A triazine hydroxamic acid derivative and its preparation method and application

Abstract

The present invention discloses a triazine hydroxamic acid derivative and a preparation method and application thereof. The general structural formula of the derivative is: The present invention constructs a class of chemical small molecules with PI3K and HDAC dual-target inhibitory activity. After tests on a variety of tumor cell lines (including colon cancer cells and breast cancer cells) and phosphatidylinositol 3-kinase (PI3K) and histone deacetylase (HDAC) inhibitory activity tests, it is proved that the derivative has good inhibitory activity on phosphatidylinositol 3-kinase (PI3K) and histone deacetylase (HDAC), and has good application prospects in the field of anti-tumor drug design and development.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a triazine hydroxamic acid derivative and a preparation method and application thereof. Background Art

[0002] Phosphatidylinositol 3-kinase (PI3K) is a lipid kinase that can phosphorylate the 3-hydroxyl group of inositol (PI) to produce inositol-3-phosphate (PIP), inositol-3,4-bisphosphate (PIP2) and inositol-3,4,5-triphosphate (PIP3). According to the differences in structure, substrate, distribution and function, PI3K is mainly divided into three categories: I, II and III. Among them, class I can be divided into two subfamilies according to its regulatory subunits: IA (consisting of dimers formed by p110α, p110β, p110δ catalytic units and p85α, p85β or p55γ regulatory subunits, divided into three subtypes of PI3Kα, PI3Kβ and PI3Kδ) and IB (PI3Kγ subtype composed of catalytic subunit p110γ and regulatory subunit p101 or p87). Class II PI3K contains only one p110-like catalytic domain and has three conformations. Class III PI3K contains only one Vps34 unit. PI3Kα and PI3Kβ are widely expressed in tissues, while PI3Kδ and PI3Kγ are mainly expressed in immune cells and hematopoietic cells. Among the various PI3K subtypes mentioned above, PI3Kα can mediate the signal transduction of EGFR and plays a particularly important role in various cancers; it has been found to mutate or amplify signals in many cancers, so it is also a subtype that has been studied more and has more corresponding inhibitors. In addition, PI3Kδ has also attracted the attention of many scientific research works due to its important physiological significance in leukemia and lymphoma. It is also the first PI3K target with approved drugs. At present, the clinical development of PI3K inhibitors mainly includes two types: combination therapy and single therapy.

[0003] Histone deacetylase (HDAC) can catalyze the removal of acetylated lysine, allowing the positively charged lysine residues to be tightly entangled with the negatively charged DNA phosphate backbone, thereby inhibiting the expression of tumor suppressor genes. A total of 18 HDACs have been found in mammals, which are divided into four subtypes based on their homology with yeast, including type I (HDAC1, HDAC2, HDAC3, HDAC8), type II A (HDAC4, HDAC5, HDAC7, HDAC9), type II B (HDAC6, HDAC10) and type IV (HDAC11) belonging to Zn 2+ Ion-dependent HDACs; type III HDACs include deacetylases SIRT1-SIRT7, which belong to NAD +As a mature target for the development of anti-tumor drugs, many HDAC inhibitors have entered clinical research, and five HDAC inhibitors (Vorinostat, Romidep sin, Belinostat, Panobinostat, Chidamide) have been launched on the market.

[0004] Since PI3K activation in tumor cells is often accompanied by Ras-MEK pathway activation and PI3K downstream oncogene mutations, and single PI3K inhibition will further cause activation of multiple compensatory pathways in tumor cells, resulting in limited therapeutic effects of PI3K inhibitors, combination therapy is an effective method to make PI3K inhibitors suitable for more tumor types. However, combination therapy has disadvantages such as complex pharmacokinetics, possible drug interactions, and more toxic side effects. Single small molecules with multi-target inhibitory activity can effectively avoid these problems, and the development of multi-target inhibitors has become a common drug development strategy.

[0005] It has been reported that the combination of PI3K inhibitors and histone deacetylase (HDAC) inhibitors has a synergistic anti-tumor effect, and CUDC-101 and CUDC-907 have entered clinical trials as potential dual-target drugs for PI3K and HDAC, which shows that the HDAC-based dual-target drug design strategy is feasible and has great potential for developing more PI3K and HDAC multi-target inhibitors with novel structures and excellent activities. Summary of the invention

[0006] The main purpose of the present invention is to provide a triazine hydroxamic acid derivative having good inhibitory activity on phosphatidylinositol 3-kinase and histone deacetylase, and a preparation method and application thereof.

[0007] To achieve the above object, the present invention provides a triazine hydroxamic acid derivative, the general structural formula of the derivative is shown in Formula I:

[0008]

[0009] in:

[0010] R1 is selected from any one of hydrogen atom, halogen, -CH3, -CF3, -CHF2, -CH2F;

[0011] X is selected from any one of carbon atom, -N-, -O-, and -S-;

[0012] Y is selected from -O-, -S-, -CH2-, -C=O-, -CH2CH2-, -CHF-, -CHCl-, -CHBr-,

[0013] Any one of -CHI-, -CF2-, -CCl2-, -CBr2-, -CI2-, -CH2CF2-, -CH2CCl2-, -CH2CBr2-, -CH2CI2-, -CF2CH2-, -CCl2CH2-, -CBr2CH2-, -CI2CH2-;

[0014] G is selected from any one of a direct bond, -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2-, -CH2CH2CH2CH2CH2-, -CH2CH2CH2CH2CH2-, -CH2CH2CH2CH2CH2CH2-, -CH2CH2CH2CH2CH2CH2-, -CHF-, -CHCl-, -CHBr-, -CHI-, -CF2-, -CCl2-, -CBr2-, -CI2-, -CH2CF2-, -CH2CCl2-, -CH2CBr2-, -CH2CI2-, -CF2CH2-, -CCl2CH2-, -CBr2CH2-, -CI2CH2-, -CH=CH-, and -CH=CH-CH=CH-.

[0015] Preferably, the halogen is selected from any one of fluorine, chlorine, bromine or iodine.

[0016] The derivatives of formula I may exist in different polymorphic forms.

[0017] The present invention also provides pharmaceutically acceptable salts of the derivatives represented by the above formula I or their tautomers, which also fall within the protection scope of the present invention.

[0018] Preferably, the structural formula of the derivative is shown in any of the following numbers:

[0019]

[0020]

[0021]

[0022]

[0023] Preferably, the pharmaceutically acceptable salt is an inorganic acid salt or an organic acid salt;

[0024] The inorganic acid salt is selected from the salt formed by any one of the following inorganic acids: hydrochloric acid, sulfuric acid and phosphoric acid;

[0025] The organic acid salt is selected from the group consisting of salts formed by any one of the following organic acids: acetic acid, trifluoroacetic acid, malonic acid, citric acid and p-toluenesulfonic acid.

[0026] The present invention also provides three methods for preparing the derivatives represented by formula I.

[0027] The first preparation method comprises the following steps:

[0028] The compound represented by formula IA is reacted with hydroxylamine or hydroxylamine hydrochloride to obtain the derivative represented by formula I;

[0029]

[0030] Preferably, the reaction is carried out in the presence of a condensing agent or in the absence of a condensing agent.

[0031] Preferably, the condensing agent is benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), and the reaction is carried out in a solvent N,N-dimethylformamide (DMF) under alkaline (triethylamine) conditions at room temperature.

[0032] The second preparation method comprises the following steps:

[0033] The compound represented by formula IA and hydroxylamine having a hydroxyl group with a protective group are first reacted under the action of a condensing agent to generate a compound represented by formula IB, and then the protective group is removed from the compound represented by formula IB to obtain a derivative represented by formula I;

[0034]

[0035] Wherein, P is a protecting group for the hydroxyl group in hydroxylamine.

[0036] Preferably, the condensing agent is benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), the hydroxylamine with a hydroxyl group protection group is O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, and the reaction is carried out in a solvent N,N-dimethylformamide (DMF) at room temperature.

[0037] Preferably, the removal of the protecting group is carried out in a solvent such as tetrahydrofuran, acidically (hydrochloric acid or trifluoroacetic acid) or in a basic (sodium hydroxide) atmosphere at a temperature between 0 and 80 °C.

[0038] The third preparation method comprises the following steps:

[0039] The compound represented by formula IC is reacted with hydroxylamine or hydroxylamine hydrochloride in an alkaline solution to obtain a derivative represented by formula I;

[0040]

[0041] Wherein, E is an alkyl group.

[0042] Preferably, the alkyl group is a C1-C6 alkyl group, including but not limited to methyl, ethyl or isopropyl.

[0043] Preferably, the reaction is carried out in an aqueous solution or an organic solvent, wherein the organic solvent is selected from at least one of the following: dimethylformamide, dimethylacetamide, chloroform, dichloromethane, acetone, tetrahydrofuran, acetonitrile and methanol.

[0044] Preferably, v said base is selected from at least one of the following: aqueous ammonia, triethylamine, diisopropylethylamine, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, sodium tert-butoxide and tert-butanol.

[0045] Preferably, the reaction temperature is 20 to 80° C., and the reaction time is 0.5 to 10 hours.

[0046] In each of the above preparations, where the synthesis of intermediates and starting materials is not described, these compounds are either commercially available or can be prepared from commercially available compounds using standard methods or by extension of the methods described in the Examples herein.

[0047] The derivatives prepared by the present invention are tested by nuclear magnetic resonance and the like, which proves that the derivatives prepared are correct and are as shown in general formula I.

[0048] The present invention also provides the use of the derivative represented by formula I or its pharmaceutically acceptable salt or its tautomer in the preparation of any of the following products:

[0049] 1) Phosphatidylinositol 3-kinase and / or histone deacetylase inhibitors;

[0050] 2) Eukaryotic tumor cell proliferation inhibitors;

[0051] 3) Drugs for the prevention and / or treatment of tumors.

[0052] The present invention also provides a product, the active ingredient of which is a derivative represented by formula I or a pharmaceutically acceptable salt or a tautomer thereof; the product is any one of the following:

[0053] 1) Phosphatidylinositol 3-kinase and / or histone deacetylase inhibitors;

[0054] 2) Eukaryotic tumor cell proliferation inhibitors;

[0055] 3) Drugs for the prevention and / or treatment of tumors.

[0056] Preferably, the phosphatidylinositol 3-kinase includes the isoforms PI3Kα, PI3Kβ, PI3Kδ and PI3Kγ known in mammalian cells;

[0057] The histone deacetylase includes the isoforms HDAC1, HDAC2, HDAC3, HDAC8, HDAC4, HDAC5, HDAC7, HDAC9, HDAC6, HDAC10, and HDAC11 known in mammalian cells;

[0058] The eukaryotic organism is a mammal; the tumor cell is a cancer cell; the cancer cell is a leukemia cancer cell, a lymphoma cell, a lung cancer cell, a breast cancer cell, an ovarian cancer cell, a cervical cancer cell, a human glioma cell, a melanoma cell, a glioblastoma cell, a nasopharyngeal cancer cell, a liver cancer cell, a brain cancer cell, a pancreatic cancer cell, a uterine cancer cell, a testicular cancer cell, a skin cancer cell, a gastric cancer cell, a colon cancer cell, a bladder cancer cell or a rectal cancer cell.

[0059] Preferably, the tumor is cancer; the cancer is leukemia, lymphoma, lung cancer, melanoma, glioblastoma, cervical cancer, nasopharyngeal cancer, liver cancer, breast cancer, brain cancer, pancreatic cancer, ovarian cancer, uterine cancer, testicular cancer, skin cancer, stomach cancer, colon cancer, bladder cancer or rectal cancer.

[0060] Preferably, the leukemia cancer cells are specifically human chronic myeloid leukemia (CML) cell line K562; the lymphoma cells are specifically human histiocytic lymphoma cells U937; the lung cancer cells are specifically human lung cancer cell line HCC827; the breast cancer cells are specifically human breast cancer cells MCF-7, T47D and MDA-MB-231; the ovarian cancer cells are specifically A2780; the cervical cancer cells are specifically human cervical cancer cell line Hela; the human glioma cells are specifically U251; the melanoma cancer cells are specifically A375; the glioblastoma cells are specifically human glioblastoma cells A172 and human astrocytic glioblastoma cells U-118MG; the nasopharyngeal cancer cells are specifically nasopharyngeal cancer cell line CNE-2; and the liver cancer cells are specifically human liver cancer cells HepG2.

[0061] The phosphatidylinositol 3-kinase and / or histone deacetylase inhibitors, eukaryotic tumor cell proliferation inhibitors, and drugs for preventing and / or treating tumors prepared using the derivatives shown in formula I or their pharmaceutically acceptable salts or tautomers can be introduced into the body such as muscle, intradermal, subcutaneous, intravenous, and mucosal tissues by injection, spraying, nasal drops, eye drops, penetration, absorption, physical or chemical mediation methods; or introduced into the body after being mixed or wrapped with other substances.

[0062] When necessary, one or more pharmaceutically acceptable carriers may be added to the above products, including conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants, etc. in the pharmaceutical field.

[0063] The phosphatidylinositol 3-kinase and / or histone deacetylase inhibitors, eukaryotic tumor cell proliferation inhibitors, and drugs for preventing and / or treating tumors prepared using the derivatives shown in formula I or their pharmaceutically acceptable salts or tautomers can be prepared in various forms such as injections, tablets, powders, granules, capsules, oral liquids, ointments, creams, etc. The drugs in the above various dosage forms can be prepared according to conventional methods in the pharmaceutical field.

[0064] The beneficial effects of the present invention are embodied in:

[0065] The present invention constructs a class of chemical small molecules with dual-target inhibitory activity of PI3K and HDAC, and studies its inhibitory activity on various tumor cells. After various tumor cell line tests (including colon cancer cells and breast cancer cells) and phosphatidylinositol 3-kinase (PI3K) and histone deacetylase (HDAC) inhibitory activity tests, it is proved that the derivative shown in formula I is a potential chemical small molecule with inhibitory activity on phosphatidylinositol 3-kinase (PI3K) and histone deacetylase (HDAC). In addition, the raw materials of the compound provided by the present invention are easy to obtain, the preparation method is simple, and experiments have shown that it has good anti-cancer effects, and has good application prospects in the field of anti-tumor drug design and development. DETAILED DESCRIPTION

[0066] The present invention is described below by means of specific embodiments, but the scope of the present invention is not limited thereto.

[0067] The experimental methods described in the following examples, unless otherwise specified, are conventional methods of organic synthesis; the reagents and biological materials, unless otherwise specified, are obtained from commercial sources.

[0068] Example 1

[0069] Preparation of hydroxamic acid derivatives of triazine No. CY-1

[0070] A: Preparation of 4-(4,6-dichloro-1,3,5-triazine-2-yl)morpholine, the structural formula is as follows:

[0071]

[0072] In an ice bath, add morpholine (50mmol, 4.3ml) dropwise to a mixture of cyanuric chloride (25mmol, 5.2077g) and dichloromethane (150ml), wait for the reaction to return to room temperature and continue to react overnight, during which thin layer chromatography (TLC) is used to monitor until the raw material reaction is complete, extract with dichloromethane (DCM), wash the reaction solution with a saturated sodium chloride solution, and dry the organic phase over anhydrous sodium sulfate and spin dry to obtain a crude product. Then separate by column chromatography (V petroleum ether (PE): V ethyl acetate (EA) = 10:1) to obtain the title compound (white solid, 2.7976g, yield 47.60%).

[0073] B: Preparation of tert-butyl 4-(4-chloro-6-morpholine-1,3,5-triazine-2-yl)piperazine-1-carboxylate, the structural formula is as follows:

[0074]

[0075] The compound prepared in step A (5mmol, 1.1701g), 1-tert-butyloxycarbonylpiperazine (5mmol, 0.9856g) and DIEA (8mmol, 1.3ml) were placed in a 100ml round-bottom flask, and tetrahydrofuran (THF, 25ml) was added thereto to react overnight at room temperature. After the reaction was completed by TLC monitoring, water was added and extracted with dichloromethane (DCM). The organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain the title compound (white solid, 1.8518g, yield 96.23%), which was directly carried out to the next step without purification.

[0076] C: Preparation of tert-butyl 4-(4-(4-aminophenyl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-carboxylate, the structural formula is as follows:

[0077]

[0078] The compound obtained in step B (3mmol, 1.1452g), sodium carbonate (6mmol, 0.6492g) and 4-aminophenylboronic acid pinacol ester (6mmol, 1.3196g) were placed in a 100ml round-bottom flask, and 1,4-dioxane / water (25ml:5ml) and an appropriate amount of tetrakis(triphenylphosphine)palladium were added under the reaction conditions of heating in an oil bath at 105°C. After one night of reaction, the reaction was monitored using TLC technology, and after the reaction was stopped, the reaction was dried using a rotary evaporator to obtain the desired solvent, extracted and washed with DCM and saturated brine, the lower organic layer was collected and dried over anhydrous sodium sulfate, and then dry-loaded, separated by column chromatography, and firstly V PE :V EA =7:1 to remove impurities, and then use V PE :VEA =1:1 to give the title compound (yellow-brown solid, 0.9420 g, yield 71.11%).

[0079] D: Preparation of 4-(4-morpholinyl-6-(piperazine-1-yl)-1,3,5-triazine-2-yl)aniline, the structural formula is as follows:

[0080]

[0081] The compound prepared in step C (1.1678 g) was reacted with concentrated hydrochloric acid (5 ml) at room temperature for 7 h, during which time it was monitored by TLC. After the reaction was stopped, sodium carbonate was added to neutralize it to a pH of about 7, i.e., no bubbles emerged. The post-treatment was extracted with DCM, and the organic layer was collected after washing with saturated brine, and then dried over anhydrous sodium sulfate and spin-dried to obtain the title compound (white-brown powder, 0.7749 g, yield 85.81%).

[0082] E: Preparation of methyl 4-(4-(4-aminophenyl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-4-oxobutanoate, the structural formula is as follows:

[0083]

[0084] Put glutaric acid monomethyl ester (0.3mmol, 0.0553g) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (0.6mmol, 0.2372g) in a 25ml round-bottom flask, add 5ml DMF, accurately weigh DIEA (1.2mmol, 0.1468g) and the compound prepared in step D (0.3mmol, 0.1008g) in the above round-bottom flask, react at room temperature for 30min, monitor the reaction by TLC, add water to quench the reaction, extract with DCM, wash with saturated brine, collect the organic layer, dry with anhydrous sodium sulfate, spin dry, and analyze by column chromatography (V PE :V EA =1:1) to give the title compound (white solid, yield 32%).

[0085] F: Preparation of 4-(4-(4-aminophenyl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-N-hydroxy-4-oxobutyramide (Compound CY-1), the structural formula of which is as follows:

[0086]

[0087] The compound (1 eq) prepared in step E was dissolved in methanol, and hydroxylamine aqueous solution (15.17 M, 10 eq) and sodium methoxide methanol solution (5 M, 5 eq) were added in sequence, and the mixture was reacted at room temperature for 1 hour. After that, the reaction solvent was removed by rotary evaporator, and an appropriate amount of water was added, and the pH was adjusted to 7-8 with 3N hydrochloric acid. After standing for a while, the mixture was filtered and the filter cake was washed with a small amount of water and ether, and the title compound was obtained after drying. White powder, yield 39%, melting point 241-242°C. The compound structure is characterized as follows: 1 H NMR (400MHz, DMSO-d6) δ8.05(d,J=8.6Hz,2H),6.57(d,J=8.6Hz,2H),5.71(s,2H),3.78(s,8H),3.6 4(t,J=4.7Hz,4H),3.59(s,3H),3.51(p,J=4.8Hz,4H),2.38(q,J=7.7Hz,4H),1.77(p,J=7.4Hz,2H). 13 C NMR (101MHz, DMSO-d6) δ169.04,164.86,164.70,162.33,150.06,137.12,120.70,107.39,66.49,44.99,43.69,43.02.

[0088] Example 2

[0089] Preparation of hydroxamic acid derivatives of triazine No. CY-2

[0090] E: Preparation of methyl 5-(4-(4-(aminophenyl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)pentanoate, the structural formula is as follows:

[0091]

[0092] The compound prepared in Example 1D and methyl bromovalerate were used as raw materials, and the synthesis method was the same as that in Example 1E to obtain a white solid with a yield of 87%.

[0093] F: Preparation of 5-(4-(4-(-4-aminophenyl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-N-hydroxypentanamide, the structural formula is as follows:

[0094]

[0095] The compound prepared in Example 2E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that in Example 1F. A white powder was obtained with a yield of 28% and a melting point of 228-229°C. The compound structure is characterized as follows: 1H NMR (500MHz, DMSO-d6) δ8.05(d,J=8.5Hz,2H),6.59(d,J=8.4Hz,2H),5.70(s,2H),3.76(s,8H),3.63(t,J=4 .8Hz,4H),2.36(t,J=4.9Hz,4H),2.27(t,J=7.2Hz,2H),1.99(t,J=7.2Hz,2H),1.47(dq,J=40.7,7.4Hz,4H). 13 C NMR(126MHz,DMSO-d6)δ169.91,169.64,165.10,164.86,152.65,130.15,124 .08,113.23,66.53,57.96,53.11,49.04,43.69,43.18,32.58,26.20,23.55.

[0096] Example 3

[0097] Preparation of hydroxamic acid derivatives of triazine No. CY-3

[0098] C: Preparation of tert-butyl 4-(4-(6-aminopyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-carboxylate, the structural formula is as follows:

[0099]

[0100] Example 1B and 2-aminopyridine-5-boronic acid pinacol ester were used as raw materials, and the synthesis method was the same as that of Example 1C to obtain a white solid with a yield of 61%.

[0101] D: Preparation of 5-(4-morpholinyl-6-(piperazine-1-yl)-1,3,5-triazine-2-yl)pyridin-2-amine, the structural formula is as follows:

[0102]

[0103] Example 3C was used as the raw material and the synthesis method was the same as that of Example 1D to obtain a white solid with a yield of 91%.

[0104] E: Preparation of 5-(4-(4-(6-aminopyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-5-oxopentanoic acid methyl ester, the structural formula is as follows:

[0105]

[0106] Example 3D and monomethyl glutarate were used as raw materials, and the synthesis method was the same as that of Example 1E. White solid, yield 96%.

[0107] F: Preparation of 5-(4-(4-(6-aminopyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-N-hydroxy-5-oxopentylamide, the structural formula is as follows:

[0108]

[0109] Example 3E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials, and the synthesis method was the same as that of Example 1F. White powder, yield 48%, melting point 178-179°C. The compound structure is characterized by: 1 H NMR (500MHz, DMSO-d6) δ10.49(s,1H),8.89(d,J=2.3Hz,1H),8.21(dd,J=8.8,2.4Hz,1H),6.57(s,2H),6.49(d,J=8.8Hz,1H),3.7 7(s,8H),3.63(t,J=4.8Hz,4H),3.50(dt,J=11.5,5.1Hz,4H),2.34(t,J=7.5Hz,2H),2.02(t,J=7.3Hz,2H),1.74(t,J=7.3Hz,2H). 13 CNMR(126MHz,DMSO)δ170.92,169.45,169.08,164.84,164.79,162.34,150.06,137. 14,120.70,107.44,66.49,45.10,43.71,43.36,42.97,41.28,32.16,32.06,21.36.

[0110] Example 4

[0111] Preparation of hydroxamic acid derivatives of triazine No. CY-4

[0112] E: Preparation of 6-(4-(4-(6-aminopyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-6-oxohexanoic acid ethyl ester, the structural formula is as follows:

[0113]

[0114] Example 3D and monoethyl adipate were used as raw materials and the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 89%.

[0115] F: Preparation of 6-(4-(4-(6-aminopyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-N-hydroxy-6-oxohexanamide (compound), the structural formula of which is as follows:

[0116]

[0117] Example 4E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 38% and a melting point of 230-231°C. The compound structure is characterized as follows: 1 H NMR (400MHz, DMSO-d6) δ10.29(s,1H),8.90(d,J=2.2Hz,1H),8.22(dd,J=8.8,2.3Hz,1H),6.55(s,2H),6.48(d,J=8.8Hz,1H),3.7 8(s,8H),3.64(t,J=4.7Hz,4H),3.53(d,J=5.0Hz,4H),2.35(t,J=6.8Hz,2H),1.98(t,J=6.6Hz,2H),1.52(dq,J=8.4,4.4Hz,4H). 13 CNMR(101MHz,DMSO-d6)δ171.19,169.44,169.08,164.84,164.78,162.36,150.10,137.13,120.65, 107.40,66.50,43.70,40.60,40.39,40.19,39.98,39.77,39.56,39.35,32.58,32.51,25.34,24.79.

[0118] Example 5

[0119] Preparation of hydroxamic acid derivatives of triazine No. CY-5

[0120] E: Preparation of methyl 4-(4-(6-aminopyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)butanoate, the structural formula is as follows:

[0121]

[0122] Example 3D and methyl bromobutyrate were used as raw materials and the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 89%.

[0123] F: Preparation of 4-(4-(6-aminopyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-N-hydroxybutyramide (compound), the structural formula of which is as follows:

[0124]

[0125] Example 5E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 59% and a melting point of 202-204°C. The compound structure is characterized as follows: 1 H NMR (400MHz, DMSO-d6) δ10.45(s,1H),8.87(s,1H),8.20(d,J=8.1Hz,1H),6.52(s,2H),6.48(d,J=8.7Hz,1H),3.76( s,8H),3.64(d,J=4.8Hz,4H),2.38(d,J=5.0Hz,4H),2.27(t,J=7.3Hz,2H),2.00(t,J=7.3Hz,2H),1.72–1.59(m,2H). 13 C NMR (101 MHz, DMSO-d6) δ 169.64, 168.98, 164.86, 164.62, 162.26, 149.95, 137.14, 120.79, 107.44, 66.49, 57.66, 53.01, 43.67, 43.16, 40.52, 40.31, 40.10, 39.89, 39.68, 39.47, 39.26, 30.70, 22.81. Example 6

[0126] Preparation of hydroxamic acid derivatives of triazine No. CY-6

[0127] E: Preparation of methyl 5-(4-(4-(6-aminopyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)pentanoate, the structural formula is as follows:

[0128]

[0129] Example 3D and methyl bromovalerate were used as raw materials and the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 81%.

[0130] F: Preparation of 5-(4-(4-(6-aminopyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-N-hydroxypentanamide, the structural formula is as follows:

[0131]

[0132] Example 6E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 43% and a melting point of 160-161°C. The compound structure is characterized as follows: 1HNMR (400MHz, DMSO-d6) δ10.38(s,1H),8.90(d,J=10.4Hz,1H),8.21(d,J=8.7Hz,1H),6.57(d,J=13.3Hz,2H),6.46(d,J=8.8Hz,1 H),3.80(d,J=23.2Hz,8H),3.64(s,4H),2.37(s,4H),2.29(d,J=7.2Hz,2H),1.97(t,J=7.4Hz,2H),1.47(dt,J=37.8,7.8Hz,4H). 13 C NMR(101MHz,DMSO-d6)δ169.50,168.96,164.84,164.60,162.32,150.04,137.11,120.70,107.37,88.94, 66.50,57.92,53.10,43.66,43.12,40.57,40.36,40.15,39.94,39.73,39.52,39.31,32.57,26.21,23.53.

[0133] Example 7

[0134] Preparation of hydroxamic acid derivatives of triazine No. CY-7

[0135] E: Preparation of methyl 6-(4-(4-(6-aminopyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)hexanoate, the structural formula is as follows:

[0136]

[0137] Example 3D and methyl bromohexanoate were used as raw materials and the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 78%.

[0138] F: Preparation of 6-(4-(4-(6-aminopyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-N-hydroxyhexanamide, the structural formula is as follows:

[0139]

[0140] Example 7E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 56% and a melting point of 173-174°C. The compound structure is characterized as follows: 1H NMR(500MHz,DMSO-d6)δ10.42(s,1H),8.87(d,J=2.3Hz,1H),8.20(dd,J=8.8, 2.4Hz,1H),6.53(s,2H),6.47(d,J=8.7Hz,1H),3.79(d,J=31.0Hz,8H),3.63( t,J=4.7Hz,4H),2.37(t,J=5.0Hz,4H),2.26(t,J=7.3Hz,2H),1.95(t,J=7.4H z,2H),1.55–1.47(m,2H),1.43(t,J=7.5Hz,2H),1.23(dd,J=15.0,7.2Hz,2H). 13 C NMR(126MHz,DMSO-d6)δ169.59,169.01,164.90,164.64,162.30,149.99,137.10 ,120.82,107.41,66.50,58.27,53.12,43.69,43.18,32.71,26.99,26.40,25.53.

[0141] Example 8

[0142] Preparation of hydroxamic acid derivatives of triazine No. CY-8

[0143] C: Preparation of tert-butyl 4-(4-(2-aminopyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-carboxylate, the structural formula is as follows:

[0144]

[0145] Example 1B and 2-aminopyrimidine-5-boronic acid were used as raw materials and the synthesis method was the same as Example 1C to obtain a white solid with a yield of 87%.

[0146] D: Preparation of 5-(4-morpholinyl-6-(piperazine-1-yl)-1,3,5-triazine-2-yl)pyrimidin-2-amine, the structural formula is as follows:

[0147]

[0148] Example 8C was used as the raw material and the synthesis method was the same as that of Example 1D to obtain a white solid with a yield of 97%.

[0149] E: Preparation of 5-(4-(4-(2-aminopyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-5-oxopentanoic acid methyl ester, the structural formula is as follows:

[0150]

[0151] Using Example 8D and monomethyl glutarate as raw materials, the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 85%.

[0152] F: Preparation of 5-(4-(4-(2-aminopyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-N-hydroxy-5-oxopentylamide, the structural formula is as follows:

[0153]

[0154] Example 8E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 63% and a melting point of 220-221°C. The compound structure is characterized as follows: 1 H NMR (400MHz, DMSO-d6) δ10.38(s,1H),9.06(s,2H),8.68(s,1H),7.27(s,2H),3.79(s,8H),3.65(t,J=4. 7Hz, 4H), 3.52 (q, J = 6.6, 6.2Hz, 4H), 2.35 (t, J = 7.3Hz, 2H), 2.01 (t, J = 7.3Hz, 2H), 1.74 (t, J = 7.3Hz, 2H). 13 C NMR (101MHz, DMSO-d6) δ170.87,169.34,167.71,165.26,164.59,159.05,118.93,66.49,32.13,32.05,21.33.

[0155] Example 9

[0156] Preparation of hydroxamic acid derivatives of triazine No. CY-9

[0157] E: Preparation of 6-(4-(4-(2-aminopyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-6-oxohexanoic acid ethyl ester, the structural formula is as follows:

[0158]

[0159] Using Example 8D and monoethyl adipate as raw materials, the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 83%.

[0160] F: Preparation of 6-(4-(4-(2-aminopyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-N-hydroxy-6-oxohexanamide, the structural formula is as follows:

[0161]

[0162] Example 9E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 55% and a melting point of 198-199°C. The compound structure is characterized as follows: 1 HNMR (400MHz, DMSO-d6) δ10.48(s,1H),9.05(s,2H),7.29(s,2H),3.79(d,J=24.7Hz,8H),3.64(d,J =4.7Hz,4H),3.53–3.50(m,4H),2.35(t,J=6.8Hz,2H),1.98(t,J=6.7Hz,2H),1.50(q,J=5.9Hz,4H). 13 CNMR(101MHz,DMSO-d6)δ171.21,169.50,167.66,165.21,164.57,164.52,159.06,118. 88,66.49,40.51,40.30,40.09,39.88,39.67,39.46,39.26,32.53,32.49,25.32,24.74.

[0163] Example 10

[0164] Preparation of hydroxamic acid derivatives of triazine No. CY-10

[0165] E: Preparation of methyl 4-(4-(2-aminopyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)butanoate, the structural formula is as follows:

[0166]

[0167] Example 8D and methyl 4-bromobutyrate were used as raw materials and the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 80%.

[0168] F: Preparation of 4-(4-(2-aminopyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-N-hydroxybutyramide, the structural formula is as follows:

[0169]

[0170] Example 10E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 62% and a melting point of 223-224°C. The compound structure is characterized as follows:1 H NMR (400MHz, DMSO-d6) δ10.37(s,1H),9.04(d,J=1.2Hz,2H),8.66(s,1H),7.25(s,2H),3.77(s,8H),3.64( t,J=4.5Hz,4H),2.39(t,J=4.8Hz,4H),2.29(t,J=7.3Hz,2H),2.00(t,J=7.4Hz,2H),1.68(t,J=7.3Hz,2H). 13 C NMR (101MHz, DMSO-d6) δ169.52,167.63,165.23,164.66,164.40,158.98,119.03,66.50,57.64,52.99,40.41,30.68.

[0171] Embodiment 11

[0172] Preparation of hydroxamic acid derivatives of triazine No. CY-11

[0173] E: Preparation of methyl 5-(4-(4-(2-aminopyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)pentanoate, the structural formula is as follows:

[0174]

[0175] Example 8D and methyl 5-bromovalerate were used as raw materials and the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 79%.

[0176] F: Preparation of 5-(4-(4-(2-aminopyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-N-hydroxypentanamide, the structural formula is as follows:

[0177]

[0178] Example 11E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 48% and a melting point of 177-178°C. The compound structure is characterized as follows: 1 H NMR(400MHz,DMSO-d6)δ10.40(s,1H),9.04(s,2H),8.67(s,1H),7.25(s,2H),3.78(s,8H) ,3.64(t,J=4.6Hz,4H),2.37(d,J=37.0Hz,6H),1.98(t,J=7.0Hz,2H),1.55–1.42(m,4H).13 CNMR(101MHz,DMSO-d6)δ169.50,167.65,165.23,164.67,164.43,159.00,71.92,66.48,52.90,40.62,32.54,23.49.

[0179] Example 12

[0180] Preparation of hydroxamic acid derivatives of triazine No. CY-12

[0181] E: Preparation of methyl 6-(4-(4-(2-aminopyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)hexanoate, the structural formula is as follows:

[0182]

[0183] Example 8D and methyl bromohexanoate were used as raw materials and the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 82%.

[0184] F: Preparation of 6-(4-(4-(2-aminopyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-yl)-N-hydroxyhexanamide, the structural formula is as follows:

[0185]

[0186] Example 12E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 49% and a melting point of 198-199°C. The compound structure is characterized as follows: 1 H NMR (400MHz, DMSO-d6) δ10.35(s,1H),9.04(s,2H),8.66(s,1H),7.25(s,2H),3.80(d,J=24.0Hz,8H),3.64(t,J= 4.6Hz, 4H), 2.36 (d, J = 43.9Hz, 6H), 1.95 (t, J = 7.3Hz, 2H), 1.48 (dt, J = 22.1, 7.5Hz, 4H), 1.27 (d, J = 18.3Hz, 2H).

[0187] Embodiment 13

[0188] Preparation of hydroxamic acid derivatives of triazine No. CY-13

[0189] C: Preparation of tert-butyl 4-(4-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-carboxylate, the structural formula is as follows:

[0190]

[0191] Example 1B and 2-amino-4-(trifluoromethyl)pyridine-5-boronic acid pinacol ester were used as raw materials and the synthesis method was the same as that of Example 1C to obtain a white solid with a yield of 91%.

[0192] D: Preparation of 5-(4-morpholinyl-6-(piperazine-1-yl)-1,3,5-triazine-2-yl)-4-(trifluoromethyl)pyridin-2-amine, the structural formula is as follows:

[0193]

[0194] Example 13C was used as the raw material and the synthesis method was the same as that of Example 1D to obtain a white solid with a yield of 96%.

[0195] E: Preparation of methyl 5-(4-(4-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)pentanoate, the structural formula is as follows:

[0196]

[0197] Example 13D and 5-bromovalerate monomethyl ester were used as raw materials and the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 88%.

[0198] F: Preparation of 5-(4-(4-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-N-hydroxypentanamide, the structural formula is as follows:

[0199]

[0200] Example 13E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 72% and a melting point of 182-183°C. The compound structure is characterized as follows: 1H NMR (500MHz, DMSO-d6) δ10.40(s,1H),8.58(s,1H),6.95(s,2H),6.81(s,1H),3.75(d,J=11.1Hz,8H),3.62(t,J=4.8Hz ,4H),2.36(t,J=5.3Hz,4H),2.28(t,J=7.2Hz,2H),1.96(t,J=7.2Hz,2H),1.51(p,J=7.1Hz,2H),1.42(q,J=7.7Hz,2H). 13 C NMR(126MHz,DMSO-d6)δ169.86,169.45,164.64,164.36,161.60,152.90,124.64,11 9.30,105.13,66.42,60.22,57.88,53.00,43.65,32.57,26.18,23.52,21.23,14.55.

[0201] Embodiment 14

[0202] Preparation of hydroxamic acid derivatives of triazine No. CY-14

[0203] E: Preparation of 5-(4-(4-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-5-oxopentanoic acid methyl ester, the structural formula is as follows:

[0204]

[0205] Using Example 13D and monomethyl glutarate as raw materials, the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 86%.

[0206] F: Preparation of 5-(4-(4-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-N-hydroxy-5-oxopentylamide, the structural formula is as follows:

[0207]

[0208] Example 14E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 65% and a melting point of 143-144°C. The compound structure is characterized as follows: 1H NMR(500MHz,DMSO-d6)δ10.39(d,J=2.8Hz,1H),8.60(s,1H),6.97(s,2H),6.83(s,1H),3.82–3.71(m,8H), 3.66–3.61(m,4H),3.51(s,4H),2.34(t,J=7.3Hz,2H),2.01(dd,J=8.7,5.7Hz,2H),1.74(q,J=7.3Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ170.90,170.88,169.92,169.33,164.59,164.54,162.76,161.66 ,152.99,119.13,105.18,66.42,53.78,43.67,38.71,36.25,32.12,32.04,31.23,21.31.

[0209] Embodiment 15

[0210] Preparation of hydroxamic acid derivatives of triazine No. CY-15

[0211] C: Preparation of tert-butyl 4-(4-(6-amino-5-(trifluoromethyl)pyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-carboxylate, the structural formula is as follows:

[0212]

[0213] Example 1B and 2-amino-3-(trifluoromethyl)pyridine-5-boronic acid pinacol ester were used as raw materials and the synthesis method was the same as that of Example 1C to obtain a white solid with a yield of 94%.

[0214] D: Preparation of 5-(4-morpholinyl-6-(piperazine-1-yl)-1,3,5-triazine-2-yl)-4-(trifluoromethyl)pyridin-2-amine, the structural formula is as follows:

[0215]

[0216] Example 15C was used as the raw material and the synthesis method was the same as that of Example 1D to obtain a white solid with a yield of 92%.

[0217] E: Preparation of methyl 5-(4-(4-(6-amino-5-(trifluoromethyl)pyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)pentanoate, the structural formula is as follows:

[0218]

[0219] Example 15D and methyl 4-bromovalerate were used as raw materials and the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 78%.

[0220] F: Preparation of 5-(4-(4-(6-amino-5-(trifluoromethyl)pyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-N-hydroxypentanamide, the structural formula is as follows:

[0221]

[0222] Example 15E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 43% and a melting point of 203-204°C. The compound structure is characterized as follows: 1 H NMR (500MHz, DMSO-d6) δ10.40(s,1H),9.10(d,J=2.1Hz,1H),8.71(s,1H),8.47(d,J=2.1Hz,1H),7.11(s,2H),3.90–3.69(m ,8H),3.65(t,J=4.7Hz,4H),2.46–2.33(m,4H),2.29(t,J=7.2Hz,2H),1.98(t,J=7.2Hz,2H),1.47(dq,J=39.3,7.5Hz,4H). 13 C NMR (126MHz, DMSO-d6) δ169.51,167.75,164.72,164.47,157.75,153.52,134.89,133.57,132.75,132.51,132. 49,131.98,131.90,129.27,129.17,125.64,123.48,120.42,66.47,57.89,53.05,43.69,32.56,26.20,23.52.

[0223] Example 16

[0224] Preparation of hydroxamic acid derivatives of triazine No. CY-16

[0225] E: Preparation of 5-(4-(4-(6-amino-5-(trifluoromethyl)pyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-5-oxopentanoic acid methyl ester, the structural formula is as follows:

[0226]

[0227] Example 15D and monomethyl glutarate were used as raw materials and the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 76%.

[0228] F: Preparation of 5-(4-(4-(6-amino-5-(trifluoromethyl)pyridin-3-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-N-hydroxy-5-oxopentylamide, the structural formula is as follows:

[0229]

[0230] Example 16E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 55% and a melting point of 213-214°C. The compound structure is characterized as follows: 1 H NMR (500MHz, DMSO-d6) δ10.37(d,J=1.8Hz,1H),9.11(d,J=2.2Hz,1H),8.69(d,J=1.7Hz,1H),8.48(d,J=2.1Hz,1H),7.12(s,2H),3.81(d,J= 23.3Hz,8H),3.65(t,J=4.9Hz,5H),3.53(s,4H),3.17(d,J=5.2Hz,2H),2.35(t,J=7.4Hz,2H),2.01(t,J=7.4Hz,2H),1.74(p,J=7.4Hz,2H). 13 C NMR (126 MHz, DMSO-d6) δ 170.85, 169.33, 167.84, 164.69, 164.64, 157.80, 153.60, 134.94, 125.63, 120.30, 66.48, 49.06, 40.47, 32.13, 32.04, 21.32. Example 17

[0231] Preparation of hydroxamic acid derivatives of triazine No. CY-17

[0232] C: Preparation of tert-butyl 4-(4-(2-amino-4-(trifluoromethyl)pyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazine-1-carboxylate, the structural formula is as follows:

[0233]

[0234] Example 1B and 2-amino-4-trifluoromethylpyrimidine-5-boronic acid pinacol ester were used as raw materials and the synthesis method was the same as that of Example 1C to obtain a white solid with a yield of 95%.

[0235] D: Preparation of 4-(4-morpholinyl-6-(piperazine-1-yl)-1,3,5-triazine-2-yl)-4-(trifluoromethyl)pyrimidin-2-amine, the structural formula is as follows:

[0236]

[0237] Example 17C was used as the raw material and the synthesis method was the same as that of Example 1D to obtain a white solid with a yield of 96%.

[0238] E: Preparation of 5-(4-(4-(2-amino-4-(trifluoromethyl)pyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-5-oxopentanoic acid methyl ester, the structural formula of which is as follows:

[0239]

[0240] Using Example 17D and monomethyl glutarate as raw materials, the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 75%.

[0241] F: Preparation of 5-(4-(4-(2-amino-4-(trifluoromethyl)pyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-N-hydroxy-5-oxopentylamide, the structural formula is as follows:

[0242]

[0243] Example 17E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 48% and a melting point of 190-191°C. The compound structure is characterized as follows: 1 H NMR (400MHz, DMSO-d6) δ8.90(s,1H),7.68(s,2H),5.17(s,1H),3.85–3.73(m,8H),3.63(t,J= 4.7Hz, 4H), 3.51 (s, 4H), 2.33 (t, J = 7.5Hz, 2H), 1.97 (t, J = 7.3Hz, 2H), 1.72 (t, J = 7.4Hz, 2H). 13 C NMR (101MHz, DMSO-d6) δ171.10,168.88,168.57,164.49,163.57,163.11,118.63,66.39,32.29,21.58.

[0244] Embodiment 18

[0245] Preparation of hydroxamic acid derivatives of triazine No. CY-18

[0246] E: Preparation of methyl 5-(4-(4-(2-amino-4-(trifluoromethyl)pyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)pentanoate, the structural formula is as follows:

[0247]

[0248] Using Example 1B and methyl bromovalerate as raw materials, the synthesis method was the same as that of Example 1E to obtain a white solid with a yield of 67%.

[0249] F: Preparation of 5-(4-(4-(2-amino-4-(trifluoromethyl)pyrimidin-5-yl)-6-morpholinyl-1,3,5-triazine-2-yl)piperazin-1-yl)-N-hydroxypentanamide, the structural formula is as follows:

[0250]

[0251] Example 18E, hydroxylamine aqueous solution (15.17M, 10eq) and sodium methoxide methanol solution (5M, 5eq) were used as raw materials and the synthesis method was the same as that of Example 1F. A white powder was obtained with a yield of 56% and a melting point of 125-126°C. The compound structure is characterized as follows: 1 H NMR (400MHz, DMSO-d6) δ10.48(s,1H),8.87(s,1H),7.66(s,2H),3.90–3.71(m,8H),3.61(t,J=4.7Hz, 4H), 2.42 (d, J = 5.9Hz, 4H), 2.32 (t, J = 7.1Hz, 2H), 1.98 (t, J = 7.0Hz, 2H), 1.48 (dt, J = 23.5, 7.5Hz, 4H). 13 C NMR (101MHz, DMSO-d6) δ169.55,168.52,164.53,164.27,163.54,163.06,118.72,66.39,57.69,52.80,43.66,42.92,32.51,25.93,23.46.

[0252] MTT assay for cell proliferation inhibition activity

[0253] The in vitro cell proliferation inhibition experiment was performed using the MTT method, and the following four cell lines were used: human colorectal cancer cell lines HT-29 and HCT116, and human breast cancer cells HCC1937 and MDA-MB-231.

[0254] HT-29, HCT116, HCC1937, and MDA-MB-231 are adherent cells and were routinely cultured at 37°C and 5% CO2 using RPIM-1640 culture medium containing 10% fetal bovine serum.

[0255] The specific operations are as follows:

[0256] First, the compounds (ie, samples) prepared in Examples CY1-CY18 were respectively prepared into DMSO (dimethyl sulfoxide) solutions with a compound concentration of 5 mmol / L, and then the obtained solutions were diluted in a gradient manner to obtain a series of sample solutions with a concentration gradient.

[0257] Next, cells in the logarithmic phase were taken and counted using a hemocytometer. Tumor cells were counted at 6-8×10 per well. 3 The cells were inoculated in a 96-well plate at a density of 100 μL per well, and the volume of culture medium per well was 99 μL. Drugs were added 12-16 hours after plating (drugs were added after cells adhered to the wall). 1 μL of the compound solution to be tested was added to each well to make the final concentration of the compound the set concentration value. Four replicate wells were set for each concentration. IC 50 Eight concentration gradients were set during the test. Two positive drug groups (HDAC inhibitor SAHA or PI3K inhibitor BKM120, PQR309 were added, respectively), a control group (1 μL DMSO was added) and a blank group (1 μL DMSO was added) were set up at the same time; after the compound and cells were co-cultured for 72 hours, MTT PBS solution (5 mg / mL, 10 μL / well) was added to the experimental group and the control group, and sterile culture was continued in the incubator for 4 hours; the 96-well plate was centrifuged and the culture medium was removed (adherent cells do not need to be centrifuged), DMSO (100 μL / well) was added, and the 96-well plate was shaken on a micro-oscillator for 5 minutes, and the OD value at 490 nm was tested using an enzyme reader. Finally, the inhibition rate (Inh%) of the compound on tumor cell proliferation at different concentrations was calculated based on the OD value, and then the IC was obtained by drawing a concentration-inhibition rate curve. 50 value.

[0258] The inhibition rate was calculated as follows: Inh% = (OD 490 - Experimental group OD 490 ) / (control group OD 490 -Blank group OD 490 )×100%.

[0259] After calculation, the in vitro cell proliferation inhibitory activity of the compounds prepared by the present invention was obtained, and the results are shown in Table 1. It can be seen that compound CY1, compound CY2 and SAHA have good proliferation inhibitory activity on the tested tumor cells. In particular, in breast cancer cells MDA-MB-231, compound CY1 and compound CY2 have better activity than the listed drug SAHA. Although the activity of most other compounds and the listed drug BKM120 on tumor cell proliferation is not as ideal as compound CY1, compound CY2 and SAHA, the IC values ​​of some cell lines are better than those of other compounds. 50 Less than 50 μM. In conclusion, the tested compounds can inhibit tumor cell proliferation and have the potential to be developed as anti-tumor drugs.

[0260] Table 1 In vitro tumor cell proliferation inhibitory activity of the compounds prepared in each example

[0261]

[0262] Note: IC 50 It indicates the half inhibitory concentration.

[0263] PI3K enzyme inhibition activity assay

[0264] The inhibitory activity of compounds CY1-CY18 on PI3Kα was detected using the PI3Kα colorimetric assay kit (purchased from Invitrogen, PV4788), and the PI3Kα inhibitor PI103 was used as a control. The experimental steps refer to the instructions of the kit, mainly including: 1) Prepare a series of diluents and prepare the source plate: dilute the compound with a concentration of 100% DMSO to 100 times the maximum inhibitor concentration required for the reaction to achieve the final reaction effect. Use this solution to extract and concentrate the sample to obtain a concentrate. Transfer 100 μL of the dilution to a 96-well plate; inject 100 μL of 100% DMSO into the 96-well plate respectively, without adding any compound or enzyme control, and mark the plate as the source plate. Transfer 40 μL of the compound from the source plate to a new 384-well plate as the intermediate plate; use Echo to transfer 50 nL of the compound in 100% DMSO to the assay plate. 2) Prepare 2× Kinase Solution: Prepare Kinase Solution in 1× Kinase Buffer at 2x the final concentration of each reagent in the assay. Add 2.5μL 1× Kinase Buffer to each well of the assay plate, except for the control wells without enzyme, and shake. 3) Prepare 2× Substrate Solution: Prepare PIP2 Substrate and ATP Substrate Solutions, using 2x the final concentration of each reagent in 1× Kinase Reaction Buffer. Inject 2.5uL of substrate solution / well into the assay plate and shake to ensure sufficient reaction. 4) Perform Kinase Reaction: Cover the assay plate and incubate at room temperature for 1 hour. 5) Kinase Assay: Equilibrate ADP-Glo ​​Reagent to room temperature. Stop the reaction by adding 5μL ADP-Glo ​​Reagent to each well. Mix briefly with centrifuge, shake slowly on a shaker, and equilibrate for 120 minutes. Add 10uL Kinase Assay Reagent to each well, shake for 1 minute, and read luminescence on a plate reader after equilibration for 30 minutes. 6) Curve Fitting: Copy the RLU values ​​from Envision. The RLU values ​​were converted to percentage inhibition values ​​and the data were fitted in XLFit excel plug-in version 5.4.0.8 to obtain IC 50 Value. The formula is: Y = Bottom + (Top-Bottom) / (1 + (IC 50 / X)^HillSlope). The activity test results are shown in Table 2. It can be seen that except for CY2 and CY18, the inhibition rate of PI3Kα at 1 μM is less than 0%, especially compounds CY9, CY10, CY12, CY13 and CY20 have PI3Kα inhibitory activity comparable to that of PI103. Therefore, most of the compounds are good PI3Kα inhibitors.

[0265] Table 2 PI3Kα enzyme inhibitory activity of the compounds prepared in each example

[0266]

[0267] HDAC enzyme inhibition activity assay

[0268] HDAC1, a subtype of the histone deacetylase family, was used as the research object to test the inhibitory activity of the compounds on histone deacetylase. Each compound was set up with 7 concentration gradients and single wells, and the marketed HDAC inhibitor SAHA was used as a control. First, the compound was dissolved in the reaction buffer, and then a certain volume of buffer solution containing histone deacetylase was added. After incubation at room temperature for 15 minutes, trypsin and acetylated peptide buffer solution were added as reaction substrates to start the deacetylation reaction. At the same time, the compound concentration and enzyme content reached the set value. After gently mixing for 60 seconds and incubating at room temperature, the kinetic parameters of the reaction under certain excitation light and emission light wavelengths were recorded within 1 hour. The inhibitory activity of the compound on the enzyme was obtained by comparison with the negative control group (no inhibitor group), and the half inhibitory concentration (IC50) of the compound on histone deacetylase was calculated. 50 The results are shown in Table 3. It can be seen that except for compound CY5 and compound CY12, the IC 50 Greater than 1000nM, IC values ​​of other compounds for HDAC1 50 Less than 1000nM. The above results show that the compounds prepared by the present invention have good HDAC1 and HDAC6 inhibitory activity. Based on the experimental results of Example 2 and Example 3, most of the compounds are good PI3K and HDAC dual-target inhibitors, and can effectively inhibit tumor cell proliferation, and have the potential to be developed as anti-tumor drugs.

[0269] Table 3 HDAC1 enzyme inhibitory activity of the compounds prepared in each example

[0270]

[0271] Note: IC 50 It indicates the half inhibitory concentration.

[0272] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A triazine hydroxamic acid derivative or a pharmaceutically acceptable salt thereof, characterized in that: The general structural formula of the derivative is shown in Formula I: in: R1 is selected from any one of hydrogen atom and -CF3; X is selected from -N-; Y is selected from any one of -CH2-, -C(O)-, -CH2CH2-; G is selected from any one of -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2-, and -CH2CH2CH2CH2CH2-.

2. The triazine hydroxamic acid derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The structural formula of the derivative is shown in any of the following numbers:

3. The triazine hydroxamic acid derivative or a pharmaceutically acceptable salt thereof according to claim 1 or 2, characterized in that: The pharmaceutically acceptable salt is an inorganic acid salt or an organic acid salt; The inorganic acid salt is selected from the salt formed by any one of the following inorganic acids: hydrochloric acid, sulfuric acid and phosphoric acid; The organic acid salt is selected from the group consisting of salts formed by any one of the following organic acids: acetic acid, trifluoroacetic acid, malonic acid, citric acid and p-toluenesulfonic acid.

4. The method for preparing a triazine hydroxamic acid derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: The steps include: The compound represented by formula IA is reacted with hydroxylamine or hydroxylamine hydrochloride to obtain the derivative represented by formula I; or: The compound represented by formula IA and hydroxylamine having a hydroxyl group with a protective group are first reacted under the action of a condensing agent to generate a compound represented by formula IB, and then the protective group is removed from the compound represented by formula IB to obtain a derivative represented by formula I; Wherein, P is a protecting group for the hydroxyl group in hydroxylamine.

5. The method for preparing a triazine hydroxamic acid derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: The steps include: The compound represented by formula IC is reacted with hydroxylamine or hydroxylamine hydrochloride in an alkaline solution to obtain a derivative represented by formula I; Wherein, E is an alkyl group.

6. Use of the derivative according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof in the preparation of any of the following products: 1) Phosphatidylinositol 3-kinase and / or histone deacetylase inhibitors; 2) Eukaryotic tumor cell proliferation inhibitor.

7. The use according to claim 6, characterized in that: The phosphatidylinositol 3-kinases include the isoforms PI3Kα, PI3Kβ, PI3Kδ, and PI3Kγ known in mammalian cells; The histone deacetylase includes the subtypes HDAC1, HDAC2, HDAC3, HDAC8, HDAC4, HDAC5, HDAC7, HDAC9, HDAC6, HDAC10, and HDAC11 known in mammalian cells; The cells include human breast cancer cells MDA-MB-231.

8. A product, characterized in that The active ingredient is a derivative as described in any one of claims 1 to 3 or a pharmaceutically acceptable salt or a tautomer thereof; the product is any one of the following: 1) Phosphatidylinositol 3-kinase and / or histone deacetylase inhibitors; 2) Eukaryotic tumor cell proliferation inhibitor.

9. The product according to claim 8, characterized in that The phosphatidylinositol 3-kinases include the isoforms PI3Kα, PI3Kβ, PI3Kδ, and PI3Kγ known in mammalian cells; The histone deacetylase includes the subtypes HDAC1, HDAC2, HDAC3, HDAC8, HDAC4, HDAC5, HDAC7, HDAC9, HDAC6, HDAC10, and HDAC11 known in mammalian cells; The cells include human breast cancer cells MDA-MB-231.

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