WO2022100586A1 - Crystal form of oxetane-containing spirocyclic compound, preparation method therefor and use thereof - Google Patents

Crystal form of oxetane-containing spirocyclic compound, preparation method therefor and use thereof Download PDF

Info

Publication number
WO2022100586A1
WO2022100586A1 PCT/CN2021/129643 CN2021129643W WO2022100586A1 WO 2022100586 A1 WO2022100586 A1 WO 2022100586A1 CN 2021129643 W CN2021129643 W CN 2021129643W WO 2022100586 A1 WO2022100586 A1 WO 2022100586A1
Authority
WO
WIPO (PCT)
Prior art keywords
crystal form
compound
formula
etoac
angles
Prior art date
Application number
PCT/CN2021/129643
Other languages
French (fr)
Chinese (zh)
Inventor
李翼
刘宁
姚婷
于涛
吴成德
陈曙辉
Original Assignee
南京明德新药研发有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 南京明德新药研发有限公司 filed Critical 南京明德新药研发有限公司
Publication of WO2022100586A1 publication Critical patent/WO2022100586A1/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • 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

Definitions

  • the present invention relates to a crystal form of an oxetane-containing spiro compound, a preparation method and an application thereof, in particular to a crystal form of a compound of formula (I), a preparation method and an application thereof.
  • Ras/Raf/MEK/ERK pathway is a classic mitogen activated protein kinase (MAPK) signaling cascade, which is involved in the signaling of various growth factors, cytokines, mitogens and hormone receptors after activation Transduction is one of the most important signal transduction pathways controlling cell growth, differentiation and survival.
  • MAPK mitogen activated protein kinase
  • Extracellular regulated protein kinases are major players and downstream key nodes in the Ras/Raf/MEK/ERK pathway, and their functions can be found in many human cancers. overactive. As the terminal signaling kinase of this pathway, ERK has not yet been found to have drug resistance mutations. Therefore, drugs targeting ERK kinase are expected to overcome the drug resistance problem after upstream target inhibitor treatment and become a more potential therapeutic strategy. But so far, research on ERK inhibitors is still in the clinical stage, and no ERK inhibitors have been approved for marketing as drugs. In conclusion, there is an urgent need to develop safe and effective ERK inhibitor drugs to meet the needs of tumor treatment.
  • ERKs Extracellular regulated protein kinases
  • the present invention provides Form A of the compound of formula (I), whose X-ray powder diffraction (XRPD) pattern has characteristic diffraction peaks at the following 2 ⁇ angles: 8.16 ⁇ 0.20°, 17.39 ⁇ 0.20° and 23.88 ⁇ 0.20°.
  • the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2 ⁇ angles: 8.16 ⁇ 0.20°, 13.57 ⁇ 0.20°, 16.95 ⁇ 0.20°, 17.39 ⁇ 0.20°, 19.37 ⁇ 0.20 °, 22.22 ⁇ 0.20°, 23.88 ⁇ 0.20° and 25.85 ⁇ 0.20°.
  • the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2 ⁇ angles: 8.16 ⁇ 0.20°, 13.57 ⁇ 0.20°, 14.51 ⁇ 0.20°, 16.95 ⁇ 0.20°, 17.39 ⁇ 0.20 °, 19.37 ⁇ 0.20°, 22.22 ⁇ 0.20°, 23.88 ⁇ 0.20° and 25.85 ⁇ 0.20°.
  • the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2 ⁇ angles: 8.16 ⁇ 0.20°, 9.99 ⁇ 0.20°, 13.57 ⁇ 0.20°, 14.51 ⁇ 0.20°, 15.61 ⁇ 0.20 degrees degrees ° and 37.72 ⁇ 0.20°.
  • the X-ray powder diffraction pattern of the above-mentioned A crystal form has characteristic diffraction peaks at the following 2 ⁇ angles: 8.16°, 9.99°, 13.57°, 14.51°, 15.61°, 16.95°, 17.39°, 18.27° , 19.37°, 19.71°, 21.40°, 21.97°, 22.22°, 23.88°, 25.02°, 25.85°, 26.50°, 27.36°, 28.72°, 30.39°, 31.19°, 31.53°, 31.97°, 32.57°, 33.05 ° and 37.72°.
  • the X-ray powder diffraction pattern of the above-mentioned Form A has characteristic diffraction peaks at the following 2 ⁇ angles: 8.16 ⁇ 0.20°, 17.39 ⁇ 0.20°, and/or 23.88 ⁇ 0.20°, and/or 13.57 ⁇ 0.20°, and/or 16.95 ⁇ 0.20°, and/or 19.37 ⁇ 0.20°, and/or 22.22 ⁇ 0.20°, and/or 25.85 ⁇ 0.20°, and/or 9.99 ⁇ 0.20°, and/or 14.51 ⁇ 0.20° , and/or 15.61 ⁇ 0.20°, and/or 18.27 ⁇ 0.20°, and/or 19.71 ⁇ 0.20°, and/or 21.40 ⁇ 0.20°, and/or 21.97 ⁇ 0.20°, and/or 25.02 ⁇ 0.20°, and /or 26.50 ⁇ 0.20°, and/or 27.36 ⁇ 0.20°, and/or 28.72 ⁇ 0.20°, and/or 30.39 ⁇ 0.20°, and/or 31
  • the XRPD pattern of the above-mentioned crystal form A is shown in FIG. 1 .
  • the differential scanning calorimetry curve of the differential scanning calorimetry curve has an onset of an endothermic peak at 225.5 ⁇ 3°C.
  • the DSC spectrum of the above-mentioned crystal form A is shown in FIG. 2 .
  • thermogravimetric analysis curve (TGA) of the above-mentioned crystal form A has a weight loss of 2.41% at 200 ⁇ 3°C.
  • the TGA spectrum of the above-mentioned A crystal form is shown in FIG. 3 .
  • the present invention also provides a method for preparing the crystal form A of the compound of formula (I), comprising:
  • the organic solvent is ethanol, MeOH, ACN, acetone, EtOAc, dioxane, toluene, H 2 O, THF:H 2 O (v/v, 1:1), EtOH:H 2 O (v/v, 2:1), Acetone:EtOAc (v/v, 1:1), DMF:EtOAc (v/v, 5:95), DMF: H2O (v/v, 5:1) 95), acetone: H2O (v/v, 95:5), dioxane: H2O (v/v, 97:3), THF, DCM:EtOAc (v/v, 2:1) .
  • the crystal form A of the compound of the present invention is stable, less affected by light, heat and humidity, and has a broad prospect for medicine; the crystal form A of the compound of formula (I) can significantly inhibit tumor growth under the administration dose, showing a dose-dependent manner; There was no significant weight loss, and the tolerance was good.
  • the intermediate compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by their combination with other chemical synthesis methods, and those skilled in the art.
  • Well-known equivalents, preferred embodiments include, but are not limited to, the examples of the present invention.
  • the solvent used in the present invention is commercially available.
  • the structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction method (SXRD), the cultured single crystal is collected by Bruker D8 venture diffractometer, the light source is CuK ⁇ radiation, and the scanning mode is: After scanning and collecting relevant data, the crystal structure was further analyzed by the direct method (Shelxs97), and the absolute configuration could be confirmed.
  • SXRD single crystal X-ray diffraction method
  • the cultured single crystal is collected by Bruker D8 venture diffractometer
  • the light source is CuK ⁇ radiation
  • the scanning mode is: After scanning and collecting relevant data, the crystal structure was further analyzed by the direct method (Shelxs97), and the absolute configuration could be confirmed.
  • DCM dichloromethane
  • DMF N,N-dimethylformamide
  • DMSO dimethyl sulfoxide
  • EtOH for ethanol
  • MeOH for methanol
  • 2-MeTHF 2-methyl Tetrahydrofuran
  • Dioxane for dioxane
  • ACN for acetonitrile
  • Toluene for toluene
  • Acetone for acetone
  • EtOAc for ethyl acetate
  • THF tetrahydrofuran
  • FIG. 5 Tumor growth curve of human colon cancer HCT-116 model animal after administration of solvent and compound A crystal form of formula (I) respectively;
  • Fig. 6 Body weight change rate (%) of human colon cancer HCT-116 xenograft tumor model animals during administration.
  • the XRPD test was performed using an X'Pert3 X-ray diffractometer from PANalytical. The test parameters are shown in Table 2.
  • the DSC spectra were collected on a TA 2500 differential scanning calorimeter, and the test parameters are shown in Table 3.
  • TGA was collected on a TA 5500 thermogravimetric analyzer, and the test parameters are shown in Table 4.
  • Dynamic moisture adsorption (DVS) curves were collected on a DVS Intrinsic instrument from SMS (Surface Measurement Systems), UK. The relative humidity at 25°C was corrected for the deliquescence points of lithium chloride (LiCl), magnesium nitrate [Mg( NO3 ) 2 ] and potassium chloride (KCl). The test parameters are shown in Table 5.
  • Hygroscopic classification ⁇ W% deliquescence Absorbs enough water to form a liquid Very hygroscopic ⁇ W% ⁇ 15% hygroscopic 15%> ⁇ W% ⁇ 2% slightly hygroscopic 2%> ⁇ W% ⁇ 0.2% No or almost no hygroscopicity ⁇ W% ⁇ 0.2%
  • ⁇ W% represents the moisture absorption weight gain of the test product at 25°C/80%RH.
  • the synthetic route is as follows:
  • step 1
  • the crude product was obtained by concentration under reduced pressure.
  • Dioxane (500 mL) and water (2.5 L) were added to the crude product, stirred at 25° C. for 0.5 hours, and the filter cake was collected by filtration.
  • the filter cake was dissolved in ethanol (2000 mL), stirred at 25° C. for 1 hour, and the filter cake was collected by filtration.
  • the filter cake was dissolved in ethanol (2000 mL), stirred at 75° C. for 1 hour, cooled to 25° C., and the filter cake was collected by filtration.
  • the crude product was dissolved in dioxane (500 mL) and stirred for 10 minutes, then deionized water (2.5 L) was added to continue stirring for 0.5 hour, and the filter cake was collected by filtration.
  • the filter cake was dissolved in deionized water (1 L) and stirred for 1 hour, and the filter cake was collected by filtration.
  • the filter cake was dissolved in absolute ethanol (2L), stirred at 25°C for 1 hour, and the filter cake was collected by filtration.
  • the filter cake was dissolved in absolute ethanol (2L), stirred at 85°C for 1 hour, filtered to collect the filter cake, and the filter cake was vacuum dried at 45°C to obtain the compound of formula (I).
  • test sample marked S1-condition-time is used for content and related substance detection; the test sample marked S2-condition-time is used as a preparation sample, and the test sample marked S3-condition-time is used for XRPD detection .
  • Table 9 shows the results of the solid stability experiment of crystal form A.
  • HCT-116 cells Human colon cancer HCT-116 cells were cultured in monolayer in vitro, and the culture conditions were McCoy's 5a medium plus 10% fetal bovine serum, and cultured in a 5% CO2 incubator at 37°C. Conventional digestion treatment with trypsin-EDTA was passaged three times a week. When the cell saturation is 80%-90% and the number reaches the requirement, the cells are collected, counted, and seeded;
  • Vehicle group 5% DMSO+95% (10% HP- ⁇ -CD).
  • Test compound group weigh the quantitative test compound into a dispensing bottle, add a corresponding volume of DMSO and vortex to obtain a clear solution, add a corresponding volume of 10% HP- ⁇ -CD and vortex to obtain a clear solution solution. Compounds were formulated every three days.
  • Tumor diameter was measured twice a week with a vernier caliper.
  • TGI (%) The tumor-inhibitory efficacy of the compounds was evaluated by TGI (%).
  • the crystal form of compound A of formula (I) can significantly inhibit tumor growth under the administration dose, showing a dose-dependent manner; the body weight of the animals does not decrease significantly during the administration process, and the tolerance is good.

Abstract

Provided are a crystal form of an oxetane-containing spirocyclic compound, a preparation method therefor and the use thereof. The present invention particularly relates to a crystal form of a compound of formula (I), a preparation method therefor and the use thereof.

Description

一种含氧杂环丁烷的螺环类化合物的晶型、制备方法及其应用A kind of oxetane-containing spiro compound crystal form, preparation method and application thereof
本申请主张如下优先权：This application claims the following priority:
CN202011252031.8，申请日2020.11.11。CN202011252031.8, application date 2020.11.11.
技术领域technical field
本发明涉及一种含氧杂环丁烷的螺环类化合物的晶型、制备方法及其应用，具体涉及式(I)化合物晶型、制备方法及其应用。The present invention relates to a crystal form of an oxetane-containing spiro compound, a preparation method and an application thereof, in particular to a crystal form of a compound of formula (I), a preparation method and an application thereof.
背景技术Background technique
Ras/Raf/MEK/ERK通路是一条经典的有丝***原活化蛋白激酶(mitogen activated protein kinase,MAPK)信号级联通路，参与各种生长因子、细胞因子、丝裂原以及激素受体活化后的信号转导，是控制细胞生长、分化和存活最重要的信号转导途径之一。The Ras/Raf/MEK/ERK pathway is a classic mitogen activated protein kinase (MAPK) signaling cascade, which is involved in the signaling of various growth factors, cytokines, mitogens and hormone receptors after activation Transduction is one of the most important signal transduction pathways controlling cell growth, differentiation and survival.
研究表明，突变或扩增引起的Ras/Raf/MEK/ERK通路异常活化是多种癌症发生的决定因素。在人类肿瘤中，RAS突变发生率约为22％，BRAF突变发生率约为7％，MEK突变发生率约为1％，因此，该通路上的关键节点蛋白已成为癌症治疗的重要靶点(Cancer Discov.2019,9,329-341)。目前，已有多个BRAF抑制剂和MEK1/2抑制剂，以及它们的联用方案，被美国FDA批准用于黑色素瘤、BRAFV600E突变型非小细胞肺癌等癌症的治疗。然而，使用这些上游节点的BRAF和MEK抑制剂后，由于突变或通路重新激活，会快速导致耐药性问题，极大地限制了它们的临床应用。Studies have shown that abnormal activation of Ras/Raf/MEK/ERK pathway caused by mutation or amplification is a determinant of various cancers. In human tumors, the incidence of RAS mutations is about 22%, the incidence of BRAF mutations is about 7%, and the incidence of MEK mutations is about 1%. Therefore, key node proteins on this pathway have become important targets for cancer therapy ( Cancer Discov. 2019, 9, 329-341). At present, a number of BRAF inhibitors and MEK1/2 inhibitors, as well as their combination regimens, have been approved by the US FDA for the treatment of melanoma, BRAFV600E mutant non-small cell lung cancer and other cancers. However, the use of BRAF and MEK inhibitors at these upstream nodes can rapidly lead to drug resistance problems due to mutation or pathway reactivation, greatly limiting their clinical application.
细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)，特别是ERK1和ERK2激酶，是Ras/Raf/MEK/ERK通路的主要参与者和下游关键节点，在许多人类的癌症中都可发现它们的过度激活。ERK作为该通路的末端信号激酶，目前尚未发现有耐药突变，因此，靶向ERK激酶的药物有望克服上游靶点抑制剂治疗后产生的耐药性问题，成为更具潜力的治疗策略。但迄今为止，关于ERK抑制剂的研究仍处于临床阶段，还没有ERK抑制剂作为药物批准上市。综上所述，迫切需要研发出安全、有效的ERK抑制剂药物满足肿瘤治疗的需要。Extracellular regulated protein kinases (ERKs), especially ERK1 and ERK2 kinases, are major players and downstream key nodes in the Ras/Raf/MEK/ERK pathway, and their functions can be found in many human cancers. overactive. As the terminal signaling kinase of this pathway, ERK has not yet been found to have drug resistance mutations. Therefore, drugs targeting ERK kinase are expected to overcome the drug resistance problem after upstream target inhibitor treatment and become a more potential therapeutic strategy. But so far, research on ERK inhibitors is still in the clinical stage, and no ERK inhibitors have been approved for marketing as drugs. In conclusion, there is an urgent need to develop safe and effective ERK inhibitor drugs to meet the needs of tumor treatment.
发明内容SUMMARY OF THE INVENTION
本发明提供了式(Ⅰ)化合物的A晶型，其X射线粉末衍射(XRPD)图谱在下列2θ角处具有特征衍射峰：8.16±0.20°、17.39±0.20°和23.88±0.20°。The present invention provides Form A of the compound of formula (I), whose X-ray powder diffraction (XRPD) pattern has characteristic diffraction peaks at the following 2θ angles: 8.16±0.20°, 17.39±0.20° and 23.88±0.20°.
Figure PCTCN2021129643-appb-000001
Figure PCTCN2021129643-appb-000001
本发明的一些方案中，上述A晶型的X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.16±0.20°、13.57±0.20°、16.95±0.20°、17.39±0.20°、19.37±0.20°、22.22±0.20°、23.88±0.20°和25.85±0.20°。In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 8.16±0.20°, 13.57±0.20°, 16.95±0.20°, 17.39±0.20°, 19.37±0.20 °, 22.22±0.20°, 23.88±0.20° and 25.85±0.20°.
本发明的一些方案中，上述A晶型的X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.16±0.20°、13.57±0.20°、14.51±0.20°、16.95±0.20°、17.39±0.20°、19.37±0.20°、22.22±0.20°、23.88±0.20°和25.85±0.20°。In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 8.16±0.20°, 13.57±0.20°, 14.51±0.20°, 16.95±0.20°, 17.39±0.20 °, 19.37±0.20°, 22.22±0.20°, 23.88±0.20° and 25.85±0.20°.
本发明的一些方案中，上述A晶型的X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.16±0.20°、9.99±0.20°、13.57±0.20°、14.51±0.20°、15.61±0.20°、16.95±0.20°、17.39±0.20°、18.27±0.20°、19.37±0.20°、19.71±0.20°、21.40±0.20°、21.97±0.20°、22.22±0.20°、23.88±0.20°、25.02±0.20°、25.85±0.20°、26.50±0.20°、27.36±0.20°、28.72±0.20°、30.39±0.20°、31.19±0.20°、31.53±0.20°、31.97±0.20°、32.57±0.20°、33.05±0.20°和37.72±0.20°。In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 8.16±0.20°, 9.99±0.20°, 13.57±0.20°, 14.51±0.20°, 15.61±0.20 degrees degrees ° and 37.72±0.20°.
本发明的一些方案中，上述A晶型的X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.16°、9.99°、13.57°、14.51°、15.61°、16.95°、17.39°、18.27°、19.37°、19.71°、21.40°、21.97°、22.22°、23.88°、25.02°、25.85°、26.50°、27.36°、28.72°、30.39°、31.19°、31.53°、31.97°、32.57°、33.05°和37.72°。In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A crystal form has characteristic diffraction peaks at the following 2θ angles: 8.16°, 9.99°, 13.57°, 14.51°, 15.61°, 16.95°, 17.39°, 18.27° , 19.37°, 19.71°, 21.40°, 21.97°, 22.22°, 23.88°, 25.02°, 25.85°, 26.50°, 27.36°, 28.72°, 30.39°, 31.19°, 31.53°, 31.97°, 32.57°, 33.05 ° and 37.72°.
本发明的一些方案中，上述A晶型的X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.16±0.20°，17.39±0.20°，和/或23.88±0.20°，和/或13.57±0.20°、和/或16.95±0.20°、和/或19.37±0.20°、和/或22.22±0.20°，和/或25.85±0.20°，和/或9.99±0.20°，和/或14.51±0.20°，和/或15.61±0.20°，和/或18.27±0.20°，和/或19.71±0.20°，和/或21.40±0.20°，和/或21.97±0.20°，和/或25.02±0.20°，和/或26.50±0.20°，和/或27.36±0.20°，和/或28.72±0.20°，和/或30.39±0.20°，和/或31.19±0.20°，和/或31.53±0.20°，和/或31.97±0.20°，和/或32.57±0.20°，和/或33.05±0.20°，和/或37.72±0.20°。In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned Form A has characteristic diffraction peaks at the following 2θ angles: 8.16±0.20°, 17.39±0.20°, and/or 23.88±0.20°, and/or 13.57± 0.20°, and/or 16.95±0.20°, and/or 19.37±0.20°, and/or 22.22±0.20°, and/or 25.85±0.20°, and/or 9.99±0.20°, and/or 14.51±0.20° , and/or 15.61±0.20°, and/or 18.27±0.20°, and/or 19.71±0.20°, and/or 21.40±0.20°, and/or 21.97±0.20°, and/or 25.02±0.20°, and /or 26.50±0.20°, and/or 27.36±0.20°, and/or 28.72±0.20°, and/or 30.39±0.20°, and/or 31.19±0.20°, and/or 31.53±0.20°, and/or 31.97±0.20°, and/or 32.57±0.20°, and/or 33.05±0.20°, and/or 37.72±0.20°.
本发明的一些方案中，上述A晶型，其XRPD图谱如附图1所示。In some solutions of the present invention, the XRPD pattern of the above-mentioned crystal form A is shown in FIG. 1 .
本发明的一些方案中，上述A晶型的XRPD图谱解析数据如表1所示。In some solutions of the present invention, the XRPD pattern analysis data of the above-mentioned A crystal form is shown in Table 1.
表1 式(I)化合物A晶型的XRPD解析数据Table 1 XRPD analysis data of the crystal form of compound A of formula (I)
Figure PCTCN2021129643-appb-000002
Figure PCTCN2021129643-appb-000002
在本发明的一些方案中，上述A晶型，其差示扫描量热曲线在225.5±3℃有一个吸热峰的起始点。In some embodiments of the present invention, for the above-mentioned crystal form A, the differential scanning calorimetry curve of the differential scanning calorimetry curve has an onset of an endothermic peak at 225.5±3°C.
在本发明的一些方案中，上述A晶型，其DSC图谱如附图2所示。In some embodiments of the present invention, the DSC spectrum of the above-mentioned crystal form A is shown in FIG. 2 .
在本发明的一些方案中，上述A晶型的热重分析曲线(TGA)在200±3℃时失重达2.41％。In some embodiments of the present invention, the thermogravimetric analysis curve (TGA) of the above-mentioned crystal form A has a weight loss of 2.41% at 200±3°C.
在本发明的一些方案中，上述A晶型的TGA图谱如图3所示。In some embodiments of the present invention, the TGA spectrum of the above-mentioned A crystal form is shown in FIG. 3 .
本发明还提供了式(Ⅰ)化合物的A晶型的制备方法，包含：The present invention also provides a method for preparing the crystal form A of the compound of formula (I), comprising:
1)式(I)化合物，加入有机溶剂；1) compound of formula (I), adding organic solvent;
2)在25～50℃温度下搅拌2天；2) Stir for 2 days at a temperature of 25 to 50 °C;
3)过滤收集固体，置于真空干燥箱中(45℃)干燥16小时。3) The solid was collected by filtration, and dried in a vacuum drying oven (45° C.) for 16 hours.
本发明的一些方案中，上述有机溶剂为乙醇、MeOH、ACN、丙酮、EtOAc、二氧六环、甲苯、H 2O、THF:H 2O(v/v，1:1)、EtOH:H 2O(v/v，2:1)、丙酮:EtOAc(v/v，1:1)、DMF:EtOAc(v/v，5:95)、DMF:H 2O(v/v，5:95)、丙酮:H 2O(v/v，95:5)、二氧六环:H 2O(v/v，97:3)、THF、DCM:EtOAc(v/v，2:1)。 In some solutions of the present invention, the organic solvent is ethanol, MeOH, ACN, acetone, EtOAc, dioxane, toluene, H 2 O, THF:H 2 O (v/v, 1:1), EtOH:H 2 O (v/v, 2:1), Acetone:EtOAc (v/v, 1:1), DMF:EtOAc (v/v, 5:95), DMF: H2O (v/v, 5:1) 95), acetone: H2O (v/v, 95:5), dioxane: H2O (v/v, 97:3), THF, DCM:EtOAc (v/v, 2:1) .
技术效果technical effect
本发明化合物的A晶型稳定、受光热湿度影响小、成药前景广阔；式(I)化合物A晶型在给药剂量下，能显著抑制肿瘤生长，呈现剂量依赖性；给药过程中动物的体重未见明显下降，耐受性好。The crystal form A of the compound of the present invention is stable, less affected by light, heat and humidity, and has a broad prospect for medicine; the crystal form A of the compound of formula (I) can significantly inhibit tumor growth under the administration dose, showing a dose-dependent manner; There was no significant weight loss, and the tolerance was good.
定义和说明Definition and Explanation
除非另有说明，本文所用的下列术语和短语旨在含有下列含义。一个特定的短语或术语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文出现商品名时，旨在指代其对应的商品或其活性成分。Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular phrase or term should not be considered indeterminate or unclear without a specific definition, but should be understood in its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commercial product or its active ingredient.
本发明的中间体化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。The intermediate compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by their combination with other chemical synthesis methods, and those skilled in the art. Well-known equivalents, preferred embodiments include, but are not limited to, the examples of the present invention.
本发明具体实施方式的化学反应是在合适的溶剂中完成的，所述的溶剂须适合于本发明的化学变化及其所需的试剂和物料。为了获得本发明的化合物，有时需要本领域技术人员在已有实施方式的基础上对合成步骤或者反应流程进行修改或选择。The chemical reactions of specific embodiments of the present invention are carried out in suitable solvents suitable for the chemical changes of the present invention and their required reagents and materials. In order to obtain the compounds of the present invention, it is sometimes necessary for those skilled in the art to modify or select the synthetic steps or reaction schemes on the basis of the existing embodiments.
下面会通过实施例具体描述本发明，这些实施例并不意味着对本发明的任何限制。The present invention will be specifically described below through examples, which do not imply any limitation to the present invention.
本发明所使用的所有溶剂是市售的，无需进一步纯化即可使用。All solvents used in the present invention are commercially available and used without further purification.
本发明所使用的溶剂可经市售获得。The solvent used in the present invention is commercially available.
本发明的化合物可以通过本领域技术人员所熟知的常规方法来确认结构，如果本发明涉及化合物的绝对构型，则该绝对构型可以通过本领域常规技术手段予以确证。例如单晶X射线衍射法(SXRD)，把培养出的单晶用Bruker D8 venture衍射仪收集衍射强度数据，光源为CuKα辐射，扫描方式：
Figure PCTCN2021129643-appb-000003
扫描，收集 相关数据后，进一步采用直接法(Shelxs97)解析晶体结构，便可以确证绝对构型。 The structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction method (SXRD), the cultured single crystal is collected by Bruker D8 venture diffractometer, the light source is CuKα radiation, and the scanning mode is:
Figure PCTCN2021129643-appb-000003
After scanning and collecting relevant data, the crystal structure was further analyzed by the direct method (Shelxs97), and the absolute configuration could be confirmed.
本发明采用下述缩略词：DCM代表二氯甲烷；DMF代表N，N-二甲基甲酰胺；DMSO代表二甲亚砜；EtOH代表乙醇；MeOH代表甲醇；2-MeTHF代表2-甲基四氢呋喃；Dioxane代表二氧六环；ACN代表乙腈；Toluene代表甲苯；Acetone代表丙酮；EtOAc代表乙酸乙酯；THF代表四氢呋喃；H 2O代表水。 The following abbreviations are used in the present invention: DCM for dichloromethane; DMF for N,N-dimethylformamide; DMSO for dimethyl sulfoxide; EtOH for ethanol; MeOH for methanol; 2-MeTHF for 2-methyl Tetrahydrofuran; Dioxane for dioxane; ACN for acetonitrile; Toluene for toluene; Acetone for acetone; EtOAc for ethyl acetate; THF for tetrahydrofuran;
化合物依据本领域常规命名原则或者使用
Figure PCTCN2021129643-appb-000004
软件命名，市售化合物采用供应商目录名称。 Compounds are named according to conventional nomenclature in the art or are used
Figure PCTCN2021129643-appb-000004
Software naming, commercially available compounds use supplier catalog names.
附图说明Description of drawings
图1：式(Ⅰ)化合物A晶型的XRPD图谱；Figure 1: XRPD pattern of the crystal form of compound A of formula (I);
图2：式(Ⅰ)化合物A晶型的DSC图谱；Figure 2: DSC spectrum of the crystal form of compound A of formula (I);
图3：式(Ⅰ)化合物A晶型的TGA图谱；Figure 3: TGA spectrum of the crystal form of compound A of formula (I);
图4：式(Ⅰ)化合物A晶型的DVS图谱；Figure 4: DVS spectrum of the crystal form of compound A of formula (I);
图5：人结肠癌HCT-116模型动物在分别给予溶剂和式(Ⅰ)化合物A晶型后的肿瘤生长曲线；Figure 5: Tumor growth curve of human colon cancer HCT-116 model animal after administration of solvent and compound A crystal form of formula (I) respectively;
图6：人结肠癌HCT-116异种移植瘤模型动物在给药过程中的体重变化率(％)。Fig. 6: Body weight change rate (%) of human colon cancer HCT-116 xenograft tumor model animals during administration.
本发明X射线粉末衍射(XRPD)仪器信息和方法X-ray Powder Diffraction (XRPD) Instrument Information and Methods of the Invention
XRPD测试使用PANalytical(帕纳科)公司的X’Pert3型X射线衍射仪。测试参数如表2所示。The XRPD test was performed using an X'Pert3 X-ray diffractometer from PANalytical. The test parameters are shown in Table 2.
表2：XRPD测试参数Table 2: XRPD Test Parameters
Figure PCTCN2021129643-appb-000005
Figure PCTCN2021129643-appb-000005
本发明差示扫描量热(DSC)仪器信息及方法Differential Scanning Calorimetry (DSC) Instrument Information and Methods of the Invention
DSC图谱在TA 2500差示扫描量热仪上采集，测试参数如表3所示。The DSC spectra were collected on a TA 2500 differential scanning calorimeter, and the test parameters are shown in Table 3.
表3：DSC测试参数Table 3: DSC Test Parameters
参数parameter 设定值set value
方法method 线性升温Linear heating
参数parameter 设定值set value
样品盘sample tray 铝盘，压盖Aluminium pan, gland
温度范围temperature range 25～350℃25～350℃
扫描速率(℃/分钟)Scan rate (℃/min) 1010
保护气体Protective gas 氮气nitrogen
本发明热重分析(TGA)仪器信息与方法Thermogravimetric Analysis (TGA) Instrument Information and Methods of the Invention
TGA在TA 5500热重分析仪上采集，测试参数如表4所示。TGA was collected on a TA 5500 thermogravimetric analyzer, and the test parameters are shown in Table 4.
表4：TGA测试参数Table 4: TGA Test Parameters
参数parameter 设定值set value
方法method 线性升温Linear heating
样品盘sample tray 铝盘，开盖Aluminium tray, open lid
温度范围temperature range 室温～350℃Room temperature～350℃
扫描速率(℃/分钟)Scan rate (℃/min) 1010
保护气体Protective gas 氮气nitrogen
本发明动态蒸汽吸附分析(DVS)仪器信息及方法Dynamic Vapor Sorption Analysis (DVS) Instrument Information and Method of the Invention
动态水分吸附(DVS)曲线在英国SMS(Surface Measurement Systems)公司的DVS Intrinsic仪器上采集。在25℃时的相对湿度用氯化锂(LiCl)，硝酸镁[Mg(NO 3) 2]和氯化钾(KCl)的潮解点校正。测试参数如表5所示。 Dynamic moisture adsorption (DVS) curves were collected on a DVS Intrinsic instrument from SMS (Surface Measurement Systems), UK. The relative humidity at 25°C was corrected for the deliquescence points of lithium chloride (LiCl), magnesium nitrate [Mg( NO3 ) 2 ] and potassium chloride (KCl). The test parameters are shown in Table 5.
表5：DVS测试参数Table 5: DVS Test Parameters
Figure PCTCN2021129643-appb-000006
Figure PCTCN2021129643-appb-000006
引湿性评价分类如表6所示：The hygroscopicity evaluation classification is shown in Table 6:
表6：引湿性分类Table 6: Hygroscopicity Classification
吸湿性分类Hygroscopic classification ΔW％ΔW%
潮解deliquescence 吸收足量水分形成液体Absorbs enough water to form a liquid
极具吸湿性Very hygroscopic ΔW％≥15％ΔW%≥15%
有吸湿性hygroscopic 15％>ΔW％≥2％15%>ΔW%≥2%
略有吸湿性slightly hygroscopic 2％>ΔW％≥0.2％2%>ΔW%≥0.2%
无或几乎无吸湿性No or almost no hygroscopicity ΔW％<0.2％ΔW%<0.2%
注：ΔW％表示受试品在25℃/80％RH下的吸湿增重。Note: ΔW% represents the moisture absorption weight gain of the test product at 25℃/80%RH.
具体实施方式Detailed ways
为了更好的理解本发明的内容，下面结合具体实施例来做进一步的说明，但具体的实施方式并不是对本发明的内容所做的限制。In order to better understand the content of the present invention, further description will be given below in conjunction with specific embodiments, but the specific embodiments do not limit the content of the present invention.
实施例1：式(I)化合物的制备Example 1: Preparation of compounds of formula (I)
合成路线如下：The synthetic route is as follows:
Figure PCTCN2021129643-appb-000007
Figure PCTCN2021129643-appb-000007
步骤1：step 1:
将化合物A(200g,768.91mmol,100％纯度,1eq)和化合物B(191.40g,922.70mmol,121.91mL,99.057％纯度,1.2eq)加到反应釜中，随后加入N,N-二甲基甲酰胺(2000mL)。开启搅拌，加入碳酸铯(375.79g,1.15mol,1.5eq)。置换氮气，在25℃搅拌12小时后，将反应液缓慢倾倒至水(10L)中，搅拌0.5小时，过滤收集滤饼。然后将滤饼用乙醇(2000mL)悬浮搅拌0.5小时，过滤收集滤饼，滤饼在45℃下真空干燥得到化合物C。 1HNMR(DMSO-d 6,400MHz)δ7.51(s,1H),7.3-7.4(m,3H),7.2-7.3(m,1H),4.94(s,2H),4.87(d,J＝7.4Hz,2H),4.74(d,J＝7.4Hz,2H)。 Compound A (200 g, 768.91 mmol, 100% purity, 1 eq) and compound B (191.40 g, 922.70 mmol, 121.91 mL, 99.057 % purity, 1.2 eq) were added to the reaction kettle, followed by N,N-dimethyl Formamide (2000 mL). Stirring was turned on and cesium carbonate (375.79 g, 1.15 mol, 1.5 eq) was added. Nitrogen was replaced, and after stirring at 25° C. for 12 hours, the reaction solution was slowly poured into water (10 L), stirred for 0.5 hour, and the filter cake was collected by filtration. Then, the filter cake was suspended and stirred with ethanol (2000 mL) for 0.5 hours, the filter cake was collected by filtration, and the filter cake was vacuum-dried at 45° C. to obtain compound C. 1 HNMR(DMSO-d 6 , 400MHz)δ7.51(s,1H),7.3-7.4(m,3H),7.2-7.3(m,1H),4.94(s,2H),4.87(d,J= 7.4Hz, 2H), 4.74 (d, J=7.4Hz, 2H).
步骤2：Step 2:
将化合物C(100g,259.96mmol,100％纯度,1eq)和化合物D(163.79g,870.87mmol,200.23mL,3.35eq)加入反应釜中，随后加入四氢呋喃(1000mL)。置换氮气，降温至-5℃，将异丙基氯化镁氯化锂复合物溶 液(1.3M,299.95mL,1.5eq)缓慢滴加至反应釜中，滴加过程中控制温度在0℃～3℃。滴加完毕后，在0℃下搅拌60分钟后，加入MeOH(24.99g,779.88mmol,31.56mL,3eq)淬灭反应得到粗品E。粗品E直接用于下一步反应。Compound C (100 g, 259.96 mmol, 100% purity, 1 eq) and compound D (163.79 g, 870.87 mmol, 200.23 mL, 3.35 eq) were added to the autoclave, followed by tetrahydrofuran (1000 mL). Replace nitrogen gas, cool down to -5°C, slowly add isopropylmagnesium chloride lithium chloride complex solution (1.3M, 299.95mL, 1.5eq) into the reaction kettle, and control the temperature during the dropwise addition at 0°C～3°C . After the completion of the dropwise addition, after stirring at 0° C. for 60 minutes, MeOH (24.99 g, 779.88 mmol, 31.56 mL, 3 eq) was added to quench the reaction to obtain crude product E. Crude product E was used directly in the next reaction.
步骤3：Step 3:
将2-甲基四氢呋喃(1000mL)和水(1000mL)加到反应釜中，随后依次加入F(78.45g,259.96mmol,98.785％纯度,1eq)、乙酸钾(76.54g,779.87mmol,3eq)和二苯基膦二茂铁二氯化钯(5.71g,7.80mmol,0.03eq)。置换氮气后，缓慢升温至75℃，然后滴加粗品E(90.88g,259.96mmol,1eq)。滴加完毕后，75℃下反应30分钟。反应完毕，减压浓缩得到粗品，将加入二氧六环(500mL)和水(2.5L)加入到粗品中，25℃拌0.5小时，过滤收集滤饼。滤饼用乙醇(2000mL)溶解，25℃搅拌1小时，过滤收集滤饼。滤饼再用乙醇(2000mL)溶解，在75℃搅拌1小时，冷却至25℃，过滤收集滤饼。滤饼再用二氯甲烷(3000mL)溶解，加入巯基硅胶(10g)，45℃搅拌12小时，冷却至25℃，过滤，滤液减压浓缩得到G。 1H NMR(DMSO-d 6,400MHz):δ(ppm)9.02(s,1H),8.04(s,1H),7.33-7.43(m,3H),7.27-7.32(m,1H),5.00(s,2H),4.96(d,J＝7.3Hz,2H),4.82(d,J＝7.3Hz,2H),3.46(s,3H),2.70(s,3H)。 2-Methyltetrahydrofuran (1000mL) and water (1000mL) were added to the reaction kettle, followed by F (78.45g, 259.96mmol, 98.785% purity, 1eq), potassium acetate (76.54g, 779.87mmol, 3eq) and Diphenylphosphinoferrocene palladium dichloride (5.71 g, 7.80 mmol, 0.03 eq). After replacing with nitrogen, the temperature was slowly raised to 75° C., and then crude product E (90.88 g, 259.96 mmol, 1 eq) was added dropwise. After the dropwise addition, the reaction was carried out at 75°C for 30 minutes. After the reaction was completed, the crude product was obtained by concentration under reduced pressure. Dioxane (500 mL) and water (2.5 L) were added to the crude product, stirred at 25° C. for 0.5 hours, and the filter cake was collected by filtration. The filter cake was dissolved in ethanol (2000 mL), stirred at 25° C. for 1 hour, and the filter cake was collected by filtration. The filter cake was dissolved in ethanol (2000 mL), stirred at 75° C. for 1 hour, cooled to 25° C., and the filter cake was collected by filtration. The filter cake was dissolved in dichloromethane (3000 mL), mercapto silica gel (10 g) was added, stirred at 45°C for 12 hours, cooled to 25°C, filtered, and the filtrate was concentrated under reduced pressure to obtain G. 1 H NMR (DMSO-d 6 , 400MHz): δ(ppm) 9.02(s, 1H), 8.04(s, 1H), 7.33-7.43(m, 3H), 7.27-7.32(m, 1H), 5.00( s, 2H), 4.96 (d, J=7.3 Hz, 2H), 4.82 (d, J=7.3 Hz, 2H), 3.46 (s, 3H), 2.70 (s, 3H).
步骤4：Step 4:
将G(100g,210.10mmol,100％纯度,1eq)和H(40.81g,420.20mmol,2eq)加到反应釜中，随后加入四氢呋喃(1000mL)和二氯甲烷(1000mL)。置换氮气，降温至-5℃。将六甲基二硅基胺基锂(1M,399.19mL,1.9eq)滴加到反应瓶中，控制温度在0℃搅拌2小时。向反应液中加入离子水(1L)搅拌10分钟后，45℃减压浓缩得粗品。粗品用二氧六环(500mL)溶解搅拌10分钟，然后加入去离子水(2.5L)继续搅拌0.5小时，过滤收集滤饼。滤饼用去离子水(1L)溶解搅拌1小时，过滤收集滤饼。滤饼用无水乙醇(2L)溶解，25℃搅拌1小时，过滤收集滤饼。滤饼再用无水乙醇(2L)溶解，85℃搅拌1小时，过滤收集滤饼，滤饼在45℃真空干燥得到式(I)化合物。 1H NMR(DMSO-d 6,400MHz):δ(ppm)9.47(s,1H),8.45(s,1H),7.80(s,1H),7.33-7.42(m,4H),7.29(m,J＝7.0Hz,1H),6.34(d,J＝1.6Hz,1H),4.98(s,2H),4.95(d,J＝7.4Hz,2H),4.75(d,J＝7.3Hz,2H),3.72(s,3H),2.46(s,3H)。 G (100 g, 210.10 mmol, 100% pure, 1 eq) and H (40.81 g, 420.20 mmol, 2 eq) were added to the autoclave, followed by tetrahydrofuran (1000 mL) and dichloromethane (1000 mL). The nitrogen was replaced, and the temperature was lowered to -5°C. Lithium hexamethyldisilazide (1M, 399.19mL, 1.9eq) was added dropwise to the reaction flask, and the temperature was controlled and stirred at 0°C for 2 hours. After adding ionized water (1 L) to the reaction solution, stirring for 10 minutes, it was concentrated under reduced pressure at 45°C to obtain a crude product. The crude product was dissolved in dioxane (500 mL) and stirred for 10 minutes, then deionized water (2.5 L) was added to continue stirring for 0.5 hour, and the filter cake was collected by filtration. The filter cake was dissolved in deionized water (1 L) and stirred for 1 hour, and the filter cake was collected by filtration. The filter cake was dissolved in absolute ethanol (2L), stirred at 25°C for 1 hour, and the filter cake was collected by filtration. The filter cake was dissolved in absolute ethanol (2L), stirred at 85°C for 1 hour, filtered to collect the filter cake, and the filter cake was vacuum dried at 45°C to obtain the compound of formula (I). 1 H NMR (DMSO-d 6 , 400MHz): δ(ppm) 9.47(s, 1H), 8.45(s, 1H), 7.80(s, 1H), 7.33-7.42(m, 4H), 7.29(m, J=7.0Hz, 1H), 6.34(d, J=1.6Hz, 1H), 4.98(s, 2H), 4.95(d, J=7.4Hz, 2H), 4.75(d, J=7.3Hz, 2H) , 3.72(s, 3H), 2.46(s, 3H).
实施例2：式(I)化合物A晶型的制备Example 2: Preparation of the crystal form of compound A of formula (I)
取5g式(I)化合物，分别用100mL表7中所示的各溶剂，在相应温度下搅拌2天，过滤收集固体。固体置于真空干燥箱中(45℃)干燥16小时，得式(I)化合物的A晶型。Take 5 g of the compound of formula (I), respectively use 100 mL of each solvent shown in Table 7, stir at the corresponding temperature for 2 days, and collect the solid by filtration. The solid was dried in a vacuum drying oven (45° C.) for 16 hours to obtain crystal form A of the compound of formula (I).
表7Table 7
Figure PCTCN2021129643-appb-000008
Figure PCTCN2021129643-appb-000008
Figure PCTCN2021129643-appb-000009
Figure PCTCN2021129643-appb-000009
实施例3：式(I)化合物A晶型的吸湿性研究Example 3: Study on Hygroscopicity of Crystal Form of Compound A of Formula (I)
1.实验方法：1. Experimental method:
1)取式(I)化合物A晶型(10～30mg)置于DVS样品盘内进行测试；1) Take the compound A crystal form (10-30 mg) of formula (I) and place it in the DVS sample tray for testing;
2)DVS测试前、后的样品分别测试XRPD。2) The samples before and after the DVS test were tested for XRPD respectively.
2.实验结果：2. Experimental results:
式(Ⅰ)所示化合物A晶型的DVS如图4所示。The DVS of the crystalline form of Compound A represented by formula (I) is shown in Figure 4 .
结论：DVS结果显示，样品在25℃/80％RH条件下吸湿增重0.03％，样品几乎无引湿性。Conclusion: The DVS results show that the sample has a hygroscopic weight gain of 0.03% under the condition of 25℃/80%RH, and the sample has almost no hygroscopicity.
实施例4：式(I)化合物A晶型的稳定性实验Example 4: Stability test of the crystal form of compound A of formula (I)
1.实验目的：1. Experimental purpose:
对式(I)化合物A晶型进行影响因素(高温、高湿及光照)和加速条件下(40℃/75％RH及60℃/75％RH) 稳定性的考察，评估A晶型的固体稳定性。The influence factors (high temperature, high humidity and light) and the stability under accelerated conditions (40°C/75%RH and 60°C/75%RH) were investigated for the crystal form of compound A of formula (I), and the solid of crystal form A was evaluated. stability.
2.实验方法：2. Experimental method:
1)分别精密称取式(I)化合物A晶型约10mg置于干燥洁净的玻璃瓶中，称2份，分别标记为S1-条件-时间和S2-条件-时间，再称取约20mg置于干燥洁净的玻璃瓶中，标记为S3-条件-时间，摊成薄薄一层，作为供试样品，放置于影响因素试验条件下(60℃，25℃/92.5％RH，光照，光照对照)和加速条件下(40℃/75％RH和60℃/75％RH)，其样品为完全暴露放样。60℃，25℃/92.5％RH，光照，光照对照在5天、10天取样分析，加速条件在1个月、2个月、3个月取样分析，分析方法如表8所示。1) Precisely weigh about 10 mg of the crystal form of compound A of formula (I) and place it in a dry and clean glass bottle, weigh 2 parts, respectively labelled as S1-condition-time and S2-condition-time, and then weigh about 20 mg and place it in a dry and clean glass bottle. In a dry and clean glass bottle, marked as S3-condition-time, spread out into a thin layer, as the test sample, placed under the influence factor test conditions (60℃, 25℃/92.5%RH, light, light control ) and accelerated conditions (40°C/75%RH and 60°C/75%RH), the samples are fully exposed. 60°C, 25°C/92.5% RH, light, and light control were sampled and analyzed at 5 days and 10 days, and the accelerated conditions were sampled and analyzed at 1 month, 2 months, and 3 months. The analysis methods are shown in Table 8.
表8Table 8
Figure PCTCN2021129643-appb-000010
Figure PCTCN2021129643-appb-000010
2)在考察时间点，将相应的供试样品取出，用瓶盖盖好，0天的样品从冰箱中取出，待样品恢复至室温后进行分析。标记为S1-条件-时间的供试品用于含量和有关物质检测；标记为S2-条件-时间的供试品用作备样，标记为S3-条件-时间的供试品用于XRPD检测。2) At the time of investigation, take out the corresponding test sample, cover it with a bottle cap, take out the 0-day-old sample from the refrigerator, and analyze it after the sample has returned to room temperature. The test sample marked S1-condition-time is used for content and related substance detection; the test sample marked S2-condition-time is used as a preparation sample, and the test sample marked S3-condition-time is used for XRPD detection .
3.实验结果：3. Experimental results:
A晶型固体稳定性实验结果如表9所示。Table 9 shows the results of the solid stability experiment of crystal form A.
表9Table 9
Figure PCTCN2021129643-appb-000011
Figure PCTCN2021129643-appb-000011
Figure PCTCN2021129643-appb-000012
Figure PCTCN2021129643-appb-000012
*光照(总照度可见光＝1.2×10 6Lux·hr/近紫外总能量＝200w·hr/m 2，敞口)；**与可见光+紫外同时。 *Illumination (total illuminance visible light=1.2×10 6 Lux·hr/near-ultraviolet total energy=200w·hr/m 2 , open); **simultaneous with visible light+ultraviolet.
结论：式(I)化合物A晶型具有良好的稳定性。Conclusion: The crystal form of compound A of formula (I) has good stability.
实施例5：式(I)化合物A晶型在小鼠HCT116模型药效研究Example 5: Pharmacodynamic study of compound A crystal form of formula (I) in mouse HCT116 model
1.实验目的：1. Experimental purpose:
使用人结肠癌HCT-116细胞皮下异种移植肿瘤裸小鼠模型，评价式(I)化合物A晶型的抗肿瘤作用。Using human colon cancer HCT-116 cell subcutaneous tumor xenograft nude mouse model, the antitumor effect of compound A of formula (I) crystalline form was evaluated.
2.实验动物：2. Experimental animals:
种属：小鼠Species: mouse
品系：BALB/c裸小鼠Strain: BALB/c nude mice
周龄：6-8周龄Age: 6-8 weeks old
性别：雄性gender: male
体重：18-22克Weight: 18-22 grams
供应商：北京维通利华实验动物有限公司Supplier: Beijing Weitong Lihua Laboratory Animal Co., Ltd.
动物合格证号：20170011005645Animal certificate number: 20170011005645
3.实验内容：3. Experiment content:
1)实验细胞及培养：人结肠癌HCT-116细胞体外单层培养，培养条件为McCoy's 5a培养基中加10％胎牛血清，37℃的5％CO2孵箱培养。一周三次用胰酶-乙二胺四乙酸进行常规消化处理传代。当细胞饱和度为80％-90％，数量到达要求时，收取细胞，计数，接种；1) Experimental cells and culture: Human colon cancer HCT-116 cells were cultured in monolayer in vitro, and the culture conditions were McCoy's 5a medium plus 10% fetal bovine serum, and cultured in a 5% CO2 incubator at 37°C. Conventional digestion treatment with trypsin-EDTA was passaged three times a week. When the cell saturation is 80%-90% and the number reaches the requirement, the cells are collected, counted, and seeded;
2)肿瘤组织接种及分组：0.2mL(5×10 6个)HCT-116细胞皮下接种于每只小鼠的右侧腋下，肿瘤平均体积达到114mm 3时，将动物随机分为四组，开始给药。实验分组和给药方案见表10： 2) Tumor tissue inoculation and grouping: 0.2 mL (5×10 6 cells) of HCT-116 cells were subcutaneously inoculated into the right armpit of each mouse. When the average tumor volume reached 114 mm 3 , the animals were randomly divided into four groups. Start dosing. The experimental grouping and dosing schedule are shown in Table 10:
表10 实验动物分组及给药方案Table 10 Experimental animal grouping and dosing schedule
Figure PCTCN2021129643-appb-000013
Figure PCTCN2021129643-appb-000013
3)实验动物日常观察：本实验方案的拟定及任何修改均通过了实验动物管理与使用委员会(IACUC)的评估核准。实验动物的使用及福利遵照国际实验动物评估和认可委员会(AAALAC)的规定执行。每天监测动物的健康状况及死亡情况，例行检查包括观察肿瘤生长和药物治疗对动物日常行为表现的影响如行为活动，摄食摄水量(仅目测)，体重变化(每周测量两次体重)，外观体征或其它不正常情况。基于各组动物数量记录了组内动物死亡数和副作用。3) Daily observation of experimental animals: The formulation of this experimental protocol and any modifications have been evaluated and approved by the Laboratory Animal Care and Use Committee (IACUC). The use and welfare of experimental animals were carried out in accordance with the regulations of the International Association for the Assessment and Accreditation of Laboratory Animals (AAALAC). The health status and death of the animals were monitored daily. Routine examinations included observing the effects of tumor growth and drug treatment on the animals' daily behaviors such as behavioral activities, food and water intake (visual observation only), body weight changes (weight measurement twice a week), Physical signs or other abnormalities. Animal deaths and side effects within groups were recorded based on the number of animals in each group.
4)受试物的配制4) Preparation of test substance
a)溶媒组：5％DMSO+95％(10％HP-β-CD)。a) Vehicle group: 5% DMSO+95% (10% HP-β-CD).
b)待测化合物组：称量定量的受试化合物于配药瓶内，加入相应体积的DMSO后涡旋，得到澄清溶液,在加入相应体积的10％HP-β-CD后涡旋，得到澄清溶液。化合物三天配制一次。b) Test compound group: weigh the quantitative test compound into a dispensing bottle, add a corresponding volume of DMSO and vortex to obtain a clear solution, add a corresponding volume of 10% HP-β-CD and vortex to obtain a clear solution solution. Compounds were formulated every three days.
5)肿瘤测量和实验指标：5) Tumor measurement and experimental indicators:
a)每周两次用游标卡尺测量肿瘤直径。肿瘤体积的计算公式为：TV＝1/2×a×b 2，a和b分别表示肿瘤的长径和短径； a) Tumor diameter was measured twice a week with a vernier caliper. The calculation formula of tumor volume is: TV=1/2×a×b 2 , a and b represent the long and short diameters of the tumor, respectively;
b)化合物的抑瘤疗效用TGI(％)评价。TGI(％)，反映肿瘤生长抑制率。TGI(％)的计算：TGI(％)＝{[1-(某处理组给药结束时平均瘤体积－该处理组开始给药时平均瘤体积)]/(溶剂对照组治疗结束时平均瘤体积－溶剂对照组开始治疗时平均瘤体积)}×100％。b) The tumor-inhibitory efficacy of the compounds was evaluated by TGI (%). TGI (%), reflecting tumor growth inhibition rate. Calculation of TGI (%): TGI (%)={[1-(average tumor volume at the end of administration of a certain treatment group - average tumor volume at the beginning of administration of this treatment group)]/(average tumor volume at the end of treatment in the solvent control group Volume - the average tumor volume of the solvent control group at the beginning of treatment)} × 100%.
4.实验结果：4. Experimental results:
1)如表11和图5所示，在人结肠癌HCT-116细胞皮下异种移植肿瘤裸小鼠模型上，口服给药至第19天，式(I)化合物A晶型三个剂量组(7.5mg/kg，15mg/kg和30mg/kg)具有明显的抑制肿瘤生长作用，呈现剂量依赖性，它们的TGI分别为66％，71％和93％。1) As shown in Table 11 and Figure 5, on the nude mouse model of human colon cancer HCT-116 cell subcutaneous xenograft tumor, orally administered to the 19th day, formula (I) Compound A crystal form three dose groups ( 7.5mg/kg, 15mg/kg and 30mg/kg) had a significant inhibitory effect on tumor growth in a dose-dependent manner, and their TGIs were 66%, 71% and 93%, respectively.
2)实验动物的体重作为间接测定药物毒性的参考指标。如图6所示，给药至第19天时，溶剂对照组和(I)化合物A晶型三个剂量组所有动物的体重均未有明显下降，无发病或死亡现象。2) The body weight of experimental animals is used as a reference index for indirect determination of drug toxicity. As shown in Figure 6, on the 19th day after administration, the body weight of all animals in the solvent control group and (I) three dose groups of Compound A crystal form did not decrease significantly, and there was no morbidity or death.
表11：小鼠HCT116模型体内药效实验结果Table 11: In vivo efficacy test results of mouse HCT116 model
组别 group 药物drug TGITGI
22 A晶型(7.5mg/kg,PO,QD)Form A (7.5mg/kg, PO, QD) 66％66%
33 A晶型(15mg/kg,PO,QD)Form A (15mg/kg, PO, QD) 71％71%
44 A晶型(30mg/kg,PO,QD)Form A (30mg/kg, PO, QD) 93％93%
结论：式(I)化合物A晶型在给药剂量下，能显著抑制肿瘤生长，呈现剂量依赖性；给药过程中动物的体重未见明显下降，耐受性好。Conclusion: The crystal form of compound A of formula (I) can significantly inhibit tumor growth under the administration dose, showing a dose-dependent manner; the body weight of the animals does not decrease significantly during the administration process, and the tolerance is good.

Claims (11)

  1. 式(Ⅰ)化合物的A晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.16±0.20°、17.39±0.20°和23.88±0.20°Form A of the compound of formula (I), its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.16±0.20°, 17.39±0.20° and 23.88±0.20°
    Figure PCTCN2021129643-appb-100001
    Figure PCTCN2021129643-appb-100001
  2. 根据权利要求1所述的A晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.16±0.20°、13.57±0.20°、16.95±0.20°、17.39±0.20°、19.37±0.20°、22.22±0.20°、23.88±0.20°和25.85±0.20°。The crystal form A according to claim 1, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.16±0.20°, 13.57±0.20°, 16.95±0.20°, 17.39±0.20°, 19.37±0.20 °, 22.22±0.20°, 23.88±0.20° and 25.85±0.20°.
  3. 根据权利要求2所述的A晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.16±0.20°、13.57±0.20°、14.51±0.20°、16.95±0.20°、17.39±0.20°、19.37±0.20°、22.22±0.20°、23.88±0.20°和25.85±0.20°。The crystal form A according to claim 2, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.16±0.20°, 13.57±0.20°, 14.51±0.20°, 16.95±0.20°, 17.39±0.20 °, 19.37±0.20°, 22.22±0.20°, 23.88±0.20° and 25.85±0.20°.
  4. 根据权利要求3所述的A晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：8.16°、9.99°、13.57°、14.51°、15.61°、16.95°、17.39°、18.27°、19.37°、19.71°、21.40°、21.97°、22.22°、23.88°、25.02°、25.85°、26.50°、27.36°、28.72°、30.39°、31.19°、31.53°、31.97°、32.57°、33.05°和37.72°。The crystal form A according to claim 3, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.16°, 9.99°, 13.57°, 14.51°, 15.61°, 16.95°, 17.39°, 18.27° , 19.37°, 19.71°, 21.40°, 21.97°, 22.22°, 23.88°, 25.02°, 25.85°, 26.50°, 27.36°, 28.72°, 30.39°, 31.19°, 31.53°, 31.97°, 32.57°, 33.05 ° and 37.72°.
  5. 根据权利要求4所述的A晶型，其XRPD图谱基本如附图1所示。Form A according to claim 4, its XRPD pattern is basically as shown in accompanying drawing 1.
  6. 根据权利要求1～5任意一项所述的A晶型，其差示扫描量热曲线在225.5±3℃有一个吸热峰的起始点。The crystal form A according to any one of claims 1 to 5, wherein the differential scanning calorimetry curve has an endothermic peak starting point at 225.5±3°C.
  7. 根据权利要求6所述的A晶型，其DSC图谱如附图2所示。Form A according to claim 6, its DSC spectrum is shown in accompanying drawing 2.
  8. 根据权利要求1～5任意一项所述的A晶型，其热重分析曲线在200.0±3℃时失重达2.41％。The crystal form A according to any one of claims 1 to 5, whose thermogravimetric analysis curve has a weight loss of 2.41% at 200.0±3°C.
  9. 根据权利要求8所述的A晶型，TGA图谱如图3所示。Form A according to claim 8, the TGA spectrum is shown in Figure 3.
  10. 式(Ⅰ)化合物的A晶型的制备方法，包含：The preparation method of the A crystal form of the compound of formula (I), comprising:
    1)式(I)化合物，加入有机溶剂；1) compound of formula (I), adding organic solvent;
    2)在25～50℃温度下搅拌2天；2) Stir for 2 days at a temperature of 25 to 50 °C;
    3)过滤收集固体，置于真空干燥箱中45℃干燥16小时，3) The solid was collected by filtration, placed in a vacuum drying oven at 45°C for drying for 16 hours,
    其中式(Ⅰ)化合物为
    Figure PCTCN2021129643-appb-100002
    wherein the compound of formula (I) is
    Figure PCTCN2021129643-appb-100002
  11. 根据权利要求10所述的式(Ⅰ)化合物A晶型的制备方法，其中，所述有机溶剂为乙醇、MeOH、ACN、丙酮、EtOAc、二氧六环、甲苯、H 2O、THF:H 2O(v/v，1:1)、EtOH:H 2O(v/v，2:1)、丙酮:EtOAc(v/v，1:1)、DMF:EtOAc(v/v，5:95)、DMF:H 2O(v/v，5:95)、丙酮:H 2O(v/v，95:5)、二氧六环:H 2O(v/v，97:3)、THF或DCM:EtOAc(v/v，2:1)。 The method for preparing crystal form of compound A of formula (I) according to claim 10, wherein the organic solvent is ethanol, MeOH, ACN, acetone, EtOAc, dioxane, toluene, H 2 O, THF:H 2 O (v/v, 1:1), EtOH: H2O (v/v, 2:1), Acetone:EtOAc (v/v, 1:1), DMF:EtOAc (v/v, 5:1) 95), DMF: H 2 O (v/v, 5:95), acetone: H 2 O (v/v, 95:5), dioxane: H 2 O (v/v, 97:3) , THF or DCM:EtOAc (v/v, 2:1).
PCT/CN2021/129643 2020-11-11 2021-11-09 Crystal form of oxetane-containing spirocyclic compound, preparation method therefor and use thereof WO2022100586A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202011252031.8 2020-11-11
CN202011252031 2020-11-11

Publications (1)

Publication Number Publication Date
WO2022100586A1 true WO2022100586A1 (en) 2022-05-19

Family

ID=81602135

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/129643 WO2022100586A1 (en) 2020-11-11 2021-11-09 Crystal form of oxetane-containing spirocyclic compound, preparation method therefor and use thereof

Country Status (1)

Country Link
WO (1) WO2022100586A1 (en)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008153947A2 (en) * 2007-06-07 2008-12-18 Amgen Inc. Heterocyclic compounds as raf kinase modulators
CN103189369A (en) * 2010-09-01 2013-07-03 吉利德康涅狄格有限公司 Pyridinones/pyrazinones, method of making, and method of use thereof
CN103201277A (en) * 2010-09-01 2013-07-10 吉利德康涅狄格有限公司 Pyridazinones, method of making, and method of use thereof
WO2013130976A1 (en) * 2012-03-01 2013-09-06 Array Biopharma Inc. Serine/threonine kinase inhibitors
CN107074874A (en) * 2014-12-22 2017-08-18 伊莱利利公司 Erk inhibitor
WO2020228817A1 (en) * 2019-05-15 2020-11-19 南京明德新药研发有限公司 Erk inhibitor and use thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008153947A2 (en) * 2007-06-07 2008-12-18 Amgen Inc. Heterocyclic compounds as raf kinase modulators
CN103189369A (en) * 2010-09-01 2013-07-03 吉利德康涅狄格有限公司 Pyridinones/pyrazinones, method of making, and method of use thereof
CN103201277A (en) * 2010-09-01 2013-07-10 吉利德康涅狄格有限公司 Pyridazinones, method of making, and method of use thereof
WO2013130976A1 (en) * 2012-03-01 2013-09-06 Array Biopharma Inc. Serine/threonine kinase inhibitors
CN107074874A (en) * 2014-12-22 2017-08-18 伊莱利利公司 Erk inhibitor
WO2020228817A1 (en) * 2019-05-15 2020-11-19 南京明德新药研发有限公司 Erk inhibitor and use thereof

Similar Documents

Publication Publication Date Title
US10457666B2 (en) Stable crystal form of tipiracil hydrochloride and crystallization method for the same
WO2020125747A1 (en) Crystal form and amorphous form of mek inhibitor and applications thereof
TWI765451B (en) Spirocyclic compounds as erk inhibitor and application thereof
TWI758999B (en) Thiazololactam compound as erk inhibitor and application thereof
JP2018537497A (en) Pyrido [1,2-a] pyrimidone analogs, their crystalline forms, their intermediates, and methods for their production
WO2023160727A1 (en) Use of indazole compound for treating psoriasis
WO2022171044A1 (en) Oxa-azaspiro compound, salt form thereof and crystal form thereof
WO2022100586A1 (en) Crystal form of oxetane-containing spirocyclic compound, preparation method therefor and use thereof
CN103476742A (en) New crystal form VII of agomelatine, preparation method and use thereof and pharmaceutical composition containing same
WO2016101867A1 (en) Α-crystal form of naproxen imatinib p-toluene sulfonate, preparation method thereof, and pharmaceutical composition containing same
WO2019149254A1 (en) Crystal form of small molecule immune compound, preparation method therefor and pharmaceutical composition containing same
WO2022063229A1 (en) Salt of arylaminoquinazoline-containing compound, and preparation method therefor and use thereof
WO2020147838A1 (en) Salt of egfr inhibitor, crystal form, and preparation method therefor
CN101083983A (en) Indole derivatives with antitumor activity
WO2019206268A1 (en) Crystal form of c-met inhibitor and salt form thereof and preparation method therefor
TWI825811B (en) Thiazololactam-spiroheterocyclic compound and application thereof
WO2022135580A1 (en) Crystal form of pyridopyrrole compound, and preparation method therefor and use thereof
WO2022253186A1 (en) Crystal form of dimethyl substituted thiazolopyrrolone compound and preparation method therefor
WO2022048545A1 (en) Crystal form of pyridopyrimidine compound
EP4365171A1 (en) Crystal form of triazolopyridine-substituted indazole compound and preparation method therefor
WO2022063325A1 (en) Crystal form of pyridinylphenyl compound and preparation method therefor
WO2023185638A1 (en) Crystal form of quinoline derivative and preparation method therefor
CN108658945A (en) A kind of Antitubulin（VDA-1）A crystal forms
WO2023072263A1 (en) 5-substituted pyridine-2(1h)-ketone compound and use thereof
CN113929663A (en) AZD 9291-2-indole formate and preparation method thereof

Legal Events

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

Ref document number: 21891106

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21891106

Country of ref document: EP

Kind code of ref document: A1

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 10/11/2023)