjected to X-ray powder diffraction testing according to the following method.
表1-1 XRD测试参数
Table 1-1 XRD test parameters
2.化合物的DSC测试2. DSC test of compounds
DSC图谱在TA Instruments discovery DSC 2500和DSC 250差式扫描量热仪上采集，化合物I的三羟甲基氨基甲烷盐的测试参数如表2-1所示，化合物I的三乙醇胺盐的测试参数如表2-2所示。DSC spectra were collected on TA Instruments discovery DSC 2500 and DSC 250 differential scanning calorimeters. The test parameters of the trishydroxymethylaminomethane salt of compound I are shown in Table 2-1. The test parameters of the triethanolamine salt of compound I As shown in Table 2-2.
表2-1 DSC测试参数
Table 2-1 DSC test parameters
表2-2 DSC测试参数
Table 2-2 DSC test parameters
3.化合物的TGA测试3. TGA testing of compounds
TGA图谱在TA Instruments discovery TGA 55和TGA 550热重分析仪上采集，化合物I的三羟甲基氨基甲烷盐的测试参数如表3-1所示，化合物I的三乙醇胺盐的测试参数如表3-2所示。 TGA spectra were collected on TA Instruments discovery TGA 55 and TGA 550 thermogravimetric analyzers. The test parameters of the trishydroxymethylaminomethane salt of compound I are shown in Table 3-1, and the test parameters of the triethanolamine salt of compound I are shown in the table. As shown in 3-2.
表3-1 TGA测试参数
Table 3-1 TGA test parameters
表3-2 TGA测试参数
Table 3-2 TGA test parameters
4.化合物的XRD测试的具体峰值表征结果4. Specific peak characterization results of XRD testing of compounds
化合物I的三羟甲基氨基甲烷盐晶型A的X-射线粉末衍射图谱(XRD)图1所示。具体峰值如表4所示。The X-ray powder diffraction pattern (XRD) of the trishydroxymethylaminomethane salt Form A of Compound I is shown in Figure 1. The specific peak values are shown in Table 4.
表4

Table 4

化合物I的三羟甲基氨基甲烷盐晶型C的X-射线粉末衍射图谱(XRD)图3所示。具体峰值如表5所示。The X-ray powder diffraction pattern (XRD) of Form C of the trishydroxymethylaminomethane salt of Compound I is shown in Figure 3. The specific peak values are shown in Table 5.
表5

table 5

化合物I的三羟甲基氨基甲烷盐晶型D的X-射线粉末衍射图谱(XRD)图5所示。具体峰值如表6所示。The X-ray powder diffraction pattern (XRD) of the trishydroxymethylaminomethane salt crystal form D of Compound I is shown in Figure 5. The specific peak values are shown in Table 6.
表6

Table 6

化合物I的三乙醇胺盐晶型A的X-射线粉末衍射图谱(XRD)图7所示。具体峰值如表7所示。The X-ray powder diffraction pattern (XRD) of the triethanolamine salt crystal form A of Compound I is shown in Figure 7. The specific peak values are shown in Table 7.
表7


Table 7


5、化合物I的药用盐稳定性数据5. Stability data of pharmaceutical salts of Compound I
取样品分别在40℃+75％RH、30℃+60％RH条件下进行试验，HPLC检测纯度(百分数表示)，实验结果见表10。Samples were taken and tested under the conditions of 40°C + 75% RH and 30°C + 60% RH respectively. The purity was detected by HPLC (expressed as a percentage). The experimental results are shown in Table 10.
供试品溶液制备方法及HPLC检测纯度条件见表8、表9；The test solution preparation method and HPLC purity testing conditions are shown in Table 8 and Table 9;
表8供试品溶液制备方法
Table 8 Preparation method of test solution
表9 HPLC检测纯度条件

Table 9 HPLC purity detection conditions

表10化合物1和不同种类的盐及晶型在不同条件下的化学稳定性(采用HPLC测定纯度)
Table 10 Chemical stability of compound 1 and different types of salts and crystal forms under different conditions (purity determined by HPLC)
结论：化合物1的三羟甲基氨基甲烷盐晶型A具有较好的化学稳定性。Conclusion: The trishydroxymethylaminomethane salt form A of compound 1 has good chemical stability.
6、溶解度测试6. Solubility test
按照中国药典2020年版的方法测试受试化合物的溶解度，结果见表11。The solubility of the test compound was tested according to the method of the Chinese Pharmacopoeia 2020 edition. The results are shown in Table 11.
表11化合物I游离态及药用盐溶解度数据
Table 11 Compound I free state and pharmaceutical salt solubility data
结论：在pH6.8，化合物I的三羟甲基氨基甲烷盐晶型A溶解度相比游离态有显著提升。Conclusion: At pH 6.8, the solubility of the trishydroxymethylaminomethane salt crystal form A of compound I is significantly improved compared to the free state.
生物测试例Biological test examples
1.HEK293/CRE-luc/GLP-1R细胞活性1.HEK293/CRE-luc/GLP-1R cell activity
细胞：HEK293/CRE-luc/GLP-1R Cells: HEK293/CRE-luc/GLP-1R
细胞培养基：DMEM+10％FBS+400μg/ml G418+100μg/ml Hygromycin BCell culture medium: DMEM+10% FBS+400μg/ml G418+100μg/ml Hygromycin B
冻存液：90％FBS,10％(V/V)DMSOFreezing solution: 90% FBS, 10% (V/V) DMSO
检测buffer：DMEM+1％FBSDetection buffer: DMEM+1%FBS
实验步骤Experimental steps
细胞用DMEM培养基+10％FBS+400μg/ml G418+100μg/ml Hygromycin B于37℃CO2培养箱中培养，3-4天传代一次。Cells were cultured in DMEM medium + 10% FBS + 400 μg/ml G418 + 100 μg/ml Hygromycin B in a 37°C CO2 incubator, and passaged once every 3-4 days.
细胞铺板：胰酶消化调整细胞密度为1.67×105cells/mL；384孔板化合物中每孔接种细胞60μL(10000cells/孔)；设置NC孔(阴性对照)，背景孔(无细胞)。培养箱孵育约18±2h。Cell plating: trypsinize and adjust the cell density to 1.67×10 5 cells/mL; inoculate 60 μL of cells into each well of the 384-well plate compound (10000 cells/well); set NC wells (negative control) and background wells (no cells). Incubate in the incubator for approximately 18±2h.
化合物用检测buffer梯度稀释，检测浓度0.01nM～1000nM。Compounds are gradient diluted with detection buffer, with detection concentrations ranging from 0.01nM to 1000nM.
将细胞培养板取出，随后从细胞中吸出全部上清。用1X PBS轻洗2遍。Remove the cell culture plate and aspirate all supernatant from the cells. Wash gently 2 times with 1X PBS.
将稀释好的化合物加入384孔板中(10μL/孔)，每个浓度设置3个复孔。NC孔加10μL检测buffer，密封37℃孵育6h。Add the diluted compound into a 384-well plate (10 μL/well), and set 3 duplicate wells for each concentration. Add 10 μL of detection buffer to the NC well, seal and incubate at 37°C for 6 hours.
将孔板取出，使细胞平衡至室温(至少15min)，随后从细胞中吸出全部上清液。Remove the well plate, allow the cells to equilibrate to room temperature (at least 15 min), and then aspirate all supernatant from the cells.
样品孔中加入10μL/孔Reagent，在室温下孵育5min使细胞裂解。Add 10μL/well to the sample well Reagent, incubate at room temperature for 5 minutes to lyse cells.
用BMG酶标仪读取检测结果。Read the test results with a BMG microplate reader.
数据处理data processing
计算平均背景值。
计算诱导倍数(Fold of induction,FI)＝(诱导孔数值–背景值)/(阴性对照孔数值–背
景值)。Calculate the average background value.
Calculate the fold of induction (FI) = (value of induction wells – background value)/(value of negative control wells – background value).
用Graphpad Prim 8.0软件采用四参数拟合分析，计算样品的EC50数值。Four-parameter fitting analysis was used using Graphpad Prim 8.0 software to calculate the EC 50 value of the sample.
统计分析Statistical Analysis
所有结果都统计平均数和标准误(Mean±SEM)，使用Graphpad Prism软件进行统计分析。具体的数据以图表形式呈现。P<0.05被认为具有统计学差异。All results were calculated as mean ± SEM, and statistical analysis was performed using Graphpad Prism software. Specific data are presented in chart form. P<0.05 was considered statistically significant.
结论：化合物I对GLP-1受体有良好的激动作用，其EC50值小于10nM。Conclusion: Compound I has a good agonistic effect on GLP-1 receptor, and its EC 50 value is less than 10 nM.
2.化合物对hERG钾离子通道作用2. Effect of compounds on hERG potassium ion channel
实验平台：电生理手动膜片钳***Experimental platform: electrophysiology manual patch clamp system
细胞系：稳定表达hERG钾离子通道的中国仓鼠卵巢(CHO)细胞Cell line: Chinese hamster ovary (CHO) cells stably expressing hERG potassium channel
实验方法：稳定表达hERG钾通道的CHO(Chinese Hamster Ovary)细胞，在室温下试验采用全细胞膜片钳技术记录hERG钾通道电流。玻璃微电极由玻璃电极毛胚(BF150-86-10，Sutter)经拉制仪拉制而成，灌注电极内液后的尖端电阻为2-5MΩ左右，将玻璃微电极***放大器探头即可连接至膜片钳放大器。钳制电压和数据记录由pClamp 10 软件通过电脑控制和记录，采样频率为10kHz，滤波频率为2kHz。在得到全细胞记录后，细胞钳制在-80mV，诱发hERG钾电流(I hERG)的步阶电压从-80mV给予一个2s的去极化电压到+20mV，再复极化到-50mV，持续1s后回到-80mV。每10s给予此电压刺激，确定hERG钾电流稳定后(至少1分钟)开始给药过程。化合物每个测试浓度的给药时间为至少1分钟，每个浓度至少测试2个细胞(n≥2)。Experimental method: CHO (Chinese Hamster Ovary) cells stably expressing hERG potassium channels were tested at room temperature using whole-cell patch clamp technology to record hERG potassium channel currents. The glass microelectrode is drawn from a glass electrode blank (BF150-86-10, Sutter) by a drawing instrument. The tip resistance after infusion of the electrode liquid is about 2-5MΩ. The glass microelectrode can be connected by inserting it into the amplifier probe. to the patch clamp amplifier. Clamp voltage and data recording by pClamp 10 The software controls and records through the computer, with a sampling frequency of 10kHz and a filtering frequency of 2kHz. After obtaining the whole-cell recording, the cells were clamped at -80mV, and the step voltage of induced hERG potassium current (I hERG ) was given a 2s depolarization voltage from -80mV to +20mV, and then repolarization to -50mV for 1s. then returns to -80mV. This voltage stimulation was given every 10 s, and the administration process was started after confirming that the hERG potassium current was stable (at least 1 minute). Compounds were administered for at least 1 minute at each concentration tested, and at least 2 cells (n ≥ 2) were tested at each concentration.
数据处理：data processing:
数据分析处理采用pClamp 10，GraphPad Prism 5和Excel软件。不同化合物浓度对hERG钾电流(-50mV时诱发的hERG尾电流峰值)的抑制程度用以下公式计算：Inhibition％＝[1–(I/Io)]×100％。Data analysis and processing used pClamp 10, GraphPad Prism 5 and Excel software. The degree of inhibition of hERG potassium current (peak hERG tail current induced at -50mV) at different compound concentrations was calculated using the following formula: Inhibition% = [1 – (I/Io)] × 100%.
其中，Inhibition％代表化合物对hERG钾电流的抑制百分率，I和Io分别表示在加药后和加药前hERG钾电流的幅度。Among them, Inhibition% represents the inhibition percentage of the hERG potassium current by the compound, and I and Io represent the amplitude of the hERG potassium current after and before the addition of the drug, respectively.
化合物IC50使用GraphPad Prism 5软件通过以下方程拟合计算得出：
Y＝Bottom+(Top-Bottom)/(1+10^((LogIC50-X)×HillSlope))Compound IC 50 was calculated by fitting the following equation using GraphPad Prism 5 software:
Y＝Bottom+(Top-Bottom)/(1+10^((LogIC 50 -X)×HillSlope))
其中，X为供试品检测浓度的Log值，Y为对应浓度下抑制百分率，Bottom和Top分别为最小和最大抑制百分率。Among them, X is the Log value of the detected concentration of the test product, Y is the inhibition percentage at the corresponding concentration, Bottom and Top are the minimum and maximum inhibition percentages respectively.
结论：化合物I具有较小的hERG抑制作用,IC50>40μM。Conclusion: Compound I has a small inhibitory effect on hERG, with IC 50 >40 μM.
3.小鼠肝微粒体稳定性试验3. Mouse liver microsome stability test
在37℃条件下，1μM待测化合物与小鼠肝微粒体(0.5mg/mL)辅以NADPH再生体系孵育5、10、20、30和60分钟后，采用LC-MS/MS方法检测所产生的样品中待测化合物浓度。通过对相应的各时间点与化合物的剩余百分比进行计算，得到化合物在小鼠肝微粒体溶液中的半衰期(T1/2)和固有清除率(CLint(mic))。At 37°C, 1 μM of the compound to be tested was incubated with mouse liver microsomes (0.5 mg/mL) supplemented with NADPH regeneration system for 5, 10, 20, 30 and 60 minutes, and then the LC-MS/MS method was used to detect the produced The concentration of the compound to be measured in the sample. By calculating the remaining percentage of the compound at each corresponding time point, the half-life (T 1/2 ) and intrinsic clearance rate (CL int(mic) ) of the compound in mouse liver microsome solution were obtained.
生物测试结果：
Biological test results:
结论：本发明化合物具有较好的小鼠肝微粒体稳定性。例如化合物I、跟对照化合物相 比在小鼠肝微粒体中清除率显著趋缓，半衰期显著延长。化合物I与化合物DZ-6的T1/2比值大于2。Conclusion: The compound of the present invention has good stability in mouse liver microsomes. For example, compound I, compared with the control compound Compared with mouse liver microsomes, the clearance rate was significantly slower and the half-life was significantly longer. The T 1/2 ratio of compound I to compound DZ-6 is greater than 2.
4.人肝微粒体稳定性试验4. Human liver microsome stability test
在37℃条件下，1μM待测化合物与人肝微粒体(0.5mg/mL)辅以NADPH再生体系孵育5、10、20、30和60分钟后，采用LC-MS/MS方法检测所产生的样品中待测化合物浓度。通过对相应的各时间点与化合物的剩余百分比进行计算，得到化合物在人肝微粒体溶液中的半衰期(T1/2)和固有清除率(CLint(mic))。At 37°C, 1 μM of the compound to be tested was incubated with human liver microsomes (0.5 mg/mL) supplemented with NADPH regeneration system for 5, 10, 20, 30 and 60 minutes, and the LC-MS/MS method was used to detect the produced The concentration of the compound to be measured in the sample. By calculating the remaining percentage of the compound at each corresponding time point, the half-life (T 1/2 ) and intrinsic clearance rate (CL int(mic) ) of the compound in human liver microsomal solution were obtained.
生物测试结果：
Biological test results:
结论：本发明化合物具有较好的人肝微粒体稳定性。例如化合物1跟对照化合物相比在人肝微粒体中清除率显著趋缓，半衰期显著延长。化合物1与化合物DZ-6的T1/2比值大于3。Conclusion: The compound of the present invention has good stability in human liver microsomes. For example, compared with the control compound, the clearance rate of compound 1 in human liver microsomes was significantly slower and the half-life was significantly longer. The T 1/2 ratio of compound 1 to compound DZ-6 is greater than 3.
5.药代动力学测试5.Pharmacokinetic testing
小鼠药代动力学评价Mouse pharmacokinetic evaluation
雄性C57小鼠(购自湖南斯莱克景达实验动物有限公司，许可证号：SCXK(湘)2019-0004)，20-25g，禁食过夜。实验当天18只雄性C57小鼠分为2组，每组9只，分别静脉和灌胃给予剂量为5mg/kg和20mg/kg的受试化合物，于给药前及给药后5,15,30min,1,2,4,6,8和24h经眼眶静脉丛取血0.1ml，置于EDTAK2离心管中，于6000rpm，4℃离心5min后，收集血浆。血浆样品加入含内标(***，500ng/mL)的乙腈溶液，涡漩后于10000rpm离心10min，取上清进行LC-MS/MS分析。采用WinNonlin Version 8.0(Pharsight,Mountain View,CA)药动学软件中的非房室模型计算药动学参数。Male C57 mice (purchased from Hunan Slack Jingda Experimental Animal Co., Ltd., license number: SCXK (Hunan) 2019-0004), 20-25g, fasted overnight. On the day of the experiment, 18 male C57 mice were divided into 2 groups, with 9 mice in each group. Test compounds at doses of 5 mg/kg and 20 mg/kg were administered intravenously and orally respectively before and after administration. 5,15, 0.1 ml of blood was taken from the orbital venous plexus at 30 minutes, 1, 2, 4, 6, 8 and 24 hours, placed in an EDTAK2 centrifuge tube, centrifuged at 6000 rpm and 4°C for 5 minutes, and then the plasma was collected. The plasma sample was added with an acetonitrile solution containing internal standard (dexamethasone, 500ng/mL), vortexed and centrifuged at 10000rpm for 10min, and the supernatant was taken for LC-MS/MS analysis. Pharmacokinetic parameters were calculated using the non-compartmental model in WinNonlin Version 8.0 (Pharsight, Mountain View, CA) pharmacokinetic software.
结论：本发明的化合物具有良好的口服性能，例如化合物1在C57小鼠的生物利用度＞25％。Conclusion: The compounds of the present invention have good oral properties. For example, the bioavailability of compound 1 in C57 mice is >25%.
食蟹猴药代动力学评价 Evaluation of pharmacokinetics in cynomolgus monkeys
雄性食蟹猴(饲养于苏州莱奥生物技术有限公司)，体重3～5kg，禁食过夜(14～18h)。实验当天6只雄性食蟹猴分为2组，每组3只，分别静脉和灌胃给予剂量为2mg/kg和10mg/kg的受试化合物，于给药前及给药后5,15,30min,1,2,4,6,8,10,12和24h经四肢静脉取血1ml，置于EDTAK2离心管中，于6000rpm，4℃离心5min后，收集血浆。血浆样品加入含内标(***，500ng/mL)的乙腈溶液，涡漩后于10000rpm离心10min，取上清进行LC-MS/MS分析。采用WinNonlin Version 8.0(Pharsight,Mountain View,CA)药动学软件中的非房室模型计算药动学参数。Male cynomolgus monkeys (raised in Suzhou Leo Biotechnology Co., Ltd.), weighing 3 to 5 kg, were fasted overnight (14 to 18 hours). On the day of the experiment, 6 male cynomolgus monkeys were divided into 2 groups, with 3 monkeys in each group. The test compounds were administered intravenously and intragastrically at doses of 2 mg/kg and 10 mg/kg respectively, before and after administration. 5,15, 1 ml of blood was taken from the veins of the limbs at 30 minutes, 1, 2, 4, 6, 8, 10, 12 and 24 hours, placed in EDTAK2 centrifuge tubes, centrifuged at 6000 rpm and 4°C for 5 minutes, and then the plasma was collected. The plasma sample was added with an acetonitrile solution containing internal standard (dexamethasone, 500ng/mL), vortexed and centrifuged at 10000rpm for 10min, and the supernatant was taken for LC-MS/MS analysis. Pharmacokinetic parameters were calculated using the non-compartmental model in WinNonlin Version 8.0 (Pharsight, Mountain View, CA) pharmacokinetic software.
结论：本发明化合物具有良好的代谢性能。例如化合物I在猴体内清除率低，半衰期较长，具有药物作用时间更长、给药频率更低的优势。Conclusion: The compounds of the present invention have good metabolic properties. For example, Compound I has a low clearance rate and a long half-life in monkeys, which has the advantages of longer drug action time and lower administration frequency.
6.细胞色素P450同工酶抑制性测试6. Cytochrome P450 isoenzyme inhibition test
实验目的：测定受试化合物对人肝微粒体细胞色素P450同工酶(CYP1A2、CYP2C19、CYP2D6和CYP3A4)活性的抑制作用。Experimental purpose: To determine the inhibitory effect of the test compound on the activity of human liver microsomal cytochrome P450 isoenzymes (CYP1A2, CYP2C19, CYP2D6 and CYP3A4).
实验操作：将孵育浓度为0,0.03,0.1,0.3,1,3,10,30和100μM的受试化合物分别与含5个酶混合底物的人肝微粒体溶液(肝微粒体浓度：0.1mg/mL)在37℃水浴中预孵育10min后，加入辅酶因子(NADPH)启动反应；于37℃水浴中继续孵育10min后，加入含内标的冰乙腈溶液终止反应，振荡1min，于4℃、10000rpm条件下离心10min后取上清液进行LC/MS/MS分析，检测各CYP酶底物所产生的代谢物的量。Experimental operation: Test compounds with incubation concentrations of 0, 0.03, 0.1, 0.3, 1, 3, 10, 30 and 100 μM were respectively mixed with human liver microsome solution containing 5 enzyme mixed substrates (liver microsome concentration: 0.1 mg/mL) in a 37°C water bath for 10 minutes, add coenzyme factor (NADPH) to start the reaction; after continuing to incubate in a 37°C water bath for 10 minutes, add an ice-cold acetonitrile solution containing the internal standard to terminate the reaction, shake for 1 minute, and incubate at 4°C, After centrifugation for 10 minutes at 10,000 rpm, the supernatant was taken for LC/MS/MS analysis to detect the amount of metabolites produced by each CYP enzyme substrate.
用各代谢物的生成速率体现各CYP同工酶的活性。设定不加受试物的溶剂对照孵育体系中各同工酶的活性为100％，将含不同浓度受试物时代谢物的生成速率与溶剂对照样品代谢产物的生成速率的比值作为各同工酶的剩余活性百分比。以剩余活性百分比为纵坐标，抑制剂浓度为横坐标，用Graphpad prism 5.0对数据进行作图并计算受试物对各CYP同工酶的IC50值。The activity of each CYP isoenzyme is reflected by the production rate of each metabolite. Set the activity of each isoenzyme in the solvent control incubation system without the test substance to 100%, and take the ratio of the generation rate of metabolites when containing the test substance at different concentrations to the generation rate of the metabolites of the solvent control sample as each The remaining activity percentage of the enzyme. Taking the remaining activity percentage as the ordinate and the inhibitor concentration as the abscissa, use Graphpad prism 5.0 to plot the data and calculate the IC 50 value of the test substance for each CYP isoenzyme.
结论：本发明的化合物对细胞色素P450同工酶抑制活性弱，例如化合物I对CYP2C19、CYP1A2、CYP2D6和CYP3A4-M抑制活性IC50＞50μM。Conclusion: The compounds of the present invention have weak inhibitory activity against cytochrome P450 isoenzymes. For example, the inhibitory activity of compound I against CYP2C19, CYP1A2, CYP2D6 and CYP3A4-M is IC 50 >50 μM.
7.食蟹猴中的静脉糖耐量实验7. Intravenous glucose tolerance test in cynomolgus monkeys
实验动物：雄性食蟹猴(饲养于中美冠科生物技术(太仓)有限公司)，体重6.5-7.5Kg，共6只。Experimental animals: male cynomolgus monkeys (raised at Sino-American Crown Biotechnology (Taicang) Co., Ltd.), weighing 6.5-7.5Kg, a total of 6 animals.
实验过程：在整个实验期间，每次采血(ivGTT操作)前，实验动物需过夜禁食，约16小时，按照如下表10-1进行动物分组及给药。Experimental process: During the entire experimental period, experimental animals need to fast overnight for about 16 hours before each blood collection (ivGTT operation). The animals are grouped and administered according to the following table 10-1.
静脉葡萄糖耐量试验(ivGTT)：动物麻醉后先采集基值血，然后开始静脉推注，接着输注。输注15min后开始采集给糖前血糖基值，输注30min后开始给与50％的葡萄糖溶液(0.25g/kg,0.5mL/kg)。整个输注时长为1.0hr。 Intravenous glucose tolerance test (ivGTT): After the animal is anesthetized, baseline blood is first collected, then intravenous bolus injection is started, and then infusion is started. 15 minutes after the infusion, the baseline blood glucose value before glucose administration was collected, and 30 minutes after the infusion, a 50% glucose solution (0.25 g/kg, 0.5 mL/kg) was administered. The total infusion duration is 1.0hr.
分别于给药前、-15min(给糖前15min)、0min(给糖前)、给糖后2、5、7、10、20、30min采血，采集后的血液常温放置至少30分钟，然后在常温下离心，3500g/分钟，离心10分钟，取上清液分装，置于2～8℃直至血糖和胰岛素分析。所有的实验数据均采用GraphPad Prism 9.3.1进行分析。Blood was collected before administration, -15min (15min before sugar administration), 0min (before sugar administration), and 2, 5, 7, 10, 20, and 30min after sugar administration. The collected blood was left at room temperature for at least 30 minutes, and then Centrifuge at room temperature at 3500 g/min for 10 minutes, aliquot the supernatant, and store at 2 to 8°C until blood glucose and insulin analysis. All experimental data were analyzed using GraphPad Prism 9.3.1.
表10-1分组及给药
Table 10-1 Grouping and administration
注：*静脉输注采用Harvard注射泵匀速进行。溶媒为5％聚乙二醇400(PEG400)+95％【20％磺丁基-β-环糊精(SEBCD)】，短暂超声后，溶媒完全溶解后，一次性注射器过滤器过滤后，用于配置药品；现配现用，并调整pH至7.0-7.4。Note: *Intravenous infusion is performed at a constant speed using a Harvard syringe pump. The solvent is 5% polyethylene glycol 400 (PEG400) + 95% [20% sulfobutyl-β-cyclodextrin (SEBCD)]. After a short period of ultrasound, the solvent is completely dissolved. After filtering with a disposable syringe filter, use For preparing medicines; prepare and use immediately, and adjust the pH to 7.0-7.4.
试验结果见表10-2：The test results are shown in Table 10-2:
表10-2血糖和胰岛素分析结果
Table 10-2 Blood glucose and insulin analysis results
结论：化合物I可以促进胰岛素的分泌、改善糖耐量和提高葡萄糖清除率。Conclusion: Compound I can promote insulin secretion, improve glucose tolerance and increase glucose clearance rate.
8.Caco2渗透性测试8.Caco2 Penetration Test
试验使用单层Caco-2细胞，在96孔Transwell板中采用三平行孵育。将含有本发明化合物(2μM)或对照化合物：化合物DZ-6、地高辛(10μM)、纳多洛尔(2μM)和美托 洛尔(2μM)的转运缓冲溶液(HBSS，10mM HEPES，pH 7.4±0.05)加入顶端侧或基底侧的给药端孔中。对应接收端孔中加入含DMSO的转运缓冲溶液。在37±1℃条件下孵育2小时后，取出细胞板并从顶端和底端各取出适量样品至新的96孔板中。随后加入含内标的乙腈沉淀蛋白。使用LC MS/MS分析样品并测定本发明化合物和对照化合物的浓度。浓度数据用于计算从单层细胞顶端侧向基底侧、以及基底侧向顶端转运的表观渗透系数，从而计算外排率。用荧光黄的渗漏评价孵育2小时后单层细胞的完整性。The assay uses a monolayer of Caco-2 cells incubated in triplicate in a 96-well Transwell plate. Compounds containing compounds of the invention (2 μM) or control compounds: compound DZ-6, digoxin (10 μM), nadolol (2 μM) and metoprolol A transport buffer solution (HBSS, 10mM HEPES, pH 7.4±0.05) of lorol (2 μM) was added to the administration port hole on the apical or basal side. Add DMSO-containing transport buffer solution to the corresponding receiving port hole. After incubating for 2 hours at 37±1°C, remove the cell plate and take appropriate amounts of samples from the top and bottom ends into a new 96-well plate. Acetonitrile containing internal standard was then added to precipitate the protein. Samples were analyzed using LC MS/MS and the concentrations of compounds of the invention and control compounds were determined. Concentration data were used to calculate apparent permeability coefficients for transport from the apical side to the basal side, and from the basal side to the apical side of the cell monolayer, and thus the efflux rate. The integrity of the cell monolayer after 2 hours of incubation was assessed by leakage of Lucifer Yellow.
表11-1.本发明化合物的Caco2测试结果
Table 11-1. Caco2 test results of the compounds of the present invention
结论：化合物I为低渗透化合物，外排率低于对照化合物DZ-6。 Conclusion: Compound I is a low-penetration compound, and its efflux rate is lower than that of the control compound DZ-6.

Claims (25)

  1. 式(I)所示化合物的可药用盐及其溶剂化物，
    Pharmaceutically acceptable salts and solvates of compounds represented by formula (I),
    所述可药用盐选自三羟甲基氨基甲烷盐或三乙醇胺盐。The pharmaceutically acceptable salt is selected from trishydroxymethylaminomethane salt or triethanolamine salt.
  2. 根据权利要求1所述的可药用盐及其溶剂化物，式(I)所示化合物：可药用盐的摩尔比为1:0.9～1:1.1。According to the pharmaceutically acceptable salt and its solvate according to claim 1, the molar ratio of the compound represented by formula (I): the pharmaceutically acceptable salt is 1:0.9~1:1.1.
  3. 根据权利要求1所述的可药用盐及其溶剂化物，式(I)所示化合物：可药用盐的摩尔比为1:1。According to the pharmaceutically acceptable salt and its solvate according to claim 1, the molar ratio of the compound represented by formula (I): the pharmaceutically acceptable salt is 1:1.
  4. 根据权利要求1所述的可药用盐及其溶剂化物，溶剂化物为甲醇。The pharmaceutically acceptable salt and solvate thereof according to claim 1, wherein the solvate is methanol.
  5. 一种式(I)所示化合物的三羟甲基氨基甲烷盐晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：8.97°±0.2°、9.32°±0.2°、9.92°±0.2°、10.68°±0.2°、17.50°±0.2°、20.17°±0.2°。A trishydroxymethylaminomethane salt crystal form A of a compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 8.97°±0.2°, 9.32 °±0.2°, 9.92°±0.2°, 10.68°±0.2°, 17.50°±0.2°, 20.17°±0.2°.
  6. 根据权利要求5所述的晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：8.97°±0.2°、9.32°±0.2°、9.92°±0.2°、10.68°±0.2°、15.07°±0.2°、17.50°±0.2°、17.71°±0.2°、19.95°±0.2°、20.17°±0.2°、24.27°±0.2°、26.81°±0.2°。According to the crystal form A of claim 5, using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 8.97°±0.2°, 9.32°±0.2°, 9.92°±0.2° , 10.68°±0.2°, 15.07°±0.2°, 17.50°±0.2°, 17.71°±0.2°, 19.95°±0.2°, 20.17°±0.2°, 24.27°±0.2°, 26.81°±0.2°.
  7. 根据权利要求5所述的晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：6.02°±0.2°、8.97°±0.2°、9.32°±0.2°、9.92°±0.2°、10.68°±0.2°、12.03±0.2°、13.77°±0.2°、15.07°±0.2°、16.18°±0.2°、17.50°±0.2°、17.71°±0.2°、18.27°±0.2°、18.70°±0.2°、19.15°±0.2°、19.95°±0.2°、20.17°±0.2°、20.90°±0.2°、21.89°±0.2°、22.35°±0.2°、24.05°±0.2°、24.27°±0.2°、24.65°±0.2°、25.03°±0.2°、25.54°±0.2°、26.81°±0.2°、28.03°±0.2°。According to the crystal form A of claim 5, using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 6.02°±0.2°, 8.97°±0.2°, 9.32°±0.2° , 9.92°±0.2°, 10.68°±0.2°, 12.03±0.2°, 13.77°±0.2°, 15.07°±0.2°, 16.18°±0.2°, 17.50°±0.2°, 17.71°±0.2°, 18.27° ±0.2°, 18.70°±0.2°, 19.15°±0.2°, 19.95°±0.2°, 20.17°±0.2°, 20.90°±0.2°, 21.89°±0.2°, 22.35°±0.2°, 24.05°±0.2 °, 24.27°±0.2°, 24.65°±0.2°, 25.03°±0.2°, 25.54°±0.2°, 26.81°±0.2°, 28.03°±0.2°.
  8. 根据权利要求5所述的晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱如图1所示。According to the crystal form A of claim 5, Cu-Kα radiation is used, and its X-ray powder diffraction pattern is shown in Figure 1.
  9. 一种式(I)所示化合物的三羟甲基氨基甲烷盐晶型C，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：10.55°±0.2°、13.69°±0.2°、19.41°±0.2°、20.70±0.2°。A trishydroxymethylaminomethane salt crystal form C of a compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 10.55°±0.2°, 13.69 °±0.2°, 19.41°±0.2°, 20.70±0.2°.
  10. 根据权利要求9所述的晶型C，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：10.55°±0.2°、10.92°±0.2°、13.69°±0.2°、17.98°±0.2°、19.41° ±0.2°、20.70±0.2°、21.97±0.2°、22.31±0.2°、26.44±0.2°。According to the crystal form C of claim 9, using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 10.55°±0.2°, 10.92°±0.2°, 13.69°±0.2° ,17.98°±0.2°,19.41° ±0.2°, 20.70±0.2°, 21.97±0.2°, 22.31±0.2°, 26.44±0.2°.
  11. 根据权利要求9所述的晶型C，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：5.44°±0.2°、8.12°±0.2°、10.17°±0.2°、10.55°±0.2°、10.92°±0.2°、11.56°±0.2°、13.08°±0.2°、13.69°±0.2°、14.49°±0.2°、15.26°±0.2°、17.06°±0.2°、17.78°±0.2°、17.98°±0.2°、18.41°±0.2°、19.41°±0.2°、20.70±0.2°、21.25±0.2°、21.97±0.2°、22.31±0.2°、23.27±0.2°、24.23±0.2°、25.24±0.2°、25.70±0.2°、26.44±0.2°、27.49±0.2°、28.71±0.2°。According to the crystal form C of claim 9, using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 5.44°±0.2°, 8.12°±0.2°, 10.17°±0.2° , 10.55°±0.2°, 10.92°±0.2°, 11.56°±0.2°, 13.08°±0.2°, 13.69°±0.2°, 14.49°±0.2°, 15.26°±0.2°, 17.06°±0.2°, 17.78 °±0.2°, 17.98°±0.2°, 18.41°±0.2°, 19.41°±0.2°, 20.70±0.2°, 21.25±0.2°, 21.97±0.2°, 22.31±0.2°, 23.27±0.2°, 24.23± 0.2°, 25.24±0.2°, 25.70±0.2°, 26.44±0.2°, 27.49±0.2°, 28.71±0.2°.
  12. 根据权利要求9所述的晶型C，使用Cu-Kα辐射，其X-射线粉末衍射图谱如图3所示。According to the crystal form C of claim 9, using Cu-Kα radiation, its X-ray powder diffraction pattern is shown in Figure 3.
  13. 一种式(I)所示化合物的三羟甲基氨基甲烷盐的晶型D，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：5.55°±0.2°、10.46°±0.2°、11.15°±0.2°、13.12°±0.2°、13.98°±0.2°、19.66°±0.2°、22.47°±0.2°。Crystalline form D of the trishydroxymethylaminomethane salt of a compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 5.55°±0.2°, 10.46°±0.2°, 11.15°±0.2°, 13.12°±0.2°, 13.98°±0.2°, 19.66°±0.2°, 22.47°±0.2°.
  14. 根据权利要求13所述的晶型D，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：5.55°±0.2°、8.34±0.2°、10.46°±0.2°、11.15°±0.2°、11.50±0.2°、13.12°±0.2°、13.98°±0.2°、19.66°±0.2°、20.80±0.2°、22.08±0.2°、22.47°±0.2°、23.26±0.2°、26.73±0.2°。According to the crystal form D of claim 13, using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 5.55°±0.2°, 8.34°±0.2°, 10.46°±0.2°, 11.15°±0.2°, 11.50±0.2°, 13.12°±0.2°, 13.98°±0.2°, 19.66°±0.2°, 20.80±0.2°, 22.08±0.2°, 22.47°±0.2°, 23.26±0.2°, 26.73±0.2°.
  15. 根据权利要求13所述的晶型D，使用Cu-Kα辐射，其X-射线粉末衍射图谱如图5所示。According to the crystal form D of claim 13, Cu-Kα radiation is used, and its X-ray powder diffraction pattern is shown in Figure 5.
  16. 一种式(I)所示化合物的三乙醇胺盐晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：10.42°±0.2°、12.68°±0.2°、14.80°±0.2°、16.37°±0.2°、16.94°±0.2°、23.39°±0.2°。The triethanolamine salt crystal form A of a compound represented by formula (I) uses Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 10.42°±0.2°, 12.68°±0.2° , 14.80°±0.2°, 16.37°±0.2°, 16.94°±0.2°, 23.39°±0.2°.
  17. 根据权利要求16所述的晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：8.42°±0.2°、10.42°±0.2°、12.68°±0.2°、13.28°±0.2°、14.80°±0.2°、16.37°±0.2°、16.94°±0.2°、18.04°±0.2°、22.26°±0.2°、23.39°±0.2°、24.98°±0.2°、25.55°±0.2°。According to the crystal form A of claim 16, using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 8.42°±0.2°, 10.42°±0.2°, 12.68°±0.2° , 13.28°±0.2°, 14.80°±0.2°, 16.37°±0.2°, 16.94°±0.2°, 18.04°±0.2°, 22.26°±0.2°, 23.39°±0.2°, 24.98°±0.2°, 25.55 °±0.2°.
  18. 根据权利要求16所述的晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱在以下2θ位置具有特征衍射峰：4.21±0.2°、6.30°±0.2°、8.42°±0.2°、10.42°±0.2°、12.68°±0.2°、13.28°±0.2°、14.35°±0.2°、14.80°±0.2°、15.11°±0.2°、16.10°±0.2°、16.37°±0.2°、16.94°±0.2°、17.32°±0.2°、18.04°±0.2°、18.72°±0.2°、19.83°±0.2°、20.96°±0.2°、21.23°±0.2°、21.39°±0.2°、21.70°±0.2°、21.85°±0.2°、22.26°±0.2°、22.61°±0.2°、23.39°±0.2°、24.22°±0.2°、24.55°±0.2°、24.98°±0.2°、25.55°±0.2°、26.48°±0.2°、28.07°±0.2°、29.90°±0.2°。According to the crystal form A of claim 16, using Cu-Kα radiation, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 4.21±0.2°, 6.30°±0.2°, 8.42°±0.2°, 10.42°±0.2°, 12.68°±0.2°, 13.28°±0.2°, 14.35°±0.2°, 14.80°±0.2°, 15.11°±0.2°, 16.10°±0.2°, 16.37°±0.2°, 16.94° ±0.2°, 17.32°±0.2°, 18.04°±0.2°, 18.72°±0.2°, 19.83°±0.2°, 20.96°±0.2°, 21.23°±0.2°, 21.39°±0.2°, 21.70°±0.2 °, 21.85°±0.2°, 22.26°±0.2°, 22.61°±0.2°, 23.39°±0.2°, 24.22°±0.2°, 24.55°±0.2°, 24.98°±0.2°, 25.55°±0.2°, 26.48°±0.2°, 28.07°±0.2°, 29.90°±0.2°.
  19. 根据权利要求16所述的晶型A，使用Cu-Kα辐射，其X-射线粉末衍射图谱如图7 所示。According to the crystal form A of claim 16, using Cu-Kα radiation, its X-ray powder diffraction pattern is shown in Figure 7 shown.
  20. 一种式(I)所示化合物的可药用盐的制备方法，其中，所述方法包括：以式(I)所示化合物和酸成盐的步骤。A method for preparing a pharmaceutically acceptable salt of a compound represented by formula (I), wherein the method includes the step of forming a salt from a compound represented by formula (I) and an acid.
  21. 根据权利要求20所述的制备方法，其中，所用溶剂选自C1-6卤代烷烃类溶剂、C2-6酯类溶剂、C2-6醚类溶剂、C1-6醇类溶剂或水中的一种或多种。The preparation method according to claim 20, wherein the solvent used is selected from C 1-6 halogenated alkane solvents, C 2-6 ester solvents, C 2-6 ether solvents, C 1-6 alcohol solvents or water of one or more.
  22. 根据权利要求21所述的制备方法，其中，所用溶剂选自二氯甲烷、1,2-二氯乙烷、乙酸乙酯、甲醇、乙醇、异丙醇、丙醇、***、四氢呋喃和水中的一种或多种。The preparation method according to claim 21, wherein the solvent used is selected from the group consisting of dichloromethane, 1,2-dichloroethane, ethyl acetate, methanol, ethanol, isopropyl alcohol, propanol, ether, tetrahydrofuran and water. one or more.
  23. 一种药物组合物，其中，所述药物组合物含有治疗有效量的权利要求1～4任意一项所述的化合物的可药用盐及其溶剂化物或者权利要求5-22任意一项所述的晶型、及药学上可接受的载体或赋形剂。A pharmaceutical composition, wherein the pharmaceutical composition contains a therapeutically effective amount of a pharmaceutically acceptable salt and a solvate of the compound described in any one of claims 1 to 4 or any one of claims 5 to 22 The crystal form, and pharmaceutically acceptable carrier or excipient.
  24. 根据权利要求23所述的药物组合物，治疗有效量以游离碱形式计算为1-600mg。According to the pharmaceutical composition of claim 23, the therapeutically effective amount is 1-600 mg in free base form.
  25. 权利要求1～4任意一项所述的化合物的可药用盐及其溶剂化物或者权利要求5-19任意一项所述的晶型或者权利要求23-24任一项所述的药物组合物在制备用于治疗糖尿病或糖尿病相关的疾病的药物中的应用。 Pharmaceutically acceptable salts and solvates of the compounds described in any one of claims 1 to 4 or crystal forms described in any one of claims 5 to 19 or pharmaceutical compositions described in any one of claims 23 to 24 Use in the preparation of medicaments for the treatment of diabetes or diabetes-related diseases.
PCT/CN2023/101590 2022-06-23 2023-06-21 Salt of thieno[2,3-d]imidazole compound, crystal form thereof and pharmaceutical use thereof WO2023246839A1 (en)

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WO2021244645A1 (en) * 2020-06-04 2021-12-09 杭州先为达生物科技有限公司 Five-membered heteroaromatic imidazole compound and use thereof
WO2021249492A1 (en) * 2020-06-10 2021-12-16 南京明德新药研发有限公司 Methyl-substituted benzobisoxazole compound and use thereof
CN113853371A (en) * 2019-04-12 2021-12-28 上海齐鲁锐格医药研发有限公司 GLP-1R agonists and uses thereof
CN114591296A (en) * 2020-12-02 2022-06-07 海思科医药集团股份有限公司 GLP-1R agonist and application thereof in medicine
WO2022135572A1 (en) * 2020-12-25 2022-06-30 四川海思科制药有限公司 Five-membered ring derivative and medical use thereof

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CN113853371A (en) * 2019-04-12 2021-12-28 上海齐鲁锐格医药研发有限公司 GLP-1R agonists and uses thereof
WO2021244645A1 (en) * 2020-06-04 2021-12-09 杭州先为达生物科技有限公司 Five-membered heteroaromatic imidazole compound and use thereof
WO2021249492A1 (en) * 2020-06-10 2021-12-16 南京明德新药研发有限公司 Methyl-substituted benzobisoxazole compound and use thereof
CN114591296A (en) * 2020-12-02 2022-06-07 海思科医药集团股份有限公司 GLP-1R agonist and application thereof in medicine
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