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WO2023083292A1 - Sel de composé de pyrrolidine, forme cristalline de celui-ci et son procédé de préparation - Google Patents

Sel de composé de pyrrolidine, forme cristalline de celui-ci et son procédé de préparation Download PDF

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Publication number
WO2023083292A1
WO2023083292A1 PCT/CN2022/131313 CN2022131313W WO2023083292A1 WO 2023083292 A1 WO2023083292 A1 WO 2023083292A1 CN 2022131313 W CN2022131313 W CN 2022131313W WO 2023083292 A1 WO2023083292 A1 WO 2023083292A1
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compound
formula
crystal form
ether
pharmaceutically acceptable
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Chinese (zh)
Inventor
胡越
刘成祥
冯爱娟
薛黎婷
杨桂梅
陈平
古鹏
唐任宏
任晋生
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Jiangsu Simcere Pharmaceutical Co Ltd
Simcere Pharmaceutical Co Ltd
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Jiangsu Simcere Pharmaceutical Co Ltd
Simcere Pharmaceutical Co Ltd
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Priority to CN202280069728.2A priority Critical patent/CN118251392A/zh
Publication of WO2023083292A1 publication Critical patent/WO2023083292A1/fr
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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
    • A61K31/435—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
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/472—Non-condensed isoquinolines, e.g. papaverine
    • A61K31/4725—Non-condensed isoquinolines, e.g. papaverine containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems

Definitions

  • the disclosure belongs to the field of medicine, and relates to pharmaceutically acceptable salts of pyrrolidine compounds, crystals of the pharmaceutically acceptable salts, and corresponding preparation methods and applications.
  • Estrogen (E2) and estrogen alpha receptor (ER ⁇ ) are important drivers of breast cancer development. More than 2/3 of breast cancer patients express ER transcription factors, and in most ER-positive patients, ER is still a key driver even in tumors that progress after early endocrine therapy, so ER is A major target for breast cancer therapy (Pharmacology & Therapeutics 186 (2016) 1–24).
  • the purpose of endocrine therapy is to reduce the activity of ER.
  • SERMs selective estrogen receptor modulators
  • tamoxifen tamoxifen
  • Aromatase inhibitors by inhibiting the conversion of androgen into estrogen, reduce the level of estrogen in the body; and selective estrogen receptor down-regulators, such as fulvestrant (fulvestrant), not only as ER Antagonists inhibit its activity and also induce ER protein degradation.
  • fulvestrant fulvestrant
  • Fulvestrant is the first and only SERD drug clinically approved for the treatment of postmenopausal patients with ER-positive, metastatic breast cancer after progression on tamoxifen or an aromatase inhibitor.
  • a number of research data show that patients treated with fulvestrant have not completely degraded ER in vivo.
  • intramuscular injection caused obvious reactions such as pain, swelling, and redness at the injection site, and the absorption was slow and the exposure in vivo was limited. And other characteristics limit its clinical application, so patients with ER-positive breast cancer urgently need new treatment options.
  • PCT/CN2021/093736 (application date May 14, 2021) describes a compound N-((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)-6-((1S, 3R)-3-methyl-2-(2,2,2-trifluoroethyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-1- Base) pyridin-3-amine (compound of formula (I)), the compound of formula (I) is a selective estrogen receptor down-regulator, and the in vivo and in vitro models show that the compound of (I) has a wide range of inhibitory effects on ER-positive cells.
  • the pharmaceutically acceptable salt is selected from sulfate, phosphate, benzoate, succinate, adipate, fumarate, L-malate and citrate.
  • the present disclosure also provides the crystal form C of the adipate salt of the compound of the above formula (I).
  • the X-ray powder diffraction pattern represented by the diffraction angle 2 ⁇ of the crystal form C there are 4.46 ⁇ 0.2°, 6.30 ⁇ 0.2°, 6.30 ⁇ 0.2°, There are diffraction peaks at 8.90 ⁇ 0.2° and 19.10 ⁇ 0.2°.
  • the present disclosure also provides the crystal form B of the benzoate salt of the compound of the above formula (I).
  • the X-ray powder diffraction pattern represented by the diffraction angle 2 ⁇ of the crystal form B there are 7.19 ⁇ 0.2°, 10.01 ⁇ 0.2°, There are diffraction peaks at 11.39 ⁇ 0.2° and 18.78 ⁇ 0.2°.
  • the present disclosure also provides the crystal form A of the succinate salt of the compound of the above formula (I).
  • the X-ray powder diffraction pattern represented by the diffraction angle 2 ⁇ of the crystal form A at 7.13 ⁇ 0.2°, 9.30 ⁇ 0.2°, 11.29 There are diffraction peaks at ⁇ 0.2°, 15.51 ⁇ 0.2°, and 19.89 ⁇ 0.2°.
  • the present disclosure also provides a method for preparing a pharmaceutically acceptable salt of the compound of formula (I), the preparation method comprising the step of reacting the compound of formula (I) with a corresponding acid to form a salt.
  • the present disclosure also provides a method for preparing the above crystal form C, comprising the following steps:
  • the present disclosure also provides a method for preparing the above crystal form B, comprising the following steps:
  • the present disclosure also provides a method for preparing the above crystal form A, comprising the following steps:
  • the present disclosure also provides a pharmaceutical composition, which comprises a pharmaceutically acceptable salt of the compound of formula (I) or the above crystal form, and pharmaceutically acceptable auxiliary materials.
  • the present disclosure also provides the use of the pharmaceutically acceptable salt of the compound of the above formula (I), the above crystal form or the above pharmaceutical composition in the preparation of a drug for preventing or treating estrogen receptor-related diseases.
  • Fig. 1 is the NOESY spectrum of the compound of formula (I).
  • Fig. 2 is the XRPD spectrum of the crystal form C of adipate salt of the compound of formula (I).
  • Fig. 3 is the DSC spectrum of the crystal form C of adipate salt of the compound of formula (I).
  • Fig. 4 is the TGA spectrum of the crystal form C of adipate salt of the compound of formula (I).
  • Fig. 5 is the XRPD spectrum of the benzoic acid salt B crystal form of the compound of formula (I).
  • Fig. 6 is the DSC spectrum of the crystal form B of the benzoate salt of the compound of formula (I).
  • Fig. 7 is the TGA spectrum of the benzoic acid salt B crystal form of the compound of formula (I).
  • Fig. 8 is the XRPD spectrum of the crystal form A of succinate salt of the compound of formula (I).
  • Fig. 9 is the DSC spectrum of the crystal form A of the compound succinate of formula (I).
  • Fig. 10 is the TGA spectrum of the crystal form A of succinate salt of the compound of formula (I).
  • Fig. 11 is a three-dimensional structural ellipsoid diagram of crystal form A of succinate salt of the compound of formula (I).
  • Fig. 12 is a packing diagram of the unit cell structure of the crystal form A of the compound succinate of formula (I).
  • Fig. 13 is a picture of anti-tumor growth effect of compound succinate of formula (I) on MCF-7 xenograft subcutaneous xenograft tumor mouse model.
  • Fig. 14 is a graph showing the change in body weight of mice with MCF-7 xenogeneic xenograft tumors transplanted with succinate of the compound of formula (I).
  • Fig. 15 is the effect diagram of anti-tumor growth of compound succinate of formula (I) on T47D xenograft subcutaneous xenograft tumor mouse model.
  • Fig. 16 is a graph showing the change in body weight of T47D xenograft subcutaneously transplanted tumor mice with succinate of the compound of formula (I).
  • Fig. 17 The survival curve of the compound succinate of formula (I) on MCF-7 heterogeneous brain orthotopic tumor-bearing mice.
  • Fig. 18 is a graph showing the survival curve of the compound of formula (I) on MCF-7 xenogeneic brain orthotopic tumor-bearing mice.
  • Fig. 19 is a graph showing the change in body weight of MCF-7 heterogeneous brain orthotopic tumor-bearing mice by the compound of formula (I).
  • the present disclosure provides a pharmaceutically acceptable salt of a compound of formula (I) with excellent physical and chemical properties and/or pharmaceutical properties and crystals thereof,
  • the present disclosure provides a pharmaceutically acceptable salt of the compound represented by formula (I), wherein the pharmaceutically acceptable salt is selected from sulfate, phosphate, benzoate, succinate, adipate, fumaric acid salt, L-malate and citrate, preferably benzoate, succinate, adipate, fumarate, L-malate and citrate, more preferably benzoate, succinate salts and adipates.
  • the molar ratio of the compound of formula (I) to the acid molecule is about 0.5-2.
  • the molar ratio of the compound of formula (I) to the acid molecule is about 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3 , 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, and 2.0.
  • the molar ratio of the compound of formula (I) to the acid molecule is about 0.5, 1.0, 1.5 and 2.0.
  • the succinate salt of the compound of formula (I) above is represented by formula (II), wherein, x is selected from 0.5-2,
  • the benzoate salt of the compound of formula (I) above is represented by formula (III), wherein, y is selected from 0.5 to 2,
  • the adipate salt of the compound of formula (I) above is represented by formula (IV), wherein, z is selected from 0.5-2,
  • x, y, z are independently selected from 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, and 2.0.
  • x, y, z are independently selected from 0.8, 0.9, 1.0, 1.1 and 1.2.
  • x, y, z are independently selected from 1.0.
  • the present disclosure also provides a method for preparing a pharmaceutically acceptable salt of the compound of formula (I), comprising: the step of forming a salt of the compound shown in formula (I) with an acid, wherein the acid is selected from sulfuric acid, phosphoric acid, benzoic acid, succinic acid, Adipic acid, fumaric acid, L-malic acid and citric acid, preferably benzoic acid, succinic acid, adipic acid, fumaric acid, L-malic acid and citric acid, more preferably benzoic acid, succinic acid and adipic acid.
  • the acid is selected from sulfuric acid, phosphoric acid, benzoic acid, succinic acid, Adipic acid, fumaric acid, L-malic acid and citric acid, preferably benzoic acid, succinic acid, adipic acid, fumaric acid, L-malic acid and citric acid, more preferably benzoic acid, succinic acid and adipic acid.
  • the solvent used in the salt-forming reaction described in the present disclosure is selected from cyclohexane, n-heptane, toluene, chloroform, ether, isopropyl ether, methyl tert-butyl ether, tetrahydrofuran, ethylene glycol dimethyl ether, acetone, methyl acetate , ethyl acetate, isopropyl acetate, butyl acetate, methanol, ethanol, isopropanol, n-butanol and acetonitrile, preferably isopropyl ether, methyl tert-butyl ether, methanol, isopropanol and tetrahydrofuran at least one of the
  • the method for preparing a pharmaceutically acceptable salt of the compound of formula (I) further includes one or a combination of solvent volatilization, stirring crystallization, filtration, drying, etc.
  • the present disclosure also provides a crystalline form of the adipate salt of the compound of formula (I).
  • the crystalline form of the adipate salt of the compound of formula (I) is an anhydrate.
  • the present disclosure provides the C crystal form of the adipate salt of the compound of formula (I), the C crystal form, in the X-ray powder diffraction pattern represented by the diffraction angle 2 ⁇ , at 4.46 ⁇ 0.2°, 6.30 ⁇ 0.2°, There are diffraction peaks at 8.90 ⁇ 0.2° and 19.10 ⁇ 0.2°.
  • the crystal form C is at 4.46 ⁇ 0.2°, 6.30 ⁇ 0.2°, 7.03 ⁇ 0.2°, 8.90 ⁇ 0.2°, 16.45 ⁇ 0.2 °, 17.78 ⁇ 0.2°, 19.10 ⁇ 0.2° and 21.03 ⁇ 0.2° have diffraction peaks.
  • the crystal form C is at 4.46 ⁇ 0.2°, 6.30 ⁇ 0.2°, 7.03 ⁇ 0.2°, 8.90 ⁇ 0.2°, 10.65 ⁇ 0.2 °, 11.95 ⁇ 0.2°, 12.55 ⁇ 0.2°, 13.34 ⁇ 0.2°, 14.20 ⁇ 0.2°, 16.02 ⁇ 0.2°, 16.45 ⁇ 0.2°, 17.78 ⁇ 0.2°, 18.33 ⁇ 0.2°, 19.10 ⁇ 0.2°, 19.77 ⁇ 0.2 °, 20.12 ⁇ 0.2°, 21.03 ⁇ 0.2°, 21.62 ⁇ 0.2°, 24.30 ⁇ 0.2°, 25.97 ⁇ 0.2°, 27.39 ⁇ 0.2° and 27.98 ⁇ 0.2° have diffraction peaks.
  • the crystal form C is at 4.46 ⁇ 0.2°, 6.30 ⁇ 0.2°, 7.03 ⁇ 0.2°, 8.90 ⁇ 0.2°, 10.65 ⁇ 0.2 °, 11.95 ⁇ 0.2°, 12.55 ⁇ 0.2°, 12.96 ⁇ 0.2°, 13.34 ⁇ 0.2°, 14.20 ⁇ 0.2°, 15.20 ⁇ 0.2°, 16.02 ⁇ 0.2°, 16.45 ⁇ 0.2°, 17.78 ⁇ 0.2°, 18.33 ⁇ 0.2 °, 19.10 ⁇ 0.2°, 19.77 ⁇ 0.2°, 20.12 ⁇ 0.2°, 21.03 ⁇ 0.2°, 21.62 ⁇ 0.2°, 22.44 ⁇ 0.2°, 22.52 ⁇ 0.2°, 22.54 ⁇ 0.2°, 23.35 ⁇ 0.2°, 23.98 ⁇ 0.2
  • the X-ray powder diffraction pattern of the crystal form C is substantially consistent with that in FIG. 2 .
  • the DSC spectrum of the crystal form C has an endothermic peak at 149.89 ⁇ 5°C.
  • the DSC spectrum of Form C is substantially the same as that of FIG. 3 .
  • the present disclosure also provides a method for preparing the crystal form C of adipate salt of the compound of formula (I), comprising the step of reacting the compound of formula (I) with adipic acid to form a salt.
  • the preparation method of the above crystal form C includes the following steps:
  • the solvent in step (1-a) and step (2-a) is selected from cyclohexane, n-heptane, toluene, chloroform, diethyl ether, isopropyl ether, methyl tert-butyl ether, tetrahydrofuran , at least one of ethylene glycol dimethyl ether, acetone, methyl acetate, ethyl acetate, isopropyl acetate, butyl acetate, methanol, ethanol, isopropanol, n-butanol and acetonitrile, preferably isopropyl ether, At least one of methyl tert-butyl ether, methanol, isopropanol and tetrahydrofuran, more preferably isopropanol.
  • the molar ratio of the compound of formula (I) to adipic acid is 1:2-2:1, preferably 1:1.
  • the step (1-a) is performed under heating.
  • the step (1-a) is carried out at a heating temperature of about 40-80°C, preferably 60°C.
  • the step (4-a) is carried out at 15-30°C, preferably at 20-25°C.
  • the present disclosure also provides a crystalline form of the benzoate salt of the compound of formula (I).
  • the crystalline form of the benzoate salt of the compound of formula (I) is an anhydrate.
  • the present disclosure provides the crystal form B of the benzoate salt of the compound of formula (I).
  • the crystal form B is at 7.19 ⁇ 0.2°, 10.01 ⁇ 0.2°, 11.39 ⁇
  • the X-ray powder diffraction pattern represented by the diffraction angle 2 ⁇ of the B crystal form there are ° has a diffraction peak.
  • the X-ray powder diffraction pattern represented by the diffraction angle 2 ⁇ of the B crystal form at 7.19 ⁇ 0.2°, 10.01 ⁇ 0.2°, 11.39 ⁇ 0.2°, 18.78 ⁇ 0.2°, 19.01 ⁇ 0.2° , 19.66 ⁇ 0.2°, 19.91 ⁇ 0.2°, 20.09 ⁇ 0.2° have diffraction peaks.
  • the X-ray powder diffraction pattern represented by the diffraction angle 2 ⁇ of the B crystal form at 4.69 ⁇ 0.2°, 6.34 ⁇ 0.2°, 7.19 ⁇ 0.2°, 10.01 ⁇ 0.2°, 11.39 ⁇ 0.2 °, 18.78 ⁇ 0.2°, 19.01 ⁇ 0.2°, 19.66 ⁇ 0.2°, 19.91 ⁇ 0.2° and 20.09 ⁇ 0.2° have diffraction peaks.
  • the X-ray powder diffraction pattern represented by the diffraction angle 2 ⁇ of the B crystal form at 4.69 ⁇ 0.2°, 6.34 ⁇ 0.2°, 7.19 ⁇ 0.2°, 10.01 ⁇ 0.2°, 11.11 ⁇ 0.2 °, 11.39 ⁇ 0.2°, 14.78 ⁇ 0.2°, 18.78 ⁇ 0.2°, 19.01 ⁇ 0.2°, 19.25 ⁇ 0.2°, 19.66 ⁇ 0.2°, 19.91 ⁇ 0.2°, 20.09 ⁇ 0.2°, 20.80 ⁇ 0.2°, 21.66 ⁇ 0.2 ° has a diffraction peak.
  • the X-ray powder diffraction pattern represented by the diffraction angle 2 ⁇ of the B crystal form at 4.69 ⁇ 0.2°, 6.34 ⁇ 0.2°, 7.19 ⁇ 0.2°, 9.83 ⁇ 0.2°, 10.01 ⁇ 0.2° , 11.11 ⁇ 0.2°, 11.39 ⁇ 0.2°, 14.78 ⁇ 0.2°, 18.78 ⁇ 0.2°, 19.01 ⁇ 0.2°, 19.25 ⁇ 0.2°, 19.66 ⁇ 0.2°, 19.91 ⁇ 0.2°, 20.09 ⁇ 0.2°, 20.80 ⁇ 0.2° , There is a diffraction peak at 21.66 ⁇ 0.2°.
  • the X-ray powder diffraction pattern represented by the diffraction angle 2 ⁇ of the B crystal form at 4.69 ⁇ 0.2°, 4.91 ⁇ 0.2°, 6.34 ⁇ 0.2°, 7.19 ⁇ 0.2°, 9.37 ⁇ 0.2 °, 9.83 ⁇ 0.2°, 10.01 ⁇ 0.2°, 10.32 ⁇ 0.2°, 11.11 ⁇ 0.2°, 11.39 ⁇ 0.2°, 12.06 ⁇ 0.2°, 12.69 ⁇ 0.2°, 13.52 ⁇ 0.2°, 14.07 ⁇ 0.2°, 14.36 ⁇ 0.2 °, 14.78 ⁇ 0.2°, 17.30 ⁇ 0.2°, 18.78 ⁇ 0.2°, 19.01 ⁇ 0.2°, 19.25 ⁇ 0.2°, 19.66 ⁇ 0.2°, 19.91 ⁇ 0.2°, 20.09 ⁇ 0.2°, 20.80 ⁇ 0.2°, 21.15 ⁇ 0.2 °, 21.66 ⁇ 0.2°, 22.35 ⁇ 0.2°, 23.57 ⁇ 0.2°, 23.70 ⁇ 0.2°, 24.63 ⁇ 0.2°, 25.68 ⁇ 0.2°, 27.05 ⁇ 0.2° have diffraction peaks.
  • the X-ray powder diffraction pattern of the crystal form B is substantially consistent with FIG. 5 .
  • the DSC spectrum of the crystal form B has an endothermic peak at 150.08 ⁇ 5°C.
  • the DSC spectrum of Form B is substantially consistent with FIG. 6 .
  • the present disclosure also provides a preparation method of the crystal form B of the benzoate salt of the compound of formula (I), comprising the step of reacting the compound of formula (I) with benzoic acid to form a salt.
  • the preparation method of the above-mentioned B crystal form includes the following steps:
  • the solvent in step (1-b) is selected from cyclohexane, n-heptane, toluene, chloroform, diethyl ether, isopropyl ether, methyl tert-butyl ether, tetrahydrofuran, ethylene glycol dimethyl ether , acetone, methyl acetate, ethyl acetate, isopropyl acetate, butyl acetate, methanol, ethanol, isopropanol, n-butanol and acetonitrile, preferably isopropyl ether, methyl tert-butyl ether, At least one of methanol, isopropanol and tetrahydrofuran, more preferably isopropyl ether.
  • the molar ratio of the compound of formula (I) to benzoic acid is 1:2-2:1, preferably 1:1.
  • the step (1-b) is carried out at 15-30°C, preferably at 20-25°C.
  • the stirring in step (3-b) is carried out under heating conditions, preferably the heating temperature is about 40-68°C, more preferably 60-68°C.
  • the preparation method of the benzoate B crystal form of the compound of formula (I) also includes the following steps:
  • the solvent in step (5-b) is selected from at least one of isopropyl ether, methyl tert-butyl ether, methanol, isopropanol and tetrahydrofuran, preferably isopropyl ether.
  • the step (6-b) is carried out at 15-30°C, preferably at 20-25°C.
  • the present disclosure also provides a crystalline form of the succinate salt of the compound of formula (I).
  • the crystalline form of the succinate salt of the compound of formula (I) is an anhydrate.
  • the present disclosure provides the crystal form A of succinate salt of the compound of formula (I).
  • the crystal form A is at 7.13 ⁇ 0.2°, 9.30 ⁇ 0.2°, 11.29 ⁇ 0.2 °, 15.51 ⁇ 0.2°, 19.89 ⁇ 0.2° have diffraction peaks.
  • the crystal form A is at 7.13 ⁇ 0.2°, 9.30 ⁇ 0.2°, 11.29 ⁇ 0.2°, 15.51 ⁇ 0.2°, 17.10 ⁇ 0.2 °, 19.89 ⁇ 0.2°, 20.26 ⁇ 0.2°, 20.91 ⁇ 0.2°, 21.51 ⁇ 0.2°, 22.69 ⁇ 0.2°, 23.58 ⁇ 0.2°, 25.42 ⁇ 0.2° have diffraction peaks.
  • the crystal form A is at 5.01 ⁇ 0.2°, 7.13 ⁇ 0.2°, 9.30 ⁇ 0.2°, 10.09 ⁇ 0.2°, 11.29 ⁇ 0.2 °, 11.73 ⁇ 0.2°, 13.75 ⁇ 0.2°, 14.30 ⁇ 0.2°, 15.18 ⁇ 0.2°, 15.51 ⁇ 0.2°, 16.00 ⁇ 0.2°, 17.10 ⁇ 0.2°, 18.25 ⁇ 0.2°, 18.69 ⁇ 0.2°, 19.89 ⁇ 0.2 °, 20.26 ⁇ 0.2°, 20.91 ⁇ 0.2°, 21.12 ⁇ 0.2°, 21.51 ⁇ 0.2°, 22.69 ⁇ 0.2°, 23.05 ⁇ 0.2°, 23.58 ⁇ 0.2°, 24.58 ⁇ 0.2°, 25.42 ⁇ 0.2°, 26.66 ⁇ 0.2 °, 27.24 ⁇ 0.2° and 29.47 ⁇ 0.2° have diffraction peaks.
  • the crystal form A is at 5.01 ⁇ 0.2°, 7.13 ⁇ 0.2°, 9.30 ⁇ 0.2°, 10.09 ⁇ 0.2°, 11.29 ⁇ 0.2 °, 11.73 ⁇ 0.2°, 13.75 ⁇ 0.2°, 14.30 ⁇ 0.2°, 15.18 ⁇ 0.2°, 15.51 ⁇ 0.2°, 16.00 ⁇ 0.2°, 17.10 ⁇ 0.2°, 18.25 ⁇ 0.2°, 18.69 ⁇ 0.2°, 19.89 ⁇ 0.2 °, 20.26 ⁇ 0.2°, 20.91 ⁇ 0.2°, 21.12 ⁇ 0.2°, 21.51 ⁇ 0.2°, 22.69 ⁇ 0.2°, 23.05 ⁇ 0.2°, 23.58 ⁇ 0.2°, 24.58 ⁇ 0.2°, 25.42 ⁇ 0.2°, 26.66 ⁇ 0.2 °, 27.24 ⁇ 0.2°, 28.65 ⁇ 0.2°, 29.47 ⁇ 0.2°, 30.90 ⁇ 0.2°, 32.31 ⁇ 0.2°, 34.45 ⁇ 0.2° have diffraction peaks.
  • the X-ray powder diffraction pattern of Form A is substantially consistent with FIG. 8 .
  • the DSC spectrum of the crystal form A has an endothermic peak at 186.82 ⁇ 5°C.
  • the DSC spectrum of Form A is substantially consistent with FIG. 9 .
  • the present disclosure also provides a method for preparing the succinate A crystal form of the compound of formula (I), comprising the following steps:
  • the solvent in step (1-c) is selected from cyclohexane, n-heptane, toluene, chloroform, diethyl ether, isopropyl ether, methyl tert-butyl ether, tetrahydrofuran, ethylene glycol dimethyl ether , acetone, methyl acetate, ethyl acetate, isopropyl acetate, butyl acetate, methanol, ethanol, isopropanol, n-butanol and acetonitrile, preferably isopropyl ether, methyl tert-butyl ether, At least one of methanol, isopropanol and tetrahydrofuran, more preferably isopropyl ether.
  • the molar ratio of the compound of formula (I) to succinic acid is 1:2-2:1, preferably 1:1.
  • the step (3-c) is carried out at 15-30°C, preferably at 20-25°C.
  • the reaction time of the step (3-c) is 1-48 hours, preferably 3-24 hours, more preferably 8-16 hours.
  • preparation method of the succinate A crystal form of the compound of formula (I) also includes the following steps:
  • the solvent in step (5-c) is at least one selected from methanol, ethanol, isopropanol, tetrahydrofuran and water, preferably a mixed solvent of ethanol/water.
  • the volume ratio of ethanol/water is about 1:1 ⁇ 50:1, preferably 2:1 ⁇ 20:1; in some specific embodiments of the present disclosure, the volume ratio of ethanol/water is about 10 :1, 9:1, 8:1, 7:1, 6:1, 5:1 or 4:1.
  • the step (6-c) is carried out at 15-30°C, preferably at 20-25°C.
  • the stirring crystallization time of the step (6-c) is 0.5-48 hours, preferably 1-12 hours, more preferably 2-4 hours.
  • the present disclosure also provides a pharmaceutical composition, which comprises a pharmaceutically acceptable salt of the compound of formula (I) above or the crystal form above and pharmaceutically acceptable excipients.
  • the present disclosure also relates to the use of the pharmaceutically acceptable salt of the compound of formula (I) above or the above crystal form, or the above pharmaceutical composition in the preparation of drugs for preventing or treating estrogen receptor-related diseases.
  • the present disclosure relates to the use of a pharmaceutically acceptable salt of the compound of formula (I) above or the above crystal form, or the above pharmaceutical composition in the prevention or treatment of estrogen receptor-related diseases.
  • the present disclosure relates to a pharmaceutically acceptable salt of the compound of the above formula (I) or the above crystal form, or the above pharmaceutical composition for preventing or treating estrogen receptor-related diseases.
  • the present disclosure also relates to a method for treating estrogen receptor-related diseases, the method comprising administering to an individual a therapeutically effective dose of the pharmaceutically acceptable salt of the compound of formula (I) or the above-mentioned crystal form, the above-mentioned pharmaceutical composition, or the above-mentioned Pharmaceutically acceptable salts of the compound of formula (I) or pharmaceutical preparations of the above crystal forms.
  • the present disclosure also relates to the use of the pharmaceutically acceptable salt of the compound of the above formula (I) or the above crystal form, or the above pharmaceutical composition in the preparation of drugs for preventing or treating tumors.
  • the present disclosure relates to the use of a pharmaceutically acceptable salt of the compound of formula (I) or the above crystal form, or the above pharmaceutical composition in the prevention or treatment of tumors.
  • the present disclosure relates to a pharmaceutically acceptable salt of the compound of formula (I) or the above crystal form, or the above pharmaceutical composition for preventing or treating tumors.
  • the present disclosure also relates to a method for treating tumors, the method comprising administering to an individual a therapeutically effective dose of a pharmaceutically acceptable salt of the above-mentioned compound of formula (I) or the above-mentioned crystal form, the above-mentioned pharmaceutical composition or a compound containing the above-mentioned formula (I) of the present disclosure Pharmaceutically acceptable salts or pharmaceutical preparations of the above crystal forms.
  • the estrogen receptor-related diseases include but are not limited to tumors.
  • the estrogen receptor-related disease or tumor is breast cancer.
  • the estrogen receptor-related disease or tumor is ER-positive breast cancer.
  • the estrogen receptor-related disease or tumor is brain metastases from ER-positive breast cancer.
  • the individual is a cancer patient.
  • the individual is a breast cancer patient.
  • the individual is a patient with ER-positive breast cancer.
  • the individual is a patient with brain metastases from ER-positive breast cancer.
  • the pharmaceutically acceptable salts of the compounds of formula (I) and their crystals provided by the present disclosure have at least one of the advantages of good water solubility, high purity, good stability, and low hygroscopicity;
  • benzoate, succinate or adipate and crystal forms thereof of the compound of formula (I) can also effectively inhibit the generation of isomers, and/or have better thermal stability;
  • the pharmaceutically acceptable salt of the compound of formula (I) or its crystal form provided by the present disclosure also has at least one of the advantages of simple preparation method, mild crystallization conditions, high crystallinity, good solubility, good stability, and difficult to absorb moisture, etc., It is suitable for preparing the desired pharmaceutical composition.
  • the daily dosage of the pharmaceutically acceptable salt of the compound of formula (I) of the present disclosure is 0.01 mg/kg to 200 mg/kg body weight, preferably 0.05 mg/kg to 50 mg/kg body weight, more preferably 0.1 mg/kg to 30 mg/kg body weight. kg body weight, in single or divided doses.
  • Typical routes of administration of the pharmaceutically acceptable salts of the present disclosure or their crystalline forms or their polymorphs, or their polymorphic compositions, or their pharmaceutical compositions include, but are not limited to, oral, rectal, topical , inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration.
  • the pharmaceutical composition is in oral form.
  • the pharmaceutical compositions can be formulated by mixing the active compounds with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present application to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, etc. for oral administration to patients.
  • Solid oral compositions can be prepared by conventional methods of mixing, filling or tabletting. It can be obtained, for example, by mixing the active compound with solid excipients, optionally milling the resulting mixture, adding other suitable excipients if desired, and processing the mixture into granules to obtain tablets Or the core of the sugar coating.
  • Suitable auxiliary materials include but are not limited to: binders, diluents, disintegrants, lubricants, glidants, sweeteners or flavoring agents, etc.
  • the room temperature mentioned in the present disclosure refers to 20 ⁇ 5°C.
  • the range “m ⁇ n” described in the present disclosure represents an abbreviated representation of any combination of real numbers between m and n, where both m and n are real numbers.
  • the numerical range “0.5 ⁇ 2” includes but is not limited to 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0 .
  • the "X-ray powder diffraction pattern" described in this disclosure is measured using Cu-K ⁇ radiation.
  • the crystalline forms disclosed in the present disclosure are substantially pure crystals.
  • the term "substantially pure” as used herein means that the crystalline form has a purity of at least 85 wt%, preferably at least 90 wt%, more preferably at least 95 wt%.
  • the peak positions of their XRPD patterns are generally similar, and the relative intensity error may be large. It should also be pointed out that in the identification of mixtures, due to the decrease of content and other factors, some diffraction lines will be missing. At this time, there is no need to rely on all the diffraction peaks observed in high-purity samples, and even one diffraction peak may also affect the given is characteristic for a given crystal.
  • the "2 ⁇ or 2 ⁇ angle" mentioned in the present disclosure refers to the diffraction angle, ⁇ is the Bragg angle, and the unit is ° or degree; the error range of each diffraction peak 2 ⁇ is ⁇ 0.20°.
  • Differential scanning calorimetry or DSC in this disclosure refers to the measurement of the temperature difference and heat flow difference between the sample and the reference object during the heating or constant temperature of the sample to characterize all the physical changes related to thermal effects and Chemical changes, to obtain the phase transition information of the sample.
  • the drying temperature described in the present disclosure is generally 20°C-100°C, preferably 25°C-70°C, more preferably 40°C-60°C, and can be dried under normal pressure or reduced pressure. Preferably, drying is performed under reduced pressure.
  • treating means administering a compound or formulation described herein to improve or eliminate a disease or one or more symptoms associated with the disease, and includes:
  • prevention means administering a compound or formulation described herein to prevent a disease or one or more symptoms associated with the disease, and includes:
  • a disease or disease state from occurring in a subject (eg, a mammal), particularly when such a subject is susceptible to the disease state but has not yet been diagnosed as having the disease state.
  • terapéuticaally effective dose means (i) treating a particular disease, condition or disorder, (ii) alleviating, ameliorating or eliminating one or more symptoms of a particular disease, condition or disorder, or (iii) delaying the The amount of a compound of the disclosure required for the onset of one or more symptoms of a particular disease, condition or disorder as described.
  • the amount of a compound of the disclosure that constitutes a “therapeutically effective dose” will vary depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by one skilled in the art. Based on its own knowledge and this disclosure.
  • pharmaceutically acceptable excipients refers to those excipients that have no obvious stimulating effect on the organism and will not impair the biological activity and performance of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and/or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water and the like.
  • mammals include mammals and non-mammals.
  • mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates (such as chimpanzees and other apes and monkeys); livestock such as cattle, horses, sheep, goats, pigs; domesticated animals , such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs.
  • non-human mammals include, but are not limited to, birds, fish, and the like.
  • Sample pan aluminum pan, non-sealed gland
  • Mobile phase A 0.01mol/L ammonium chloride solution (adjust the pH to 10.0 with ammonia water)
  • XRPD peak positions and/or intensities for a given crystalline form of the same compound will vary within error.
  • the 2 ⁇ values allow for an appropriate margin of error.
  • the margin of error is represented by " ⁇ ".
  • 5.92 ⁇ 0.2° 2 ⁇ represents a range of about 6.12 to 5.72.
  • suitable error ranges for XRPD diffraction angles (2 ⁇ ) may be about ⁇ 0.20°, ⁇ 0.15°, ⁇ 0.10° , ⁇ 0.05° or less.
  • the term “substantially identical” or “substantially as shown” means comprising at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 90% Or a pattern of diffraction peaks with at least 99% of the diffraction angles within a standard deviation of ⁇ 0.2° 2 ⁇ .
  • measurements of a DSC thermogram for a given crystalline form of the same compound will vary within a margin of error.
  • Single-peak peak values (expressed in degrees Celsius) allow for a modest margin of error.
  • the margin of error is represented by " ⁇ ".
  • thermal transition temperatures and melting points are typically within about ⁇ 5.0°C in consecutive analyses.
  • the peak value of "186.82 ⁇ 5°C” indicates that it is in the range of 181.82 to 191.82.
  • suitable error ranges for unimodal peaks may be ⁇ 5.0, ⁇ 4.0, ⁇ 3.0, ⁇ 2.0 or less.
  • Lux (lux, legal symbol lx) is the unit of luminance.
  • the luminous flux obtained by an object uniformly illuminated by light is 1 lumen on an area of 1 square meter, its illuminance is 1 lux.
  • NMR nuclear magnetic resonance
  • MS mass spectroscopy
  • Example 1 N-((S)(1-(3-fluoropropyl)pyrrolidin-3-yl)-6-((1S,3R)-3-methyl-2-(2,2,2 Synthesis of -trifluoroethyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)pyridin-3-amine (formula (I) compound)
  • Step 3 Synthesis of (R)-1-(1H-indol-3-yl)-N-(2,2,2-trifluoroethyl)propane-2-amine
  • Step 4 (1S,3R)-1-(5-Bromopyridin-2-yl)-3-methyl-2-(2,2,2-trifluoroethyl)-2,3,4,9- Synthesis of Tetrahydro-1H-pyrido[3,4-b]indole
  • Step 5 N-((S)-1-(3-fluoropropyl)pyrrolidin-3-yl)-6-(((1S,3R)-3-methyl-2-(2,2,2 Synthesis of -trifluoroethyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)pyridin-3-amine (compound of formula (I))
  • the NOESY spectrum (Fig. 1) shows that the methyl hydrogen on the 3-position of the compound of formula (I) and the hydrogen on the 1-position have a significant NOE effect, which proves that both are on the same side, and the pyridyl group on the 1-position and the hydrogen on the 3-position
  • the relative configuration of the methyl group on the 6-membered piperidine ring is trans, and the absolute configuration of the carbon atom at the 3-position is known as R, so the absolute configuration of the carbon atom at the 1-position is S.
  • Embodiment 2 the preparation of formula (I) compound sulfate
  • Embodiment 3 the preparation of formula (I) compound phosphate
  • Embodiment 4 the preparation of formula (I) compound fumarate
  • Embodiment 5 the preparation of formula (I) compound L-malate
  • Embodiment 6 the preparation of formula (I) compound citrate
  • Embodiment 7 the preparation of formula (I) compound adipate C crystal form
  • Embodiment 8 the preparation of formula (I) compound benzoate B crystal form
  • the XRPD spectrum of the crystalline solid is shown in Figure 5, its DSC spectrum is shown in Figure 6, and the TGA spectrum is shown in Figure 7. Its DSC endothermic peak peak is at 150.08 Around °C, the XRPD diffraction peak positions are shown in Table 2 below.
  • Embodiment 9 the preparation of formula (I) compound succinate
  • Embodiment 10 Preparation of crystal form A of compound succinate of formula (I)
  • Example 10-1 The succinate obtained in Example 9 was dissolved in methanol, and concentrated under reduced pressure to obtain a foamy solid. Weigh 500 mg of foamy solid, add ethanol (4.5 ml) and water (0.5 ml) into a 10 ml single-necked bottle, stir at room temperature, the solid dissolves, keep stirring for 2 hours to precipitate a solid, filter it with suction, and dry the obtained solid in vacuum at room temperature for 3 hours to obtain 0.22 g Crystal form A of the succinate salt of the compound of formula (I). The obtained solid was confirmed by X-ray powder diffraction spectrum. The XRPD spectrum of the crystalline solid is shown in Figure 8, its DSC spectrum is shown in Figure 9, and its TGA spectrum is shown in Figure 10. As shown in Table 3 below.
  • the crystal form A of compound succinate of formula (I) can also be prepared by the following examples: add 100 mg of the above-mentioned foamy solid and 500 ⁇ L of solvent (see Table 4) to a 5 ml centrifuge tube, stir at room temperature for 1-24 hours, and filter out solid and dried under vacuum at room temperature. See Table 4 for details.
  • Example solvent crystal form 10-2 isopropyl ether Form A 10-3 methyl tert-butyl ether Form A
  • Embodiment 11 Research on single crystal of compound succinate of formula (I)
  • a single crystal of the succinate of the compound of formula (I) can be prepared by the following method: take about 10 mg of the succinate of the compound of the formula (I) (prepared in Example 9) in a 3 mL sample bottle, add 1500 ⁇ L of isopropyl ether and 900 ⁇ L tetrahydrofuran, after ultrasonic dissolution, seal the bottle mouth with a parafilm and pierce a small hole in the parafilm with a needle, leave it at room temperature for 3 days, and precipitate crystals, and the obtained single crystal sample is detected by X-powder diffraction as A crystal form.
  • the single crystal belongs to the tetragonal crystal system and the space group I 4.
  • the stoichiometric formula is C 30 H 37 F 4 N 5 O 4 , and the calculated molecular weight of the asymmetric unit is 607.64.
  • Experimental method Accurately weigh about 10 mg of the sample to be tested, put it in a 10 ml centrifuge tube, add 1 ml of water, and shake in a water bath constant temperature shaker (37 ° C, 150 rpm). If the sample is completely dissolved, continue to add 10 mg of the sample and shake for about After 24 hours, take the supernatant, perform HPLC detection after appropriate dilution with acetonitrile, and determine its concentration by the external standard method.
  • Test example 2 investigation test of isomer impurity of formula (I) compound and pharmaceutically acceptable salt thereof
  • Test method formula (I) compound succinate (embodiment 10-1, A crystal form), formula (I) compound adipate (embodiment 7, C crystal form), formula (I) compound (implementation Example 1) are packaged with two layers of polyethylene and one layer of pharmaceutical composite film, focusing on the changes in the content of isomer impurities of the compound of formula (I) under conditions such as high temperature, light, high humidity, and acceleration.
  • the results are shown in Table 7 shown.
  • the isomer impurities are isomer 1 and isomer 2, respectively, and the structures are shown below. N.D means the impurity was not detected.
  • Test method the formula (I) compound succinate (embodiment 10-1, A crystal form), the formula (I) compound benzoate (embodiment 8, B crystal form), the formula (I) compound hexadiene Salt (embodiment 7, C crystal form) all adopts two layers of polyethylene, one layer of pharmaceutical composite film packing, investigates the change of impurity amount and impurity total amount under conditions such as high temperature, illumination, acceleration and high humidity, the result As shown in Table 8.
  • formula (I) compound succinate, formula (I) compound benzoate and formula (I) compound adipate all can control total impurity content preferably, especially formula (I) compound succinic acid
  • the salt and the adipate salt of the compound of formula (I) perform well in both aspects of total impurity content and impurity growth rate under conditions such as high temperature, light, acceleration and high humidity.
  • Test Example 4 Rat Pharmacokinetic Evaluation of Compounds of Formula (I) and Pharmaceutically Acceptable Salts thereof
  • MC methylcellulose
  • Acetonitrile was purchased from Merck (USA).
  • formula (I) compound benzoate, formula (I) compound succinate and formula (I) compound adipate have achieved more excellent PK characteristics, and have good oral bioavailability .
  • MCF-7, CAMA-1, HCC1500, T47D, and SK-BR-3 cells were purchased from ATCC, and EFM-19 and MCF-7 ESR1 Y537S mutant strains were purchased from Kebai Biotech.
  • the cell culture conditions refer to ATCC or Kebai product instructions, the medium is purchased from Invitrogen, and the serum is purchased from Gibico.
  • MCF-7 medium is DMEM+10%FBS+0.01mg/ml human insulin (Yisheng)+1% non-essential amino acid.
  • CAMA-1 medium is EMEM+10% FBS.
  • HCC1500 and EFM-19 medium are RPMI-1640+10% FBS.
  • T47D medium is RPMI-1640+10% HI-FBS+2 unit/mL bovine insulin (Solarbio).
  • MCF-7 ESR1 Y537S medium is MEM + 10% FBS + 1mM sodium pyruvate + 1% non-essential amino acids.
  • SK-BR-3 medium is McCoy's 5A+10% FBS. Fulvestrant was purchased from MCE.
  • the crystal form A of succinate salt of the compound of formula (I) was prepared according to the method in Example 10-1.
  • Use Echo650 (Beckman) to transfer 120nL of 3-fold diluted compound to the cell plate, add 40 ⁇ L of corresponding medium, after 7 days of compound treatment, add CellTiter-Glo (Promega) to detect cell viability, the cell well is used as 0% inhibition control, 0.1 ⁇ M fluorine Wells treated with fulvestrant were used as 100% inhibition control (wells treated with 2 ⁇ M fulvestrant for MCF-7 ESR1 Y537S were used as 100% inhibition control, and medium wells were used as 100% inhibition control for SK-BR-3). Using IDBS XLfit to perform 4-parameter fitting to calculate IC 50 and maximum inhibition rate data.
  • the succinate A crystal form of the compound of formula (I) showed strong proliferation inhibitory effects on different ER-positive breast cancer cell lines, with IC 50 ⁇ 1nM, and the IC 50 for proliferation inhibition of ESR1 Y537S mutant strains was 27.6nM, SK-BR-3 has no inhibitory effect on the proliferation of ER-negative cells at the highest concentration of 2 ⁇ M.
  • Test example 6 formula (I) compound succinate A crystal form to MCF-7 and MCF-7 ESR1 Y537S mutant strain ER protein
  • MCF-7 was purchased from ATCC, and the MCF-7 ESR1 Y537S mutant strain was purchased from Kebai Biotech.
  • the cell culture conditions refer to ATCC or Kebai product instructions, the medium is purchased from Invitrogen, and the serum is purchased from Gibico.
  • MCF-7 medium is DMEM+10%FBS+0.01mg/ml human insulin (Yisheng)+1% non-essential amino acid.
  • CAMA-1 medium is EMEM+10% FBS.
  • MCF-7 ESR1 Y537S medium is MEM + 10% FBS + 1mM sodium pyruvate + 1% non-essential amino acids. Fulvestrant was purchased from MCE.
  • the crystal form A of succinate salt of the compound of formula (I) was prepared according to the method in Example 10-1.
  • IRDye 800 goat anti-rabbit IgG antibody and IRDye 680 goat anti-mouse IgG antibody (LI-COR) and incubate in the dark for 45 minutes, wash off the secondary antibody, invert 1200 rpm and centrifuge for 1 minute to remove residual liquid, use Sapphire RGBNIR (Azure) detects 800nM and 680nM signals.
  • Use AzureSpot for fluorescence quantitative analysis calculate Chanel 800(ER)/Chanel 680(GAPDH) value, cell wells as 0% inhibition control, MCF-7 use 0.1 ⁇ M fulvestrant treatment wells as 100% inhibition control, MCF-7 ESR1 Y537S wells were treated with 2 ⁇ M fulvestrant as 100% degradation.
  • IDBS XLfit was used to perform 4-parameter fitting to calculate DC 50 and maximum degradation rate data.
  • Human breast cancer MCF-7 cells ECACC, 86012803
  • EMEM medium ATCC, Cat No.: 30-2003
  • Fetal bovine serum ExCell; Cat No.: FND500
  • Double antibody Gibco, Cat No.: 15240-062
  • Fulvestrant injection (Fulvestrant): AstraZeneca, 250mg/5mL/bottle
  • mice Female, 6-8 weeks old, weighing about 18-22 grams, were purchased from Shanghai Lingchang Biotechnology Co., Ltd., and the mice were raised in an SPF-grade environment. Each cage Separate exhaust ventilation was provided and all animals had free access to a standard certified commercial laboratory diet and water ad libitum.
  • Cell culture human breast cancer MCF-7 cell line was cultured in vitro, and the culture conditions were 10% fetal bovine serum and 1% double antibody in EMEM (cell culture medium), 37° C., 5% CO 2 incubator. Routine digestion with 0.25% trypsin-EDTA digestion solution was performed twice a week for subculture. When the confluence of the cells is 80%-90% and the number reaches the requirement, the cells are collected and counted.
  • Cell inoculation Inoculate 0.2ml/(containing 1 ⁇ 107 ) MCF-7 cell suspension (DPBS: Matrigel, volume ratio 1:1) subcutaneously on the right back of each mouse, and inoculate 17 ⁇ -estradiol tablets were subcutaneously inoculated two days before. On the 6th day after cell inoculation, when the average tumor volume reached 173.65 mm 3 , group administration began, and random grouping was administered according to the tumor volume. The day of group administration was Day 0 (Day 0).
  • the administration dose of formula (I) compound succinate (Example 10-1, A crystal form) is 0.3, 1, 3 or 10 mg/kg (calculated as free base), orally administered (PO), Once-daily dosing (QD) x21 times.
  • the dosage of fulvestrant (Fulvestrant) is 250 mg/kg, subcutaneous injection (SC), once a week (QW) x 4 times. 8 mice per group.
  • Tumor diameters were measured twice a week with vernier calipers.
  • Mouse body weights were measured twice a week.
  • TGI (%) [(1-(average tumor volume at the end of administration of a certain treatment group-average tumor volume at the beginning of administration of this treatment group)/(average tumor volume at the end of treatment of the solvent control group-at the beginning of treatment of the solvent control group Mean tumor volume)] x 100%.
  • mice have more than 10% weight loss, it has nothing to do with the dose of the test product formula (I) compound succinate, and the body weight of the mice in the test drug group is not particularly significant compared with the vehicle control group. Considering that the model is affected by the estrogen patch and may affect the state and body weight of the mice, the weight loss of these individual mice has nothing to do with the test product formula (I) compound succinate. No mouse died in each group of the experiment.
  • test results show that on the 20th day (Day 20) after the start of administration, subcutaneous injection of Fulvestrant 250 mg/kg into human breast cancer MCF-7 tumor-bearing mice once a week has a significant effect on the tumor growth of the MCF-7 xenograft tumor model. Weak inhibitory effect (P ⁇ 0.05).
  • formula (I) compound succinate shows significantly better than reference substance Fulvestrant (250mg/kg, SC) at a dose of 1mg/kg and above (PO, QD). , QW) antitumor activity (P ⁇ 0.0001).
  • Test Example 8 Pharmacodynamic study of compound succinate of formula (I) on human ER-positive breast cancer T47D xenograft subcutaneous xenograft tumor mouse model
  • T47D Human breast cancer xxT47D cells are T47D (ATCC, HTB-133) cells that have been passaged twice in vivo
  • RPMI-1640 culture medium Gibco, Cat No.: C22400500BT
  • Bovine insulin Yuanye, Cat.No.:11070-73-8
  • Double antibody Gibco, Cat No.: 15240-062
  • Fulvestrant injection (Fulvestrant): AstraZeneca, 250mg/5mL/bottle
  • mice Female, 6-8 weeks old, weighing about 18-22 grams, were purchased from Shanghai Lingchang Biotechnology Co., Ltd., and the mice were raised in an SPF-grade environment. Each cage Separate exhaust ventilation was provided and all animals had free access to a standard certified commercial laboratory diet and water ad libitum.
  • Cell culture Human breast cancer xxT47D cells were cultured in vitro. The culture conditions were RPMI-1640 medium plus 10% fetal bovine serum, 0.2 Units/mL bovine insulin, 1% double antibody, and cultured in a 5% CO 2 incubator at 37°C. Routine digestion with trypsin-EDTA was performed twice a week for passaging. When the confluence of the cells is 80%-90% and the number reaches the requirement, the cells are collected and counted.
  • Cell inoculation Inoculate 0.2mL (containing 1 ⁇ 107 ) xxT47D cell suspension (DPBS: Matrigel, volume ratio 1:1) subcutaneously on the right back of each mouse, and two days before cell inoculation Subcutaneous inoculation of 17 ⁇ -estradiol tablets. On the 6th day after cell inoculation, when the average volume of the tumor reached 137.09 mm 3 , group administration began, and the group was randomly assigned according to the tumor volume, and the day of group administration was Day 0 (Day 0).
  • DPBS Matrigel, volume ratio 1:1
  • the dosage of the compound succinate of formula (I) (Example 10-1, A crystal form) is 0.3, 1 or 3mg/kg, administered orally (PO), administered once a day (QD) x28 Second-rate.
  • the dosage of fulvestrant (Fulvestrant) is 250 mg/kg, subcutaneous injection (SC), once a week (QW) x 4 times. 8 mice per group.
  • Tumor diameters were measured twice a week with vernier calipers.
  • Mouse body weights were measured twice a week.
  • TGI (%) [(1-(average tumor volume at the end of administration of a certain treatment group-average tumor volume at the beginning of administration of this treatment group)/(average tumor volume at the end of treatment of the solvent control group-at the beginning of treatment of the solvent control group Mean tumor volume)] x 100%.
  • test results showed that on the 28th day (Day 28) after the start of administration, subcutaneous injection of 250mg/kg Fulvestrant to human breast cancer T47D tumor-bearing mice once a week showed a certain anti-tumor effect compared with the vehicle control group.
  • the effect of growth (P ⁇ 0.0001).
  • formula (I) compound succinate shows significantly better than reference substance Fulvestrant (250mg/kg, SC, QW) under the dose (PO, QD) of 1mg/kg and above. ) antitumor activity (P ⁇ 0.0001).
  • Human breast cancer MCF-7 cells ATCC, HTB-22
  • EMEM medium ATCC, Cat No.: 30-2003
  • Fetal bovine serum Gibco, Cat No.: 10099-141C
  • DPBS Hyclone, Cat. No.: SH30256.01
  • Fulvestrant injection (Fulvestrant): AstraZeneca, 250mg/5mL/bottle
  • mice Female, 6-7 weeks old, weighing about 17-23 grams, were purchased from Beijing Weitongda Biotechnology Co., Ltd., and the mice were raised in an SPF-grade environment, and each cage was sent separately All animals had free access to a standard certified commercial laboratory diet and water ad libitum.
  • Cell culture In vitro culture of human breast cancer MCF-7 cell line, the culture conditions are 10% fetal bovine serum, 10 ⁇ g/ml recombinant human insulin and 1% P/S in EMEM (cell culture medium), 37 ° C, 5% CO 2 incubators. Passage with routine digestion with 0.25% trypsin-EDTA once or twice a week. When the number reaches the requirement and the cells are in the logarithmic growth phase, the cells are collected and counted.
  • MCF-7 0.015ml/(containing 2 ⁇ 10 6 ) MCF-7 cell suspension was inoculated in the brain of each mouse (1mm in front of bregma, 2mm on the right side of the raphe, 3mm below the dura mater plane of the skull) , and 17 ⁇ -estradiol tablets were subcutaneously inoculated three days before cell inoculation. On the 8th day after cell inoculation, group administration began, and animals were randomly divided into groups according to body weight, and the day of grouping was Day 0.
  • the dosage of the compound succinate of formula (I) (Example 10-1, crystal form A) is 3, 10 or 30 mg/kg, administered orally (PO), administered once a day (QD), Dosing continued until Day 60.
  • the dosage of fulvestrant (Fulvestrant) was 250mg/kg, subcutaneously injected (SC), once a week (QW), and continued until Day 60. 8 mice per group.
  • mice were weighed twice a week.
  • the antitumor efficacy of compounds was evaluated by the median survival time.
  • Test Example 10 Research on the effect of compounds of formula (I) on the degradation of estrogen receptors in MCF7 cells
  • the purpose of this experiment is to measure the degradation activity of the compound of formula (I) on the endogenously expressed estrogen receptor in MCF7 cells, and evaluate the activity of the compound according to DC 50 and maximum degradation efficiency.
  • MCF7 cells (ATCC, HTB-22) were cultured with DMEM (Gibco, 11995-065) complete medium containing 10% fetal bovine serum. On the first day of the experiment, MCF7 cells were seeded in a 384-well plate at a density of 3000 cells/well using complete medium, and cultured in a 5% CO 2 cell incubator at 37°C.
  • DMEM Gibco, 11995-065
  • the compound to be tested was dissolved in DMSO with a storage concentration of 10mM, diluted with Echo 550 (Labcyte Inc.) and added to the cell culture plate, the initial concentration of the compound to be tested was 100nM, 3-fold serial dilution, 9 concentration points, set A blank control containing 0.5% DMSO was used, and a double-well control was set at each concentration point.
  • DMSO fetal sulfate
  • Echo 550 Echo 550
  • the wells treated with 0.1 ⁇ M fulvestrant were used as the 100% degradation control, and the degradation rate at each concentration point was calculated.
  • XlLfit was used to analyze the processing data, and the degradation activity DC50 and the maximum degradation rate Imax of the compound were calculated. See Table 15 for data analysis.
  • Table 15 formula (I) compound is to MCF7 intracellular estrogen receptor degradation activity
  • Test Example 11 Study on the inhibitory effect of the compound of formula (I) on the proliferation of MCF7 cells
  • the purpose of this experiment is to determine the inhibitory effect of the compound of formula (I) on the proliferation of MCF7 cells in vitro, and to evaluate the activity of the compound according to IC 50 .
  • MCF7 cells (ATCC, HTB-22) were cultured with DMEM (Gibco, 11995-065) complete medium containing 10% fetal bovine serum. On the first day of the experiment, MCF7 cells were seeded in a 384-well plate at a density of 500 cells/well using complete medium, and cultured overnight at 37° C. in a 5% CO 2 cell incubator. The next day, add the compound to be tested for drug treatment, and use Echo550 (Labcyte Inc.) to dilute the compound solution with a storage concentration of 10 mM and transfer it to each cell culture well.
  • DMEM Gibco, 11995-065
  • Echo550 Echo550
  • the initial concentration of the compound to be tested in the cells is 100nM, 3-fold serial dilution, 9 concentration points, a blank control containing 0.3% DMSO was set, and double-well controls were set at each concentration point.
  • Test Example 12 Apparent Solubility of Compound of Formula (I) in Phosphate Buffered Saline at pH 7.4
  • the compounds to be tested were prepared according to the methods described.
  • the control drug progesterone was purchased from Sigma.
  • Phosphate buffer with a pH value of 7.4 was prepared by our laboratory.
  • Acetonitrile and methanol were purchased from Fisher.
  • Other reagents were purchased from the market.
  • Test Example 13 The blood-brain barrier (BBB) penetration ability of the compound of formula (I) in rats
  • Drugs can pass through the blood-brain barrier of animals and have sufficient exposure in the brain is the key to the effectiveness of drugs on brain metastases. Therefore, by measuring drug concentrations in plasma and brain tissue after administration to animals, the effect of drugs in the brain Distribution, and then judge whether the drug can inhibit tumor growth in the brain orthotopic model.
  • SD female rats were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.
  • MC methylcellulose
  • acetonitrile was purchased from Merck (USA).
  • PBS phosphate buffered saline
  • Rat brain tissue samples were first homogenized with 4 times the mass volume of PBS homogenate. Take 20 ⁇ L of brain tissue homogenate sample, add 20 ⁇ L of blank mouse plasma to dilute and mix, then add 600 ⁇ L of acetonitrile solvent (including internal standard compound) to precipitate protein, vortex for 5 min, centrifuge (14,000 rpm) for 5 min, supernatant with Diluted 2 times with 0.1% (v/v) FA in water, and carried out quantitative detection on LC-MS/MS system (AB Sciex Triple Quad 6500+).
  • the compound of formula (I) exhibits excellent blood-brain barrier penetration ability, and the drug exposure in rat brain tissue is relatively high.
  • the BBB test results are as follows:
  • Test Example 14 Growth inhibition experiment of the compound of formula (I) on MCF-7 mouse subcutaneous tumor model
  • Human breast cancer MCF-7 cells ATCC, HTB-22
  • EMEM medium ATCC, Cat No.: 30-2003
  • Fetal bovine serum Gbico; Cat No.:1099-141C
  • Pen Strep (Pen Strep): Gibco, Cat No.: 15240-122
  • D-PBS phosphate buffered saline without calcium and magnesium ions
  • mice Female, 6-7 weeks old, weighing about 19-28 grams, were purchased from Beijing Weitongda Biotechnology Co., Ltd., and the mice were raised in an SPF-grade environment, and each cage was sent separately All animals had free access to a standard certified commercial laboratory diet and water ad libitum.
  • Cell culture In vitro culture of human breast cancer MCF-7 cell line, the culture conditions are 10% fetal bovine serum, 1% Pen Strep, 10 ⁇ g/ml recombinant human insulin in EMEM (cell culture medium), 37°C, 5% CO 2 incubator. Routine digestion with 0.25% trypsin-EDTA digestion solution once a week for passage. When the cell saturation is 80%-90% and the number reaches the requirement, collect the cells and count them.
  • Cell inoculation Subcutaneously inoculate 0.1ml/(containing 1 ⁇ 10 7 ) MCF-7 cell suspension (D-PBS: Matrigel, volume ratio 1:1) on the right back of each mouse, and inoculate In the first four days, 17 ⁇ -estradiol tablets were subcutaneously inoculated. On the 24th day after cell inoculation, the drugs were randomly divided into groups according to the tumor volume, and the grouping day was Day 0 (Day 0).
  • the dosage of the compound of formula (I) is 1, 3 or 10 mg/kg, administered orally (PO), administered once a day (QD) x 3 weeks. There were 8 mice in the vehicle group and 6 mice in the administration group.
  • Tumor diameters were measured twice a week with vernier calipers.
  • Mouse body weights were measured twice a week.
  • TGI (%) [(1-(average tumor volume at the end of administration of a certain treatment group-average tumor volume at the beginning of administration of this treatment group)/(average tumor volume at the end of treatment of the solvent control group-at the beginning of treatment of the solvent control group Mean tumor volume)] x 100%.
  • the compound of formula (I) has a significant inhibitory effect on tumor growth (P ⁇ 0.01) at 1 mg/kg, 3 mg/kg, or 10 mg/kg orally administered once a day, and has It has a good dose-response relationship, and it has the effect of shrinking tumors at the doses of 3mg/kg and 10mg/kg.
  • Oral administration of the compound of formula (I) once a day at 10 mg/kg has a significant inhibitory effect on tumor growth (P ⁇ 0.01), and has the effect of shrinking tumors.
  • the compound of formula (I) did not significantly affect the body weight of mice at the doses tried.
  • Test Example 15 Inhibitory experiment of the compound of formula (I) on the growth of mouse MCF-7 brain orthotopic tumor model
  • Human breast cancer MCF-7 cells ATCC, HTB-22
  • EMEM medium ATCC, Cat No.: 30-2003
  • Fetal bovine serum Gibco, Cat.No.:1099-141C
  • Pen Strep (Pen Strep): Gibco, Cat No.: 15240-122
  • Micro injection pump KDS, Cat No.: Legato130
  • mice Female, 6-8 weeks old, weighing about 17-29 grams, were purchased from Beijing Weitongda Biotechnology Co., Ltd., and the mice were raised in an SPF-grade environment, and each cage was sent separately All animals had free access to a standard certified commercial laboratory diet and water ad libitum.
  • Cell culture In vitro culture of human breast cancer MCF-7 cell line, the culture conditions are 10% fetal bovine serum, 1% Pen Strep, 10 ⁇ g/ml recombinant human insulin in EMEM (cell culture medium), 37°C, 5% CO 2 incubator. Routine digestion with 0.25% trypsin-EDTA digestion solution was performed twice a week for subculture. When the cell saturation is 80%-90% and the number reaches the requirement, collect the cells and count them.
  • MCF-7 15 ⁇ l/(containing 2 ⁇ 10 6 ) MCF-7 cell suspension was inoculated into the mouse skull using a brain localizer, a micro-injection pump and a miniature hand-held cranial drill, and 17 ⁇ was subcutaneously inoculated three days before cell inoculation - Estradiol tablets. On the 8th day after cell inoculation, mice were randomly divided into groups according to body weight for administration, and the grouping day was Day 0 (Day 0).
  • Fulvestrant (Fulvestrant, AstraZeneca) dosage is 250mg/kg, subcutaneous injection (SC), administration (QW) once a week, the dosage of formula (I) compound is 30mg/kg , administered orally (PO), administered once a day (QD).
  • SC subcutaneous injection
  • QW administration
  • PO administration
  • QD administration
  • mice There were 11 mice in the vehicle group and 8 mice in the administration group. All groups continued administration until the mice died, were euthanized due to poor condition or the experiment ended.
  • mice The body weight of the mice was measured twice a week, and the survival status of the mice was observed.
  • mice in the Fulvestrant group continued to decrease, and the survival status of the mice was not significantly different from that of the solvent control group (median survival period, vehicle 29 days in the control group and 29.5 days in the Fulvestrant group).
  • the mice in the compound group of formula (I) (30 mg/kg administered orally once a day) had a stable body weight and no abnormal state. Until the end of the experiment, the mice in the compound group of formula (I) did not die.
  • the compound It has a significant inhibitory effect on MCF-7 brain orthotopic tumor model mice, and the survival period of mice is significantly prolonged (P ⁇ 0.01).

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  • Medicinal Chemistry (AREA)
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  • Life Sciences & Earth Sciences (AREA)
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  • Public Health (AREA)
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  • General Chemical & Material Sciences (AREA)
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  • Epidemiology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne un sel pharmaceutiquement acceptable d'un composé de formule (I), un cristal de celui-ci, son procédé de préparation et son utilisation. Le sel pharmaceutiquement acceptable est choisi parmi sulfate, phosphate, benzoate, succinate, adipate, fumarate, L-malate et citrate, de préférence benzoate, succinate et adipate.
PCT/CN2022/131313 2021-11-12 2022-11-11 Sel de composé de pyrrolidine, forme cristalline de celui-ci et son procédé de préparation Ceased WO2023083292A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019223715A1 (fr) * 2018-05-23 2019-11-28 江苏恒瑞医药股份有限公司 Dérivé de benzopipéridine ou d'hétéroarylpipéridine, son procédé de préparation et son utilisation médicale
CN111499614A (zh) * 2019-01-31 2020-08-07 江苏恒瑞医药股份有限公司 四氢异喹啉类衍生物、其制备方法及其在医药上的应用
WO2021228210A1 (fr) * 2020-05-15 2021-11-18 江苏先声药业有限公司 Composé pyrrolidine et son utilisation
WO2021249533A1 (fr) * 2020-06-12 2021-12-16 江苏先声药业有限公司 Composé modulateur du récepteur des oestrogènes et son utilisation
WO2022166983A1 (fr) * 2021-02-08 2022-08-11 贝达药业股份有限公司 Dérivé d'hétéroaryloxypipéridine, composition pharmaceutique de celui-ci et son utilisation

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019223715A1 (fr) * 2018-05-23 2019-11-28 江苏恒瑞医药股份有限公司 Dérivé de benzopipéridine ou d'hétéroarylpipéridine, son procédé de préparation et son utilisation médicale
CN111499614A (zh) * 2019-01-31 2020-08-07 江苏恒瑞医药股份有限公司 四氢异喹啉类衍生物、其制备方法及其在医药上的应用
WO2021228210A1 (fr) * 2020-05-15 2021-11-18 江苏先声药业有限公司 Composé pyrrolidine et son utilisation
WO2021249533A1 (fr) * 2020-06-12 2021-12-16 江苏先声药业有限公司 Composé modulateur du récepteur des oestrogènes et son utilisation
WO2022166983A1 (fr) * 2021-02-08 2022-08-11 贝达药业股份有限公司 Dérivé d'hétéroaryloxypipéridine, composition pharmaceutique de celui-ci et son utilisation

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