A compound for inhibiting myeloid blast activity and its preparation method and application

Abstract

The present invention belongs to the field of pharmaceutical preparation technology and relates to a compound that inhibits the activity of myeloblasts, as well as its preparation method and application. Compound 1 undergoes a Suzuki coupling reaction with a boronic acid compound in the presence of a first catalyst and a first base to produce Compound 2; Compound 2 undergoes an esterification reaction in the presence of a second base to produce Compound 3; Compound 3 undergoes a condensation reaction with 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one hydrochloride in the presence of a condensing agent and a third base to produce Compound I. Compound I provided herein is a novel compound that, in biological testing, has demonstrated strong inhibitory activity against myeloblasts. Myeloblast proliferation can cause brain tumors in children. Therefore, Compound I is expected to be further developed into a drug for treating childhood brain tumors. #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparation, and relates to a compound for inhibiting the activity of myeloid blasts, and a preparation method and application thereof. Background Art

[0002] Medulloblastoma (MB) is an embryonic malignant tumor originating from early neural progenitor cells in the cerebellum. It is a type of primitive neuroectodermal tumor and a common intracranial tumor in children, ranking second among intracranial tumors in children. It is classified as grade IV by the WHO and is highly malignant. The disease is common in children aged 3 to 7 years. Medulloblastoma mainly affects the cerebellar vermis and has a strong tendency to spread into the cerebrospinal fluid. It is a common central nervous system malignant tumor in children with high invasiveness, high recurrence rate, poor prognosis, and serious impact on children's health and quality of life.

[0003] Medulloblastoma is histologically classified into classic and desmoplastic types. Current common treatments include surgery, chemotherapy, and radiotherapy. However, many patients still die from the disease. Even if they survive, they will be left with various sequelae, including cognitive deficits and endocrine disorders. Therefore, people urgently need to improve strategies and methods for treating medulloblastoma.

[0004]

[0005] The development and progression of medulloblastoma involve alterations in signaling pathways that regulate cell proliferation. UNC1999 and EPZ6438 are EZH inhibitors that have demonstrated efficacy in medullary blastoma (MB), effectively inhibiting medulloblastoma growth. However, their main drawbacks are poor water solubility and lack of selectivity for cancer cells versus normal cells, leading to severe toxic side effects. For these reasons, further research is needed to identify novel and effective small-molecule compounds that can overcome these limitations. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to address the deficiencies of the existing technology and provide a compound for inhibiting the activity of medulloblasts, a preparation method thereof, and an application thereof, so as to solve the problem that there is a lack of effective drugs for treating medulloblastoma on the market. The product provided by the present invention can effectively inhibit the growth of medulloblasts and can be used to prepare drugs for treating medulloblastoma-related diseases.

[0007] In order to solve the above technical problems, the technical solutions adopted by the present invention are as follows:

[0008] The present invention discloses a compound for inhibiting myeloblast activity as shown in formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs;

[0009]

[0010] Furthermore, the present invention discloses a method for preparing the above-mentioned compound for inhibiting the activity of myeloblasts, comprising the following steps:

[0011] Compound 1 undergoes a Suzuki coupling reaction with a boronic acid compound in the presence of a first catalyst and a first base to obtain compound 2; compound 2 undergoes an esterification reaction in the presence of a second base to obtain compound 3; compound 3 undergoes a condensation reaction with 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one hydrochloride in the presence of a condensing agent and a third base to obtain compound 1;

[0012]

[0013] In some embodiments, the boronic acid compound is 3-(methoxycarbonyl)phenylboronic acid or methyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate; the first catalyst includes but is not limited to any one or a combination of tetrakistriphenylphosphine palladium, di(tri-tert-butylphosphine) palladium, 1,1′-bisdiphenylphosphinodiferrocenepalladium dichloride, tris(dibenzylideneacetone)dipalladium and palladium acetate; the first base includes but is not limited to any one or a combination of sodium carbonate, potassium carbonate and sodium bicarbonate; the molar ratio of compound 1 to the boronic acid compound, the first catalyst and the first base is 1:1.5:0.08:3; the Suzuki coupling reaction is carried out under the protection of an inert gas.

[0014] In some embodiments, preferably, the boronic acid compound is 3-(methoxycarbonyl)phenylboronic acid or 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid methyl ester; the first catalyst is tetrakistriphenylphosphine palladium; the first base is sodium carbonate; and the molar ratio of the compound 1 to the boronic acid compound, the first catalyst, and the first base is 1:1.5:0.08:3.

[0015] Wherein, the solvent used in the Suzuki coupling reaction includes but is not limited to any one or a combination of dioxane, water, DMF, ethanol, ethylene glycol dimethyl ether and toluene. Preferably, the solvent used in the Suzuki coupling reaction includes but is not limited to a mixed solvent of dioxane and water, a mixed solvent of DMF and water, a mixed solvent of ethanol and water, a mixed solvent of ethylene glycol dimethyl ether and water, or a mixed solvent of toluene and water. Further preferably, the solvent used in the Suzuki coupling reaction is a mixed solvent of dioxane and water in any proportion. Further preferably, the solvent used in the Suzuki coupling reaction is a mixed solvent of dioxane and water in a volume ratio of 10:3. There is no special requirement for the amount of solvent used in the Suzuki coupling reaction, and the raw material solvent or dispersion can be uniform.

[0016] The inert gas is preferably nitrogen or argon, more preferably nitrogen; the Suzuki coupling reaction is a reflux reaction, and the reaction temperature is determined by the solvent used, and the reaction temperature can be such that the solvent refluxes.

[0017] In some embodiments, the second base includes but is not limited to any one or a combination of sodium hydroxide, potassium hydroxide, potassium carbonate and sodium carbonate; and the esterification reaction is carried out at room temperature.

[0018] In some embodiments, preferably, the second base is sodium hydroxide; and the esterification reaction is carried out at room temperature.

[0019] The second base exists in the form of a solution. Unless otherwise specified, the solvent in the solution is water. The amount of the second base used is excessive, and the amount of the second base used is at least twice the molar amount of compound 2.

[0020] The solvent used in the esterification reaction includes but is not limited to any one or a combination of methanol, DMSO, DMF, DCM, acetonitrile, n-butanol, dioxane and tetrahydrofuran. Preferably, the solvent used in the esterification reaction is a mixture of methanol and tetrahydrofuran in any proportion. Further preferably, the solvent used in the esterification reaction is a mixture of methanol and tetrahydrofuran in a volume ratio of 4:1. There is no special requirement for the amount of the solvent used in the esterification reaction, and the raw material solvent can be evenly dispersed.

[0021] In some embodiments, the condensing agent includes but is not limited to any one or a combination of 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, benzotriazole-1-tris(trimethylamino)-hexafluorophosphate, 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate, N,N-dicyclohexylcarbodiimide and O-benzotriazole-tetramethyluronium hexafluorophosphate; the third base includes but is not limited to any one or a combination of triethylamine, diethylamine, diisopropylethylamine, N-methylmorpholine, potassium carbonate and sodium carbonate; the molar ratio of the compound 3 to 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one hydrochloride, the condensing agent and the third base is 1:1.5:4:5; the condensation reaction is carried out at room temperature.

[0022] In some embodiments, preferably, the condensing agent is a mixture of 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride in any proportion, further preferably, the condensing agent is a mixture of 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride in a molar ratio of 1:1; the third base is N-methylmorpholine; the molar ratio of the compound 3 to 3-(aminomethyl)-4,6-dimethylpyridine-2(1H)-one hydrochloride, the condensing agent, and the third base is 1:1.5:4:5; and the condensation reaction is carried out at room temperature.

[0023] Among them, 1-hydroxybenzotriazole is abbreviated as HOBt; 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is abbreviated as EDCI; benzotriazole-1-tris(trimethylamino)-hexafluorophosphate is abbreviated as BOP; 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate is abbreviated as HATU; N,N-dicyclohexylcarbodiimide is abbreviated as DCC; O-benzotriazole-tetramethyluronium hexafluorophosphate is abbreviated as HBTU.

[0024] The solvent used in the condensation reaction includes but is not limited to any one or a combination of DMSO, dichloromethane and DMF. Preferably, the solvent used in the condensation reaction is DMSO. There is no special requirement for the amount of the solvent used in the condensation reaction, and the raw material solvent can be evenly dispersed.

[0025] Furthermore, the present invention discloses a pharmaceutical composition comprising a compound for inhibiting myeloblast activity as shown in Formula I according to claim 1 or any one of its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and one or more pharmaceutically acceptable carriers.

[0026] Specifically, the dosage form of the pharmaceutical composition is one or more of subcutaneous injection, intradermal injection, spray, powder aerosol, external solution, lotion, liniment, ointment, plaster, paste, patch, granule, tablet, capsule, and liquid preparation.

[0027] The compound shown in the above-mentioned formula I that inhibits the activity of myeloblasts or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the use of the above-mentioned pharmaceutical composition in the preparation of anti-tumor drugs is also within the scope of protection of the present invention.

[0028] The use of the compound shown in the above-mentioned formula I for inhibiting the activity of medulloblastoma or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above-mentioned pharmaceutical composition in the preparation of a drug for treating medulloblastoma is also within the scope of protection of the present invention.

[0029] The use of the compound of Formula I for inhibiting medulloblastoma activity or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above-mentioned pharmaceutical composition in the preparation of a drug for treating medulloblastoma-related diseases is also within the scope of protection of the present invention.

[0030] Specifically, the disease related to medulloblastoma is a childhood brain tumor.

[0031] Unless otherwise indicated, the term "pharmaceutically acceptable salt" as used herein refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid. When the compound of the present invention is acidic, its corresponding salt can be readily prepared from inorganic or organic bases. Salts derived from such inorganic bases include salts of aluminum, ammonium, calcium, copper (copper and cuprous), iron, ferrous, lithium, magnesium, manganese (manganese and manganous), potassium, sodium, zinc, and the like. Preferred are salts of ammonium, calcium, magnesium, potassium, and sodium. Salts prepared from organic bases include primary, secondary, and tertiary amines derived from natural and synthetic sources. Pharmaceutically acceptable organic non-toxic bases that can form salts include arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, dicyclohexylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When the compound of the present invention is basic, its corresponding salt can be easily prepared from inorganic or organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, and the like.

[0032] The term "solvate" refers to a complex of variable stoichiometry formed by a solute (i.e., a compound of Formula I) or a pharmaceutically acceptable salt thereof and a solvent that does not interfere with the biological activity of the solute. Examples of solvents include, but are not limited to, water, ethanol, and acetic acid. When the solvent is water, the solvate is referred to as a hydrate. Hydrates include, but are not limited to, hemihydrates, monohydrates, monosesquihydrates, dihydrates, and trihydrates.

[0033] In the context of pharmaceutical compositions, the term "composition" includes a product comprising an active ingredient and an inert ingredient (pharmaceutically acceptable excipient) constituting a carrier, as well as any product resulting directly or indirectly from the combination, complexation, or aggregation of two or more ingredients, or the decomposition of one or more ingredients, or from other types of reactions or interactions of one or more ingredients. Thus, the pharmaceutical compositions of the present invention include any composition prepared by mixing a compound of Formula I, other active ingredients, and a pharmaceutically acceptable excipient.

[0034] Pharmaceutical composition of the present invention comprises the compound shown in the formula I as active ingredient (or its pharmaceutically acceptable salt or solvate), pharmaceutically acceptable carrier and optional other therapeutic ingredients or adjuvant.Pharmaceutical composition includes the composition that is applicable to oral, rectal, local and parenteral (including subcutaneous, intramuscular and intravenous) administration, although the most suitable approach depends on specific main body, the character and the severity of the disease to which active ingredient is given in any particular case.Pharmaceutical composition can be prepared by any method known to the field of pharmacy.

[0035] The active ingredient can be administered orally in solid dosage forms such as capsules, tablets, lozenges, troches, granules, and powders, or in liquid dosage forms such as elixirs, syrups, emulsions, dispersions, and suspensions. The active ingredient can also be administered parenterally in sterile liquid dosage forms such as dispersions, suspensions, or solutions. Other dosage forms that can be used to administer the active ingredient include subcutaneous injections, intradermal injections, topical solutions, lotions, liniments, pastes, ointments, creams, drops, transdermal patches, or powders for topical administration; ophthalmic solutions or suspensions, i.e., eye drops, for administration to the eye; sprays or powder compositions for inhalation or intranasal administration, or creams, ointments, sprays, or suppositories for rectal or vaginal administration. Gelatin capsules contain the active ingredient and a powdered carrier, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to prepare compressed tablets. Tablets and capsules can be prepared into sustained-release products to provide the sustained release of medicine within a few hours. Compressed tablets can be coated with sugar or film to cover any unpleasant taste and protect the tablet from air, or can be enteric-coated for selective disintegration in the gastrointestinal tract. Liquid dosage forms for oral administration can include coloring agents and flavoring agents to increase patient acceptance. Generally speaking, water, suitable oil, saline, dextrose (glucose) aqueous solution and related sugar solutions and glycols such as propylene glycol or polyethylene glycol are suitable carriers of parenteral solutions. Solutions for parenteral administration preferably include water-soluble salts of active ingredients, suitable stabilizers and the buffer substances used as needed. Antioxidants such as sodium bisulfite, sodium sulfite or ascorbic acid, alone or in combination, are suitable stabilizers. Citric acid and its salts and sodium EDTA can also be used. In addition, parenteral solutions can also include preservatives, such as benzalkonium chloride, methylparaben or propylparaben and chlorobutanol. For inhalation administration, compound of the present invention can be easily delivered in aerosol form from pressurized packaging or aerosol.The compound can also be delivered in the powder form prepared, and the powder composition can be sucked with the help of blowing into a powder inhaler device.The preferred delivery system for sucking is metered dose inhalation (MDI) aerosol, which can be formulated into a suspension or a solution of the compound of formula I, II in a suitable propellant, the propellant being fluorocarbon or hydrocarbon for example.For eye administration, ophthalmic preparations can be prepared with a solution or a suspension of the appropriate weight percent of the compound of formula I in a suitable eye carrier, thereby keeping the compound in contact with the surface of the eye enough time so that the compound is infiltrated into the cornea and the inner area of ​​the eyes.

[0036] Useful pharmaceutical dosage forms for administering the compounds of the present invention include, but are not limited to, hard and soft gelatin capsules, tablets, parenteral injection solutions, and oral suspensions.

[0037] When the compounds of the present invention are administered stepwise or in combination with other therapeutic agents, the same dosage forms as described above can be used. When the drugs are administered in a physical combination, the dosage form and route of administration should be selected based on the compatibility of the combined drugs. The compounds of the present invention can be administered as the sole active ingredient or in combination with a second active ingredient, including an active ingredient known to increase erythropoietin levels in a patient.

[0038] Beneficial effects:

[0039] The 3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-((1,2-dihydro-4,6-dimethyl-2-oxopyridin-3-yl)methyl)benzamide (i.e., Compound I) provided in this application is a new compound. Biological tests have shown that it has strong inhibitory activity against myeloblasts, and the proliferation of myeloblasts can cause childhood brain tumors. Therefore, Compound I is expected to be further developed into a drug for treating childhood brain tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, and the above and / or other advantages of the present invention will become more apparent.

[0041] Figure 1 is the H NMR spectrum of compound I.

[0042] Figure 2 is the mass spectrum of compound 1. DETAILED DESCRIPTION

[0043] The present invention can be better understood according to the following examples. However, it is easy for those skilled in the art to understand that the contents described in the examples are only used to illustrate the present invention, and should not and will not limit the present invention described in detail in the claims.

[0044] The experimental methods described in the following examples are conventional methods unless otherwise specified; the reagents and materials are commercially available unless otherwise specified.

[0045] Example 1:

[0046]

[0047] (1) Synthesis of methyl 3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)benzoate (Compound 2)

[0048] 3-Bromo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 1, 204.8 mg, 0.8 mmol), methyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (314.5 mg, 1.2 mmol), Pd(PPh3)4 (74 mg, 0.064 mmol), sodium carbonate (254 mg, 2.4 mmol) and dioxane-water mixed solvent (10 mL-3 mL) were mixed and stirred under nitrogen. The Suzuki coupling reaction was carried out under reflux under the protection of gas for 6 hours. After the reaction, the reaction solution was cooled and concentrated to dryness, and water (50 mL) was added, shaken, and extracted with dichloromethane (200 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by flash column chromatography on silica gel (eluted with dichloromethane: methanol in a volume ratio of 40:1). After concentration, a light yellow solid 3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)benzoic acid methyl ester (namely, compound 2, with a molecular formula of C 16 H17N5O2), 160 mg, 0.51 mmol, yield 64.2%.

[0049] Compound 2 was detected by mass spectrometry, EI-MS MS (m / z): 312.1 [M+H] + .

[0050] The H NMR spectrum data of compound 2 are: 1 H-NMR (CDCl3, 400MHz): δ8.43 (s, 2H), 8.19 (d, J=8.0Hz, 1H), 7.97 (d, J=8.0Hz, 1H), 7.67 (t, J=8.0Hz, 1H), 5.53 (br, s, 2H), 5.26~5.23 (m, 1H), 4.00 (s, 3H), 1.66 (d, J=8.0Hz, 6H).

[0051] (2) Synthesis of 3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)benzoic acid (Compound 3)

[0052] Methyl 3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)benzoate (140 mg, 0.45 mmol, prepared in step (1)) was dissolved in a mixed solvent of methanol (4 mL) and THF (1 mL), and a 3N aqueous sodium hydroxide solution (0.75 mL, 2.25 mmol) was added. The mixture was stirred at room temperature for esterification reaction. The reaction was allowed to proceed overnight. After the reaction was completed, the reaction solution was concentrated to dryness under reduced pressure, water (10 mL) was added, and the mixture was stirred. 1N HCl was slowly added dropwise to adjust the pH to 3-4. A solid precipitated, which was filtered, washed with water, and dried to obtain 3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)benzoic acid as a yellow solid (denoted as compound 3, with a molecular formula of C 15 H 15 N5O2), 110 mg, 0.37 mmol, yield 82.2%.

[0053] Compound 3 was detected by mass spectrometry, EI-MS MS (m / z): 298.1 [M+H] + .

[0054] (3) Synthesis of 3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-((1,2-dihydro-4,6-dimethyl-2-oxopyridin-3-yl)methyl)benzamide (Compound I)

[0055] 3-(4-Amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)benzoic acid (Compound 3, 90 mg, 0.30 mmol, prepared in step (2)), 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one hydrochloride (85 mg, 0.45 mmol), HOBt (81 mg, 0.60 mmol) and EDCI (115 mg, 0.60 mmol) were suspended in DMSO (3 mL) and N- Methylmorpholine (152 mg, 1.5 mmol) was stirred at room temperature for condensation reaction. The reaction was continued overnight. After the reaction was completed, water (20 mL) was slowly added to the reaction solution. Solid precipitated and stirred for half an hour. The mixture was filtered, washed with water, and dried to obtain a brown solid 3-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-((1,2-dihydro-4,6-dimethyl-2-oxopyridin-3-yl)methyl)benzamide (denoted as compound 1, with a molecular formula of C 23 H 25 N7O2), 81 mg, 0.19 mmol, yield 63.3%.

[0056] Compound I was detected by mass spectrometry, EI-MS MS (m / z): 432.4 [M+H] + .

[0057] 1 H-NMR (DMSO-d6, 400MHz): δ11.50 (s, 1H), 8.52 (s, 1H), 8.27 (s, 1H), 8.14 (s, 1H), 7.95 (d, J=8.0Hz, 1H), 7.78 (d, J=8.0Hz, 1H), 7.62 (d, J=8.0Hz, 1H), 5.89 (s, 1H), 5.10~5.08 (m, 1H), 4.35 (s, 2H), 2.21 (s, 3H), 2.14 (s, 3H), 1.52 (d, J=8.0Hz, 6H).

[0058] The H NMR spectrum of compound I is as follows Figure 1 As shown, the mass spectrum of compound I is as shown Figure 2 shown.

[0059] Example 2: Effect of Compound I on Medulloblastoma Proliferation

[0060] 1. Cell culture

[0061] Primary medulloblastoma cells were dissected from the brains of Patch1-deficient mice (purchased from Jackson Laboratory, USA), digested with papain, and centrifuged through Percoll to obtain a single cell suspension. 2 x 10^5 cells were plated onto 24-well slides. After two hours of complete attachment, 500 μL of culture medium (B27 medium containing 1 mM sodium pyruvate, 2 mM L-glutamine, and 1% penicillin / streptomycin) was added to each well and incubated at 37°C, 5% CO2 for 48 hours. 3 x 10^5 cells were seeded into 96-well plates in 100 μL of culture medium. After 4-6 hours of complete attachment, the culture medium was removed and 100 μL of culture medium containing various drug concentrations (prepared in "2. Dosing Plan") was added. Culture was continued for another 48 hours.

[0062] 2. Dosing plan

[0063] UNC1999, purchased from Shanghai Bidex Pharmaceuticals, 98%.

[0064] The compounds (UNC1999 and compound I prepared in Example 1) were dissolved in a small amount of DMSO and then prepared into a 100 μM stock solution in culture medium (B27 medium containing 1 mM sodium pyruvate, 2 mM L-glutamine, and 1% penicillin / streptomycin). The stock solution was then diluted to five concentrations of 500 nM, 1 μM, 5 μM, 10 μM, and 20 μM, with three replicate wells set for each concentration.

[0065] At the same time, the control group was added with equal volume of DMSO, and the blank control group was added with equal volume of culture medium and no cells.

[0066] 3. CCK-8 absorbance detection

[0067] After culturing myeloid blasts for 48 hours, 10 μl of CCK-8 was added to each well of a 96-well plate. The plates were incubated at 37°C for 3 hours, and the absorbance was measured at 450 nm. The inhibition rate was determined, and the concentration required to inhibit 50% cell growth, IC, was calculated. 50 .

[0068] 4. Data analysis: All data were analyzed using GraphPad Prism 6.0.

[0069] 5. Test results

[0070] Table 1 IC of different drugs inhibiting myeloid blasts 50

[0071] <![CDATA[IC 50 ]]> Compound I 5.75nM UNC1999 2.12μM

[0072] The results are shown in Table 1. The IC 50 The IC value of the control substance UNC1999 is 5.75 nM. 50 The test results show that the compound I provided by the present invention has a good effect of inhibiting the growth of myeloblasts, and its activity in inhibiting myeloblasts is 368 times that of the reference substance UNC1999.

[0073] The present invention provides a compound for inhibiting myeloblast activity, as well as a method for its preparation and application. Numerous methods and approaches exist for implementing this technical solution. The foregoing merely represents a preferred embodiment of the present invention. It should be noted that those skilled in the art may make various improvements and modifications without departing from the principles of the present invention, and such improvements and modifications are also within the scope of protection of the present invention. Any components not specified in this embodiment may be implemented using existing technologies.

Claims

1. A compound or a pharmaceutically acceptable salt thereof for inhibiting myeloblast activity as shown in formula I; 2. The method for preparing the compound for inhibiting myeloblast activity according to claim 1, characterized in that: The steps include: Compound 1 undergoes a Suzuki coupling reaction with a boronic acid compound in the presence of a first catalyst and a first base to obtain compound 2; compound 2 undergoes an esterification reaction in the presence of a second base to obtain compound 3; compound 3 undergoes a condensation reaction with 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one hydrochloride in the presence of a condensing agent and a third base to obtain compound 1; The condensing agent is any one or a combination of 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, benzotriazole-1-tris(trimethylamino)-hexafluorophosphate, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N-dicyclohexylcarbodiimide and O-benzotriazole-tetramethyluronium hexafluorophosphate; the third base is any one or a combination of triethylamine, diethylamine, diisopropylethylamine, N-methylmorpholine, potassium carbonate and sodium carbonate; the molar ratio of the compound 3 to 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one hydrochloride, the condensing agent and the third base is 1:1.5:4:5; and the condensation reaction is carried out at room temperature.

3. The preparation method according to claim 2, characterized in that The boric acid compound is 3-(methoxycarbonyl)phenylboronic acid or 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid methyl ester; the first catalyst is any one or a combination of tetrakistriphenylphosphine palladium, di(tri-tert-butylphosphine) palladium, 1,1'-bisdiphenylphosphinodichloropalladium, tris(dibenzylideneacetone)dipalladium and palladium acetate; the first base is any one or a combination of sodium carbonate, potassium carbonate and sodium bicarbonate; the molar ratio of the compound 1 to the boric acid compound, the first catalyst and the first base is 1:1.5:0.08:3; and the Suzuki coupling reaction is carried out under the protection of an inert gas.

4. The preparation method according to claim 2, characterized in that The second base is any one of sodium hydroxide, potassium hydroxide, potassium carbonate and sodium carbonate, or a combination thereof; and the esterification reaction is carried out at room temperature.

5. A pharmaceutical composition, characterized in that It comprises the compound for inhibiting myeloblast activity as shown in formula I of claim 1 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

6. The pharmaceutical composition according to claim 5, characterized in that The dosage form of the pharmaceutical composition is one or more of subcutaneous injection, intradermal injection, spray, ointment, granule, tablet, capsule, and liquid preparation.

7. Use of the compound for inhibiting myeloblast activity of formula I according to claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 5 in the preparation of an anti-tumor drug.

8. Use of the compound of formula I or a pharmaceutically acceptable salt thereof for inhibiting medulloblastoma activity according to claim 1, or the pharmaceutical composition according to claim 5, in the preparation of a drug for treating medulloblastoma.

9. Use of the compound or pharmaceutically acceptable salt of formula I for inhibiting medulloblastoma activity according to claim 1, or the pharmaceutical composition according to claim 5, in the preparation of a drug for treating diseases related to medulloblastoma.

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