A diacylpyrrole class histone deacetylase 6 inhibitor and uses thereof

Abstract

The application belongs to the field of pharmaceutical chemistry, and specifically discloses a diacyl pyrrole class of histone deacetylase 6 inhibitor and purposes thereof, wherein the compound is a compound as shown in a general formula (I) or a pharmaceutically acceptable salt thereof. The application further discloses a pharmaceutical composition containing the compound, which can be used for preventing or treating clinical conditions related to HDAC1 or HDAC6. Compared with reported clinical selective HDAC6 inhibitors, the HDAC6 inhibitor in the patent has significantly improved enzyme activity and cell activity.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical and chemical engineering, specifically relating to a diacylpyrrole histone deacetylase 6 inhibitor and its uses. Background Technology

[0002] Histone deacetylases (HDACs) are a class of proteases whose acetylation and deacetylation of histones in chromatin are key steps in regulating gene expression. Abnormal gene expression is the molecular biological basis for tumors and some genetic and metabolic diseases. The degree of histone acetylation is coordinated and controlled by histone acetyltransferases (HATs) and histone deacetylases (HDACs). When HDACs are overexpressed and recruited by transcription factors, it leads to abnormal repression of specific genes, resulting in tumors and other diseases.

[0003] HDAC is a large family of enzymes, currently known to comprise four main classes and 18 different subtypes. Class I includes four subtypes: HDAC1, 2, 3, and 8; Class II includes six subtypes: HDAC4, 5, 6, 7, 9, and 10 (of which 4, 5, 7, and 9 belong to IIa, and 6 and 10 belong to IIb); Class IV consists of only one subtype, HDAC11, which shares some homology with the first two classes; and Class III includes seven subtypes, SIRT1-7, which do not share structural homology with the first three classes.

[0004] According to publicly available clinical results, selective HDAC1,2,3 inhibitors such as MS275 (Entinostat), Chidamide (marketed), and CXD101, although having significantly fewer toxic side effects compared to Pan HDAC inhibitors such as vorinostat, romedixin, belistat, and pabistat, have still been observed to cause serious adverse reactions in clinical trials. These include hematological adverse reactions, systemic adverse reactions such as fatigue and fever, gastrointestinal adverse reactions, metabolic and nutritional system adverse reactions, and other adverse reactions such as dizziness and rash. Summary of the Invention

[0005] In view of the shortcomings of the existing technology, the purpose of this invention is to provide a selective HDAC6 inhibitor that reduces off-target toxicity and enhances the therapeutic window.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] On the one hand, the present invention provides compounds as shown in Formula I or pharmaceutically acceptable salts thereof:

[0008]

[0009] Or its pharmaceutically acceptable salts or solvates, wherein,

[0010] R 1 R 2 Each can be independently represented by hydrogen, alkyl, alkenyl, alkynyl, aryl, aralkyl, or Het;

[0011] Q 1 It is selected from aryl, aralkyl, or Het;

[0012] Q 2 It is selected from alkyl or aryl groups;

[0013] The alkyl group is a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms, or a cyclic saturated hydrocarbon group having 3-6 carbon atoms, or a cyclic saturated hydrocarbon group having 3-6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms.

[0014] The alkoxy group is a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms; or a cyclic saturated hydrocarbon group having 3-6 carbon atoms; or a cyclic saturated hydrocarbon group having 3-6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms; wherein each carbon atom may optionally be substituted with oxygen.

[0015] Alkoxyalkyl refers to an alkoxy group connected to an alkyl group as defined above;

[0016] Alkenyl and alkynyl are straight-chain or branched unsaturated hydrocarbon groups containing double or triple bonds and having 1-6 carbon atoms;

[0017] The aryl group is a carbon ring selected from phenyl, naphthyl, acenaphthel or tetrahydronaphthyl, each of which is optionally substituted by 1, 2 or 3 substituents, each substituent being independently selected from hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxyl, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl or Het.

[0018] Arylalkyl and diarylalkyl are aryl groups linked to alkyl groups as defined above;

[0019] Het is a monocyclic heterocyclic compound selected from pyrrole, pyrazolyl, imidazolyl, furanyl, thiophene, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, or pyridazinyl; or selected from quinolinyl, quinoxolinyl, indolyl, benzimidazolyl, benzoxazolyl, benzoisoxazolyl, benzothiazolyl, benzoisothiazolyl, benzofuranyl, benzothiaphenyl, 2,3-dihydrobenzo[1,4] A bicyclic heterocycle of dioxanehexenyl or benzo[1,3]dioxanepentenyl; or a monocyclic saturated hydrocarbon group of 3-6 carbon atoms or a bicyclic saturated hydrocarbon group of 6-12 carbon atoms, wherein the carbon atoms on the ring are optionally substituted by 1-4 O, S, N or NH atoms; each monocyclic or bicyclic ring is optionally substituted by 1, 2 or 3 substituents, each substituent being optionally selected from halogen, haloalkyl, hydroxyl, alkyl or alkoxy groups;

[0020] Halogens are substituents selected from fluorine, chlorine, bromine or iodine;

[0021] The haloalkyl group is a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms, or a cyclic saturated hydrocarbon group having 3-6 carbon atoms, or a cyclic saturated hydrocarbon group having 3-6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms; wherein one or more carbon atoms are replaced by one or more halogen atoms.

[0022] As a preferred technical solution, among which...

[0023] R 1 R 2 Each can independently represent hydrogen, alkyl, aryl, or Het;

[0024] Q 1 It is selected from aryl;

[0025] Q 2 It is selected from alkyl or aryl groups.

[0026] As a preferred technical solution, among which...

[0027] R 1 It represents hydrogen or phenyl, and the substituents include, but are not limited to, alkyl, methoxy, and halogen;

[0028] R 2 It is a hydroxyl group;

[0029] Q 1 It is phenyl;

[0030] Q 2 It is selected from phenyl or a straight chain with 4-5 carbon atoms.

[0031] In another aspect, the present invention provides a pharmaceutical composition comprising any of the compounds described above or a pharmaceutically acceptable carrier.

[0032] In another aspect, the present invention provides the use of the above-described compounds in the preparation of medicaments for the prevention or treatment of clinical conditions related to HDAC6.

[0033] In the specific implementation plan, diseases associated with HDAC1 or HDAC6 can include lung cancer, melanoma, liver cancer, kidney cancer, leukemia, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, testicular cancer, breast cancer, bladder cancer, gallbladder cancer, myelodysplastic syndrome, lymphoma, esophageal cancer, gastrointestinal cancer, astrocytoma, neuroblastoma, glioma, schwannoma, mesothelioma, non-insulin-dependent diabetes mellitus, and autoimmune diseases.

[0034] In another aspect, the present invention provides a method for preparing the above-mentioned compound.

[0035]

[0036] All compounds of the present invention can be prepared by the above or similar preparation methods. The appropriate raw materials can be selected according to the different substituents and their positions.

[0037] Due to the adoption of the above technical solutions, the beneficial effects of the present invention include:

[0038] (1) It selectively acts on HDAC6 isoenzyme, avoiding interference with other HDAC subtypes (such as class I HDACs) and reducing toxic side effects caused by extensive inhibition (such as bone marrow suppression, gastrointestinal reactions, etc.).

[0039] (2) Compared with the reported clinically selective HDAC6 inhibitors, the HDAC6 inhibitors in this patent have significantly improved enzyme activity and cell activity. Detailed Implementation

[0040] The following description will elaborate on the specific aspects, characteristics, and advantages of the aforementioned compounds, methods, and pharmaceutical compositions, making the content of this invention readily apparent. It should be understood that the detailed descriptions and examples described below are specific embodiments and are for reference only. After reading this description, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by this application.

[0041] The starting materials used in the examples of this invention are known and commercially available, and can also be synthesized using or according to methods known in the art.

[0042] Example 1: Synthesis of 5-benzoyl-1-benzyl-N-(7-(hydroxyamino)-7-oxohepyl)-1H-pyrrole-2-carboxamide (KI-38)

[0043]

[0044] (1) Synthesis of methyl 4-benzoyl-1H-pyrrole-2-carboxylate

[0045] Anhydrous AlCl3 (2.00 g, 14.99 mmol) was added to a 200 mL two-necked flask, the air was purged three times and protected with argon, anhydrous dichloromethane (50 mL) was added at -20 °C, and the mixture was stirred for 30 min. Then, benzoyl chloride (0.86 mL, 7.49 mmol) solution was slowly added dropwise. After 1 h, methyl 2-pyrrolecarboxylate (0.85 g, 6.81 mmol) was added. The temperature was then gradually increased to 40 °C for another 1 h. After 6 h, the reaction was detected by TLC to be complete. 6 mL of water was slowly added dropwise. After the reaction was completely quenched, 14 mL of water was added, and the mixture was extracted with DCM (60 mL × 3). The organic phases were combined and purified by concentrated column chromatography under reduced pressure (petroleum ether: ethyl acetate = 4:1) to obtain the target compound in 95% yield.

[0046] (2) Synthesis of methyl 4-benzoyl-1-benzyl-1H-pyrrole-2-carboxylate

[0047] Methyl 4-benzoyl-1H-pyrrole-2-carboxylate (0.69 g, 3.00 mmol), methyl iodoforme (0.51 g, 3.60 mmol), and NaH (0.11 g, 4.50 mmol) were placed in a 100 mL round-bottom flask, and 12 mL of DMF was added. The mixture was reacted at room temperature for 8–12 h, and the reaction was monitored by TLC until complete. Excess NaH was quenched by adding 5 mL of ammonium chloride solution, and 75 mL of water was added. The mixture was then extracted with ethyl acetate (150 mL × 3). The organic phases were combined, concentrated under reduced pressure, and purified by column chromatography (petroleum ether:ethyl acetate = 8:1) to obtain the target compound as a white solid in 72% yield.

[0048] (3) Synthesis of 4-benzoyl-1-benzyl-1H-pyrrole-2-carboxylic acid

[0049] 0.42 g (1.50 mmol) of methyl 4-benzoyl-1-benzyl-1H-pyrrole-2-carboxylate and 0.24 g (6.00 mmol) of NaOH were weighed into a 100 mL round-bottom flask. 30 mL of methanol was added, and the mixture was reacted at 90 °C for 16 h. The reaction was confirmed to be complete by TLC. The solvent was removed by concentration under reduced pressure. 10 mL of water was added, and 30 mL of 30% hydrochloric acid was added dropwise to adjust the pH to acidic. A white flocculent precipitate was formed. The filter cake was collected by suction filtration and dried to obtain a white solid with a yield of 80%.

[0050] (4) Synthesis of methyl 7-(4-benzoyl-1-benzyl-1H-pyrrole-2-carbamoyl) heptanoate

[0051] Compound 5a (1.57 g, 5.86 mmol) and HATU (2.44 g, 6.44 mmol) were weighed into a 200 mL two-necked reaction flask under ice bath conditions. DMF (30 mL) was added to dissolve the compound, followed by the addition of DIPEA (4.59 mL, 26.37 mmol). Once the solution temperature dropped to 0 °C, methyl 4-aminomethylbenzoate (1.06 g, 6.44 mmol) was added, and the reaction was carried out for 8 h. TLC was used to confirm the completeness of the reaction. Water (150 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (250 mL × 3). The organic phases were combined, concentrated under reduced pressure, and purified by column chromatography (petroleum ether:ethyl acetate = 1:2) to obtain the target compound as a white solid, with a yield of 81%.

[0052] (5) Synthesis of 5-benzoyl-1-benzyl-N-(7-(hydroxyamino)-7-oxohepyl)-1H-pyrrole-2-carboxamide

[0053] Hydroxylamine hydrochloride (1.70 g, 24.46 mmol) was added to a flask containing 9 mL of methanol, and potassium hydroxide (1.70 g, 30.30 mmol) was added to another flask containing 18 mL of methanol. The two solutions were stirred until dissolved, mixed, and placed in an ice bath at 0°C for 30 min. The mixture was then filtered. Methyl 7-(4-benzoyl-1-benzyl-1H-pyrrole-2-carboxamido) heptanoate (0.25 g, 0.69 mmol) was added to the filtrate. The mixture was reacted in an ice bath for 6 h. After the reaction was confirmed to be complete by TLC, methanol was removed by vacuum distillation. The product was dissolved in water (10 mL), and the pH was adjusted to 7-8 with acetic acid. Ethyl acetate (30 mL × 3) was added for extraction. The organic phases were combined and purified by column chromatography (dichloromethane:methanol = 20:1) to obtain the target compound as a pale yellow solid with a yield of 61.8% and a melting point of 151.7-153.1°C. 1H NMR (400 MHz, DMSO-d6) δ 10.35 (s, 1H), 8.69 (s, 1H), 8.30 (t, J = 5.7 Hz, 1H), 7.87 – 7.68 (m, 3H), 7.58 (dt, J = 34.3, 7.4 Hz, 3H), 7.27 (dq, J = 14.5, 7.0 Hz, 4H), 7.17 (d, J = 6.9 Hz, 2H), 5.65 (s, 2H), 3.12 (q, J = 6.6 Hz, 2H), 1.92 (t, J = 7.3 Hz, 2H), 1.50 – 1.36 (m, 4H), 1.21(dq, J = 7.7, 4.1 Hz, 4H). HR-MS (ESI, m / z): Calcd for 448.2230. (C 26 H 30 N3O4 + [M+H] + ). Found 448.2230.

[0054] Example 2: Synthesis of 5-benzoyl-N-(7-(hydroxyamino)-7-oxohepyl)-1-(3-methoxybenzyl)-1H-pyrrole-2-carboxamide (KI-39)

[0055]

[0056] White solid, yield 63.5%, melting point 153.1~154.8 ℃. 1H NMR (400 MHz, DMSO-d6) δ10.35 (s, 1H), 8.68 (s, 1H), 8.32 (t, J = 5.7 Hz, 1H), 7.84 – 7.73 (m, 3H), 7.62 (t, J = 7.4 Hz, 1H), 7.54 (t, J = 7.5 Hz, 2H), 7.27 (d, J = 1.9 Hz, 1H), 7.21 (t, J = 7.9 Hz, 1H), 6.81 (d, J = 8.1 Hz, 1H), 6.77 – 6.70 (m, 2H), 5.62(s, 2H), 3.69 (s, 3H), 3.13 (q, J = 6.7 Hz, 2H), 1.92 (t, J = 7.4 Hz, 2H), 1.50 – 1.38 (m, 4H), 1.22 (q, J = 3.6 Hz, 4H). HR-MS (ESI, m / z): Calcd for478.2336. (C 27 H 32 N3O5 + [M+H] + ). Found 478.2334.

[0057] Example 3: Synthesis of 5-benzoyl-1-(4-fluorobenzyl)-N-(7-(hydroxyamino)-7-oxohepyl)-1H-pyrrole-2-carboxamide (KI-44)

[0058]

[0059] White solid, yield 57.1%, melting point 119.9~121.9 ℃. 1H NMR (400 MHz, DMSO-d6) δ11.39 (s, 1H), 10.35 (s, 1H), 8.69 (s, 1H), 8.13 (dt, J = 11.3, 5.7 Hz, 1H), 7.77 (d, J = 1.8 Hz, 1H), 7.47 – 7.39 (m, 4H), 7.27 – 7.05 (m, 4H), 6.92 (d,J = 2.4 Hz, 1H), 5.61 (s, 2H), 3.09 (q, J = 6.6 Hz, 2H), 2.01 – 1.84 (m, 2H),1.51 – 1.35 (m, 4H), 1.24 – 1.18 (m, 4H). HR-MS (ESI, m / z): Calcd for466.2136. (C 26 H 29 FN3O4 + [M+H] + ). Found 466.2135.

[0060] Example 4: Synthesis of 5-benzoyl-N-(7-(hydroxyamino)-7-oxohepyl)-1-(4-(trifluoromethyl)benzyl)-1H-pyrrole-2-carboxamide (KI-52)

[0061]

[0062] Pale yellow solid, yield 55%, melting point 138.1 ~139.8 ℃. 1 H NMR (400 MHz, DMSO-d6) δ11.41 (s, 1H), 10.32 (s, 1H), 8.66 (s, 1H), 8.14 (dt, J = 11.4, 5.9 Hz, 1H), 7.80 (s, 1H), 7.66 (t, J = 8.7 Hz, 2H), 7.45 (dd, J = 15.7, 5.2 Hz, 4H), 7.25 (dd, J = 18.3, 8.0 Hz, 2H), 6.98 (s, 1H), 5.73 (s, 2H), 3.06 (t, J = 6.7 Hz,2H), 1.90 (t, J = 7.3 Hz, 2H), 1.39 (dt, J = 29.7, 7.2 Hz, 4H), 1.18 (s, 4H).HR-MS (ESI, m / z): Calcd for 516.2104. (C27 H 29 F3N3O4 + [M+H] + ). Found 516.2104.

[0063] Example 5: Synthesis of 5-benzoyl-1-(4-chlorobenzyl)-N-(7-(hydroxyamino)-7-oxohepyl)-1H-pyrrole-2-carboxamide (KI-56)

[0064]

[0065] A pale yellow solid, yield 59.5%, melting point 142.2~143.7 ℃. 1 H NMR (400 MHz, DMSO-d6) δ11.98 (s, 1H), 10.94 (s, 1H), 9.28 (s, 1H), 8.70 (s, 1H), 8.05 – 7.95 (m,4H), 7.94 – 7.86 (m, 3H), 7.69 (dd, J = 19.2, 8.1 Hz, 2H), 7.56 – 7.51 (m,1H), 6.19 (s, 2H), 3.74 (dd, J = 5.3, 2.2 Hz, 2H), 3.66 (t, J = 6.8 Hz, 2H), 2.05 – 1.91 (m, 4H), 1.80 – 1.72 (m, 4H). 13 C NMR (101 MHz, dmso) δ 169.25,160.94, 150.14, 138.23, 137.54, 131.86, 128.90, 128.83, 128.72, 128.63,128.42, 128.37, 128.22, 128.03, 124.67, 120.34, 114.12, 48.71, 38.46, 32.34,29.16, 28.41, 26.23, 25.18. HR-MS (ESI, m / z): Calcd for 482.1841. (C 26 H 29 ClN3O4 + [M+H] + ). Found 482.1841.

[0066] Example 6: Synthesis of 5-benzoyl-N-(7-(hydroxyamino)-7-oxohepyl)-1-(4-methoxybenzyl)-1H-pyrrole-2-carboxamide (KI-64)

[0067]

[0068] A pale yellow solid, yield 61.9%, melting point 158.5~159.9 ℃. 1 H NMR (400 MHz, DMSO-d6) δ10.34 (s, 1H), 8.67 (s, 1H), 8.28 (t, J = 5.7 Hz, 1H), 7.83 – 7.73 (m, 3H), 7.62 (t, J = 7.3 Hz, 1H), 7.54 (t, J = 7.4 Hz, 2H), 7.24 (d, J = 1.9 Hz, 1H), 7.19 (d, J = 8.7 Hz, 2H), 6.85 (d, J = 8.7 Hz, 2H), 5.56 (s, 2H), 3.70 (s,3H), 3.17 – 3.12 (m, 2H), 1.93 (t, J = 7.3 Hz, 2H), 1.44 (dd, J = 13.1, 5.9Hz, 4H), 1.23 (s, 4H). HR-MS (ESI, m / z): Calcd for 478.2336. (C 27 H 32 N3O5 + [M+H] + ). Found 478.2333.

[0069] Example 7: Synthesis of 5-benzoyl-1-(2-chlorobenzyl)-N-(7-(hydroxyamino)-7-oxohepyl)-1H-pyrrole-2-carboxamide (KI-51)

[0070]

[0071] Pale yellow solid, yield 66.4%, melting point 152.1~153.9 ℃. 1H NMR (400 MHz, DMSO-d6) δ11.39 (s, 1H), 10.32 (s, 1H), 8.58 (s, 1H), 8.25 – 8.02 (m, 1H), 7.84 – 7.58(m, 1H), 7.51 – 7.39 (m, 5H), 1.90 (t, J = 7.3 Hz, 2H), 1.38 (dt, J = 27.5, 5.4 Hz, 4H), 1.22 –1.09 (m, 4H). HR-MS (ESI, m / z): Calcd for 482.1841. C 26 H 29 ClN3O4 + [M+H] + ). Found482.1840.

[0072] Example 8: Synthesis of 5-benzoyl-1-(3-chlorobenzyl)-N-(7-(hydroxyamino)-7-oxohepyl)-1H-pyrrole-2-carboxamide (DBY-I-180-1)

[0073]

[0074] A pale yellow solid, yield 56.2%, melting point 122.9~124.3 ℃. 1 H NMR (400 MHz, DMSO-d6) δ11.40 (s, 1H), 10.32 (s, 1H), 8.15 (q, J = 5.5 Hz, 1H), 7.78 (d, J = 1.7 Hz,1H), 7.47 – 7.44 (m, 2H), 7.43 – 7.41 (m, 3H), 7.34 – 7.28 (m, 3H), 7.16 (s,1H), 6.93 (d, J = 1.7 Hz, 1H), 5.63 (s, 2H), 1.40 (dt, J = 24.4, 6.4 Hz, 6H),1.24 – 1.14 (m, 6H). HR-MS (ESI, m / z): Calcd for 482.1841. (C 26 H29 ClN3O4 + [M+H] + ). Found 482.1841.

[0075] Example 9: Synthesis of 5-benzoyl-1-(3-chlorobenzyl)-N-(6-(hydroxyamino)-6-oxohexyl)-1H-pyrrole-2-carboxamide (DBY-I-181-1)

[0076]

[0077] Pale yellow solid, yield 63.3%, melting point 104.6~106.2 ℃. 1 H NMR (400 MHz, DMSO-d6) δ11.40 (s, 1H), 10.32 (s, 1H), 8.15 (q, J = 5.5 Hz, 1H), 7.78 (d, J = 1.7 Hz,1H), 7.47 – 7.44 (m, 2H), 7.43 – 7.41 (m, 3H), 7.34 – 7.28 (m, 3H), 7.16 (s,1H), 6.93 (d, J = 1.7 Hz, 1H), 5.63 (s, 2H), 1.40 (dt, J = 24.4, 6.4 Hz, 6H),1.24 – 1.14 (m, 6H). 13 C NMR (101 MHz, DMSO) δ 160.88, 150.02, 141.68, 137.45,133.00, 130.32, 128.62, 128.15, 126.77, 125.66, 124.61, 114.45, 114.06,32.25, 29.09, 28.34, 26.13, 25.09. HR-MS (ESI, m / z): Calcd for 468.1684.(C 25 H 27 ClN3O4 + [M+H] + ). Found 468.1684.

[0078] Example 10: Synthesis of 5-benzoyl-N-(4-(hydroxycarbamoyl)benzyl)-1-methyl-1H-pyrrole-2-carboxamide (DBY-I-164-1)

[0079]

[0080] White solid, yield 51.2%, melting point 137.3~138.9 ℃. 1 H NMR (400 MHz, DMSO-d6) δ11.35 (s, 1H), 11.16 (s, 1H), 8.71 (t, J = 6.0 Hz, 1H), 7.68 (d, J = 8.1 Hz,2H), 7.59 (s, 1H), 7.47 – 7.40 (m, 5H), 7.32 (d, J = 8.1 Hz, 2H), 7.04 – 7.01(m, 1H), 4.37 (d, J = 5.9 Hz, 2H), 3.88 (s, 3H). HR-MS (ESI, m / z): Calcd for378.1448. (C 21 H 20 N3O4 + [M+H] + ). Found 378.1446.

[0081] Example 11: Synthesis of 5-benzoyl-1-(cyclopropylmethyl)-N-(4-(hydroxycarbamoyl)benzyl)-1H-pyrrole-2-carboxamide (DBY-I-165-1)

[0082]

[0083] White solid, yield 62.5%, melting point 138.9~140.5 ℃. 1 H NMR (400 MHz, DMSO-d6) δ11.35 (s, 1H), 11.16 (s, 1H), 8.73 (t, J = 6.1 Hz, 1H), 7.68 (d, J = 8.0 Hz, 3H), 7.46 (dd, J = 6.7, 3.2 Hz, 2H), 7.44 – 7.40 (m, 3H), 7.32 (d, J = 8.2Hz, 2H), 7.01 (d, J = 1.6 Hz, 1H), 4.38 (d, J = 6.0 Hz, 2H), 4.22 (d, J = 7.1Hz, 2H), 1.23 (s, 1H), 0.42 (d, J = 9.5 Hz, 2H), 0.30 (d, J = 4.9 Hz, 2H).HR-MS (ESI, m / z): Calcd for 418.1761. (C 24 H 24 N3O4 +[M+H] + ). Found 418.1759.

[0084] Example 12: Synthesis of 5-benzoyl-1-(3-chlorobenzyl)-N-(4-(hydroxycarbamoyl)benzyl)-1H-pyrrole-2-carboxamide (DBY-I-182-1)

[0085]

[0086] Pale yellow solid, yield 58%, melting point 154.1~155.7 ℃. 1 H NMR (400 MHz, DMSO-d6) δ11.43 (s, 1H), 11.16 (s, 1H), 8.99 (s, 1H), 8.78 (dt, J = 12.2, 6.1 Hz, 1H), 7.84 (d, J = 1.6 Hz, 1H), 7.65 (dd, J = 8.2, 3.6 Hz, 2H), 7.49 – 7.38 (m,5H), 7.33 (d, J = 5.8 Hz, 2H), 7.23 – 7.19 (m, 2H), 7.06 (dd, J = 8.5, 6.8Hz, 2H), 5.65 (s, 2H), 4.35 (t, J = 6.3 Hz, 2H). HR-MS (ESI, m / z): Calcd for488.1371. (C 27 H 23 ClN3O4 + [M+H] + ). Found 488.1371.

[0087] Example 13: Synthesis of 5-benzoyl-1-(4-fluorobenzyl)-N-(4-(hydroxycarbamoyl)benzyl)-1H-pyrrole-2-carboxamide (KI-87)

[0088]

[0089] White solid, yield 62.5%, melting point 138.9~140.5 ℃. 1H NMR (400 MHz, DMSO-d6) δ11.18 (s, 1H), 8.76 (dt, J = 12.3, 6.1 Hz, 1H), 7.83 (d, J = 1.8 Hz, 1H), 7.67 (dd, J = 8.1, 3.9 Hz, 2H), 7.50 – 7.29 (m, 5H), 7.26 – 6.98 (m, 8H), 5.63 (s, 2H), 4.35 (d, J = 6.0 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 164.13,160.89, 149.94, 143.96, 143.06, 137.40, 131.76, 128.65, 128.56, 128.36,128.17, 127.54, 127.36, 126.89, 126.83, 125.35, 125.31, 124.12, 115.06,114.32, 50.73, 41.60.

[0090] Example 14: Synthesis of 5-benzoyl-N-(4-(hydroxycarbamoyl)benzyl)-1-(3-methoxybenzyl)-1H-pyrrole-2-carboxamide (ZTT-5)

[0091]

[0092] White solid, yield 64.0%, melting point 135.3~136.4 ℃. 1 H NMR (400 MHz, DMSO-d6) δ11.42 (s, 1H), 10.37 (s, 1H), 8.78 (q, J = 6.4 Hz, 2H), 7.71 – 7.63 (m, 2H),7.50 – 7.30 (m, 5H), 7.27 – 7.17 (m, 3H), 7.05 (d, J = 1.8 Hz, 1H), 6.86 –6.79 (m, 1H), 6.72 – 6.62 (m, 2H), 5.64 (s, 2H), 4.37 (s, 2H), 3.68 (s, 3H).

[0093] Example 15: Synthesis of 5-benzoyl-N-(4-(hydroxycarbamoyl)benzyl)-1-(4-(trifluoromethyl)benzyl)-1H-pyrrole-2-carboxamide (ZTT-11)

[0094]

[0095] White solid, yield 61.0%, melting point 132.1~133.2 ℃. 1 H NMR (400 MHz, DMSO-d6) δ11.15 (s, 1H), 8.99 (s, 1H), 8.76 (t, J = 6.2 Hz, 1H), 7.88 – 7.79 (m, 1H),7.70 – 7.61 (m, 4H), 7.58 – 7.45 (m, 2H), 7.45 – 7.31 (m, 3H), 7.30 – 7.16(m, 4H), 7.13 – 7.01 (m, 1H), 5.75 (s, 2H), 4.33 (s, 2H).

[0096] Example 16: Synthesis of 5-benzoyl-1-(4-chlorophenyl)-N-(4-(hydroxycarbamoyl)benzyl)-1H-pyrrole-2-carboxamide

[0097]

[0098] White solid, 70% yield, melting point 130.5~131.4 ℃. 1 H NMR (400 MHz, DMSO-d6) δ 11.13(s, 1H), 9.00 (s, 1H), 8.94 (t, J = 6.0 Hz, 1H), 7.90 (d, J = 1.9 Hz, 1H),7.84 – 7.76 (m, 2H), 7.68 (d, J = 8.3 Hz, 2H), 7.62 (d, J = 7.4 Hz, 1H), 7.55(t, J = 7.4 Hz, 2H), 7.40 (d, J = 1.8 Hz, 1H), 7.37 (d, J = 8.5 Hz, 2H), 7.24(d, J = 8.0 Hz, 2H), 7.18 (d, J = 8.5 Hz, 2H), 5.64 (s, 2H), 4.39 (d, J = 6.0Hz, 2H).

[0099] Example 17: Synthesis of 5-benzoyl-1-(cyclopropylmethyl)-N-(7-(hydroxyamino)-7-oxohepyl)-1H-pyrrole-2-carboxamide

[0100]

[0101] White solid, yield 60%, melting point 172.5~173.2 ℃. 1 H NMR (400 MHz, DMSO-d6) δ 11.32(s, 1H), 10.33 (s, 1H), 8.67 (s, 1H), 8.11 (t, J = 6.1 Hz, 1H), 7.67 – 7.58(m, 1H), 7.49 – 7.35 (m, 4H), 6.92 – 6.78 (m, 1H), 4.21 (dd, J = 11.5, 7.1Hz, 2H), 3.11 (q, J = 6.5 Hz, 2H), 1.91 (t, J = 7.2 Hz, 2H), 1.45 (d, J = 6.9Hz, 4H), 1.23 (s, 4H), 0.84 (d, J = 6.9 Hz, 1H), 0.46 – 0.38 (m, 2H), 0.33 –0.21 (m, 2H).

[0102] Example 18: Synthesis of 5-benzoyl-N-(7-(hydroxyamino)-7-oxohepyl)-1-methyl-1H-pyrrole-2-carboxamide

[0103]

[0104] White solid, yield 70.4%, melting point 176.3~177.5 ℃. 1 H NMR (400 MHz, DMSO-d6) δ11.32 (s, 1H), 10.34 (s, 1H), 8.67 (s, 1H), 8.09 (q, J = 5.4 Hz, 1H), 7.55(d, J = 1.8 Hz, 1H), 7.43 (dt, J = 7.2, 5.2 Hz, 4H), 6.90 (d, J = 1.9 Hz, 1H), 3.86 (s, 3H), 3.11 (p, J = 6.6 Hz, 2H), 1.92 (td, J = 7.4, 3.4 Hz, 2H), 1.44 (dt, J = 14.8, 7.4 Hz, 4H), 1.22 (dd, J = 9.0, 4.3 Hz, 4H).

[0105] Example 19: Synthesis of 5-benzoyl-1-benzyl-N-(4-(hydroxycarbamoyl)benzyl)-1H-pyrrole-2-carboxamide

[0106]

[0107] White solid, yield 67%, melting point 144.9~145.5 ℃. 1 H NMR (400 MHz, DMSO-d6) δ 11.20(s, 1H), 9.03 (s, 1H), 8.94 (t, J = 6.1 Hz, 1H), 7.89 (d, J = 1.9 Hz, 1H),7.83 – 7.76 (m, 2H), 7.71 – 7.58 (m, 3H), 7.54 (dd, J = 8.1, 6.7 Hz, 2H), 7.38 (d, J = 1.9 Hz, 1H), 7.34 – 7.20 (m, 5H), 7.19 – 7.11 (m, 2H), 5.67 (s, 2H), 4.40 (d, J = 6.0 Hz, 2H).

[0108] Example 20: Synthesis of 5-benzoyl-N-(4-(hydroxycarbamoyl)benzyl)-1-(4-methoxybenzyl)-1H-pyrrole-2-carboxamide

[0109]

[0110] White solid, yield 59%, melting point 128.9~130.5 ℃. 1 H NMR (400 MHz, DMSO-d6) δ 11.18(s, 1H), 9.00 (s, 1H), 8.92 (t, J = 6.1 Hz, 1H), 7.85 (d, J = 1.9 Hz, 1H),7.82 – 7.75 (m, 2H), 7.68 (d, J = 8.2 Hz, 2H), 7.65 – 7.60 (m, 1H), 7.54 (dd,J = 8.1, 6.8 Hz, 2H), 7.33 (d, J = 1.8 Hz, 1H), 7.27 (d, J = 8.1 Hz, 2H), 7.20 – 7.11 (m, 2H), 6.87 – 6.78 (m, 2H), 5.57 (s, 2H), 4.41 (d, J = 6.0 Hz, 2H), 3.72 (s, 3H).

[0111] Partial pharmacological experiments and results:

[0112] (1) Determination and results of the inhibitory activity of the target compound against HDAC

[0113] The inhibitory activity of the synthesized compounds against HDAC was determined by fluorescence resonance energy transfer (FRET), and compared with positive control drugs to screen for compounds with better activity. HDAC was obtained by purification or by purchasing kits directly.

[0114] Specific method: Add enzyme to the reaction wells and reaction buffer to the control wells. Add the sample dissolved in DMSO to the reaction wells and incubate using a non-contact nano-level ultrasonic pipetting system. Add the corresponding fluorescent substrate to each reaction well and rotate. Incubate at 30°C for 1-2 hours. Add a chromogenic reagent containing TMP26 to stop the reaction, generating fluorescence. Detect the fluorescence intensity (excitation: 490 nM, emission: 520 nM) using an EnVision multi-label microplate reader (Perkin Elmer). Read the endpoint values ​​after the color development has stabilized. Calculate the percentage (relative to the DMSO control group) and half-maximal inhibition rate using GraphPad Prism 4 software.

[0115] Table 1: IC50 of some compounds for HDAC6 / HDAC1 50 (nM)

[0116]

[0117] Table 2: Suppression activity of Example 3 against different HDAC subtypes (IC50) 50, nM)

[0118]

[0119] (2) Determination of the in vitro antitumor activity of the target compound

[0120] This experiment used the CTG method to determine the inhibitory effect of compounds on the gastric cancer cell line AGS. Experimental method: Cells in the exponential growth phase were collected and viable cell counts were performed. The cell suspension concentration was adjusted using the corresponding culture medium for each cell type. 90 µL of cell suspension was added to each well of a 96-well cell culture plate. Each tested compound was dissolved in DMSO to prepare a 10 mM or 5 mM stock solution. The solutions were then diluted 10-fold with culture medium, with two replicates per well. 10 µL of the corresponding 10-fold solution was added to each well for each cell line, resulting in a final drug concentration of 10 µM or 25 µM, with a final DMSO concentration of 0.1%–0.5% (see Compound Preparation Method and Sample Addition Design: Experimental Plate Sample Addition Design). The plates were incubated at 37 ºC in a 5% CO2 incubator for 72 h. After 72 h of drug treatment, 50 µL (1 / 2 culture volume) of pre-melted and equilibrated CTG solution to room temperature was added to each well. The mixture was shaken for 2 min using a microplate shaker, and after incubation at room temperature for 10 min, the fluorescence signal was measured using an Envision 2104 plate reader. The cell inhibition rate was calculated using the formula: (1 - V)sample / V vehicle control Calculated as (×100%). Where V... sample V represents the mean value of the drug-treated groups. vehicle control This represents the average value of the solvent control group.

[0121]

[0122] Therefore, compound 3 exhibits superior antiproliferative activity against AGS cells compared to the selective HDAC6 inhibitor ACY-1215, and comparable to the non-selective HDAC inhibitor SAHA. Furthermore, it shows no toxicity to normal human bone marrow stromal cells, making it superior to SAHA.

[0123] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. However, any modifications, alterations, and variations made by those skilled in the art without departing from the scope of the present invention based on the disclosed technical content are equivalent embodiments of the present invention. Furthermore, any modifications, alterations, and variations made to the above embodiments based on the essential technology of the present invention are still within the protection scope of the present invention.

Claims

1. A compound, characterized in that, Selected from the following structures: ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， 。 2. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable carrier.

3. Use of the compound of claim 1 in the preparation of a medicament for the prevention or treatment of clinical conditions related to HDAC1 or HDAC6.

4. The use according to claim 3, characterized in that, The disease associated with HDAC1 or HDAC6 is gastric cancer.

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