WO2023185667A1 - 一种hdac11亚型选择性抑制剂及其制备方法和应用 - Google Patents

一种hdac11亚型选择性抑制剂及其制备方法和应用 Download PDF

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WO2023185667A1
WO2023185667A1 PCT/CN2023/083623 CN2023083623W WO2023185667A1 WO 2023185667 A1 WO2023185667 A1 WO 2023185667A1 CN 2023083623 W CN2023083623 W CN 2023083623W WO 2023185667 A1 WO2023185667 A1 WO 2023185667A1
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compound
reaction
reacts
room temperature
tetrahydrofuran
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张颖杰
赵伟
张国真
柴启鹏
李顺达
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山东大学
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Definitions

  • the invention relates to the technical fields of organic compound synthesis and medical application, and in particular to an HDAC11 subtype selective inhibitor and its preparation method and application.
  • Histone deacetylase is an important class of epigenetic modification-related proteins. There are 18 subtypes of human HDAC discovered so far, among which HDAC1-11 are zinc ion-dependent metalloproteases. HDAC11 is the latest zinc ion-dependent HDAC discovered. It is distributed in both the cytoplasm and nucleus and plays a variety of physiological and pathological functions. It is worth pointing out that multiple recent studies have shown that HDAC11 has very strong deacylation activity of long-chain fatty acids (see: CaoJ.et al., Proc Natl Acad Sci USA.2019,116,5487-5492; Kutil Z.et al., ACS Chem. Biol. 2018, 13, 685-693).
  • High expression of HDAC11 is associated with hepatocellular carcinoma, myeloproliferative neoplasms, multiple myeloma, Hodgkin lymphoma, non-small cell lung cancer, glioblastoma, pituitary tumors, prostate cancer, ovarian cancer, acute lymphoblastic leukemia, etc.
  • the occurrence, development and poor prognosis of various cancers are closely related, so it is regarded as a potential cancer treatment target (see: Liu S. et al., Biomed. Pharmacother, 2020, 131, 110607; Y. et al., FEBS J. https://doi.org/10.1111/febs.15895).
  • HDAC11 histone deacetylase 11
  • HDAC11 knockout/knockdown can effectively inhibit the growth, invasion and metastasis of hepatocellular carcinoma, reduce cancer stem cell (CSC) stemness and sorafenib resistance (see: Bi L. et al., Cancer Res. 2021, 81, 2015 -2028; Wang W. et al., Front. Cell Dev. Biol. 2020, 8, 724).
  • CSC cancer stem cell
  • HDAC11 is a potential cancer therapeutic target.
  • HDAC11 is crucial for the maintenance of CSC stemness (see: Bi L. et al., Cancer Res.
  • HDAC11 isoform-selective inhibitors are also expected to solve the difficult problems of cancer drug resistance, recurrence and metastasis during cancer treatment.
  • HDAC11 selective inhibitors are also expected to be used in autoimmune diseases such as inflammation, psoriasis, rheumatoid arthritis, rheumatoid arthritis, and systemic lupus erythematosus, as well as metabolic diseases such as obesity and diabetes. Treatment of diseases (see: Liu S. et al., Biomed. Pharmacother, 2020, 131, 110607; Y. et al., FEBS J. https://doi.org/10.1111/febs.15895).
  • HDAC11 subtype selective inhibitors At present, the research and development of HDAC11 subtype selective inhibitors is in the ascendant at home and abroad, and only three compounds (FT895, SIS17, garcinol) with strong HDAC11 selective inhibitory activity have been reported (see: Y.et al., FEBS J. https://doi.org/10.1111/febs.15895), and its activity has not been fully studied.
  • the present invention provides a selective inhibitor of HDAC11 subtype.
  • the present invention also provides the preparation method and application of this type of compound.
  • Inhibitors having the structure represented by the following general formula A or B, as well as their optical isomers and pharmaceutically acceptable salts:
  • R 1 is an aromatic ring, aromatic heterocycle, substituted aromatic ring or substituted aromatic heterocycle; wherein, the substituent in the substituted aromatic ring or substituted aromatic heterocycle is selected from alkyl, aminoalkyl, halogen, haloalkyl, alkoxy , amino, amine, piperazinyl, alkylpiperazinyl, morpholinyl, alkylmorpholinyl, cyano, alkylamido or amide;
  • R 2 is hydrogen, alkyl, alkylpiperazinyl, alkylmorpholinyl or aminealkyl;
  • R 3 is hydroxyl, amino or primary amino group.
  • R 1 is benzene ring, substituted benzene ring, pyridine ring, substituted pyridine ring, pyrazine ring, substituted pyrazine ring, pyrimidine ring or substituted pyrimidine ring; wherein, the substituted benzene ring, substituted pyridine ring, substituted pyrazine ring or Substituents for the pyrimidine ring are selected from
  • R 3 is a hydroxyl group, an amino group, or an alkyl-substituted primary amino group containing 1 to 16 carbon atoms.
  • the structure of the inhibitor is one of the following:
  • the preparation method of the inhibitor in which R3 is hydroxyl in the general formula A of the inhibitor is selected from one of the following:
  • compound 4 is deprotected to form compound 16, and compound 16 is The reaction produces compound 17, and compound 17 reacts with potassium hydroxylamine to obtain compounds A51-A55.
  • the substituent R 1 is the same as the substituent R 1 in the corresponding compound A1-A13; in the reaction formula for preparing compounds A51-A55, the substituents R 1 and R 2 are the same as the substituent R 1 in the corresponding compound A51-A55.
  • the substituents R 1 and R 2 in A51-A55 are the same;
  • the substituent R 1 in the reaction formula is the same as the substituent R 1 in the corresponding compounds A14-A20;
  • eR 1 I copper iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;
  • eR 1 I copper iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;
  • the preparation method of the inhibitor in which R3 in the general formula A of the inhibitor is an amino or primary amino group is selected from one of the following:
  • compound 20 undergoes a reductive amination reaction with an aliphatic aldehyde to form compound 21, and compound 21 is deprotected to form compound A57;
  • the substituent R 1 in the reaction formula is the same as the substituent R 1 in compound A56 and compound A57, which is phenyl; in compound 21 and compound A57, n is 2.
  • compound A58 undergoes a reductive amination reaction with a fatty aldehyde to form compounds A59-A61.
  • substituents R 1 and R 2 in the reaction formula are the same as the substituents R 1 and R 2 in compounds A58-A61; in compound A59, n is 2; in compound A60, n is 15; in compound A61, n is 2;
  • the preparation method of the inhibitor in which R3 is a hydroxyl group in the general formula B of the inhibitor is selected from one of the following:
  • eR 1 I copper iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;
  • eR 1 I copper iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;
  • HDAC11 isoform selective inhibitors in the preparation of drugs for preventing or treating diseases associated with abnormal expression or activity of HDAC11.
  • the diseases related to abnormal expression or activity of HDAC11 are cancer, autoimmune diseases or metabolic diseases.
  • the cancer is liver cancer, myeloproliferative neoplasm, multiple myeloma, Hodgkin lymphoma, non-small cell lung cancer, glioblastoma, pituitary tumor, prostate cancer, ovarian cancer or acute lymphoblastic leukemia.
  • the autoimmune disease is inflammation, psoriasis, rheumatoid arthritis, rheumatoid arthritis or systemic lupus erythematosus.
  • the metabolic disease is obesity or diabetes.
  • a pharmaceutical composition for preventing or treating cancer, autoimmune diseases or metabolic diseases comprising the HDAC11 subtype selective inhibitor of the present invention or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carrier or excipient.
  • the HDAC11 subtype selective inhibitor of the present invention has a completely new structure; compared with existing inhibitors, it has potential advantages in resisting drug resistance, recurrence and metastasis of hepatocellular carcinoma.
  • the HDAC11 subtype selective inhibitor of the present invention has good inhibitory activity and subtype selectivity for HDAC11; exhibits certain anti-proliferative activity on tumor cells and has low toxicity to normal cells; and can inhibit the colony formation of hepatocellular carcinoma cells. And cancer stem cells form balls; at the same time, it can effectively inhibit the migration of hepatocellular carcinoma cells.
  • the reagents used in the examples are all commercially available; unless otherwise specified, the methods and equipment used can be based on existing technology.
  • Example 1 Preparation of compounds A1-A13 and B1-B3, taking compound A1 as an example.
  • Compound 2 NaH (150 mg, 60 wt%) was added to the flask, then 5 mL of THF was added to the flask at 0°C.
  • Compound 1 was dissolved in 10 mL of THF, and then compound 1 (1 g, 5.7 mmol) was added to the stirring solution of NaH via a syringe. After 30 minutes, TsCl (p-toluenesulfonyl chloride, 1.19 g, 6.2 mmol) was added to the solution. The resulting mixture was stirred at 20°C for 6 hours. After the reaction was completed, 100 mL of ice water was added, and the solid was precipitated and filtered.
  • Target compound A1 Weigh hydroxylamine hydrochloride (5.0g, 72mmol), dissolve it in 12mL of anhydrous methanol, and stir in an ice bath. Weigh KOH (6.06g, 108mmol), add it to 20mL of anhydrous methanol, and stir until dissolved. Add the KOH solution dropwise to the hydroxylamine hydrochloride solution under ice bath conditions, and continue stirring for 1 hour. Filter to obtain potassium hydroxylamine solution. Compound 4a (180 mg, 0.42 mmol) was dissolved in potassium hydroxylamine solution (6 mL) and stirred at room temperature for 10 hours.
  • Example 2 Preparation of compounds A14-A20 and B4-B6, taking compound A17 as an example.
  • Compound 5 Compound 1 (1g, 5.71mmol) was dissolved in 20mL DCM (dichloromethane), triethylamine (1mL) and di-tert-butyl dicarbonate (1.49g, 6.85mmol) were added, and stirred at room temperature for 5h. After the reaction, column chromatography was performed and petroleum ether/ethyl acetate (volume ratio 100/1) was used to obtain transparent oil 5 (1.4 g, yield 89%).
  • Compound 10a Weigh hydroxylamine hydrochloride (5.0 g, 72 mmol), dissolve it in 12 mL of anhydrous methanol, and stir in an ice bath. Weigh KOH (6.06g, 108mmol), add it to 20mL of anhydrous methanol, and stir until dissolved. Add the KOH solution dropwise to the hydroxylamine hydrochloride solution under ice bath conditions, and continue stirring for 1 hour. Filter to obtain potassium hydroxylamine solution. Compound 9a (130 mg, 0.30 mmol) was dissolved in potassium hydroxylamine solution (6 mL) and stirred at room temperature for 10 hours.
  • Target compound A17 To a CH 2 Cl 2 solution (5 mL) of compound 10a (434 mg, 1.0 mmol), 2 mL of TFA (trifluoroacetic acid) was added, and the mixture was stirred at room temperature for 3 hours. After the reaction, the reaction solution was washed three times with saturated Na 2 CO 3 solution, and then concentrated to obtain a residue. The residue was subjected to petroleum ether/ethyl acetate (volume ratio 100/10) column chromatography to obtain light yellow solid A17 (217 mg, yield 65%).
  • TFA trifluoroacetic acid
  • Example 3 Preparation of compounds A21-A50 and B7, taking compound A21 as an example.
  • Compound 11a Compound 1 (1.0g, 5.7mmol) was dissolved in THF (5mL), NaH (150mg, 60wt%) was slowly added in an ice bath, stirred at 0°C for 30 minutes, and then CH 3 I (1.2g, 8.4) was added dropwise mmol), moved to room temperature and reacted for 5 hours. opposite After completion, add NH 4 Cl to quench, extract with ethyl acetate, combine the organic phases, dry over anhydrous magnesium sulfate, and concentrate to obtain colorless and transparent oil 11a (0.82g, yield 77%).
  • Compound 12a Compound 11a (0.27g, 0.85mmol) was dissolved in 10mL of ultra-dry THF, n-BuLi (0.4mL, 2.5M hexane solution) was added to the solution of compound 11a at -78°C. After reacting at this temperature for 30 minutes, add iodine (0.24g, 0.95mmol) in 5 mLTHF solution. After reacting at this temperature for 30 minutes, move to room temperature and continue the reaction for 2 hours.
  • reaction is quenched with saturated ammonium chloride solution, extracted with ethyl acetate, the organic layer is washed with brine, dried with MgSO 4 , and then concentrated to obtain crude compound 12a, which is used in the next step.
  • Compound 14a Compound 13a (131 mg, 0.46 mmol) was dissolved in THF (5 mL), TBAF (0.18 g, 0.69 mmol) was added, and stirred at 70°C for 4 hours. After the reaction was completed, column chromatography was performed using petroleum ether/ethyl acetate (volume ratio 100/1) to obtain solid 14a (69 mg, yield 70%).
  • Target compound A21 Weigh hydroxylamine hydrochloride (5.00g, 72mmol), dissolve it in 12mL of anhydrous methanol, and stir in an ice bath. Weigh KOH (6.06g, 108mmol), add it to 20mL of anhydrous methanol, and stir until dissolved. Add the KOH solution dropwise to the hydroxylamine hydrochloride solution under ice bath conditions, and continue stirring for 1 hour. Filter to obtain potassium hydroxylamine solution. Compound 15a (90 mg, 0.31 mmol) was dissolved in potassium hydroxylamine solution (6 mL) and stirred at room temperature for 10 hours.
  • Compound 17a Compound 16a (0.55g, 2.0mmol) was dissolved in THF (10mL), NaH (55mg, 60wt%) was slowly added in an ice bath, stirred at 0°C for 30 minutes, and then 2-(dimethylamino)ethane was added dropwise. A solution of 4-methylbenzenesulfonate (0.61g, 2.5mmol) in 8mL THF was reacted at room temperature for 8 hours. After the reaction was completed, NH 4 Cl was added to quench, and the ethyl acetate extraction was performed.
  • Target compound A51 Weigh hydroxylamine hydrochloride (5.00g, 72mmol), dissolve it in 12mL of anhydrous methanol, and stir in an ice bath. Weigh KOH (6.06g, 108mmol), add it to 20mL of anhydrous methanol, and stir until dissolved. Add the KOH solution dropwise to the hydroxylamine hydrochloride solution under ice bath conditions, and continue stirring for 1 hour. Filter to obtain potassium hydroxylamine solution. Compound 17a (104 mg, 0.30 mmol) was dissolved in potassium hydroxylamine solution (6 mL) and stirred at room temperature for 10 hours.
  • the preparation method of compounds A52-A55 is similar to the preparation method of compound A51.
  • Compound 20a Dissolve compound 19a (0.23g, 0.65mmol) in DMSO (5mL), add TBTU (O-benzotriazole-N,N,N',N'-tetramethylurea tetrazole) under ice bath Fluoboric acid, 0.25g, 0.78mmol), TEA (triethylamine, 180uL), add 80wt% hydrazine hydrate after 30 minutes, and react at room temperature for 3 hours. After the reaction was completed, water was added to precipitate the solid, which was filtered to obtain yellow solid 20a (0.11 g, yield 45%).
  • Target compound A56 To a CH 2 Cl 2 solution (5 mL) of compound 20a (375 mg, 1.0 mmol), 2 mL of TFA was added, and the mixture was stirred at room temperature for 3 hours. After the reaction, the reaction solution was washed three times with saturated Na 2 CO 3 solution, and then concentrated to obtain a residue. The residue was subjected to petroleum ether/ethyl acetate (volume ratio 100/10) column chromatography to obtain light yellow solid A56 (220 mg, yield 80%).
  • Target compound A57 To a CH 2 Cl 2 solution (5 mL) of compound 21a (417 mg, 1.0 mmol), 2 mL of TFA was added, and the mixture was stirred at room temperature for 3 hours. After the reaction, the reaction solution was washed three times with saturated Na 2 CO 3 solution, and then concentrated to obtain a residue. The residue was subjected to petroleum ether/ethyl acetate (volume ratio 100/10) column chromatography to obtain light yellow solid A57 (238 mg, yield 75%).
  • Target compound A58 Dissolve compound 22a (0.18g, 0.65mmol) in DMSO (5mL), add TBTU (0.25g, 0.78mmol) and TEA (180uL) under ice bath, add 80wt% hydrazine hydrate after 30 minutes, and keep at room temperature Reaction takes 3 hours. After the reaction was completed, water was added to precipitate the solid, which was filtered to obtain yellow solid A58 (90 mg, yield 48%).
  • the NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d 6 ) ⁇ 9.49 (s, 1H), 7.82 (s, 1H), 7.65-7.60 (m, 3H), 7.40-7.26 (m, 4H) ,7.16-7.13(m,1H),4.49(s,2H),3.85(s,3H).
  • ESI-MS, m/z 290.3[M+H] + .
  • Example 8 Preparation of compounds A59-A61, taking compound A59 as an example.
  • the preparation method of compounds A60-A61 is similar to the preparation method of compound A59.
  • the present invention uses a fluorescence analysis method to measure the inhibitory activity of HDAC11, HDAC1 and HDAC6 of the target compound, and uses the HDAC inhibitor SAHA, which has been approved for marketing, as a positive control.
  • the target compounds A1 and B1 were selected as representatives, and their half inhibitory concentrations (IC 50 ) for HDAC1, HDAC4, HDAC6, HDAC8 and HDAC11 were measured.
  • IC 50 half inhibitory concentrations
  • Table 2 show that the IC 50 of A1 and B1 against HDAC11 are 0.031 ⁇ M and 0.17 ⁇ M respectively, which are far lower than the IC 50 against other HDAC subtypes, further confirming that A1 and B1 have good selective inhibition of HDAC11 subtypes. active.
  • Test Example 2 In vitro anti-proliferation test of tumor cells and normal cytotoxicity test of the target compound
  • Select target compound A1 as a representative to evaluate the in vitro anti-proliferative activity of this compound against human hepatocellular carcinoma cells (Huh7 and PLC/PRF/5), human erythroid leukemia cells (HEL), mouse myeloma cells P3x63Ag8.653, and Toxicity to human normal liver cells HL-7702 and human umbilical vein endothelial cells HUVEC.
  • HEL human erythroid leukemia cells
  • P3x63Ag8.653 mouse myeloma cells
  • Toxicity to human normal liver cells HL-7702
  • HUVEC human umbilical vein endothelial cells
  • Test Example 3 Target compound inhibits colony formation of hepatocellular carcinoma cells and sphere formation of cancer stem cells in vitro
  • the target compound A1 was selected as a representative to evaluate the compound's inhibitory activity on the colony formation of hepatocellular carcinoma cells and the spheroidization of cancer stem cells in vitro.
  • the results of the colony formation experiment showed that at a concentration of 2 ⁇ M, compared with the Ctrl group (dmso treatment), both compound A1 and sorafenib can effectively inhibit the formation of Huh7 colony formation of hepatocellular carcinoma cells, and A1 has a stronger inhibitory effect on Huh7 cell formation than sorafenib.
  • the ability of colonies (Figure 1), which may be related to the inhibitory effect of A1 on HCC tumor stem cells.
  • Primary spherification refers to the spheroidization experiment of Huh7 cells
  • secondary spherification refers to the spheroidization experiment performed with cancer stem cells digested from the tumor stem cell spheres after the primary spheroidization of Huh7 cells.
  • the scale bar in Figure 2 is 200 ⁇ m.
  • Test Example 4 Target compound inhibits the migration of hepatocellular carcinoma cells in vitro (scratch test)
  • the target compound A1 was selected as a representative to evaluate the compound's inhibitory activity on the migration of hepatocellular carcinoma cells in vitro.
  • the scratch test results show that after treating cells with 2 ⁇ M concentration of compounds for 12 hours, compared with the Ctrl group (dmso treatment), both compound A1 and sorafenib can effectively inhibit the migration of Huh7 cells, and A1 has a stronger effect than sorafenib. Anti-Huh7 cell migration activity.
  • HDAC11 subtype-selective inhibitors have good therapeutic prospects for cancers such as hepatocellular carcinoma, and also have the potential advantages of anti-drug resistance, anti-recurrence and anti-metastasis.

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Abstract

本发明提供一种HDAC11亚型选择性抑制剂及其制备方法和应用。本发明抑制剂是具有如下结构通式A或B所示结构的抑制剂,以及其光学异构体,药学上可接受的盐。本发明抑制剂具有抗肝细胞癌耐药、复发和转移的潜在优势,可应用于制备预防或治疗与HDAC11表达或活性异常相关的疾病的药物。

Description

一种HDAC11亚型选择性抑制剂及其制备方法和应用 技术领域
本发明涉及有机化合物合成与医药应用技术领域,特别涉及一种HDAC11亚型选择性抑制剂及其制备方法和应用。
背景技术
组蛋白去乙酰化酶(HDAC)是一类重要的表观遗传修饰相关蛋白。目前发现的人源HDAC共有18个亚型,其中HDAC1-11是锌离子依赖型金属蛋白酶。HDAC11是最晚发现的锌离子依赖型HDAC,其在细胞质和细胞核中均有分布,并发挥多种生理和病理功能。值得指出的是,最近的多项研究表明HDAC11具有非常强的去长链脂肪酸酰化活性(参见:CaoJ.et al.,Proc Natl Acad Sci USA.2019,116,5487-5492;Kutil Z.et al.,ACS Chem.Biol.2018,13,685-693)。
HDAC11的高表达与肝细胞癌、骨髓增殖性肿瘤、多发性骨髓瘤、霍奇金淋巴瘤、非小细胞肺癌、胶质母细胞瘤、垂体瘤、前列腺癌、卵巢癌、急性淋巴细胞白血病等多种癌症的发生、发展和预后不良密切相关,因此被视为一种潜在的癌症治疗靶标(参见:Liu S.et al.,Biomed.Pharmacother,2020,131,110607;Y.et al.,FEBS J.https://doi.org/10.1111/febs.15895)。例如,近年的多项研究均发现,组蛋白去乙酰化酶11(HDAC11)在肝细胞癌组织/细胞中的表达水平显著高于正常肝组织/细胞,且HDAC11高表达与索拉非尼(sorafenib)耐药和患者预后不良密切相关(参见:Bi L.et al.,Cancer Res.2021,81,2015-2028;Wang W.et al.,Front.Cell Dev.Biol.2020,8,724;Freese K.et al.,Cancers 2019,11,1587;Gong D.et al.,Am.J.Transl.Res.2019,11,983-990)。相应的,HDAC11敲除/敲减能有效抑制肝细胞癌生长和侵袭转移、降低肿瘤干细胞(CSC)干性和sorafenib耐药(参见:Bi L.et al.,Cancer Res.2021,81,2015-2028;Wang W.et al.,Front.Cell Dev.Biol.2020,8,724)。综上,HDAC11是一种潜在的癌症治疗靶标。又由于HDAC11对CSC干性的维持至关重要(参见:Bi L.et al.,Cancer Res.2021,81,2015-2028),而CSC又与肿瘤耐药、复发和转移密切相关(参见:Lytel N,et al.,Nat.Rev.Cancer 2018,18,669-680),因此HDAC11亚型选择性抑制剂还有望解决癌症治疗过程中棘手的癌症耐药、复发和转移问题。
此外,还有大量研究表明HDAC11选择性抑制剂还有望用于炎症、银屑病、风湿性关节炎、类风湿性关节炎、系统性红斑狼疮等自身免疫性疾病,以及肥胖症和糖尿病等代谢性疾病的治疗(参见:Liu S.et al.,Biomed.Pharmacother,2020,131,110607;Y.et al.,FEBS J.https://doi.org/10.1111/febs.15895)。
目前,国内外HDAC11亚型选择性抑制剂的研究开发方兴未艾,仅有3个具有较强HDAC11选择性抑制活性的化合物(FT895、SIS17、garcinol)被报道(参见:Y.et al.,FEBS J.https://doi.org/10.1111/febs.15895),且对其活性研究尚不充分。
发明内容
针对现有技术的不足,本发明提供一种HDAC11亚型选择性抑制剂,本发明还提供了该类化合物的制备方法和应用。
本发明的技术方案如下:
1.HDAC11亚型选择性抑制剂
具有如下结构通式A或B所示结构的抑制剂,以及其光学异构体,药学上可接受的盐:
其中,
R1是芳环、芳杂环、取代芳环或取代芳杂环;其中,取代芳环或取代芳杂环中的取代基选自烷基、胺烷基、卤素、卤代烷基、烷氧基、氨基、胺基、哌嗪基、烷基哌嗪基、吗啉基、烷基吗啉基、氰基、烷基酰胺基或酰胺基;
R2是氢、烷基、烷基哌嗪基、烷基吗啉基或胺烷基;
R3是羟基、氨基或伯胺基。
根据本发明优选的,
R1是苯环、取代苯环、吡啶环、取代吡啶环、吡嗪环、取代吡嗪环、嘧啶环或取代嘧啶环;其中,所述取代苯环、取代吡啶环、取代吡嗪环或取代嘧啶环的取代基选自
R2
R3是羟基、氨基、含1-16个碳原子烷基取代的伯胺基。
优选的,所述抑制剂的结构为下列之一:

2.HDAC11亚型选择性抑制剂的制备方法
抑制剂结构通式A中R3为羟基的抑制剂的制备方法选自以下之一:
(一)化合物1与对甲苯磺酰氯反应生成化合物2,化合物2与四溴化碳反应生成化合物3,化合物3与通过Sonogashira偶联反应生成化合物4,化合物4与羟胺钾反应得到化合物A1-A13;
或者,化合物4脱去保护基生成化合物16,化合物16与反应生成化合物17,化合物17与羟胺钾反应得到化合物A51-A55。
反应式如下:
其中,制备化合物A1-A13的反应式中,取代基R1与相应化合物A1-A13中的取代基R1相同;制备化合物A51-A55的反应式中,取代基R1、R2与相应化合物A51-A55中的取代基R1、R2相同;
上述反应式中的试剂和条件:
a.对甲苯磺酰氯,氢化钠,四氢呋喃,室温反应;
b.四溴化碳,二异丙基氨基锂,四氢呋喃,-78℃和室温反应;
c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
d.羟胺钾,甲醇,室温反应;
e.四丁基氟化铵;四氢呋喃;70℃反应;
f.氢化钠,四氢呋喃,室温反应;
(二)化合物1与二碳酸二叔丁酯反应生成化合物5,化合物5与四溴化碳反应生成化合物6,化合物6与三甲基硅乙炔通过Sonogashira偶联反应生成化合物7,化合物7经反应生成化合物8,化合物8与R1I通过Sonogashira偶联反应生成化合物9,化合物9与羟胺钾反应得到化合物10,化合物10脱去保护基得到化合物A14-A20。
反应式如下:
其中,反应式中取代基R1与相应化合物A14-A20中的取代基R1相同;
上述反应式中的试剂和条件:
a.二碳酸二叔丁酯,三乙胺,二氯甲烷,室温反应;
b.四溴化碳,二异丙基氨基锂,四氢呋喃,-78℃和室温反应;
c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
d.四丁基氟化铵;四氢呋喃;70℃反应;
e.R1I,碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
f.羟胺钾,甲醇,室温反应;
g.三氟乙酸,二氯甲烷,室温反应;
(三)化合物1与R2I反应生成化合物11,化合物11与单质碘反应生成化合物12,化合物12与三甲基硅乙炔通过Sonogashira偶联反应生成化合物13,化合物13经反应生成化合物14,化合物14与R1I通过Sonogashira偶联反应生成化合物15,化合物15与羟胺钾反应得到化合物A21-A50;
反应式如下:
其中,反应式中取代基R1、R2与相应化合物A21-A50中的取代基R1、R2相同;
上述反应式中的试剂和条件:
a.R2I,氢化钠,四氢呋喃,室温反应;
b.碘,正丁基锂,四氢呋喃,-78℃和室温反应;
c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
d.四丁基氟化铵;四氢呋喃;70℃反应;
e.R1I,碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
f.羟胺钾,甲醇,室温反应;
抑制剂结构通式A中R3为氨基或伯胺基的抑制剂的制备方法选自以下之一:
(四)化合物8与R1I通过Sonogashira偶联反应生成化合物18,化合物18水解生成化合物19,化合物19与水合肼缩合得到化合物20,化合物20脱去保护基生成化合物A56;
或者,化合物20与脂肪醛发生还原胺化反应生成化合物21,化合物21脱去保护基生成化合物A57;
反应式如下:
其中,反应式中取代基R1与化合物A56、化合物A57中的取代基R1相同,为苯基;化合物21、化合物A57中,n为2。
上述反应式中的试剂和条件:
a.R1I,碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
b.氢氧化钠,甲醇,50℃反应;
c.水合肼,O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸,三乙胺,二甲亚砜,室温反应;
d.三氟乙酸,二氯甲烷,室温反应;
e.脂肪醛,氰基硼氢化钠,甲醇,冰醋酸,室温反应;
(五)化合物15水解生成化合物22,化合物22与水合肼缩合得到化合物A58;
或者,化合物A58与脂肪醛发生还原胺化反应生成化合物A59-A61。
反应式如下:
其中,反应式中取代基R1、R2与化合物A58-A61中的取代基R1、R2相同;化合物A59中,n为2;化合物A60中,n为15;化合物A61中,n为2;
上述反应式中的试剂和条件:
a.氢氧化钠,甲醇,50℃反应;
b.水合肼,O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸,三乙胺,二甲亚砜,室温反应;
c.脂肪醛,氰基硼氢化钠,甲醇,冰醋酸,室温反应;
抑制剂结构通式B中R3为羟基的抑制剂的制备方法选自以下之一:
(六)化合物23与对甲苯磺酰氯反应生成化合物24,化合物24与四溴化碳反应生成化合物25,化合物25与通过Sonogashira偶联反应生成化合物26,化合物26与羟胺钾反应得到化合物B1-B3。
反应式如下:
其中,反应式中取代基R1与相应化合物B1-B3中的取代基R1相同;
上述反应式中的试剂和条件:
a.对甲苯磺酰氯,氢化钠,四氢呋喃,室温反应;
b.四溴化碳,二异丙基氨基锂,四氢呋喃,-78℃和室温反应;
c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
d.羟胺钾,甲醇,室温反应;
(七)化合物23与二碳酸二叔丁酯反应生成化合物27,化合物27与四溴化碳反应生成化合物28,化合物28与三甲基硅乙炔通过Sonogashira偶联反应生成化合物29,化合物29经反应生成化合物30,化合物30与R1I通过Sonogashira偶联反应生成化合物31,化合物31与羟胺钾反应得到化合物32,化合物32脱去保护基得到化合物B4-B6。
反应式如下:
其中,反应式中取代基R1与相应化合物B4-B6中的取代基R1相同;
上述反应式中的试剂和条件:
a.二碳酸二叔丁酯,三乙胺,二氯甲烷,室温反应;
b.四溴化碳,二异丙基氨基锂,四氢呋喃,-78℃和室温反应;
c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
d.四丁基氟化铵;四氢呋喃;70℃反应;
e.R1I,碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
f.羟胺钾,甲醇,室温反应;
g.三氟乙酸,二氯甲烷,室温反应;
(八)化合物23与R2I生成化合物33,化合物33与单质碘反应生成化合物34,化合物34与三甲基硅乙炔通过Sonogashira偶联反应生成化合物35,化合物35经反应生成化合物36,化合物36与R1I通过Sonogashira偶联反应生成化合物37,化合物37与羟胺钾反应得到化合物B7。
反应式如下:
其中,反应式中取代基R1、R2与相应化合物B7中的取代基R1、R2相同;
上述反应式中的试剂和条件:
a.R2I,氢化钠,四氢呋喃,室温反应;
b.碘,正丁基锂,四氢呋喃,-78℃和室温反应;
c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
d.四丁基氟化铵;四氢呋喃;70℃反应;
e.R1I,碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
f.羟胺钾,甲醇,室温反应;
3.HDAC11亚型选择性抑制剂的应用
HDAC11亚型选择性抑制剂在制备预防或治疗与HDAC11表达或活性异常相关的疾病的药物中的应用。
所述的与HDAC11表达或活性异常相关的疾病为癌症、自身免疫性疾病或代谢性疾病。
所述的癌症为肝癌、骨髓增殖性肿瘤、多发性骨髓瘤、霍奇金淋巴瘤、非小细胞肺癌、胶质母细胞瘤、垂体瘤、前列腺癌、卵巢癌或急性淋巴细胞白血病。
所述的自身免疫性疾病为炎症、银屑病、风湿性关节炎、类风湿性关节炎或系统性红斑狼疮。
所述的代谢性疾病为肥胖症或糖尿病。
一种预防或治疗癌症、自身免疫性疾病或代谢性疾病的药物组合物,包含本发明的HDAC11亚型选择性抑制剂或其药学上可接受的盐以及一种或多种药学上可接受的载体或赋形剂。
本发明的有益效果如下:
本发明HDAC11亚型选择性抑制剂具有全新的结构;与现有抑制剂相比,具有抗肝细胞癌耐药、复发和转移的潜在优势。本发明HDAC11亚型选择性抑制剂对于HDAC11具有较好的抑制活性和亚型选择性;对于肿瘤细胞表现出一定的抗增殖活性,且对正常细胞毒性较低;能抑制肝细胞癌细胞集落形成和肿瘤干细胞成球;同时能有效抑制肝细胞癌细胞迁移。
附图说明
图1.试验例3中化合物A1和sorafenib对Huh7细胞集落形成的抑制活性;
图2.试验例3中化合物A1和sorafenib对Huh7肿瘤干细胞成球的抑制活性;
图3.试验例4中化合物A1和sorafenib对Huh7细胞迁移的抑制活性。
具体实施方式
下面结合实施例对本发明做进一步的说明,但不限于此。
同时,实施例中所用试剂如无特殊说明,均可市购获得;所用方法和设备如无特殊说明,可按现有技术。
实施例1.化合物A1-A13、B1-B3的制备,以化合物A1为例。
合成路线:
具体合成方法和步骤如下:
化合物2:将NaH(150mg,60wt%)加入烧瓶中,然后在0℃下将5mL THF加入烧瓶中。化合物1溶于10mLTHF,然后通过注射器将化合物1(1g,5.7mmol)添加到NaH的搅拌溶液中,30分钟后,将TsCl(对甲苯磺酰氯,1.19g,6.2mmol)添加到溶液中。将所得混合物在20℃下搅拌6小时。反应完成后向其中加入100mL冰水,析出固体后过滤,干燥后EtOAc/PE(1/5,20mL)进一步打浆得白色固体2(1.76g,产率94%)。产物的核磁数据如 下:1H NMR(400MHz,DMSO-d6)δ8.23(d,J=8.3Hz,1H),8.01(d,J=3.7Hz,1H),7.90(dd,J=7.6,5.4Hz,3H),7.48(t,J=8.0Hz,1H),7.39(d,J=8.2Hz,2H),7.29(d,J=3.6Hz,1H),3.89(s,3H),2.32(s,3H)。ESI-MS,m/z=330.3[M+H]+
化合物3:在-78℃下将LDA(二异丙基氨基锂,1.2mL,2M的THF溶液)添加到化合物2(660mg,2mmol)的THF溶液(5mL)中,30分钟后滴加CBr4(797mg,2.4mmol)的THF溶液(5mL),在该温度下反应30分钟后,移至室温反应30分钟。反应完成后采用乙酸乙酯(50mL)和HCl(0.5M,40mL)萃取,用盐水清洗有机层,并在MgSO4上干燥,然后浓缩得到残渣,残渣进行柱层析,石油醚/乙酸乙酯(体积比:100/10)得白色固体3(130mg,产率16%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ8.46(d,J=8.5Hz,1H),7.93(d,J=7.7Hz,1H),7.81-7.76(m,2H),7.52(t,J=8.1Hz,1H),7.47-7.40(m,3H),3.88(s,3H),2.34(s,3H)。
化合物4a:将化合物3(340mg,0.8mmol),CuI(32mg,0.2eq)、Pd(PPh3)2Cl2(双(三苯基膦)二氯化钯,130mg,0.2eq)溶于1,4-二氧六环(4mL)中,置换氩气后添加Et3N(1.5mL)、乙炔基苯(102mg,1mmol),将所得混合物在70℃下搅拌20小时,浓缩反应液,并用硅胶柱(PE/EA=5/1)纯化,得到棕色油状物4a(280mg,产率82%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ8.46(d,J=8.5Hz,1H),7.93(d,J=7.6Hz,2H),7.79(d,J=8.4Hz,2H),7.77-7.58(m,1H),7.52(t,J=8.1Hz,2H),7.45(s,2H),7.42(d,J=8.2Hz,2H),3.88(s,3H),2.34(s,3H)。ESI-MS,m/z=428.3[M-H]-
目标化合物A1:称取盐酸羟胺(5.0g,72mmol)溶于12mL无水甲醇中,冰浴搅拌。称取KOH(6.06g,108mmol)后加入20mL无水甲醇中,搅拌至溶解。冰浴条件下将KOH溶液逐滴加入盐酸羟胺溶液中,继续搅拌1小时。过滤得到羟胺钾溶液。将化合物4a(180mg,0.42mmol)溶于羟胺钾溶液(6mL)室温搅拌10小时。反应完成后旋干液体,加水溶解均匀后,滴加1N盐酸调节pH至中性析出固体,过滤得粗品。通过反相柱层析(MeCN/H2O=50%/50%,HAc条件)纯化,得淡黄色固体A1(90mg,产率50%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ11.92(s,1H),11.02(s,1H),8.99(s,1H),7.64-7.56(m,2H),7.52-7.43(m,4H),7.36(d,J=7.2Hz,1H),7.21(t,J=7.7Hz,1H),7.13(s,1H)。ESI-MS,m/z=277.1[M+H]+
化合物A2-A13、B1-B3的制备方法与化合物A1的制备方法类似。
实施例2.化合物A14-A20、B4-B6的制备,以化合物A17为例。
合成路线:
具体合成方法和步骤如下:
化合物5:化合物1(1g,5.71mmol)溶于20mLDCM(二氯甲烷),加入三乙胺(1mL)和二碳酸二叔丁酯(1.49g,6.85mmol),室温搅拌5h。反应结束后进行柱层析,石油醚/乙酸乙酯(体积比100/1)得透明油状物5(1.4g,产率89%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ8.36(d,J=8.2Hz,1H),7.94-7.83(m,2H),7.45(t,J=8.0Hz,1H),7.20(d,J=3.7Hz,1H),3.92(s,3H),1.64(s,9H)。ESI-MS,m/z=310.1[M+Cl]-
化合物6:将化合物5(0.24g,0.87mmol)溶于5mL超干THF中,在-78℃下将LDA(0.65mL,2M的THF溶液)添加到化合物5的溶液中,在该温度下反应30分钟后,滴加CBr4(0.36g,1.1mmol)的THF溶液(5mL),在该温度下反应30分钟后,移至室温继续反应30分钟。反应完成后用乙酸乙酯和HCl(0.5M)萃取,用盐水洗涤有机层,并用MgSO4干燥,然后浓缩得到残渣,残渣用石油醚/乙酸乙酯(体积比100/10)柱层析,得淡黄色油状物6(0.1g,产率33%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ8.27(d,J=8.4Hz,1H),7.90(d,J=7.6Hz,1H),7.45(t,J=8.0Hz,1H),7.39(s,1H),3.92(s,3H),1.67(s,9H)。
化合物7:将化合物6(100mg,0.28mmol),CuI(11mg,0.05mmol)、Pd(PPh3)2Cl2(35mg,0.05mmol)溶于1,4-二氧六环(4mL)中,置换氩气后添加Et3N(2mL)、三甲基硅基乙炔(41mg,0.42mmol),将所得混合物在70℃下搅拌20小时,浓缩反应液,并用硅胶柱(PE/EA=5/1)柱层析纯化,得到棕色油状物7(70mg,产率67%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ8.35(d,J=8.4Hz,1H),7.91(d,J=7.6Hz,1H),7.53-7.48(m,2H),3.91(s,3H),1.64(s,9H),0.26(s,9H)。
化合物8:将化合物7(170mg,0.45mmol)溶于THF(5mL),加入TBAF(四丁基氟化铵,0.18g,0.69mmol),于70℃搅拌4小时。反应完成后进行柱层析,石油醚/乙酸乙酯(体积比100/1)得固体8(92mg,产率68%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ8.41(d,J=8.4Hz,1H),7.99(d,J=7.6Hz,1H),7.62-7.55(m,2H),4.86(s,1H),3.98(s,3H),1.71(s,9H)。
化合物9a:将化合物1-(4-碘苯基)-N,N-二甲基甲胺(73mg,0.28mmol),CuI(11mg,0.05mmol)、Pd(PPh3)2Cl2(35mg,0.05mmol)溶于1,4-二氧六环(4mL)中,置换氩气后添加Et3N(2mL)、化合物8(125mg,0.42mmol),将所得混合物在70℃下搅拌20小时,浓缩反应液,并用硅胶柱(PE/EA=5/1)柱层析纯化,得到棕色油状物9a(67mg,产率55%)。ESI-MS,m/z=433.4[M+H]+
化合物10a:称取盐酸羟胺(5.0g,72mmol)溶于12mL无水甲醇中,冰浴搅拌。称取KOH(6.06g,108mmol)后加入20mL无水甲醇中,搅拌至溶解。冰浴条件下将KOH溶液逐滴加入盐酸羟胺溶液中,继续搅拌1小时。过滤得到羟胺钾溶液。将化合物9a(130mg,0.30mmol)溶于羟胺钾溶液(6mL)室温搅拌10小时。反应完成后旋干液体,加水溶解均匀后,滴加1N盐酸调节pH至固体析出完全,过滤得粗品。通过反相柱层析(MeCN/H2O=50%/50%,HAc条件)纯化,得淡黄色固体10a(87mg,产率67%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ11.02(s,1H),8.97(s,1H),7.76-7.23(m,8H),3.39(s,2H),2.13(s,6H),1.63(s,9H)。
目标化合物A17:向化合物10a(434mg,1.0mmol)的CH2Cl2溶液(5mL)中,加入2mLTFA(三氟乙酸),室温搅拌3小时。反应结束后将反应液用饱和Na2CO3溶液洗涤三次,然后浓缩得到残渣,残渣经石油醚/乙酸乙酯(体积比100/10)柱层析,得淡黄色固体A17(217mg,产率65%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ11.92(s,1H),11.02(s,1H),8.97(s,1H),7.64-7.15(m,8H),3.37(s,2H),2.15(s,6H)。ESI-MS,m/z=334.3[M+H]+
化合物A14-A16、A18-A20、B4-B6的制备方法与化合物A17的制备方法类似。
实施例3.化合物A21-A50、B7的制备,以化合物A21为例。
合成路线:
具体合成方法和步骤如下:
化合物11a:化合物1(1.0g,5.7mmol)溶于THF(5mL),冰浴下缓慢加入NaH(150mg,60wt%),0℃下搅拌30分钟后逐滴加入CH3I(1.2g,8.4mmol),移至室温反应5小时。反 应完成后加入NH4Cl淬灭,乙酸乙酯萃取,合并有机相,无水硫酸镁干燥,浓缩得无色透明油状物11a(0.82g,产率77%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ7.77(dd,J=7.7,3.7Hz,2H),7.53(d,J=2.9Hz,1H),7.27(t,J=7.8Hz,1H),6.93(d,J=2.5Hz,1H),3.90(s,3H),3.85(s,3H)。ESI-MS,m/z=378.6[2M+H]+
化合物12a:将化合物11a(0.27g,0.85mmol)溶于10mL超干THF中,在-78℃下将n-BuLi(0.4mL,2.5M的己烷溶液)添加到化合物11a的溶液中,在该温度下反应30分钟后,加入碘(0.24g,0.95mmol)的5mLTHF溶液,在该温度下反应30分钟后,移至室温继续反应2小时。反应完成后用饱和氯化铵溶液淬灭反应,用乙酸乙酯萃取,盐水洗涤有机层,并用MgSO4干燥,然后浓缩得到化合物12a粗品,用于下一步反应。
化合物13a:将化合物12a粗品(160mg),CuI(20mg,0.11mmol)、Pd(PPh3)2Cl2(77mg,0.11mmol)溶于1,4-二氧六环(4mL)中,置换氩气后添加Et3N(2mL)、三甲基硅乙炔(62mg,0.63mmol),将所得混合物在70℃下搅拌20小时,浓缩反应液,并用硅胶柱(PE/EA=10/1)纯化,得到棕色油状物13a(100mg)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ7.84(s,1H),7.66(d,J=8.2Hz,1H),7.43(d,J=7.3Hz,1H),7.24(t,J=7.8Hz,1H),3.78(s,3H),3.76(s,3H),0.15(s,9H)。ESI-MS,m/z=286.1[M+H]+
化合物14a:将化合物13a(131mg,0.46mmol)溶于THF(5mL),加入TBAF(0.18g,0.69mmol),于70℃搅拌4小时。反应完成后进行柱层析,石油醚/乙酸乙酯(体积比100/1)得固体14a(69mg,产率70%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ7.88(s,1H),7.74(d,J=8.2Hz,1H),7.50(d,J=7.3Hz,1H),7.31(t,J=7.8Hz,1H),3.97(s,1H),3.85(s,3H),3.84(s,3H)。
化合物15a:将化合物碘苯(108mg,0.53mmol),CuI(20mg,0.11mmol)、Pd(PPh3)2Cl2(77mg,0.11mmol)溶于1,4-二氧六环(4mL)中,置换氩气后添加Et3N(2mL)、化合物14a(134mg,0.63mmol),将所得混合物在70℃下搅拌20小时,浓缩反应液,并用硅胶柱(PE/EA=10/1)纯化,得到棕色油状物15a(101mg,产率66%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ7.85-7.80(m,2H),7.68(dd,J=6.1Hz,2.7Hz,2H),7.52-7.47(m,3H),7.38(t,J=7.8Hz,1H),7.33(s,1H),3.94(s,3H),3.93(s,3H)。
目标化合物A21:称取盐酸羟胺(5.00g,72mmol)溶于12mL无水甲醇中,冰浴搅拌。称取KOH(6.06g,108mmol)后加入20mL无水甲醇中,搅拌至溶解。冰浴条件下将KOH溶液逐滴加入盐酸羟胺溶液中,继续搅拌1小时。过滤得到羟胺钾溶液。将化合物15a(90mg,0.31mmol)溶于羟胺钾溶液(6mL)室温搅拌10小时。反应完成后旋干液体,加水溶解均匀后,滴加1N盐酸调节pH至中性析出固体,过滤得粗品。通过反相柱层析(MeCN/H2O=50%/50%,HAc条件)纯化,得淡黄色固体A21(60mg,产率67%)。产物 的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),9.04(s,1H),7.70-7.64(m,3H),7.51-7.46(m,3H),7.41(d,J=7.2Hz,1H),7.33-7.27(m,1H),7.19(s,1H),3.91(s,3H)。ESI-MS,m/z=291.5[M+H]+
化合物A22-A50、B7的制备方法与化合物A21制备方法类似。
实施例4.化合物A51-A55的制备,以化合物A51为例。
合成路线:
具体合成方法和步骤如下:
化合物16a:将化合物4a(0.2g,0.46mmol)溶于THF(5mL),加入TBAF(0.18g,0.69mmol),于70℃搅拌4小时。反应完成后进行柱层析,石油醚/乙酸乙酯(体积比100/1)得白色固体16a(90mg,产率70%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ12.06(s,1H),7.75(d,J=7.3Hz,1H),7.55-7.42(m,3H),7.50-7.44(m,3H),7.34-7.22(m,2H),3.84(s,3H)。
化合物17a:化合物16a(0.55g,2.0mmol)溶于THF(10mL),冰浴下缓慢加入NaH(55mg,60wt%),0℃下搅拌30分钟后逐滴加入2-(二甲氨基)乙基4-甲基苯磺酸酯(0.61g,2.5mmol)的8mLTHF溶液,室温反应8小时。反应完成后加入NH4Cl淬灭,乙酸乙酯萃取,合并有机相,无水硫酸镁干燥,浓缩后用硅胶柱(PE/EA=10/1)纯化,得油状物17a(0.52g,75%)备用。ESI-MS,m/z=347.3[M+H]+
目标化合物A51:称取盐酸羟胺(5.00g,72mmol)溶于12mL无水甲醇中,冰浴搅拌。称取KOH(6.06g,108mmol)后加入20mL无水甲醇中,搅拌至溶解。冰浴条件下将KOH溶液逐滴加入盐酸羟胺溶液中,继续搅拌1小时。过滤得到羟胺钾溶液。将化合物17a(104mg,0.30mmol)溶于羟胺钾溶液(6mL)室温搅拌10小时。反应完成后旋干液体,加水溶解均匀后,滴加1N盐酸调节pH至固体析出完全,过滤得粗品。通过反相柱层析(MeCN/H2O=50%/50%,HAc条件)纯化,得淡黄色固体A51(64mg,产率61%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ11.02(s,1H),8.98(s,1H),7.78(d,J=7.4Hz,1H),7.54-7.47(m,3H),7.51-7.44(m,3H),7.30-7.24(m,2H),4.57-4.50(m,2H),2.75-2.69(m,2H),2.90(s,6H)。ESI-MS,m/z=348.6[M+H]+
化合物A52-A55的制备方法和化合物A51的制备方法类似。
实施例5.化合物A56的制备。
合成路线:
具体合成方法和步骤如下:
化合物18a:将化合物碘苯(0.11g,0.53mmol),CuI(20mg,0.11mmol)、Pd(PPh3)2Cl2(77mg,0.11mmol)溶于1,4-二氧六环(4mL)中,置换氩气后添加Et3N(2mL)、化合物8(189mg,0.63mmol),将所得混合物在70℃下搅拌20小时,浓缩反应液,并用硅胶柱(PE/EA=10/1)纯化,得到棕色油状物18a(139mg,产率70%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ8.39(d,J=8.4Hz,1H),7.95(d,J=7.5Hz,1H),7.66-7.60(m,3H),7.54(d,J=8.0Hz,1H),7.51-7.45(m,3H),3.94(s,3H),1.65(s,9H)。
化合物19a:将化合物18a(0.42g,1.12mmol)溶于甲醇中,加入2.5N NaOH(5mL),加热至50℃反应12小时。反应完成后旋干甲醇,用1N HCl调节pH至析出固体完全,过滤得黄色固体19a(0.24g,产率60%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ12.70(s,1H),7.76(d,J=7.4Hz,1H),7.51-7.49(m,3H),7.51-7.44(m,3H),7.30-7.23(m,2H),1.64(s,9H)。
化合物20a:将化合物19a(0.23g,0.65mmol)溶于DMSO(5mL)中,冰浴下加入TBTU(O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸,0.25g,0.78mmol),TEA(三乙胺,180uL),30分钟后加入80wt%水合肼,室温反应3小时。反应完毕向其中加水析出固体,过滤得黄色固体20a(0.11g,产率45%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ9.63(s,1H),7.76(d,J=7.4Hz,1H),7.58-7.48(m,3H),7.52-7.43(m,3H),7.32-7.23(m,2H),4.42(s,2H),1.65(s,9H)。
目标化合物A56:向化合物20a(375mg,1.0mmol)的CH2Cl2溶液(5mL)中,加入2mLTFA,室温搅拌3小时。反应结束后将反应液用饱和Na2CO3溶液洗涤三次,然后浓缩得到残渣,残渣经石油醚/乙酸乙酯(体积比100/10)柱层析,得淡黄色固体A56(220mg,产率80%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ11.93(s,1H),9.60(s,1H),7.61-7.23(m,7H),7.21-7.20(m,2H),4.75(s,2H)。ESI-MS,m/z=276.2[M+H]+
实施例6.化合物A57的制备。
合成路线:
具体合成方法和步骤如下:
化合物21a:化合物20a(108.9mg,0.29mmol)溶于15mL甲醇中,加入一滴冰醋酸,5分钟后加入丙醛(17mg,0.29mmol),1小时后加入NaCNBH3(55mg,0.87mmol),室温搅拌2小时。通过反相柱层析(CH3OH/H2O=90%/10%,HAc条件)纯化,得淡黄色固体21a(66mg,产率55%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ9.50(s,1H),7.60(dd,J=6.6,3.0Hz,2H),7.52-7.45(m,4H),7.36(d,J=7.3Hz,1H),7.22(t,J=7.7Hz,1H),7.07-7.01(m,1H),5.08(s,1H),2.77(t,J=7.2Hz,2H),1.36-1.45(m,2H),1.62(s,9H),0.85-0.90(m,3H)。
目标化合物A57:向化合物21a(417mg,1.0mmol)的CH2Cl2溶液(5mL)中,加入2mLTFA,室温搅拌3小时。反应结束后将反应液用饱和Na2CO3溶液洗涤三次,然后浓缩得到残渣,残渣经石油醚/乙酸乙酯(体积比100/10)柱层析,得淡黄色固体A57(238mg,产率75%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ11.90(s,1H),9.51(s,1H),7.75(d,J=7.4Hz,1H),7.59(s,3H),7.52-7.43(m,3H),7.32-7.23(m,2H),5.05(s,1H),2.76(t,J=7.2Hz,2H),1.43-1.51(m,2H),0.85-0.91(m,3H)。ESI-MS,m/z=318.42[M+H]+
实施例7.化合物A58的制备。
合成路线:
具体合成方法和步骤如下:
化合物22a:将化合物15a(0.32g,1.12mmol)溶于20mL甲醇中,加入2.5N NaOH(5mL),加热至50℃反应12小时。反应完成后旋干甲醇,用1N HCl调节pH至析出固体完全,过滤得黄色固体22a(0.21g,产率68%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ12.66(s,1H),7.83(s,1H),7.65-7.60(m,3H),7.41-7.25(m,4H),7.15-7.12(m,1H),3.81(s,3H)。
目标化合物A58:将化合物22a(0.18g,0.65mmol)溶于DMSO(5mL)中,冰浴下加入TBTU(0.25g,0.78mmol),TEA(180uL),30分钟后加入80wt%水合肼,室温反应3小时。反应完毕向其中加水析出固体,过滤得黄色固体A58(90mg,产率48%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ9.49(s,1H),7.82(s,1H),7.65-7.60(m,3H),7.40-7.26(m,4H),7.16-7.13(m,1H),4.49(s,2H),3.85(s,3H)。ESI-MS,m/z=290.3[M+H]+
实施例8.化合物A59-A61的制备,以化合物A59为例。
合成路线:
具体合成方法和步骤如下:
目标化合物A59:化合物A58(83.8mg,0.29mmol)溶于10mL甲醇中,加入一滴冰醋酸,5分钟后加入丙醛(17mg,0.29mmol),1小时后加入NaCNBH3(55mg,0.87mmol),室温搅拌2小时。通过反相柱层析(CH3OH/H2O=90%/10%,HAc条件)纯化,得淡黄色固体A59(53mg,产率55%)。产物的核磁数据如下:1H NMR(400MHz,DMSO-d6)δ9.51(s,1H),7.82(s,1H),7.65-7.60(m,3H),7.40-7.26(m,4H),7.16-7.13(m,1H),5.06(s,1H),3.85(s,3H),2.78(t,J=7.2Hz,2H),1.38-1.53(m,2H),0.83-0.93(m,3H)。ESI-MS,m/z=332.4[M+H]+
化合物A60-A61的制备方法和化合物A59的制备方法类似。
试验例1.目标化合物体外HDAC抑制活性和亚型选择性评价实验
本发明利用荧光分析法对目标化合物的HDAC11、HDAC1和HDAC6的抑制活性进行了测定,以已批准上市的HDAC抑制剂SAHA作为阳性对照。
实验结果(表1)表明,本发明的绝大多数目标化合物在0.5μM浓度下对HDAC11的抑制率均超过50%,但对HDAC1和HDAC6的抑制率小于20%,因此具有很好的HDAC11亚型选择性抑制活性。而0.5μM的SAHA对HDAC1和HDAC6的抑制作用很明显(抑制率超过50%),但对HDAC11抑制作用很弱(抑制率小于10%)。
表1.化合物体外HDAC11、HDAC1和HDAC6抑制率测试结果

选取目标化合物A1和B1为代表,测得了其对HDAC1、HDAC4、HDAC6、HDAC8和HDAC11的半数抑制浓度(IC50)。表2的结果表明,A1和B1对HDAC11的IC50分别为0.031μM和0.17μM,远远低于对其他HDAC亚型的IC50,进一步证实A1和B1具有很好的HDAC11亚型选择性抑制活性。
表2.化合物体外HDAC1、HDAC4、HDAC6、HDAC8和HDAC11的IC50测试结果
试验例2.目标化合物对肿瘤细胞体外抗增殖实验和正常细胞毒性实验
选取目标化合物A1为代表,评价该化合物对人肝细胞癌细胞(Huh7和PLC/PRF/5)、人红白细胞白血病细胞(HEL)、小鼠骨髓瘤细胞P3x63Ag8.653的体外抗增殖活性,以及对人正常肝细胞HL-7702和人脐静脉内皮细胞HUVEC的毒性。表3的结果表明,化合物A1对几种肿瘤细胞均表现出了一定的抗增殖活性,且对正常细胞毒性较低。值得指出的是,化合物A1抑制HCC细胞Huh7和PLC/PRF/5增殖的活性与临床一线肝细胞癌治疗药物sorafenib相当,且对人正常肝细胞HL-7702和人脐静脉内皮细胞HUVEC具有比sorafenib更低的细胞毒性。
表3.化合物对肿瘤细胞体外抗增殖实验和正常细胞毒性结果
试验例3.目标化合物体外抑制肝细胞癌细胞集落形成和肿瘤干细胞成球实验
选取目标化合物A1为代表,评价该化合物体外对肝细胞癌细胞集落形成和肿瘤干细胞成球的抑制活性。集落形成实验结果表明,在2μM浓度条件下,与Ctrl组(dmso处理)相比,化合物A1和sorafenib均能有效抑制肝细胞癌细胞Huh7集落形成,且A1具有比sorafenib更强的抑制Huh7细胞形成集落的能力(图1),这可能与A1对HCC肿瘤干细胞的抑制作用有关。干细胞成球实验进一步证实,在2μM浓度条件下,与Ctrl组(dmso处理)相比,化合物A1和sorafenib均能有效抑制Huh7肿瘤干细胞成球(一次成球和二次成球),且A1 抑制Huh7肿瘤干细胞成球的活性要明显优于sorafenib(图2)。注:一次成球指Huh7细胞的成球实验,二次成球指用Huh7细胞一次成球后的肿瘤干细胞球消化得到的肿瘤干细胞进行的成球实验。图2中的标尺为200μm。
试验例4.目标化合物抑制体外肝细胞癌细胞迁移实验(划痕实验)
选取目标化合物A1为代表,评价该化合物对体外肝细胞癌细胞迁移的抑制活性。划痕实验测试结果(图3)表明,用2μM浓度化合物处理细胞12h后,与Ctrl组(dmso处理)相比,化合物A1和sorafenib均能有效抑制Huh7细胞迁移,且A1具有比sorafenib更强的抗Huh7细胞迁移活性。
以上结果均提示,HDAC11亚型选择性抑制剂对肝细胞癌等癌症具有良好的治疗前景,且还具有抗耐药、抗复发和抗转移的潜在优势。

Claims (10)

  1. 一种HDAC11亚型选择性抑制剂,其特征在于,所述抑制剂为具有如下结构通式A或B所示结构的抑制剂,以及其光学异构体,药学上可接受的盐:
    其中,
    R1是芳环、芳杂环、取代芳环或取代芳杂环;其中,取代芳环或取代芳杂环中的取代基选自烷基、胺烷基、卤素、卤代烷基、烷氧基、氨基、胺基、哌嗪基、烷基哌嗪基、吗啉基、烷基吗啉基、氰基、烷基酰胺基或酰胺基;
    R2是氢、烷基、烷基哌嗪基、烷基吗啉基或胺烷基;
    R3是羟基、氨基或伯胺基。
  2. 根据权利要求1所述HDAC11亚型选择性抑制剂,其特征在于,
    R1是苯环、取代苯环、吡啶环、取代吡啶环、吡嗪环、取代吡嗪环、嘧啶环或取代嘧啶环;其中,所述取代苯环、取代吡啶环、取代吡嗪环或取代嘧啶环的取代基选自
    R2
    R3是羟基、氨基、含1-16个碳原子烷基取代的伯胺基。
  3. 根据权利要求2所述HDAC11亚型选择性抑制剂,其特征在于,所述抑制剂的结构为下列之一:

  4. 如权利要求3所述HDAC11亚型选择性抑制剂的制备方法,选自以下之一:
    (一)化合物1与对甲苯磺酰氯反应生成化合物2,化合物2与四溴化碳反应生成化合物3,化合物3与通过Sonogashira偶联反应生成化合物4,化合物4与羟胺钾反应得到化合物A1-A13;
    或者,化合物4脱去保护基生成化合物16,化合物16与反应生成化合物17,化合物17与羟胺钾反应得到化合物A51-A55;
    反应式如下:
    其中,制备化合物A1-A13的反应式中,取代基R1与相应化合物A1-A13中的取代基R1相同;制备化合物A51-A55的反应式中,取代基R1、R2与相应化合物A51-A55中的取代基R1、R2相同;
    上述反应式中的试剂和条件:
    a.对甲苯磺酰氯,氢化钠,四氢呋喃,室温反应;
    b.四溴化碳,二异丙基氨基锂,四氢呋喃,-78℃和室温反应;
    c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
    d.羟胺钾,甲醇,室温反应;
    e.四丁基氟化铵;四氢呋喃;70℃反应;
    f.氢化钠,四氢呋喃,室温反应;
    (二)化合物1与二碳酸二叔丁酯反应生成化合物5,化合物5与四溴化碳反应生成化合物6,化合物6与三甲基硅乙炔通过Sonogashira偶联反应生成化合物7,化合物7经反应生成化合物8,化合物8与R1I通过Sonogashira偶联反应生成化合物9,化合物9与羟胺钾反应得到化合物10,化合物10脱去保护基得到化合物A14-A20;
    反应式如下:
    其中,反应式中取代基R1与相应化合物A14-A20中的取代基R1相同;
    上述反应式中的试剂和条件:
    a.二碳酸二叔丁酯,三乙胺,二氯甲烷,室温反应;
    b.四溴化碳,二异丙基氨基锂,四氢呋喃,-78℃和室温反应;
    c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
    d.四丁基氟化铵;四氢呋喃;70℃反应;
    e.R1I,碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
    f.羟胺钾,甲醇,室温反应;
    g.三氟乙酸,二氯甲烷,室温反应;
    (三)化合物1与R2I反应生成化合物11,化合物11与单质碘反应生成化合物12,化合物12与三甲基硅乙炔通过Sonogashira偶联反应生成化合物13,化合物13经反应生成化 合物14,化合物14与R1I通过Sonogashira偶联反应生成化合物15,化合物15与羟胺钾反应得到化合物A21-A50;
    反应式如下:
    其中,反应式中取代基R1、R2与相应化合物A21-A50中的取代基R1、R2相同;
    上述反应式中的试剂和条件:
    a.R2I,氢化钠,四氢呋喃,室温反应;
    b.碘,正丁基锂,四氢呋喃,-78℃和室温反应;
    c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
    d.四丁基氟化铵;四氢呋喃;70℃反应;
    e.R1I,碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
    f.羟胺钾,甲醇,室温反应;
    (四)化合物8与R1I通过Sonogashira偶联反应生成化合物18,化合物18水解生成化合物19,化合物19与水合肼缩合得到化合物20,化合物20脱去保护基生成化合物A56;
    或者,化合物20与脂肪醛发生还原胺化反应生成化合物21,化合物21脱去保护基生成化合物A57;
    反应式如下:
    其中,反应式中取代基R1与化合物A56、化合物A57中的取代基R1相同,为苯基;化合物21、化合物A57中,n为2;
    上述反应式中的试剂和条件:
    a.R1I,碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
    b.氢氧化钠,甲醇,50℃反应;
    c.水合肼,O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸,三乙胺,二甲亚砜,室温反应;
    d.三氟乙酸,二氯甲烷,室温反应;
    e.脂肪醛,氰基硼氢化钠,甲醇,冰醋酸,室温反应;
    (五)化合物15水解生成化合物22,化合物22与水合肼缩合得到化合物A58;
    或者,化合物A58与脂肪醛发生还原胺化反应生成化合物A59-A61;
    反应式如下:
    其中,反应式中取代基R1、R2与化合物A58-A61中的取代基R1、R2相同;化合物A59中,n为2;化合物A60中,n为15;化合物A61中,n为2;
    上述反应式中的试剂和条件:
    a.氢氧化钠,甲醇,50℃反应;
    b.水合肼,O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸,三乙胺,二甲亚砜,室温反应;
    c.脂肪醛,氰基硼氢化钠,甲醇,冰醋酸,室温反应;
    (六)化合物23与对甲苯磺酰氯反应生成化合物24,化合物24与四溴化碳反应生成化合物25,化合物25与通过Sonogashira偶联反应生成化合物26,化合物26与羟胺钾反应得到化合物B1-B3;
    反应式如下:
    其中,反应式中取代基R1与相应化合物B1-B3中的取代基R1相同;
    上述反应式中的试剂和条件:
    a.对甲苯磺酰氯,氢化钠,四氢呋喃,室温反应;
    b.四溴化碳,二异丙基氨基锂,四氢呋喃,-78℃和室温反应;
    c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
    d.羟胺钾,甲醇,室温反应;
    (七)化合物23与二碳酸二叔丁酯反应生成化合物27,化合物27与四溴化碳反应生成化合物28,化合物28与三甲基硅乙炔通过Sonogashira偶联反应生成化合物29,化合物29经反应生成化合物30,化合物30与R1I通过Sonogashira偶联反应生成化合物31,化合物31与羟胺钾反应得到化合物32,化合物32脱去保护基得到化合物B4-B6;
    反应式如下:
    其中,反应式中取代基R1与相应化合物B4-B6中的取代基R1相同;
    上述反应式中的试剂和条件:
    a.二碳酸二叔丁酯,三乙胺,二氯甲烷,室温反应;
    b.四溴化碳,二异丙基氨基锂,四氢呋喃,-78℃和室温反应;
    c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
    d.四丁基氟化铵;四氢呋喃;70℃反应;
    e.R1I,碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
    f.羟胺钾,甲醇,室温反应;
    g.三氟乙酸,二氯甲烷,室温反应;
    (八)化合物23与R2I生成化合物33,化合物33与单质碘反应生成化合物34,化合物34与三甲基硅乙炔通过Sonogashira偶联反应生成化合物35,化合物35经反应生成化合物36,化合物36与R1I通过Sonogashira偶联反应生成化合物37,化合物37与羟胺钾反应得到化合物B7;
    反应式如下:
    其中,反应式中取代基R1、R2与相应化合物B7中的取代基R1、R2相同;
    上述反应式中的试剂和条件:
    a.R2I,氢化钠,四氢呋喃,室温反应;
    b.碘,正丁基锂,四氢呋喃,-78℃和室温反应;
    c.碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
    d.四丁基氟化铵;四氢呋喃;70℃反应;
    e.R1I,碘化亚铜,三乙胺,双(三苯基膦)二氯化钯,1,4-二氧六环,70℃反应;
    f.羟胺钾,甲醇,室温反应。
  5. 如权利要求1-3任意一项所述HDAC11亚型选择性抑制剂在制备预防或治疗与HDAC11表达或活性异常相关的疾病的药物中的应用。
  6. 根据权利要求5所述的应用,其特征在于,所述的与HDAC11表达或活性异常相关的疾病为癌症、自身免疫性疾病或代谢性疾病。
  7. 根据权利要求6所述的应用,所述的癌症为肝癌、骨髓增殖性肿瘤、多发性骨髓瘤、霍奇金淋巴瘤、非小细胞肺癌、胶质母细胞瘤、垂体瘤、前列腺癌、卵巢癌或急性淋巴细胞白血病。
  8. 根据权利要求6所述的应用,所述的自身免疫性疾病为炎症、银屑病、风湿性关节炎、类风湿性关节炎或系统性红斑狼疮。
  9. 根据权利要求6所述的应用,所述的代谢性疾病为肥胖症或糖尿病。
  10. 一种预防或治疗癌症、自身免疫性疾病或代谢性疾病的药物组合物,包含权利要求1-3任意一项所述的HDAC11亚型选择性抑制剂或其药学上可接受的盐以及一种或多种药学上可接受的载体或赋形剂。
PCT/CN2023/083623 2022-03-30 2023-03-24 一种hdac11亚型选择性抑制剂及其制备方法和应用 WO2023185667A1 (zh)

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