WO2023011416A1 - 靶向于hdac和nad合成的多靶点抑制剂及其用途 - Google Patents

靶向于hdac和nad合成的多靶点抑制剂及其用途 Download PDF

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WO2023011416A1
WO2023011416A1 PCT/CN2022/109469 CN2022109469W WO2023011416A1 WO 2023011416 A1 WO2023011416 A1 WO 2023011416A1 CN 2022109469 W CN2022109469 W CN 2022109469W WO 2023011416 A1 WO2023011416 A1 WO 2023011416A1
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acid
propylhydrazine
benzyl
carbonyl
compound
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PCT/CN2022/109469
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English (en)
French (fr)
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李晓杨
周仲仁
江余祺
岳凯瑞
徐文方
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中国海洋大学
青岛海济生物医药有限公司
青岛海洋生物医药研究院股份有限公司
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Priority to US18/293,840 priority Critical patent/US20240327418A1/en
Priority to JP2024506856A priority patent/JP2024528251A/ja
Publication of WO2023011416A1 publication Critical patent/WO2023011416A1/zh

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Definitions

  • the invention belongs to the field of biomedical technology, and in particular relates to a multi-target inhibitor targeting HDAC and NAD synthesis, pharmaceutically acceptable salts, solvates and prodrugs thereof and applications thereof.
  • multi-target anticancer drug design has been considered as an effective strategy and has attracted extensive attention in drug development.
  • multi-target drugs can simultaneously modulate the network of disease-related targets and produce synergistic effects (Proc Natl Acad Sci U S A. 2019, 116, 7129-7136). Compared with combined drugs, multi-target drugs can avoid drug-drug interactions, reduce toxic side effects, and improve patient compliance.
  • Histone deacetylases are a family of enzymes involved in the regulation of many cellular processes, including cell proliferation, apoptosis, and cytoskeleton assembly. HDACs affect cellular function through the regulation of histone and non-histone acetylation levels. This process involves a mutual balance between histone acetyltransferase (HAT) and HDAC, both of which are involved in the post-translational modification of histones (Cancer Chemoth Pharm. 2001, 48, 20-26).
  • HAT histone acetyltransferase
  • HDAC cancer Chemoth Pharm. 2001, 48, 20-26.
  • HAT and HDAC have opposing roles in acetylating and deacetylating highly conserved lysine residues on the N-terminal tails of histones, thereby altering chromatin assembly and transcriptional activity.
  • HDACs are also involved in regulating the acetylation of many non-histone proteins, such as ⁇ -tubulin and the tumor suppressor p53 (Pharmacol Res. 2021, 163, 105274; J Invest Dermatol. 2020, 140, 2009-2022).
  • HDAC has synergistic anti-tumor effects with various tumor targets, such as tubulin and heat shock protein 90 (Hsp90), which makes multi-target molecules designed based on the inhibition of HDAC widely used.
  • Hsp90 tubulin and heat shock protein 90
  • NAD + nicotinamide adenine dinucleotide
  • Nicotinamide phosphoribosyltransferase is the most representative NAMPT catalyzes nicotinamide (NAM) to generate nicotinamine mononucleotide (NMN), and regulates the level of NAD, which is an essential energy substance in mammalian cells (Nat Rev Endocrinol.2015 , 11, 535-546). NAMPT is the rate-limiting enzyme of the NAD production pathway and plays a crucial role in cell physiological activities.
  • NAD is an essential coenzyme involved in the synthesis of various essential substances in tumor cells, and NAD can significantly reduce the content of reactive oxygen species (ROS) in the environment , to protect tumor cells; 3) NAMPT plays a crucial role in angiogenesis and induces the production of vascular endothelial growth factor.
  • ROS reactive oxygen species
  • Targeting HDAC and NAD synthesis has synergistic antitumor effects.
  • Certain genotypes such as p53-deficient or mutated tumor cells have primary resistance to HDAC inhibitors, and the combination with NAD synthesis blocking drugs may have a synergistic lethal effect on these cells (Synthetic Lethality), thereby achieving Better antitumor effect. Therefore, designing multi-target inhibitors targeting HDAC and NAD synthesis is of great significance for tumor therapy.
  • the analysis of the pharmacophore of NAMPT and HDAC inhibitors shows that they have similar structural features, which provides a basis for the design of dual inhibitors.
  • the present application particularly provides a HDAC compound with multi-target inhibitory activity and pharmaceutically acceptable salts, hydrates, deuteriums, isomers, or prodrugs thereof, characterized in that the multi-target HDAC compound has the general formula I,
  • G is selected from CH 2 , NH, N(CH 2 ) n CH 3 , O or S, wherein n is 0-9;
  • X 1 is selected from CR 4 or N;
  • X 2 is selected from CR 5 or N;
  • X 4 is selected from CR 7 or N;
  • X 5 , X 6 or X 7 are independently selected from CH or N;
  • R 2 and R 3 are each independently selected from H, halogen, CH 3 , OCH 3 ;
  • R 4 , R 5 , R 6 , R 7 are independently selected from H, halogen, (C 1-2 ) alkyl, halomethyl, OH, OCH 3 , O(CH 2 ) n CH 3 , cyclopropyl Oxygen, OC(CH 3 ) 3 , OCH(CH 3 ) 2 , 5-6 membered alkoxy, NH 2 , N(CH 3 ) 2 , NH(CH 2 ) n CH 3 , CN, N 3 etc., where n is 0-9.
  • the present invention preferably defines compounds shown in general formula I-general formula V and pharmaceutically acceptable salts, hydrates, deuteriums, isomers, or prodrugs thereof,
  • Ring A is selected from the following ring systems:
  • X 1 is selected from CR 4 or N;
  • X 2 is selected from CR 5 or N;
  • X 3 is selected from CR 6 or N;
  • X 4 is selected from CR 7 or N;
  • R 4 , R 5 , R 6 and R 7 are all H;
  • R 1 is selected from H, C 1-4 alkyl, C 3-5 cycloalkyl or C 1-2 alkyl substituted C 3-5 cycloalkyl;
  • the present invention preferably defines compounds shown in formula III and pharmaceutically acceptable salts, hydrates, deuterated products, isomers, or prodrugs thereof,
  • X 1 is selected from CR 4 or N;
  • X 2 is selected from CR 5 or N;
  • X 3 is selected from CR 6 or N;
  • X 4 is selected from CR 7 or N;
  • X is selected from CH or N;
  • R 4 , R 5 , R 6 and R 7 are all H;
  • R 1 is selected from H, C 1-4 alkyl, C 3-5 cycloalkyl or C 1-2 alkyl substituted C 3-5 cycloalkyl;
  • the present invention preferably defines compounds shown in formula IV and pharmaceutically acceptable salts, hydrates, deuterated products, isomers, or prodrugs thereof,
  • Ring A is selected from the following ring systems:
  • X 2 is selected from CR 5 or N;
  • the present invention preferably defines compounds shown in formula V and pharmaceutically acceptable salts, hydrates, deuterated products, isomers, or prodrugs thereof,
  • X 3 is selected from CR 6 or N;
  • X 5 , X 6 or X 7 are independently selected from CH or N
  • R 4 , R 5 , R 6 and R 7 are all H;
  • R 1 is selected from H, C 1-4 alkyl, C 3-5 cycloalkyl or C 1-2 alkyl substituted C 3-5 cycloalkyl;
  • the compounds and salts provided herein may also include all isotopes of atoms present in the intermediate or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers.
  • the preparation of the compounds described herein may involve the protection and deprotection of various chemical groups.
  • the need for protection and deprotection and selection of appropriate protecting groups can be readily determined by those skilled in the art.
  • the chemistry of protecting groups can be found, for example, in T.W. Greene and P.G.M. Wuts, Protecting Groups in Organic Synthesis, 3rd Edition, Wiley & Sons, Inc., New York (1999).
  • the reaction can be monitored according to any suitable method known in the art.
  • product formation can be monitored by spectroscopic means such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), mass spectrometry, or chromatographic methods, Such as high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LCMS) or thin layer chromatography (TLC).
  • HPLC high performance liquid chromatography
  • LCMS liquid chromatography-mass spectrometry
  • TLC thin layer chromatography
  • Compounds can be purified by a variety of methods including high performance liquid chromatography (HPLC) and normal phase silica gel chromatography by those skilled in the art.
  • the phrase "optionally substituted” means unsubstituted or substituted.
  • substituted means that a hydrogen atom is removed and replaced by a substituent. It should be understood that substitution at a given atom is limited by valency.
  • Cn - m means an inclusive range, where n and m are integers and represent the number of carbons. Examples include C 1 - 14 and C 2 - 14 and the like.
  • C n - m alkyl alone or in combination with other terms refers to a saturated hydrocarbon group having n to m carbons, which may be linear or branched.
  • alkyl moieties include, but are not limited to, chemical groups such as: methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl; higher homologues, Such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl and 1,2,2-trimethylpropyl, etc.
  • the alkyl group comprises 1 to 14 carbon atoms, 1 to 13 carbon atoms, 1 to 12 carbon atoms, 1 to 11 carbon atoms, 1 to 10 carbon atoms, 1 to 9 carbon atoms , 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, 1 to 2 carbon atoms.
  • C nm alkoxy refers to a group of formula -O-alkyl, wherein the alkyl group has n to m carbons.
  • alkoxy groups include methoxy, ethoxy, propoxy (eg, n-propoxy and isopropoxy), tert-butoxy, and the like.
  • the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.
  • halogen refers to F, Cl, Br or I. In some embodiments, the halogen is F, Cl or Br. In some embodiments, halo is F. In some embodiments, the halogen is Cl. In some embodiments, the halogen is Br. In some embodiments, halo is I.
  • C nm haloalkyl refers to an alkyl group having from 1 halogen atom to 2s+1 halogen atoms, which may be the same or different, where "s" is the number of carbon atoms in the alkyl group number, where the alkyl group has n to m carbon atoms.
  • a haloalkyl group is only fluorinated (eg, C 1-6 fluoroalkyl).
  • the alkyl group has 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2 carbon atoms.
  • aryl refers to an aromatic hydrocarbon group, which may be monocyclic or polycyclic (eg, having 2 fused rings).
  • C nm aryl refers to an aryl group having n to m ring carbon atoms.
  • Aryl includes, for example, phenyl, naphthyl, and the like. In some embodiments, aryl groups have 6 to 10 carbon atoms. In some embodiments, aryl is substituted or unsubstituted phenyl.
  • cycloalkyl examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, or cyclohexadienyl, and the like.
  • the cycloalkyl group is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.
  • a cycloalkyl has 3-6 ring-forming carbon atoms (ie, C3-6 cycloalkyl).
  • heterocycloalkyl refers to a non-aromatic monocyclic or multicyclic heterocyclic ring having one or more ring-forming heteroatoms selected from O, N, or S. Included in heterocycloalkyl are monocyclic 4-membered, 5-membered and 6-membered heterocycloalkyl groups.
  • heterocycloalkyl groups include pyrrolidin-2-one, 1,3-isoxazolidin-2-one, pyranyl, tetrahydropyranyl, oxetanyl, azetidinyl , morpholino, thiomorpholino, piperazinyl, tetrahydrofuryl, tetrahydrothienyl, piperidinyl, pyrrolidinyl, isoxazolidinyl, isothiazolidinyl, pyrazolidinyl, oxazolidinyl group, thiazolidinyl, imidazolidinyl and azaalkyl, etc.
  • the heterocycloalkyl group can be attached through a ring carbon atom or a ring heteroatom.
  • a heterocycloalkyl group contains 0 to 3 double bonds.
  • Tautomeric forms result from the exchange of a single bond for an adjacent double bond with concomitant proton migration.
  • Tautomeric forms include prototropic tautomers, which are isomeric protonation states having the same empirical formula and overall charge. Examples of prototropic tautomers include keto-enol pairs, amide-imidic acid pairs, lactam-lactimine pairs, enamine-imine pairs, and those in which a proton can occupy a heterocyclic ring system.
  • All compounds and their pharmaceutically acceptable salts may be found together with other substances such as water and solvents (eg hydrates and solvates) or may be isolated.
  • preparation of compounds may involve the addition of acids or bases, thereby affecting, for example, the catalysis of a desired reaction or the formation of salt forms such as acid addition salts.
  • acids may be mineral acids or organic acids and include, but are not limited to, strong and weak acids.
  • Some examples of acids include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, p-toluenesulfonic acid, 4-nitrobenzoic acid, methanesulfonic acid, benzenesulfonic acid, trifluoroacetic acid, and nitric acid.
  • Some weak acids include, but are not limited to, acetic acid, propionic acid, butyric acid, benzoic acid, pyroglutamic acid, tartaric acid, valeric acid, caproic acid, heptanoic acid, caprylic acid, nonanoic acid, and capric acid.
  • the compounds and salts provided herein are substantially isolated.
  • substantially isolated is meant that the compound is at least partially or substantially separated from the environment in which it was formed or detected.
  • Partial isolation can include, for example, a composition enriched in a compound provided herein.
  • substantially separating can include comprising at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of Compositions of compounds provided herein, or salts thereof. Methods for isolating compounds and their salts are routine in the art.
  • phrases "pharmaceutically acceptable” is used herein to refer to those compounds, materials, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissues without Excessive toxicity, irritation, allergic reaction, or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • the pharmaceutically acceptable salts of the present application include, for example, the conventional non-toxic salts of the parent compound formed by non-toxic inorganic or organic acids, mainly including inorganic acid salts such as sulfuric acid, nitric acid, hydrobromic acid, phosphoric acid, hydrochloric acid, boric acid , sulfamic acid, etc.; or organic acids such as acetic acid, propionic acid, butyric acid, camphoric acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, glycolic acid, trifluoroacetic acid, adipic acid, alginic acid, 2- Hydroxypropionic acid, 2-oxopropionic acid, stearic acid, lactic acid, citric acid, oxalic acid, malonic acid, succinic acid, pyroglutamic acid, ascorbic acid, aspartic acid, phenylacetic acid, glutamic acid, benzoic acid , salicylic acid,
  • the pharmaceutically acceptable salts of the present application can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods.
  • such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of the two; usually, non-aqueous media such as Ether, ethyl acetate, alcohols (eg methanol, ethanol, isopropanol or butanol) or acetonitrile (MeCN) are preferred.
  • non-aqueous media such as Ether, ethyl acetate, alcohols (eg methanol, ethanol, isopropanol or butanol) or acetonitrile (MeCN) are preferred.
  • the cancer is a hematological cancer.
  • the hematological cancer is selected from the group consisting of leukemia and lymphoma.
  • the blood cancer is selected from the group consisting of acute myeloid leukemia (AML), chronic myeloid leukemia, B-cell lymphoma, chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma, Hairy cell leukemia, mantle cell lymphoma, Burkitt lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, extranodal marginal zone lymphoma lymphoma), activated B-cell like (ABC) diffuse large B cell lymphoma, and germinal center B-cell (GCB) diffuse large B-cell lymphoma (germinal center B cell (GCB) diffuse large B cell lymphoma).
  • AML acute myeloid leukemia
  • CLL chronic lymphocytic leuk
  • the compounds described herein may be used alone or in combination with other therapeutic agents for the treatment of diseases or conditions described herein.
  • the compounds of the present invention are combined with other antineoplastic drugs.
  • the anti-tumor drugs include, but are not limited to: cyclophosphamide, nitrogen mustard, melphalan, cycloxan, carmustine, metal platinums such as carboplatin, cisplatin, oxaliplatin, camptothecin, irinotecan , daunorubicin, doxorubicin, bleomycin, plicamycin, paclitaxel, vinorelbine, docetaxel, doxorubicin, fluorouracil, methotrexate, cytarabine, gemcitabine, EGFR inhibitors, VEGFR inhibitors, ALK inhibitors, BTK inhibitors, mTOR inhibitors, HDAC inhibitors.
  • compositions When used as a medicament, the compounds and salts provided herein can be administered in the form of pharmaceutical compositions. These compositions may be prepared as described herein or elsewhere and may be administered by a variety of routes depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration can be topical (including transdermal, epidermal, ophthalmic, and administration to mucosal membranes including intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or insufflation of a powder or aerosol). including by nebulizer; intratracheal or intranasal), oral or parenteral.
  • topical including transdermal, epidermal, ophthalmic, and administration to mucosal membranes including intranasal, vaginal, and rectal delivery
  • pulmonary e.g., by inhalation or insufflation of a powder or aerosol
  • nebulizer intratracheal or intranasal
  • Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular injection or infusion; or intracranial (eg, intrathecal or intraventricular administration).
  • Parenteral administration can be in the form of a single bolus dose or can be by, for example, a continuous infusion pump.
  • the compounds, salts, and pharmaceutical compositions provided herein are suitable for parenteral administration. In some embodiments, the compounds, salts, and pharmaceutical compositions provided herein are suitable for intravenous administration.
  • compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders.
  • Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like will be necessary or desirable.
  • compositions comprising, as an active ingredient, a compound provided herein, or a pharmaceutically acceptable salt thereof, in combination with one or more pharmaceutically acceptable carriers (eg, excipients).
  • the active ingredient will generally be mixed with an excipient, diluted by an excipient or enclosed within such a carrier which is in the form of, for example, a capsule, sachet, paper or other container.
  • the excipient acts as a diluent, it can be a solid, semi-solid or liquid material which acts as a vehicle, carrier or medium for the active ingredient.
  • Reagents and conditions (a) different amine, TBTU, TEA, DCM, yield 55%; (b) CH 3 OH/H 2 O, KOH, reflux, yield 80%; (c) 4-nitrophenyl chloroformate, TEA, DCM , yield 80%; (d) methyl 8-aminocaprylate hydrochlorid, TEA, DCM, yield 65%; (e) sodium azide, DMF, 80°C, yield 80%.
  • Reagents and conditions (a) propionaldehyde, MeOH, yield 98%; NaBH 3 CN, MeOH, concentrated hydrochloric acid, methyl orange, yield 60%; (b) (Boc) 2 O, triethylamine, EtOH, yield 85%; ( c) Pd/C, H 2 , MeOH, yield 85%; (d) trifluoroacetic anhydride, DCM, yield 85%; (e) EDCI, HOBt, TEA, DCM, yield 55%; (f) Na 2 CO 3 , MeOH, yield 70%.
  • Reagents and conditions (a) TBTU, TEA, DCM, yield 55%; (b) TFA, DCM, triethylamine, yield 85%.
  • Reagents and conditions (a) TBTU, TEA, DCM, yield 55%.
  • tert-butyl 1-propylhydrazine-1-carboxylate (11) Compound 10 (0.6 g, 2 mmol) was dissolved in methanol, and Pd/C (0.06 g) was added. Then react at room temperature under hydrogen for 4 hours. After the reaction was completed, Pd/C was removed by filtration, and the filtrate was evaporated to dryness to obtain compound 11 as a colorless oil (0.25 g, 72%).
  • reaction solution was washed with saturated NaHCO 3 (2 ⁇ 30 mL) and saturated brine (2 ⁇ 30 mL), and dried over MgSO 4 . After filtration, the solvent was evaporated to dryness and purified by flash chromatography to give a white solid powder (0.35 g, 54%).
  • Embodiment 6 is a diagrammatic representation of Embodiment 6
  • Embodiment 7 is a diagrammatic representation of Embodiment 7:
  • Embodiment 8 is a diagrammatic representation of Embodiment 8
  • Embodiment 9 is a diagrammatic representation of Embodiment 9:
  • N-(2-aminophenyl)-8-azidooctylamide (20) Using the synthetic method of 1a, using compound 8 and 1,2-diaminobenzene as raw materials, a white solid 20 was obtained in a yield of 60 %.
  • ester (23v) Dissolve compound 2d (0.29g, 1mmol) in 15mL of dichloromethane, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide under ice-cooling Hydrochloride (EDCI, 0.23 g, 1.2 mmol) and 1-hydroxybenzotriazole (HOBt at 0 °C, 0.18 g, 1.2 mmol).
  • LEE17 N-(4-(2-propylhydrazine-1-carbonyl)benzyl)benzo[b]thiophene-2-carboxamide
  • LEE19 N-(4-(2-Propylhydrazine-1-carbonyl)benzyl)thieno[3,2-c]pyridine-2-carboxamide
  • Example 44 Inhibitory activity of compounds on HDAC1,2,3.
  • HDAC buffer 15mM Tris-HCl (PH 8.0), 250 ⁇ M EDTA, 250mM NaCl, 10% glycerol.
  • Trypsin stop solution 10mg/ml trypsin, 50mM Tris-HCL (pH8.0), 100mM NaCl, 2 ⁇ M TSA.
  • Substrate Dimethyl sulfoxide, the exclusive substrate of HDAC1, HDAC2 and HDAC3, was dissolved to make a 30mM stock solution, and diluted to 300 ⁇ M with HDAC buffer, so that the content of dimethyl sulfoxide was about 1%.
  • Enzyme solution HDAC1, HDAC2 and HDAC3 were diluted 1:20 with HDAC buffer.
  • the data in the table a come from three independent experiments, the value is the mean value, the standard deviation ⁇ 10%; ND: not detected.
  • Example 45 Inhibitory activity of compounds against NAMPT.
  • the data in the table a come from three independent experiments, and the values are mean values with standard deviation ⁇ 10%.
  • Example 46 Compounds have a median growth inhibitory concentration (GI 50 ) and a median lethal concentration (LC 50 ) of tumor cells
  • the median growth inhibitory concentration (GI 50 ) and median lethal concentration (LC 50 ) were determined by the NCI method.
  • Leukemia cell lines MV4-11, HL60, PL21, KASUMI-1, MONO-MAC-1, and NB-4 were cultured in IMDM medium containing 10% fetal bovine serum, and seeded in 96-well cells at a density of 10,000/100uL culture plate overnight.
  • Select 6 wells as Tz wells add 0.125mg/mL Cell Titer-Blue dye, and read the fluorescence intensity at 560nM/590nM (excitation wavelength/emission wavelength) after incubation for 4 hours. Different concentrations of compounds were added to the remaining wells, and a 100% control group was set at the same time.
  • the fluorescence intensity of the drug-dosed group is represented by Ti
  • the fluorescence intensity of the 100% control group is represented by C
  • the fluorescence intensity before drug addition is represented by Tz.
  • the GI 50 and LC 50 values of some compounds against leukemia cell lines MV4-11, HL60, PL21, KASUMI-1, MONO-MAC-1, and NB-4 are shown in Figure 1, Table 3, Figure 2 and Table 4.
  • KF866 is a NAMPT inhibitor reported in the literature.
  • the data in the table are all from three independent experiments, the values are the average value, the standard deviation ⁇ 10%; b ND: not detected.
  • the data in the table a come from three independent experiments, and the values are mean values with standard deviation ⁇ 10%.
  • Figure 2 and Table 4 also show that HDAC and NAMPT dual inhibitors LEE12 and LEE18 have a lethal effect on p53-mutant and p53-null cell lines, and most of the compounds have achieved unexpected results compared with the compounds in Comparative Example 1 technical effect.
  • Example 47 In vivo anti-colon cancer activity of the target compound LEE17
  • Colon cancer cells HCT116 were inoculated subcutaneously in the right shoulder of nude mice, 100uL each (cell count: 1.8* 108 cells/mL), and administered in groups after one week. The tumor-bearing mice were divided into groups and administered intragastrically.
  • Relative tumor volume (RTV) Vt/Vo
  • the evaluation index of anti-tumor activity is the relative tumor proliferation rate T/C (%),
  • 5-FU 5-fluorouracil, a traditional anti-tumor chemotherapy drug
  • Oxaliplatin Oxaliplatin, the third generation platinum anti-cancer drug, is an anti-tumor chemotherapy drug.
  • Example 48 In vivo anti-leukemia activity of target compounds LEE15 and LEE16 Panobinostat: panobinostat, a broad-spectrum HDAC inhibitor on the market
  • Acute myeloid leukemia cells MV4-11 were inoculated subcutaneously in the right shoulder of nude mice, 100uL each (cell count: 1.8* 108 cells/mL), and administered in groups after one week. The grouping is as follows:
  • Test group compound LEE15, LEE16, administration agent/4mg/kg/d, administration volume: 200uL/20g each time;
  • Positive control group positive drug panobinostat, administered at a dose of 4 mg/kg/d, injected intraperitoneally.
  • Blank control group given the same volume of PBS.
  • Relative tumor volume (RTV) Vt/Vo
  • the evaluation index of anti-tumor activity is the relative tumor proliferation rate T/C (%),
  • Panobinostat a positive control drug, is a marketed broad-spectrum HDAC inhibitor.
  • Example 49 Pharmacokinetic properties of target compounds LEE15, LEE16, LEE17
  • LEE15, LEE16, LEE17 were dissolved in 40% PEG300 and 60% H2O.
  • Three rats in each group were given a single dose by oral gavage (po) 20 mg/kg and intravenous injection (iv) 5 mg/kg.
  • Blood was collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours after the administration, and t 1/2 , C 0 , AUC, Vss, CLp, MRT, C max , and t max , F% and other parameters.

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Abstract

本发明提供了一种靶向于HDAC和NAD合成的化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，及其制备和应用。具体来讲，提供了结构通式(I)所示化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药。所述的结构通式(I)的化合物为多靶点抑制剂，靶向于HDAC和NAD靶点，表现出显著的HDAC抑制活性，同时代表性化合物表现出一定的抑制NAD的活性。环E-B-L-C(O)-(NH)r-R(通式I)

Description

靶向于HDAC和NAD合成的多靶点抑制剂及其用途 技术领域
本发明属于生物医学技术领域，尤其涉及一种靶向于HDAC和NAD合成的多靶点抑制剂、其药学上可接受的盐、溶剂合物和前药及其应用。
发明背景
传统药物研究的重点是寻找对单个靶标具有高亲和力和高选择性的分子。但是，肿瘤的发生和发展依赖于多种受体或信号通路，这使得作用于单靶点的抗肿瘤药物由于代偿性抵抗而不能完全杀灭肿瘤细胞或产生耐药。为了克服这些限制，多靶点抗肿瘤药物设计已被认为是一种有效的策略，并在药物开发中引起了广泛的关注。理想地，多靶标药物可以同时调节疾病相关靶标的网络并产生协同效应(Proc Natl Acad Sci U S A.2019,116,7129-7136)。与联合用药相比，多靶点药物可避免药物-药物相互作用，减少毒副作用、提高患者依从性等。
组蛋白去乙酰化酶(Histone deacetylase,HDAC)是涉及许多细胞过程调节的酶家族，包括细胞增殖，凋亡和细胞骨架装配。HDAC通过对组蛋白和非组蛋白乙酰化水平进行调控来影响细胞功能。这一过程涉及组蛋白乙酰基转移酶(HAT)和HDAC之间的相互平衡，两者均参与组蛋白的翻译后修饰(Cancer Chemoth Pharm.2001,48,20-26)。HAT和HDAC在乙酰化和脱乙酰化组蛋白N末端尾部上高度保守的赖氨酸残基方面具有相反的作用，从而改变了染色质的组装和转录活性。HDAC还参与调节许多非组蛋白的乙酰化作用，例如α-微管蛋白和肿瘤抑制因子p53(Pharmacol Res.2021,163,105274；J Invest Dermatol.2020,140,2009-2022)。这些结果以及许多肿瘤类型中HDAC活性异常的报道表明，HDAC抑制代表了可行的抗癌策略。
鉴于HDAC在肿瘤发生中的重要作用，HDAC与多种肿瘤靶标(例如微管蛋白和热休克蛋白90(Hsp90)具有协同抗肿瘤作用，这使得以抑制HDAC为基础设计的多靶点分子被广泛研究(Eur J Med Chem,2020,208,112831)。烟酰胺腺嘌呤二核苷酸(NAD +)作为生物体内最重要的辅酶和核心代谢物，不仅广泛参与了能量代谢氧化还原反应。由于肿瘤细胞增殖迅速，具有更高的产能需求，肿瘤组织中的NAD +的生物合成也经常会上调(Nat Rev Cancer.2012,12,741-752)。烟酰胺磷酸核糖基转移酶(NAMPT)是最具代表性的代谢目标之一。NAMPT催化烟酰胺(NAM)生成烟碱胺单核苷酸(NMN)，并调节NAD的水平，NAD是哺乳动物细胞中一种必需的能量物质(Nat Rev Endocrinol.2015,11,535-546)。NAMPT是NAD产生途径的限速酶，在细胞生理活动中起着至关重要的作用。研究表明，靶向于NAD合成具有重要的抗肿瘤作用1)肿瘤细胞的NAD消耗和新陈代谢率比正常细胞高，而且它更容易 受到NAMPT抑制剂的影响；2)NAD是参与肿瘤细胞中多种必需物质合成的必需辅酶，NAD可以显着降低环境中的活性氧(ROS)含量，以保护肿瘤细胞；3)NAMPT在血管生成中起着至关重要的作用，并诱导血管内皮生长因子的产生。目前，有两个靶向于NAD合成的NAMPT抑制剂FK866和CHS-828已进入临床研究(Cancer Res.2003,63,7436-7442；Cancer Res.1999,59,5751-5757)。
靶向于HDAC和NAD合成具有协同抗肿瘤作用。某些特定基因型比如p53缺陷或者突变的肿瘤细胞对HDAC抑制剂产生原发性耐药，而和NAD合成阻断药物联用有可能会对这些细胞产生协同致死作用(Synthetic Lethality)，从而达到更好的抗肿瘤作用。因此，设计靶向于HDAC和NAD合成的多靶点抑制剂对肿瘤治疗具有重要意义。另外，对NAMPT和HDAC抑制剂的药效团进行分析，表明二者具有相似的结构特征，为设计双重抑制剂提供了基础。
发明内容
本申请特别提供一种具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，其特征在于，所述的多靶点HDAC化合物具有如通式I，
环E-B-L-C(O)-(NH)r-R(通式I)
其中环E选自
Figure PCTCN2022109469-appb-000001
r＝1、2，R选自H、C 1-4烷基、C 3-5环烷基或C 1-2烷基取代的C 3-5环烷基、
Figure PCTCN2022109469-appb-000002
更进一步的通式I化合物进一步选自通式Ⅱ、通式Ⅲ、通式Ⅳ、通式Ⅴ化合物：
Figure PCTCN2022109469-appb-000003
Figure PCTCN2022109469-appb-000004
或其药学上可接受的盐、水合物、氘代物或前药，其中：
A环选自
Figure PCTCN2022109469-appb-000005
G选自CH 2、NH、N(CH 2) nCH 3、O或S，其中n为0-9；
Figure PCTCN2022109469-appb-000006
表示与相邻并环相连的键；
---表示与B相连的键；
X 1选自CR 4或N；
X 2选自CR 5或N；
X 3选自CR 6或N；
X 4选自CR 7或N；
X 5、X 6或X 7独立地选自CH或N；
R 1选自H、C 1-C 4烷基、C 3-C 5环烷基或C 1-C 2烷基取代的C 3-C 5环烷基；
R 2和R 3各自独立地选自H、卤素、CH 3、OCH 3；
B选自
Figure PCTCN2022109469-appb-000007
Figure PCTCN2022109469-appb-000008
表示与A环相连的键；
---表示与L相连的键；
L选自由C 1-14烷基、C 1-14烷氧基、C 2-14烯基、C 2-14炔基、C 3-10环烷基、C 1-9烷基取代的C 3-10环烷基、C 1-9烷氧基取代的C 3-10环烷基、C 6-10芳基、C 1-9烷基取代的C 6-10芳基、C 1-9烷氧基取代的C 6-10芳基、(C 1-9烷基))-(C＝O)NH取代的芳基、苄基或(C 1-8烷基)-(C＝O)NH取代的苄基组成的组；
R 4、R 5、R 6、R 7独立地选自H、卤素、(C 1-2)烷基、卤代甲基、OH、OCH 3、O(CH 2) nCH 3、 环丙基氧基、OC(CH 3) 3、OCH(CH 3) 2、5-6元烷氧基、NH 2、N(CH 3) 2、NH(CH 2) nCH 3、CN、N 3等，其中n为0-9。
本发明优选定义如通式I-通式Ⅴ所示的化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，
其中，
A环选自如下环系：
Figure PCTCN2022109469-appb-000009
G选自NH、O或S；
Figure PCTCN2022109469-appb-000010
表示与相邻并环相连的键；
---表示与B相连的键；
X 1选自CR 4或N；
X 2选自CR 5或N；
X 3选自CR 6或N；
X 4选自CR 7或N；
R 4、R 5、R 6、R 7均为H；
R 1选自H、C 1-4烷基、C 3-5环烷基或C 1-2烷基取代的C 3-5环烷基；
B选自如下结构
Figure PCTCN2022109469-appb-000011
Figure PCTCN2022109469-appb-000012
表示与A环相连的键；
---表示与L相连的键；
L选自C 1-14烷基、C 1-14烷氧基、C 2-14烯基、C 2-14炔基、C 1-9烷基取代的C 3-10环烷基、C 1-9烷氧基取代的C 3-10环烷基、C 6-10芳基、C 1-9烷基取代的C 6-10芳基、C 1-9烷氧基取代的C 6-10芳基、(C 1-9烷基))-(C＝O)NH取代的芳基、苄基或(C 1-8烷基)-(C＝O)NH取代的苄基；
本发明优选定义如式Ⅲ所示的化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，
其中，
X 1选自CR 4或N；
X 2选自CR 5或N；
X 3选自CR 6或N；
X 4选自CR 7或N；
X 5选自CH或N；
R 4、R 5、R 6和R 7均为H；
R 1选自H、C 1-4烷基、C 3-5环烷基或C 1-2烷基取代的C 3-5环烷基；
B选自如下结构
Figure PCTCN2022109469-appb-000013
Figure PCTCN2022109469-appb-000014
表示与A环相连的键；
---表示与L相连的键；
L选自C 1-14烷基、C 1-14烷氧基、C 2-14烯基、C 2-14炔基、C 1-9烷基取代的C 3-10环烷基、C 1-9烷氧基取代的C 3-10环烷基、C 6-10芳基、C 1-9烷基取代的C 6-10芳基、C 1-9烷氧基取代的C 6-10芳基、(C 1-9烷基)-(C＝O)NH取代的芳基、苄基或(C 1-8烷基)-(C＝O)NH取代的苄基；
本发明优选定义如式Ⅳ所示的化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，
其中，
A环选自如下环系：
Figure PCTCN2022109469-appb-000015
G选自NH、O或S；
Figure PCTCN2022109469-appb-000016
表示与相邻并环相连的键；
---表示与B相连的键；
X 1选自CR 4或N；
X 2选自CR 5或N；
X 3选自CR 6或N；
X 4选自CR 7或N；
X 6或X 7独立地选自CH或N；
R 4、R 5、R 6、R 7均为H；
R 1选自H、C 1-4烷基、C 3-5环烷基或C 1-2烷基取代的C 3-5环烷基；
R 2和R 3均为H；
B选自如下结构
Figure PCTCN2022109469-appb-000017
Figure PCTCN2022109469-appb-000018
表示与A环相连的键；
---表示与L相连的键；
L选自C 1-14烷基、C 1-14烷氧基、C 2-14烯基、C 2-14炔基、C 1-9烷基取代的C 3-10环烷基、C 1-9烷氧基取代的C 3-10环烷基、C 6-10芳基、C 1-9烷基取代的C 6-10芳基、C 1-9烷氧基取代的C 6-10 芳基、(C 1-9烷基)-(C＝O)NH取代的芳基、苄基或(C 1-8烷基)-(C＝O)NH取代的苄基；
本发明优选定义如式Ⅴ所示的化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，
其中，
X 1选自CR 4或N；
X 2选自CR 5或N；
X 3选自CR 6或N；
X 4选自CR 7或N；
X 5、X 6或X 7独立地选自CH或N
R 4、R 5、R 6和R 7均为H；
R 1选自H、C 1-4烷基、C 3-5环烷基或C 1-2烷基取代的C 3-5环烷基；
B选自如下结构
Figure PCTCN2022109469-appb-000019
Figure PCTCN2022109469-appb-000020
表示与A环相连的键；
---表示与L相连的键；
L选自C 1-14烷基、C 1-14烷氧基、C 2-14烯基、C 2-14炔基、C 1-9烷基取代的C 3-10环烷基、C 1-9烷氧基取代的C 3-10环烷基、C 6-10芳基、C 1-9烷基取代的C 6-10芳基、C 1-9烷氧基取代的C 6-10芳基、(C 1-9烷基)-(C＝O)NH取代的芳基、苄基或(C 1-8烷基)-(C＝O)NH取代的苄基；
除非特别定义，本文中提供的化合物和盐还可以包含存在于中间体或最终化合物中的原子的所有同位素。同位素包括具有相同的原子序数但是具有不同的质量数的那些原子。
本领域技术人员将理解的是，所描述的方法不是可以合成本文中提供的化合物的排他性手段，并且可以获得合成有机反应的广泛的集合从而潜在地用于合成本文中提供的化合物。本领域技术人员知道如何选择和实施适当的合成路线。起始原料、中间体和产物的适当的合成方法可以通过参考包括例如如下的参考文献的文献来确定：杂环化学的进展，第1-107卷(Elsevier,1963-2012)；杂环化学杂志，第1-49卷(杂环化学杂志,1964-2012)；Carreira等人(编)，合成科学，第1-48卷(2001-2010)；Katritzky等人(编)，综合有机官能团转化(Pergamon Press,1996)；Katritzky等人(编)；综合有机官能团转化II(Elsevier，第2版,2004)；Katritzky等人(编)，综合杂环化学(Pergamon Press,1984)；Smith等人，高等有机化学：反应、机理和结构，第6版(Wiley,2007)；Trost等人(编)，综合有机合成(Pergamon Press,1991)。
本文中所述的化合物的制备可以涉及各种化学基团的保护和脱保护。对保护和脱保护的需要和适当的保护基团的选择可以由本领域技术人员容易地确定。可以在例如T.W.Greene 和P.G.M.Wuts，有机合成中的保护基团，第3版，Wiley&Sons,Inc.,New York(1999)中找到保护基团的化学。
可以根据本领域已知的任何适当的方法来监测反应。例如，产物形成可以通过如下来监测：光谱手段，如核磁共振光谱法(例如， 1H或 13C)、红外光谱法、分光光度法(例如，UV-可见)、质谱法，或者色谱方法，如高效液相色谱法(HPLC)、液相色谱-质谱法(LCMS)或薄层色谱法(TLC)。化合物可以由本领域技术人员通过包括高效液相色谱法(HPLC)和正相硅胶色谱法的多种多样的方法来纯化。
如本文中所使用的，短语“任选取代”意味着未取代或取代。如本文中所使用的，术语“取代”意味着将氢原子除去并且由取代基替代。应当理解的是，在给定的原子上的取代受到化合价限制。
贯穿所有定义，术语“C n- m”表示包括端点的范围，其中n和m为整数并且表示碳数。实例包括C 1- 14和C 2- 14等。
如本文中所使用的，单独或与其它术语组合使用的术语“C n- m烷基”是指可以为直链或支链的、具有n至m个碳的饱和烃基。烷基部分的实例包括但不限于例如如下的化学基团：甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基、仲丁基；高级同系物，如2-甲基-1丁基、正戊基、3-戊基、正己基和1,2,2-三甲基丙基等。在一些实施方案中，烷基包含1至14个碳原子、1至13个碳原子、1至12个碳原子、1至11个碳原子、1至10个碳原子、1至9个碳原子、1至8个碳原子、1至7个碳原子、1至6个碳原子、1至5个碳原子、1至4个碳原子、1至3个碳原子、1至2个碳原子。
如本文中所使用的，单独或与其它术语组合使用的术语“C n-m烷氧基”是指式-O-烷基的基团，其中烷基具有n至m个碳。烷氧基的实例包括甲氧基、乙氧基、丙氧基(例如，正丙氧基和异丙氧基)和叔丁氧基等。在一些实施方案中，烷基具有1至6个、1至4个、或者1至3个碳原子。
如本文中所使用的，“卤素”是指F、Cl、Br或I。在一些实施方案中，卤素为F、Cl或Br。在一些实施方案中，卤素为F。在一些实施方案中，卤素为Cl。在一些实施方案中，卤素为Br。在一些实施方案中，卤素为I。
如本文中所使用的，术语“C n-m卤代烷基”是指具有可以相同或不同的1个卤素原子至2s+1个卤素原子的烷基，其中“s”为烷基中的碳原子的个数，其中烷基具有n至m个碳原子。在一些实施方案中，卤代烷基仅被氟化(例如，C 1-6氟烷基)。在一些实施方案中，烷基具有1至14个、1至13个、1至12个、1至11个、1至10个、1至9个、1至8个、1至7个、1至6个、1至5个、1至4个、1至3个、1至2个碳原子。
如本文中所使用的，术语“芳基”是指芳香族烃基，其可以为单环的或多环的(例如，具有2个稠环)。术语“C n-m芳基”是指具有n至m个环碳原子的芳基。芳基包括例如苯基、萘基等。在一些实施方案中，芳基具有6个至10个碳原子。在一些实施方案中，芳基为取代或未取代的苯基。
如本文中所使用的，“环烷基”是指包括环化的烷基和/或烯基的非芳香族环状烃。环烷基可以包括单环或多环(例如，具有2个稠环)基团。环烷基可以具有3个、4个、5个、6个成环碳(即，C 3-6环烷基)。环烷基的成环碳原子可以任选由氧基(oxo)或硫基(sulfido)取代(例如，C(＝O)或C(＝S))。环烷基的实例包括但不限于环丙基、环丁基、环戊基、环己基、环庚基、环戊烯基、环己烯基或环己二烯基等。在一些实施方案中，环烷基选自由环丙基、环丁基、环戊基、环己基组成的组。在一些实施方案中，环烷基具有3-6个成环碳原子(即，C3-6环烷基)。
如本文中所使用的，“杂环烷基”是指具有一个或多个选自O、N或S的成环杂原子的非芳香族单环或多环杂环。包括在杂环烷基中的是单环4元、5元和6元杂环烷基。杂环烷基的实例包括吡咯烷-2-酮、1,3-异噁唑烷-2-酮、吡喃基、四氢吡喃基、氧杂环丁烷基、氮杂环丁烷基、吗啉代、硫代吗啉代、哌嗪基、四氢呋喃基、四氢噻吩基、哌啶基、吡咯烷基、异噁唑烷基、异噻唑烷基、吡唑烷基、噁唑烷基、噻唑烷基、咪唑烷基和氮杂烷基等。杂环烷基的成环碳原子和杂原子可以任选由氧基(＝O)取代。可以通过成环碳原子或成环杂原子来连接杂环烷基。在一些实施方案中，杂环烷基包含0至3个双键。
如本文中所使用的术语“化合物”意在包括所描绘的结构的所有立体异构体、几何异构体、互变异构体和同位素。除非另有说明，在本文中通过名称或结构确定为一种特定的互变异构形式的化合物旨在包括其它互变异构形式。
本文中提供的化合物还包括互变异构形式。互变异构形式由单键与相邻的双键的交换以及伴随的质子迁移引起。互变异构形式包括质子移变的互变异构体，其为具有相同的经验式和总电荷的同分异构的质子化状态。质子移变的互变异构体的实例包括酮-烯醇对，酰胺-亚氨酸对，内酰胺-内酰亚胺对，烯胺-亚胺对，以及其中质子可以占据杂环体系的两个以上的位置的环状形式，例如，1H-和3H-咪唑，1H-、2H-和4H-1,2,4-三唑，1H-和2H-异吲哚，以及1H-和2H-吡唑。互变异构形式可以处于平衡状态或者通过适当的取代立体地锁定为一种形式。
所有化合物及其药学上可接受的盐可以与例如水和溶剂等其它物质一起被发现(例如水合物和溶剂化物)，或者可以是分离的。
在一些实施方案中，化合物的制备可以涉及添加酸或碱，从而影响例如期望的反应的催 化或者例如酸加成盐等盐形式的形成。
酸的实例可以为无机酸或有机酸并且包括但不限于强酸和弱酸。酸的一些实例包括盐酸、氢溴酸、硫酸、磷酸、对甲苯磺酸、4-硝基苯甲酸、甲磺酸、苯磺酸、三氟乙酸和硝酸。一些弱酸包括但不限于乙酸、丙酸、丁酸、苯甲酸、焦谷氨酸、酒石酸、戊酸、己酸、庚酸、辛酸、壬酸和癸酸。
碱的实例包括氢氧化锂、氢氧化钠、氢氧化钾、碳酸锂、碳酸钠、碳酸钾和碳酸氢钠。强碱的一些实例包括但不限于氢氧化物、醇盐、金属氨基化合物、金属氢化物、金属二烷基酰胺和芳基胺，其中，醇盐包括甲基、乙基和叔丁基氧化物的锂盐、钠盐和钾盐；金属氨基化合物包括氨基钠、氨基钾和氨基锂；金属氢化物包括氢化钠、氢化钾和氢化锂；并且金属二烷基酰胺包括甲基、乙基、正丙基、异丙基、正丁基、叔丁基、三甲基甲硅烷基和环己基取代的酰胺的锂盐、钠盐和钾盐。
在一些实施方案中，本文中提供的化合物和盐是基本上分离的。通过“基本上分离”意指使化合物至少部分地或基本上从其形成或被检测到的环境分离。部分分离可以包括例如富含本文中提供的化合物的组合物。基本上分离可以包括包含按重量计至少约50％、至少约60％、至少约70％、至少约80％、至少约90％、至少约95％、至少约97％、或至少约99％的本文中提供的化合物或其盐的组合物。用于将化合物和它们的盐分离的方法在本领域中是常规的。
在本文中使用短语“药学上可接受的”以指如下的那些化合物、材料、组合物和/或剂型，其在合理的医学判断的范围内，适合用于与人类和动物的组织接触而没有过度的毒性、刺激、过敏反应或者其它问题或并发症，与合理的获益/风险比相称。
本申请还包括本文中所述的化合物的药学上可接受的盐。如本文中所使用的，“药学上可接受的盐”是指所公开的化合物的衍生物，其中通过将现有的酸或碱部分转化为其盐形式来修饰母体化合物。药学上可接受的盐的实例包括但不限于例如胺等碱性残基的无机酸盐或有机酸盐；和例如羧酸等酸性残基的碱金属盐或有机盐等。本申请的药学上可接受的盐包括例如由无毒的无机酸或有机酸形成的母体化合物的常规的无毒盐,主要包括无机酸盐如硫酸、硝酸、氢溴酸、磷酸、盐酸、硼酸、氨基磺酸等；或有机酸如乙酸、丙酸、丁酸、樟脑酸，癸酸、己酸、辛酸、碳酸、肉桂酸、羟基乙酸、三氟乙酸、己二酸、海藻酸、2-羟基丙酸、2-氧代丙酸、硬脂酸、乳酸、柠檬酸、草酸、丙二酸、琥珀酸、焦谷氨酸、抗坏血酸、天冬氨酸、苯乙酸、谷氨酸、苯甲酸、水杨酸、羟基马来酸、棕榈酸、肉桂酸、异丁酸、月桂酸、扁桃酸,、马来酸、富马酸、苹果酸、酒石酸、对氨基苯磺酸、2一乙酰氧基一苯甲酸、2-羟基-1,2,3-丙三酸、辛二酸、葡糖酸、葡萄糖醛酸、谷氨酸、戊二酸、甲酸、反丁烯二酸、粘酸、龙胆酸、丙酮酸、水杨酸、甲磺酸、乙基磺酸、苯甲磺酸、对甲苯磺酸、环己 基亚磺酸、羟乙基磺酸、乙烷二磺酸、4-(笏甲氧羰基氨基)丁酸、二氯乙酸、1，2-乙烷二磺酸、樟脑-10-磺酸、2，4-二羟基苯甲酸、α-酮戊二酸、1-羟基-2-萘甲酸、对乙酰氨基苯甲酸、2-羟基苯甲酸、4-氨基-2-羟基苯甲酸、全反式维甲酸、丙戊酸等。可以通过常规的化学方法从包含碱性或酸性部分的母体化合物合成本申请的药学上可接受的盐。通常，可以通过使这些化合物的游离酸或碱形式与化学计量量的适当的碱或酸在水中或在有机溶剂中或者在二者的混合物中反应来制备这样的盐；通常，非水性介质如醚、乙酸乙酯、醇类(例如，甲醇、乙醇、异丙醇或丁醇)或乙腈(MeCN)是优选的。
在一些实施方案中，疾病为癌症。在一些实施方案中，癌症选自由结直肠癌、子宫内膜癌、脑癌(例如，多形性成胶质细胞瘤)、黑色素瘤、胃癌、乳腺癌、卵巢癌、胰腺癌、肝癌、脑胶质瘤、脑内肿瘤、肾癌、前列腺癌、膀胱癌、肺癌、胰腺癌、卵巢癌、皮肤癌、上皮细胞癌、鼻咽癌、表皮细胞癌、宫颈癌、口腔癌、舌癌、人纤维肉瘤、多发性骨髓瘤和血液癌症组成的组。在一些实施方案中，癌症包括实体瘤。在一些实施方案中，癌症选自由胶质瘤、成胶质细胞瘤、非小细胞肺癌和血液癌症组成的组。
在一些实施方案中，癌症为血液癌症。在一些实施方案中，血液癌症选自由白血病和淋巴瘤组成的组。在一些实施方案中，血液癌症选自由以下组成的组：急性髓细胞白血病(AML)、慢性髓细胞样白血病、B细胞淋巴瘤、慢性淋巴细胞性白血病(CLL)、非何杰金淋巴瘤、毛细胞性白血病、套细胞淋巴瘤、伯基特淋巴瘤(Burkitt lymphoma)、小淋巴细胞性淋巴瘤、滤泡性淋巴瘤、淋巴浆细胞性淋巴瘤、结外缘区淋巴瘤(extranodal marginal zone lymphoma)、活化的B细胞样(ABC)弥漫性大B细胞淋巴瘤(activated B-cell like(ABC)diffuse large B cell lym phoma)、和生发中心B细胞(GCB)弥漫性大B细胞淋巴瘤(germinal center B cell(GCB)diffuse large B cell lymphoma)。
本发明所述的化合物可以单独使用或与其他治疗本发明所述疾病或病症的治疗剂联合使用。本发明的化合物与其他抗肿瘤药物联合。所述抗肿瘤药物包括但不限于：环磷酰胺、氮芥、马法兰、瘤可宁、卡莫斯汀、金属铂类如卡铂、顺铂、奥沙利铂、喜树碱、依立替康、柔红霉素、阿霉素、博来霉素、普卡霉素、紫杉醇、长春瑞滨、多西他赛、多柔比星、氟尿嘧啶、甲氨蝶呤、阿糖胞苷、吉西他滨、EGFR抑制剂、VEGFR抑制剂、ALK抑制剂、BTK抑制剂、mTOR抑制剂、HDAC抑制剂。
当用作药物时，可以将本文中提供的化合物和盐以药物组合物的形式给药。这些组合物可以如本文中或别处所述来制备，并且可以通过多种多样的途径给药，这取决于期望局部治疗还是全身治疗并且取决于要治疗的区域。给药可以是局部的(包括经皮、皮上、经眼以及包括鼻内递送、阴道递送和直肠递送的对黏膜给药)、经肺的(例如，通过散剂或气雾剂的吸入 或吹入，包括借助喷雾器；气管内或鼻内)、口服的或肠胃外的。肠胃外给药包括静脉内、动脉内、皮下、腹膜内、肌内注射或输注；或者颅内(例如，鞘内或心室内给药)。肠胃外给药可以为单次推注剂量(single bolus dose)的形式，或者可以例如通过连续灌注泵。
在一些实施方案中，本文中提供的化合物、盐和药物组合物适合用于肠胃外给药。在一些实施方案中，本文中提供的化合物、盐和药物组合物适合用于静脉内给药。
用于局部给药的药物组合物和制剂可以包括透皮贴剂、软膏剂、洗剂、乳膏剂、凝胶剂、滴剂、栓剂、喷雾剂、液体制剂和散剂。常规的药物载体，水性、粉末或油状基质、和增稠剂等会是必要的或期望的。
还提供包含与一种或多种药学上可接受的载体(例如，赋形剂)组合的作为活性成分的本文中提供的化合物或其药学上可接受的盐的药物组合物。在制备本文中提供的组合物时，通常将活性成分与赋形剂混合，通过赋形剂来稀释或者包封在为例如胶囊、药囊、纸或其它容器的形式的这样的载体内。当赋形剂用作稀释剂时，其可以为固体、半固体或液体材料，其起到活性成分的溶媒、载体或介质的作用。因此，组合物可以为如下形式：片剂、丸剂、散剂、锭剂、囊剂、扁囊剂、酏剂、混悬剂、乳剂、溶液剂、糖浆剂、气雾剂(作为固体或在液体介质中)、软膏剂、软的和硬的明胶胶囊剂、栓剂、无菌可注射溶液剂和无菌包装的散剂。
适当的赋形剂的一些实例包括以下而没有限制：乳糖、葡萄糖、蔗糖、山梨糖醇、甘露糖醇、淀粉、阿拉伯树胶、磷酸钙、藻酸盐、黄蓍胶、明胶、硅酸钙、微晶纤维素、聚乙烯吡咯烷酮、纤维素、水、糖浆和甲基纤维素。制剂可以另外包括以下而没有限制：润滑剂，如滑石、硬脂酸镁和矿物油；润湿剂；乳化剂和助悬剂；防腐剂，如羟基苯甲酸甲酯和羟基苯甲酸丙酯；甜味剂；调味剂；或其组合。
活性成分可以在宽的剂量范围内有效并且通常以药学上有效的量给药。然而，将理解的是，实际给予的化合物的量将通常由医师根据相关情况来确定，所述相关情况包括要治疗的病状，所选的给药途径，给予的实际化合物，个体受试者的年龄、体重和反应，和受试者的症状的严重性等。
本发明的有益效果在于基于HDAC多靶点的药物设计，为多种癌症的治疗提供新的化学实体，从而转变当前癌症治疗的方法。
附图说明
图1、部分化合物对急性髓性白血病细胞MV4-11和HL60的浓度-反应曲线。
图2、部分化合物对急性髓性白血病细胞PL21、KASUMI-1、MONO-MAC-1、NB-4的浓度-反应曲线。
图3、CZ411、FK866和它们的混合液对急性髓性白血病细胞MV4-11和HL60的浓度-反应曲 线。
图4、NMN对LEE18和LEE12导致的细胞死亡具有逆转作用。NMN为烟酰胺单核苷酸。
图5、HCT116肿瘤增长曲线和肿瘤图片。5-FU：5-氟尿嘧啶，传统的抗肿瘤化疗药物；Oxaliplatin：奥沙利铂，第3代铂类抗癌药，为抗肿瘤化疗药物。5-FU+Oxaliplatin为阳性对照。
图6、肿瘤生长曲线图。Panobinostat是上市的广谱HDAC抑制剂，为阳性对照药。
具体实施方式
将通过具体实施例的方式更详细地描述本发明。提供以下实施例用于说明性目的，并且不旨在以任何方式限制本发明。本领域技术人员将容易地认识到可以被改变或修改从而得到基本上相同的结果的多种多样的非关键参数。
一般材料和方法
所有对空气和水分不敏感的反应均在环境气氛下进行并且进行磁力搅拌。从深紫色的二苯甲酮羰游基钠(sodium benzophenone ketyl)中蒸馏四氢呋喃。干燥的DMF、干燥的DMSO。干燥的乙腈、干燥的二氯甲烷、干燥的甲苯、干燥的二氧六环购自安耐吉化学。所有对空气和水分敏感的操作均在氮气气氛下使用烘干的玻璃器皿来进行。
薄层色谱(TLC)通过预覆盖有250μm厚的硅胶60F254板的EMD TLC板来进行，并且在UV光和KMnO 4染色下通过荧光猝灭而可视化。
所有氘代溶剂均购自北京百灵威。在以下仪器上记录NMR谱图：对于 1H和 13C采集，在400MHz下运行的JEOL 400波谱仪。在以溶剂共振作为内标( 1H：CDCl 3，δ7.26；DMSO-d 6，δ2.50)，( 13C：CDCl 3，δ77.16；DMSO-d 6，δ39.52)的情况下，以ppm报告化学位移。如下报告数据：s为单峰，d为双峰，t为三重峰，q为四重峰，m为多重峰；以Hz计的耦合常数；积分；除非另有注明，碳信号为单峰。
下文中提供的实施例和制备例进一步阐明和举例说明本发明化合物及其制备方法。应当理解，下述实施例和制备例的范围并不以任何方式限制本发明的范围。
下面的合成路线描述了本发明中式II、III、IV或Ⅴ的化合物的制备，所有的原料都是通过这些路线中描述的方法、通过有机化学领域普通技术人员熟知的方法制备的或者可商购的。本发明的全部最终化合物都是通过这些路线中描述的方法或通过与其类似的方法制备的，这些方法是有机化学领域普通技术人员熟知的。这些路线中应用的全部可变因数如下文的定义或如权利要求中的定义。
本发明中式II、III、IV或Ⅴ的中间体化合物的制备如路线一和路线二所示，各取代基如发明内容部分所定义。
合成路线一：
Figure PCTCN2022109469-appb-000021
试剂与条件:(a)different amine,TBTU,TEA,DCM,yield 55％；(b)CH 3OH/H 2O,KOH,reflux,yield 80％；(c)4-nitrophenyl chloroformate,TEA,DCM,yield 80％；(d)methyl 8-aminocaprylate hydrochlorid,TEA,DCM,yield 65％；(e)sodium azide,DMF,80℃,yield 80％.
合成路线二：
Figure PCTCN2022109469-appb-000022
试剂与条件:(a)propionaldehyde,MeOH,yield 98％；NaBH 3CN,MeOH,concentrated hydrochloric acid,methyl orange,yield 60％；(b)(Boc) 2O,triethylamine,EtOH,yield 85％；(c)Pd/C,H 2,MeOH,yield 85％；(d)trifluoroacetic anhydride,DCM,yield 85％；(e)EDCI,HOBt,TEA,DCM,yield 55％；(f)Na 2CO 3,MeOH,yield 70％.
本发明中式II、III、IV或Ⅴ的化合物的制备如路线三、路线四、路线五和路线六所示，各取代基如发明内容部分所定义。
路线三：
Figure PCTCN2022109469-appb-000023
试剂与条件:(a)TBTU,TEA,DCM,yield 55％；(b)TFA,DCM,triethylamine,yield 85％.
路线四：
Figure PCTCN2022109469-appb-000024
试剂与条件:(a)TBTU,TEA,DCM,yield 55％.
路线五：
Figure PCTCN2022109469-appb-000025
试剂与条件:(a)3-ethynylpyridine,sodium ascorbate,CuSO 4,THF,H 2O,yield 85％.
路线六：
Figure PCTCN2022109469-appb-000026
试剂与条件:(a)TBTU,TEA,DCM,yield 55％；(b)EDCI,HOBt,triethylamine,DCM,yield50％；(c)TFA,DCM,triethylamine,yield 85％；(d)3-ethynylpyridine,sodium ascorbate,CuSO 4,THF,H 2O,yield 85％.
具体实施方案：
下面通过具体的实施例对本发明进行详细说明，但这些例举性实施方式的用途和目的仅用来例举本发明，并非对本发明的实际保护范围构成任何形式的任何限定，更非将本发明的保护范围局限于此。
路线一中化合物的制备：
实施例1:
Figure PCTCN2022109469-appb-000027
(E)-3-(3-(吡啶-3-基)丙烯酰胺基)丙酸甲酯(1a)的制备：将3-(吡啶-3-基)丙烯酸(0.44g，3mmol)溶于20mL二氯甲烷中，于冰浴下加入2-(1H-苯并三唑-1-基)-1,1,3,3-四甲基脲四氟硼酸(TBTU，1.05g，3.6mmol)和TEA(0.6mL，4.5mmol)。30分钟后，加入3-氨基丙酸甲酯盐酸盐(0.46g，3.3mmol)，然后加入0.6mL TEA，反应过夜。用饱和NaHCO 3(2×30mL)和饱和盐水(2×30mL)洗涤，并用MgSO 4干燥。蒸干溶剂后，通过快速色谱纯化，得到化合物1a，为白色固体粉末(0.35g，51％)。 1H NMR(600MHz,DMSO-d 6)δ8.75(d,J＝2.3Hz,1H),8.55(dd,J＝4.8,1.6Hz,1H),8.28(t,J＝5.7Hz,1H),7.98(dt,J＝8.0,2.0Hz,1H),7.49-7.42(m,2H),6.73(d,J＝15.9Hz,1H),3.62(s,3H),3.42(td,J＝6.8,5.6Hz,2H),2.55(t,J＝6.8Hz,2H).ESI-MS m/z:234.87[M+H] +。
Figure PCTCN2022109469-appb-000028
(E)-5-(3-(吡啶-3-基)丙烯酰胺基)戊酸甲酯(1b)的制备：采用1a的合成方法，以3-(吡啶-3-基)丙烯酸和5-氨基戊酸甲酯盐酸盐为原料，得到白色固体1b，收率55％。 1H NMR(600MHz,DMSO-d 6)δ8.75(d,J＝2.3Hz,1H),8.55(dd,J＝4.8,1.6Hz,1H),8.18(t,J＝5.8Hz,1H),7.98(dt,J＝7.9,2.0Hz,1H),7.48-7.42(m,2H),6.72(d,J＝15.9Hz,1H),3.59(s,3H),3.22-3.16(m,2H),2.34(t,J＝7.4Hz,2H),1.60-1.52(m,2H),1.52-1.43(m,2H).ESI-MS m/z:248.97[M+H] +。
Figure PCTCN2022109469-appb-000029
(E)-7-(3-(吡啶-3-基)丙烯酰胺基)庚酸甲酯(1c)的制备：采用1a的合成方法，以3-(吡啶-3-基)丙烯酸和7-氨基庚酸甲酯盐酸盐为原料，得到白色固体1c，收率53％。 1H NMR(600MHz,DMSO-d 6)δ8.75(d,J＝2.3Hz,1H),8.55(dd,J＝4.7,1.6Hz,1H),8.14(t,J＝5.7Hz,1H),7.97(dt,J＝7.9,2.0Hz,1H),7.50-7.41(m,2H),6.72(d,J＝15.9Hz,1H),3.58(s,3H),3.17(td,J＝7.0,5.7Hz,2H),2.34-2.26(m,2H),1.53(qd,J＝7.4,3.2Hz,2H),1.45(p,J＝7.4Hz,2H),1.30(h,J＝4.5,3.5Hz,4H).ESI-MS m/z:290.86[M+H] +。
Figure PCTCN2022109469-appb-000030
(E)-7-(3-(吡啶-3-基)丙烯酰胺基)辛酸甲酯(1d)的制备：采用1a的合成方法，以3-(吡啶-3-基)丙烯酸和8-氨基辛酸甲酯盐酸盐为原料，得到白色固体1d，收率53％。 1H NMR(600MHz,DMSO-d 6)δ8.75(d,J＝2.3Hz,1H),8.55(dd,J＝4.7,1.6Hz,1H),8.14(t,J＝5.7Hz,1H),7.97(dt,J＝7.9,2.0Hz,1H),7.47-7.42(m,2H),6.72(d,J＝15.9Hz,1H),3.58(s,3H),3.20-3.14(m,2H),2.30(t,J＝7.4Hz,2H),1.56-1.50(m,2H),1.49-1.40(m,2H),1.31-1.24(m,7H)。
Figure PCTCN2022109469-appb-000031
7-(3H-吡咯并[3,2-c]吡啶-2-甲酰胺)庚酸甲酯(6)的制备：采用1a的合成方法，以1H-吡咯并[3,2-c]吡啶-2-羧酸和7-氨基庚酸甲酯盐酸盐为原料，得到白色固体6，产率55％。 1H NMR(600MHz,DMSO-d 6)δ8.75(d,J＝2.3Hz,1H),8.55(dd,J＝4.7,1.6Hz,1H),8.14(t,J＝5.7Hz,1H),7.97(dt,J＝7.9,2.0Hz,1H),7.47-7.42(m,2H),6.72(d,J＝15.9Hz,1H),3.58(s,3H),3.17(td,J＝7.0,5.7Hz,2H),2.30(t,J＝7.4Hz,2H),1.52(t,J＝7.2Hz,2H),1.45(q,J＝7.1Hz,2H),1.31-1.24(m,7H).ESI-MS m/z:304.89[M+H] +。
Figure PCTCN2022109469-appb-000032
(E)-3-(3-(吡啶-3-基)丙烯酰胺基)丙酸(2a)的制备：将化合物1a(0.35g，1.5mmol)溶解于5mL甲醇中，然后添加2mL 3M KOH水溶液。将混合物在85℃回流2h。反应完成后在真空下蒸发MeOH。残渣用1N HCl酸化至pH 5-6，然后过滤。沉淀为相应的酸性2a，白色固体(0.28g，85％)。原料直接用于下一步，无需进一步纯化。ESI-MS m/z:219.15[M–H] -。
Figure PCTCN2022109469-appb-000033
(E)-5-(3-(吡啶-3-基)丙烯酰胺基)戊酸(2b)的制备：采用2a的合成方法，得到2b为白色固体，收率88％。 1H NMR(400MHz,DMSO-d 6)δ8.39(d,J＝13.3Hz,2H),7.59(d,J＝7.9Hz,1H),7.29(dd,J＝8.1,4.3Hz,1H),6.38(t,J＝6.0Hz,1H),5.98(t,J＝5.8Hz,1H),4.17(d,J＝5.9Hz,2H),2.97-2.92(m,2H),2.14(t,J＝7.3Hz,2H),1.45-1.24(m,8H).ESI-MS m/z: 278.18[M–H] -。
Figure PCTCN2022109469-appb-000034
(E)-7-(3-(吡啶-3-基)丙烯酰胺基)庚酸(2c)的制备：采用2a的合成方法，以化合物LL433为原料，得到白色固体2c，收率55％。 1H NMR(400MHz,DMSO-d 6)δ8.97(d,J＝2.1Hz,1H),8.73(dd,J＝5.4,1.4Hz,1H),8.46(dt,J＝8.2,1.8Hz,1H),8.29(t,J＝5.7Hz,1H),7.85(dd,J＝8.1,5.4Hz,1H),7.50(d,J＝15.9Hz,1H),6.85(d,J＝15.9Hz,1H),3.14(q,J＝6.6Hz,2H),2.26(t,J＝7.4Hz,2H),1.45(dt,J＝28.5,7.1Hz,4H),1.25(p,J＝3.6Hz,4H).ESI-MS m/z:275.24[M–H] -。
Figure PCTCN2022109469-appb-000035
(E)-8-(3-(吡啶-3-基)丙烯酰胺基)辛酸(2d)的制备：采用2a的合成方法，以化合物1d为原料，得到白色固体2d，收率87％。 1H NMR(400MHz,DMSO-d 6)δ8.97(d,J＝2.1Hz,1H),8.73(dd,J＝5.4,1.4Hz,1H),8.46(dt,J＝8.2,1.8Hz,1H),8.29(t,J＝5.7Hz,1H),7.85(dd,J＝8.1,5.4Hz,1H),7.50(d,J＝15.9Hz,1H),6.85(d,J＝15.9Hz,1H),3.14(q,J＝6.6Hz,2H),2.26(t,J＝7.4Hz,2H),1.45(dt,J＝28.5,7.1Hz,4H),1.25(p,J＝3.6Hz,4H).ESI-MS m/z:275.24[M–H] -。
Figure PCTCN2022109469-appb-000036
7-(3-(吡啶-3-基甲基)脲基)庚酸(5)的制备：采用2a的合成方法，以化合物4为原料，得到白色固体5，收率85％。 1H NMR(400MHz,DMSO-d 6)δ8.39(d,J＝13.3Hz,2H),7.59(d,J＝7.9Hz,1H),7.29(dd,J＝8.1,4.3Hz,1H),6.38(t,J＝6.0Hz,1H),5.98(t,J＝5.8Hz,1H),4.17(d,J＝5.9Hz,2H),2.97-2.92(m,2H),2.14(t,J＝7.3Hz,2H),1.45-1.24(m,8H).ESI-MS m/z:278.18[M–H] -。
Figure PCTCN2022109469-appb-000037
7-(1H-吡咯并[3,2-c]吡啶-2-甲酰胺)庚酸(7)。采用2a的合成方法，以化合物6为原料，得到白色固体7，收率85％。ESI-MS m/z:288.21[M–H] -。
Figure PCTCN2022109469-appb-000038
4-硝基苯基(吡啶-3-基甲基)氨基甲酸酯(3)的制备：将3-吡啶甲胺(0.22g，2mmol)和三乙胺(0.43g，2mmol)溶解于DCM中，然后加入4-硝基苯基氯甲酸盐(0.61g，3mmol)，在室温下搅拌5小时。反应完成后，用盐水洗涤3次并用Na 2SO 4干燥。蒸干溶剂得到化合物3，白色固体粉末(0.07g，70％产率)。 1H NMR(400MHz,DMSO-d 6)δ8.61(t,J＝6.1Hz,1H),8.52(s,1H),8.46(d,J＝4.8Hz,1H),8.26-8.19(m,2H),7.72(d,J＝8.0Hz,1H),7.43-7.38(m,2H),7.37-7.34(m,1H),4.31(d,J＝6.0Hz,2H).ESI-MS m/z:273.84[M+H] +。
Figure PCTCN2022109469-appb-000039
7-(3-(吡啶-3-基甲基)脲基)庚酸甲酯(4)的制备：向7-氨基庚酸甲酯盐酸盐(0.19g，1mmol)在二氯甲烷中的溶液中，在0℃下加入三乙胺(0.12g，1.2mmol)和化合物3(0.33g，1.2mmol)。室温下反应2小时。反应完成后，用盐水洗涤3次并用Na 2SO 4干燥。蒸干溶剂后，通过快速色谱纯化得到白色固体粉末化合物4(0.2g，70％产率)。 1H NMR(400MHz,DMSO-d 6)δ8.42(s,1H),8.39(d,J＝4.8Hz,1H),7.60(d,J＝7.8Hz,1H),7.29(dd,J＝7.8,4.8Hz,1H),6.31(t,J＝6.0Hz,1H),5.92(t,J＝5.7Hz,1H),4.17(d,J＝6.0Hz,2H),3.54(s,3H),2.94(q,J＝6.5Hz,2H),2.24(t,J＝7.4Hz,2H),1.47(p,J＝7.2Hz,2H),1.31(p,J＝6.9Hz,2H),1.21(h,J＝3.8,3.0Hz,4H).ESI-MS m/z:293.92[M+H] +。
Figure PCTCN2022109469-appb-000040
8-叠氮辛酸(8)的制备：将8-溴辛酸(0.33g，1.5mmol)溶于DMF，然后加入NaN 3(0.15g，2.25mmol)，80℃反应过夜。向该溶液中加入25mL DCM，用水洗涤溶剂5次，用Na 2SO 4干燥，蒸干溶剂后得化合物8，无色油状物(0.27g，95％收率)。 1H NMR(400MHz,DMSO-d 6)δ2.15(t,J＝7.4Hz,2H),1.46(dp,J＝14.4,7.1Hz,4H),1.28-1.19(m,6H).ESI-MS m/z:184.15[M–H] -。
路线二中化合物的制备：
实施例2：
Figure PCTCN2022109469-appb-000041
2-丙基肼-1-羧酸苄酯(9)的制备：将肼甲酸苄酯(1.66g，10mmol)溶解于50mL甲醇中，然后加入丙醛(0.61g，10.5mmol)，室温反应2小时。反应完全后，除去甲醇。将所得固体溶于30mL甲醇中，然后加入NaBH 3CN(1.2g，20mmol)和2滴浓HCl/MeOH(v:v＝1:1)溶液，反应过夜。反应完全后，蒸干溶剂，并通过快速色谱纯化粗产物，得到化合物9，为白色固体粉末(1.2g，60％)。 1H NMR(400MHz,DMSO-d 6)δ8.54(s,1H),7.36-7.23(m,5H),4.99(s,2H),4.46(s,1H),2.61(t,J＝7.1Hz,2H),1.32(h,J＝7.3Hz,2H),0.81(t,J＝7.4Hz,3H).ESI-MS m/z:108.92[M+H] +。
Figure PCTCN2022109469-appb-000042
1-(叔丁基)1-丙基肼-1,2-二羧酸2-苄基酯(10)的制备：将9(1g，5mmol)溶于50mL无水二氯甲烷中，加入三乙胺(1.5g，15mmol)和(Boc) 2O(0.22g，10mmol)。在室温下反应2小时后，用1M柠檬酸水溶液(3×100mL)和盐水(3×100mL)洗涤溶液，然后用MgSO 4干燥，蒸干溶剂后得到化合物10，为白色固体(1.3g，85％)。 1H NMR(400MHz,DMSO-d 6)δ9.45(s,1H),7.39-7.22(m,5H),5.05(s,2H),3.24(s,2H),1.44-1.27(m,11H),0.79(t,J＝7.3Hz,3H).ESI-MS m/z:308.86[M+H] +。
Figure PCTCN2022109469-appb-000043
1-丙基肼-1-羧酸叔丁酯(11)的制备：将化合物10(0.6g，2mmol)溶于甲醇中，加入Pd/C(0.06g)。然后在氢气中室温反应4小时。反应完成后，过滤除去Pd/C，蒸干滤液后，得到化合物11，为无色油状物(0.25g，72％)。 1H NMR(400MHz,DMSO-d 6)δ4.37(s,1H),3.16(t,J＝7.0Hz,2H),1.45(h,J＝14.3,7.2Hz,2H),1.36(s,9H),0.76(t,J＝7.4Hz,3H).ESI-MS m/z:174.87[M+H] +。
Figure PCTCN2022109469-appb-000044
4-((2,2,2-三氟乙酰氨基)甲基)苯甲酸(13a)的制备：在冰浴中将10mL三氟乙酸酐缓慢滴加到4-(氨基甲基)苯甲酸(3.0g，20mmol)中，滴毕在室温下反应2小时。反应完全后，加入100mL冰水淬灭反应，然后过滤，干燥滤饼，得到13a(4.43g，95％)。 1H NMR(400MHz,DMSO-d 6)δ10.05(t,J＝6.0Hz,1H),7.89(d,J＝6.4Hz,2H),7.35(d,J＝8.0Hz,2H),4.43(d,J＝6.0Hz,2H).ESI-MS m/z:246.16[M–H] -。
Figure PCTCN2022109469-appb-000045
4-((2,2,2-三氟乙酰氨基)甲基)苯甲酸(13b)的制备：采用13a的合成方法，以4-氨基苯甲酸和三氟乙酸酐为原料，得到白色固体13b，收率98％。 1H NMR(400MHz DMSO-d 6)δ12.94(s,1H),10.07(s,1H),7.94(d,J＝8.2Hz,2H),7.40(d,J＝8.2Hz,2H),4.47(d,J＝6.0Hz,2H).ESI-MS m/z:232.12[M–H] -。
Figure PCTCN2022109469-appb-000046
1-丙基-2-(4-((2,2,2-三氟乙酰氨基)甲基)苯甲酰)肼-1-羧酸叔丁酯(14a)的制备：将化合物13a(0.7g，3mmol)溶于20mL二氯甲烷中，冰浴下加入1-乙基-3-(3-二甲氨基丙基)碳二亚胺盐酸盐(EDCI·HCl，0.7g，3.6mmol)、1-羟基苯并三唑(HOBt，0.44g，3.6mmol)和三乙胺(0.6mL，4.5mmol)。30分钟后加入化合物11(0.57g，3.3mmol)，反应过夜。用饱和NaHCO 3(2×30mL)和盐水(2×30mL)洗涤反应液，合并有机相并用无水硫酸镁干燥。蒸干溶剂后通过快速色谱法纯化，得到白色固体粉末14a(0.66g，55％)。 1H NMR(400MHz,DMSO-d 6)δ10.47(s,1H),10.05(t,J＝6.1Hz,1H),7.78(dd,J＝16.9,7.8Hz,2H),7.39-7.29(m,2H),4.41(d,J＝5.9Hz,2H),3.34(s,2H),1.47-1.27(m,11H),0.84(t,J＝7.1Hz,3H).ESI-MS m/z:403.89[M+H] +。
Figure PCTCN2022109469-appb-000047
1-丙基-2-(4-(2,2,2-三氟乙酰氨基)苯甲酰)肼-1-羧酸叔丁酯(14b)的制备：采用14a的合成方法，以化合物13b和11为原料，得到白色固体14b，收率50％。 1H NMR(400MHz,DMSO-d 6)δ11.46(s,1H),10.49(s,1H),7.85(dd,J＝14.7,8.6Hz,2H),7.75(d,J＝8.4Hz,2H),1.55-1.43(m,2H),1.40-1.29(m,9H),0.84(t,J＝6.9Hz,3H).ESI-MS m/z:389.91[M+H] +。
Figure PCTCN2022109469-appb-000048
2-(4-(氨基甲基)苯甲酰基)-1-丙基肼-1-羧酸叔丁酯(15a)的制备：将化合物14a(1.2 g，3mmol)溶于20mL甲醇/水(v:v＝1:1)的溶液中，然后加入K 2CO 3(1.24g，9mmol)，室温下反应过夜。蒸干甲醇后用乙酸乙酯(2×20mL)萃取，合并有机相并用盐水(2×30mL)洗涤，用无水Na 2SO 4干燥。过滤后蒸干溶剂得到15a(0.74g，80％)。 1H NMR(400MHz,DMSO-d 6)δ10.42(s,1H),7.74(dd,J＝16.8,7.9Hz,2H),7.39(d,J＝8.0Hz,2H),3.72(s,2H),1.47(p,J＝7.3Hz,3H),1.39-1.28(m,9H),0.83(t,J＝7.0Hz,4H).ESI-MS m/z:307.94[M+H] +。
Figure PCTCN2022109469-appb-000049
2-(4-氨基苯甲酰基)-1-丙基肼-1-羧酸叔丁酯(15b)的制备：采用15a的合成方法，以化合物14b为原料，得到白色固体15b，收率82％。 1H NMR(400MHz,DMSO-d 6)δ10.00(s,1H),10.00(s,0H),7.58-7.49(m,2H),6.54-6.46(m,2H),5.68(d,J＝4.7Hz,2H),3.32-3.25(m,2H),1.46(h,J＝7.5Hz,2H),1.38-1.27(m,9H),0.82(t,J＝7.1Hz,3H).ESI-MS m/z:293.87[M+H] +。
路线三中化合物的制备
实施例3：
Figure PCTCN2022109469-appb-000050
(E)-N-(3-氧代-3-(2-丙基肼基)丙基)-3-(吡啶-3-基)丙烯酰胺(LEE1)的制备：将化合物2b(0.53g，3mmol)溶解于15mL二氯甲烷中，冰浴下加入O-苯并三氮唑-N,N,N’,N’-四甲基脲四氟硼酸(TBTU，1.05g，3.6mmol)和三乙胺(0.6mL，4.5mmol)。反应30分钟后，加入化合物11(0.57g，3.3mmol)，室温反应过夜。反应液用饱和NaHCO 3(2×30mL)和饱和盐水(2×30mL)洗涤，并用MgSO 4干燥。过滤后蒸干溶剂，通过快速色谱纯化得到白色固体粉末(0.35g，54％)。
将前一步骤的产物(0.30g，0.8mmol)溶解于15mL二氯甲烷中，然后加入三氟乙酸(TFA，0.11g，1.0mmol)后反应过夜。反应完全后，加入TEA(0.10g，1.0mmol)将pH调至8。溶液用盐水洗涤，并用Na 2SO 4上干燥。过滤后蒸干溶剂，然后经快速色谱纯化，得到白色固体粉末化合物LEE1(0.18g，85％)。 1H NMR(400MHz,DMSO-d 6)δ9.32(s,1H),8.75(d,J＝2.2Hz,1H),8.55(dd,J＝4.7,1.6Hz,1H),8.21(t,J＝5.8Hz,1H),7.97(dt,J＝8.0,2.0Hz,1H),7.50-7.40(m,2H),6.74(d,J＝15.9Hz,1H),4.79(s,1H),3.39(q,J＝6.6Hz,2H),2.60(dt,J＝18.8,6.8Hz,2H),2.27(t,J＝7.0Hz,2H),1.38(h,J＝7.3Hz,2H),0.85(t,J＝7.4Hz,3H). 13C  NMR(101MHz,DMSO-d 6)δ169.49,165.00,150.57,149.55,135.71,134.36,131.18,124.65,124.43,53.50,35.92,34.05,21.21,12.02.ESI-MS m/z:276.88[M+H] +。
实施例4：
Figure PCTCN2022109469-appb-000051
(E)-N-(5-氧代-5-(2-丙基肼基)戊基)-3-(吡啶-3-基)丙烯酰胺(LEE2)的制备：采用LEE1的合成方法，以化合物11和化合物2b为原料，得到白色固体LEE2，收率55％。 1H NMR(400MHz,DMSO-d 6)δ9.23(d,J＝5.6Hz,1H),8.75(d,J＝2.2Hz,1H),8.55(dd,J＝4.8,1.6Hz,1H),8.16(q,J＝5.8,4.4Hz,1H),7.97(dt,J＝8.0,2.0Hz,1H),7.49-7.38(m,2H),6.73(d,J＝15.9Hz,1H),4.77(d,J＝5.9Hz,1H),3.18(q,J＝6.5Hz,2H),2.61(h,J＝4.4Hz,2H),2.04(t,J＝7.2Hz,2H),1.60-1.32(m,6H),0.86(t,J＝7.4Hz,3H). 13C NMR(101MHz,DMSO-d 6)δ171.22,164.86,150.52,149.53,135.57,134.36,131.23,124.81,124.42,53.52,38.94,33.64,29.16,23.27,21.22,12.04.ESI-MS m/z:304.91[M+H] +。
实施例5：
Figure PCTCN2022109469-appb-000052
(E)-N-(7-氧代-7-(2-丙基肼基)庚基)-3-(吡啶-3-基)丙烯酰胺(LEE3)的制备：采用LEE1的合成方法，以化合物11和2c为原料，得到白色固体LEE3，收率57％。 1H NMR(400MHz,DMSO-d 6)δ9.21(d,J＝5.7Hz,1H),8.75(d,J＝2.2Hz,1H),8.55(dd,J＝4.8,1.6Hz,1H),8.14(t,J＝5.6Hz,1H),7.97(dt,J＝8.0,2.0Hz,1H),7.49-7.39(m,2H),6.73(d,J＝15.9Hz,1H),4.76(d,J＝6.2Hz,1H),3.17(q,J＝6.6Hz,2H),2.61(td,J＝6.9,4.5Hz,2H),2.01(t,J＝7.4Hz,2H),1.57-1.34(m,6H),1.28(tq,J＝14.3,8.6,8.0Hz,4H),0.86(t,J＝7.4Hz,3H). 13C NMR(101MHz,DMSO-d6)δ171.36,164.84,150.51,149.53,135.52,134.35,131.25,124.84,124.42,53.50,39.15,33.91,29.46,28.74,26.65,25.63,21.22,12.04.ESI-MS m/z:332.04[M+H] +。
实施例6：
Figure PCTCN2022109469-appb-000053
(E)-N-(8-氧代-8-(2-丙基肼基)辛基)-3-(吡啶-3-基)丙烯酰胺(LEE4)的制备：采用LEE1的合成方法，以化合物11和2d为原料，得到白色固体LEE4，收率60％。 1H NMR (400MHz,DMSO-d 6)δ10.04(s,1H),9.85(s,1H),8.71(d,J＝2.3Hz,1H),8.50(dd,J＝4.7,1.6Hz,1H),8.11(t,J＝5.6Hz,1H),7.93(dt,J＝8.0,2.0Hz,1H),7.72(d,J＝8.7Hz,2H),7.62(s,2H),7.45-7.35(m,2H),6.68(d,J＝15.9Hz,1H),3.13(q,J＝6.6Hz,2H),2.69(t,J＝7.1Hz,2H),2.29(t,J＝7.4Hz,2H),1.56(p,J＝6.8Hz,2H),1.42(h,J＝7.3Hz,4H),1.27(s,6H),0.87(t,J＝7.4Hz,3H). 13C NMR(126MHz,DMSO-d 6)δ171.55,164.84,150.41,149.39,135.50,134.48,131.24,124.81,124.47,52.86,39.14,33.65,29.50,28.87,28.83,26.79,25.38,20.05,11.73.ESI-MS m/z:346.97[M+H] +。
实施例7：
Figure PCTCN2022109469-appb-000054
1-(7-氧代-7-(2-丙基肼基)庚基)-3-(吡啶-3-基甲基)脲(LEE5)的制备：采用LEE1的合成方法，以化合物11和5为原料，得到白色固体LEE5，收率61％。 1H NMR(400MHz,DMSO-d 6)δ9.19(s,1H),8.41(d,J＝1.9Hz,1H),8.39(dd,J＝4.8,1.7Hz,1H),7.59(dt,J＝7.9,1.9Hz,1H),7.29(dd,J＝7.8,4.8,0.9Hz,1H),6.32(t,J＝6.1Hz,1H),5.93(t,J＝5.7Hz,1H),4.17(d,J＝5.9Hz,2H),2.93(q,J＝6.6Hz,2H),2.56(t,J＝7.1Hz,2H),1.95(t,J＝7.3Hz,2H),1.43(t,J＝7.2Hz,2H),1.37-1.29(m,3H),1.19(s,4H),0.81(t,J＝7.4Hz,3H)； 13C NMR(126MHz,DMSO-d 6)δ171.35,158.49,149.01,148.24,136.96,135.29,123.82,109.99,53.44,41.03,33.89,30.33,28.77,26.54,25.65,21.15,12.03.ESI-MS m/z:336.12[M+H] +。
实施例8：
Figure PCTCN2022109469-appb-000055
N-(7-氧代-7-(2-丙基肼基)庚基)-3H-吡咯并[3,2-c]吡啶-2-甲酰胺(LEE7)的制备：采用LEE1的合成方法，以化合物11和7为原料，得到白色固体LEE7，收率56％。 1H NMR(400MHz,DMSO-d 6)δ9.19(d,J＝7.9Hz,1H),8.93(s,1H),8.60(t,J＝5.7Hz,1H),8.21(d,J＝5.8Hz,1H),7.37(d,J＝5.8Hz,1H),7.25(s,1H),3.27(s,2H),2.60(t,J＝7.1Hz,2H),2.01(t,J＝7.3Hz,2H),1.59-1.46(m,4H),1.42-1.23(m,7H),0.86(t,J＝7.4Hz,3H). 13C NMR(101MHz,DMSO-d 6)δ171.36,160.86,145.13,141.64,139.89,133.77,124.88,107.83,101.93,53.49,33.91,29.52,28.77,26.66,25.64,21.22,12.05.ESI-MS m/z:346.07[M+H] +。
实施例9：
Figure PCTCN2022109469-appb-000056
8-叠氮-N'-丙辛烷酰肼(LEE6)的制备：采用1a的合成方法，以化合物8和11为原料，得到无色油状的LEE6，收率56％。ESI-MS m/z:241.98[M+H] +。
路线四中化合物的制备
实施例10：
Figure PCTCN2022109469-appb-000057
(E)-N-(2-氨基苯基)-8-(3-(吡啶-3-基)丙烯酰胺)辛酰胺(LEE8)的制备：采用1a的合成方法，以化合物2d和1,2-二氨基苯为原料，得到白色固体LEE8，收率53％。 1H NMR(400MHz,DMSO-d 6)δ9.05(s,1H),8.71(s,1H),8.51(d,J＝4.8Hz,1H),8.12(t,J＝5.7Hz,1H),7.93(d,J＝7.9Hz,1H),7.47-7.34(m,2H),7.11(d,J＝7.8Hz,1H),6.85(t,J＝7.6Hz,1H),6.74-6.63(m,2H),6.50(t,J＝7.5Hz,1H),4.82(s,1H),3.14(q,J＝6.5Hz,2H),2.27(t,J＝7.4Hz,2H),1.56(p,J＝7.0Hz,2H),1.43(p,J＝7.8Hz,2H),1.28(s,7H). 13C NMR(126MHz,DMSO-d 6)δ171.62,164.82,150.48,149.50,142.26,135.51,134.36,131.23,126.13,125.72,124.82,124.42,124.09,116.69,116.38,39.17,36.21,29.55,29.11,29.00,26.86,25.73.ESI-MS m/z:381.05[M+H] +。
实施例11：
Figure PCTCN2022109469-appb-000058
N-(2-氨基苯基)-8-叠氮辛酰胺(20)的制备：采用1a的合成方法，以化合物8与1,2-二氨基苯为原料，得到白色固体20，收率60％。 1H NMR(400MHz,DMSO-d 6)δ9.05(s,1H),7.10(d,J＝7.9Hz,1H),6.85(t,J＝7.6Hz,1H),6.67(d,J＝8.0Hz,1H),6.50(d,J＝7.5Hz,1H),4.75(s,2H),2.27(t,J＝7.4Hz,2H),1.52(dt,J＝20.1,7.0Hz,4H),1.28(s,8H).ESI-MS m/z:375.06[M+H] +。
路线五中化合物的制备
实施例12：
Figure PCTCN2022109469-appb-000059
正丙基-8-(4-(吡啶-3-基)-1H-1,2,3-三唑-1-基)辛烷酰肼(LEE10)的制备：将化合物19(60.2mg，0.25mmol)和3-乙炔基吡啶(26mg，0.25mmol)溶于THF/H2O(v:v＝2:1)的溶液中，加入抗坏血酸钠(50mg，0.25mmol)和硫酸铜(4mg，0.025mmol)，在室温下反应过夜。反应完全后，反应液用水洗涤3次，并用Na 2SO 4干燥。过滤后蒸干溶剂得到化合物LEE10，为白色固体粉末(0.07g，84％)。 1H NMR(400MHz,DMSO-d 6)δ9.19(s,1H),9.01(d,J＝2.2Hz,1H),8.68(s,1H),8.50(dd,J＝4.8,1.7Hz,1H),8.17(dt,J＝7.9,2.0Hz,1H),7.44(dd,J＝8.0,4.8Hz,1H),4.37(t,J＝7.1Hz,2H),2.55(t,J＝7.1Hz,1H),1.95(t,J＝7.3Hz,1H),1.82(p,J＝7.1Hz,2H),1.43(p,J＝7.3Hz,2H),1.35-1.16(m,8H),0.80(t,J＝7.4Hz,2H). 13C NMR(126MHz,DMSO-d 6)δ171.33,149.26,146.79,143.92,132.79,127.26,124.46,122.39,53.44,50.06,33.84,30.00,28.76,28.50,26.17,25.52,21.16,12.01.ESI-MS m/z:345.06[M+H] +。
实施例13：
Figure PCTCN2022109469-appb-000060
N-(2-氨基苯基)-8-(4-(吡啶-3-基)-1H-1,2,3-三唑-1-基)辛酰胺(LEE11)的制备：采用LEE10的合成方法，以化合物20和3-乙炔基吡啶为原料，得到化合物LEE11，为白色固体，收率76％。 1H NMR(400MHz,DMSO-d 6)δ9.08(s,1H),9.01(d,J＝2.2Hz,1H),8.68(s,1H),8.50(dd,J＝4.9,1.7Hz,1H),8.17(dt,J＝7.9,2.1Hz,1H),7.44(dd,J＝7.9,4.8Hz,1H),7.10(dd,J＝7.9,1.5Hz,1H),6.84(td,J＝7.6,1.6Hz,1H),6.67(dd,J＝8.0,1.5Hz,1H),6.49(td,J＝7.6,1.5Hz,1H),4.90(s,1H),4.39(t,J＝7.1Hz,2H),2.27(t,J＝7.4Hz,2H),1.84(p,J＝7.2Hz,2H),1.54(p,J＝7.3Hz,2H),1.40-1.20(m,6H). 13C NMR(126MHz,DMSO-d 6)δ171.60,149.24,146.78,143.92,142.20,132.82,127.27,126.11,125.69,124.47,124.11,122.40,116.70,116.39,50.08,36.16,30.03,28.93,28.62,26.21,25.65.ESI-MS m/z:379.07[M+H] +。
路线六中化合物的制备
实施例14：
Figure PCTCN2022109469-appb-000061
2-(4-((3H-吡咯[3,2-c]吡啶-2-甲酰胺)甲基)苯甲酰基)-1-丙基肼-1-羧酸叔丁酯(23c)的制备：将1H-吡咯并[3,2-c]吡啶-2-羧酸(0.48g，3mmol)溶解于15mL二氯甲烷中，于冰浴下将O-苯并三氮唑-N,N,N’,N’-四甲基脲四氟硼酸(TBTU，1.05g，3.6mmol)和三乙胺(0.6mL，4.5mmol)加入反应液。30分钟后加入化合物15a(1.0g，3.3mmol)，然后反应过夜。反应液用饱和NaHCO 3(2×30mL)和盐水(2×30mL)洗涤，并用MgSO 4干燥。过滤后蒸干溶剂，通过快速色谱纯化得到白色固体粉末(0.7g，54％)。 1H NMR(400MHz,DMSO-d 6)δ10.46(d,J＝5.5Hz,1H),9.29(t,J＝6.1Hz,1H),8.93(s,1H),8.20(d,J＝5.8Hz,1H),7.78(dd,J＝16.8,7.9Hz,2H),7.41(d,J＝8.1Hz,2H),7.36(d,J＝5.9Hz,1H),7.32(d,J＝4.0Hz,1H),4.55(d,J＝5.9Hz,2H),3.05(q,J＝7.3Hz,1H),1.53-1.42(m,2H),1.39-1.28(m,9H),0.83(t,J＝7.1Hz,3H).ESI-MS m/z:451.88[M+H] +。
2-(4-(((1H-吲哚-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-羧酸叔丁基酯(23g)的制备：采用23c的合成方法，以2-吲哚甲酸和化合物15a为原料，得到白色固体23g，收率56％。 1H NMR(400MHz,DMSO-d 6)δ11.60(s,1H),10.46(d,J＝7.5Hz,1H),9.09(t,J＝6.0Hz,1H),7.78(dd,J＝16.6,7.8Hz,2H),7.58(d,J＝8.0Hz,1H),7.40(dd,J＝8.2,3.6Hz,3H),7.15(d,J＝2.6Hz,2H),7.00(t,J＝7.5Hz,1H),4.53(d,J＝5.9Hz,2H),1.51-1.41(m,1H),1.34(s,9H),0.84(q,J＝7.1,6.7Hz,3H).ESI-MS m/z:451.05[M+H] +。
Figure PCTCN2022109469-appb-000062
2-(4-((苯并呋喃-2-甲酰胺)甲基)苯甲酰基)-1-丙基肼-1-羧酸叔丁酯(23a)的制备：采用23c的合成方法，以2-苯并呋喃甲酸和化合物15a为原料，得到白色固体23a，收率56％。 1H NMR(400MHz,DMSO-d 6)δ10.47(d,J＝5.4Hz,1H),9.36(t,J＝6.2Hz,1H),7.82-7.72(m,3H),7.63(d,J＝8.3Hz,1H),7.56(s,1H),7.42(t,J＝7.2Hz,3H),7.30(t,J＝7.5Hz,1H),4.51(d,J＝5.9Hz,2H),1.46(q,J＝7.1Hz,2H),1.39-1.28(m,9H),0.83(t,J＝7.0Hz,3H).ESI-MS m/z:451.08[M+H] +。
Figure PCTCN2022109469-appb-000063
2-(4-((苯并噻吩-2-甲酰胺)甲基)苯甲酰基)-1-丙基肼-1-羧酸叔丁酯(23b)的制备：采用23c的合成方法，以苯并噻吩-2-羧酸和化合物15a为原料，得到白色固体23b，收率53％。 1H  NMR(400MHz,DMSO-d 6)δ10.47(d,J＝5.9Hz,1H),9.38(t,J＝6.0Hz,1H),8.11(s,1H),8.05-7.95(m,1H),7.96-7.87(m,1H),7.78(dd,J＝16.9,7.9Hz,2H),7.47-7.37(m,5H),4.51(d,J＝5.8Hz,2H),1.46(q,J＝7.1Hz,2H),1.39-1.29(m,6H),0.83(t,J＝7.3Hz,3H).ESI-MS m/z:467.98[M+H] +。
Figure PCTCN2022109469-appb-000064
2-(4-((3H-吡咯并[2,3-c]吡啶-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-羧酸叔丁基酯(23e)的制备：采用23c的合成方法，以1H-吡咯并[2，3-c]吡啶-2-羧酸和化合物15a为原料，得到白色固体23e，产率为49％。 1H NMR(400MHz,DMSO-d 6)δ12.11(s,1H),10.46(s,1H),9.32(s,1H),8.76(s,1H),8.10(d,J＝5.5Hz,1H),7.85-7.72(m,2H),7.59(d,J＝5.6Hz,1H),7.41(d,J＝8.1Hz,2H),7.19(s,1H),4.55(d,J＝5.9Hz,2H),1.46(d,J＝7.2Hz,2H),1.28(s,9H),0.82(d,J＝7.0Hz,3H).ESI-MS m/z:452.07[M+H] +。
Figure PCTCN2022109469-appb-000065
2-(4-(((呋喃[3,2-c]吡啶-2-羧酰胺基)甲基)苯甲酰基)-1-丙基羟嗪-1-羧酸叔丁酯(23d)的制备：采用23c的合成方法，以苯并[b]噻吩-2-羧酸和化合物15a为原料，得到呈白色固体状的23d，产率为53％。 1H NMR(400MHz,DMSO-d 6)δ10.46(s,1H),9.49(t,J＝6.1Hz,1H),9.05(s,1H),8.55(d,J＝5.8Hz,1H),7.83-7.70(m,3H),7.68(s,1H),7.40(d,J＝8.2Hz,2H),4.51(d,J＝6.0Hz,2H),1.52-1.42(m,2H),1.39-1.28(m,9H),0.83(t,J＝7.2Hz,3H).ESI-MS m/z:453.08[M+H] +。
Figure PCTCN2022109469-appb-000066
2-(4-((5-溴-1H-吲哚-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-甲酸叔丁酯(23j)的制备:采用23c的合成方法，以5-溴吲哚-2-羧酸和化合物15a为原料，得到呈白色固体状的23j，产率为58％。
Figure PCTCN2022109469-appb-000067
2-(4-(((5-氟-1H-吲哚-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-甲酸叔丁酯(23l)的制备：采用23c的合成方法，以5-氟吲哚-2-羧酸和化合物15a为原料，得到呈白色固体状的23l，产率为53％。
Figure PCTCN2022109469-appb-000068
2-(4-((6-(二甲基氨基)-1H-吲哚-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-羧酸叔丁基酯(23k)的制备：采用23c的合成方法，以6-(二甲氨基)-1H-吲哚-2-羧酸和化合物15a为原料，得到呈白色固体状的23k，产率为50％。
Figure PCTCN2022109469-appb-000069
2-(4-((5-氯-1H-吲哚-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-甲酸叔丁酯(23m)的制备：采用23c的合成方法，以5-氯吲哚-2-羧酸和化合物15a为原料，得到呈白色固体状的23m，产率为51％。
Figure PCTCN2022109469-appb-000070
2-(4-(((5-甲氧基苯并呋喃-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-羧酸叔丁酯(23n)的制备：采用23c的合成方法，以5-甲氧基苯并呋喃-2-甲酸和化合物15a为原料，得到呈白色固体状的23n，产率为47％。
Figure PCTCN2022109469-appb-000071
2-(4-((5-氟苯并呋喃-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-甲酸叔丁酯(23o)的 制备：采用23c的合成方法，以5-氟苯并呋喃-2-羧酸和化合物15a为原料，得到呈白色固体状的23o，产率为59％。
Figure PCTCN2022109469-appb-000072
2-(4-((5-溴苯并呋喃-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-甲酸叔丁酯(23p)的制备：采用23c的合成方法，以5-溴苯并呋喃-2-羧酸和化合物15a为原料，得到呈白色固体状的23p，产率为52％。
Figure PCTCN2022109469-appb-000073
2-(4-(((5-甲氧基-1H-吲哚-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-甲酸叔丁酯(23q)的制备：采用23c的合成方法，以5-甲氧基吲哚-2-甲酸和化合物15a为原料，得到呈白色固体状的23q，产率为49％。
Figure PCTCN2022109469-appb-000074
2-(4-((5-氯苯并呋喃-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-甲酸叔丁酯(23r)的制备：采用23c的合成方法，以5-氯苯并呋喃-2-羧酸和化合物15a为原料，得到呈白色固体状的23r，产率为46％。
Figure PCTCN2022109469-appb-000075
2-(4-(((5-(丙-2-炔-1-基氧基)苯并呋喃-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-羧酸叔丁基酯(23s)的制备：采用23c的合成方法，以5-(丙-2-炔-1-基氧基)苯并呋喃-2-羧酸和化合物15a为原料，得到呈白色固体状的23s，产率为50％。
Figure PCTCN2022109469-appb-000076
2-(4-(((5-羟基苯并呋喃-2-羧酰胺基)甲基)苯甲酰基)-1-丙基肼-1-羧酸叔丁酯(23u)的制备：采用23c的合成方法，以5-羟基苯并呋喃-2-羧酸和化合物15a为原料，得到呈白色固体状的23u，产率为58％。
Figure PCTCN2022109469-appb-000077
(E)-1-丙基-2-(4-(((8-(3-(吡啶-3-基)丙烯酰胺基)辛酰胺基)甲基)苯甲酰基)肼-1-羧酸酯(23v)的制备：将化合物2d(0.29g，1mmol)溶于15mL二氯甲烷中，于冰浴下加入1-(3-二甲基氨基丙基)-3-乙基碳二亚胺盐酸盐(EDCI，0.23g，1.2mmol)和1-羟基苯并三唑(在0℃的HOBt，0.18g，1.2mmol)。30分钟后，加入15a(0.37mg，1.2mmol)和三乙胺(0.17mL，1.2mmol)，室温反应过夜。用盐水(2×30mL)洗涤反应液，并用MgSO 4干燥。过滤后蒸干溶剂，通过快速色谱纯化，得到化合物23v，为白色固体粉末(0.3g，51％)。 1H NMR(400MHz,DMSO-d 6)δ9.81(s,1H),8.71(d,J＝2.2Hz,1H),8.51(dd,J＝4.8,1.6Hz,1H),8.11(t,J＝5.7Hz,1H),7.95(dt,J＝8.0,1.9Hz,1H),7.45-7.36(m,2H),6.68(d,J＝15.9Hz,1H),3.21(s,3H),3.16-3.09(m,3H),2.01(t,J＝7.3Hz,2H),1.56-1.15(m,25H),0.78(t,J＝7.4Hz,3H).ESI-MS m/z:579.96[M+H] +。
Figure PCTCN2022109469-appb-000078
(E)-1-丙基-2-(4-(7-(3-(吡啶-3-基)丙烯酰胺基)庚酰胺基)苯甲酰基)叔丁基肼(羧酸)(23x)的制备：采用23v的合成方法，以化合物2c和15b为原料，得到为白色固体的23x，产率为53％。 1H NMR(400MHz,DMSO-d 6)δ10.36(d,J＝3.6Hz,1H),10.12(s,1H),8.72(d,J＝2.2Hz,1H),8.52(dd,J＝4.8,1.6Hz,1H),8.15(t,J＝5.7Hz,1H),7.96(dd,J＝8.0,2.0Hz,1H),7.69-7.67(m,4H),7.49(d,J＝1.3Hz,1H),6.69(d,J＝15.9Hz,1H),3.13(t,J＝6.5Hz,2H),2.32-2.26(m,2H),1.45(d,J＝7.2Hz,2H),1.41-1.26(m,13H),1.22(t,J＝6.2Hz,4H),0.83(t,J＝6.1Hz,3H).ESI-MS m/z:552.06[M+H] +。
Figure PCTCN2022109469-appb-000079
(E)-1-丙基-2-(4-(8-(3-(吡啶-3-基)丙烯酰胺基)辛酰胺基)苯甲酰基)肼叔丁基-1- 羧酸盐(23w)的制备：采用23v的合成方法，以化合物2d和15b为原料，得到呈白色固体状的23w，产率为51％。 1H NMR(400MHz,DMSO-d 6)δ10.34(s,1H),10.09(s,1H),8.71(s,1H),8.50(s,1H),8.11(t,J＝5.7Hz,1H),7.93(d,J＝7.9Hz,1H),7.74(d,J＝9.7Hz,2H),7.65(s,2H),7.42-7.37(m,3H),6.68(d,J＝15.9Hz,1H),3.13(q,J＝6.7Hz,3H),2.30(t,J＝7.4Hz,2H),1.56(t,J＝7.1Hz,2H),1.52-1.34(m,8H),1.33-1.16(m,13H),0.84(t,J＝8.4Hz,3H).ESI-MS m/z:566.04[M+H] +。
Figure PCTCN2022109469-appb-000080
2-(4-(8-叠氮基八氨基)苯甲酰基)-1-丙基肼-1-羧酸叔丁基酯(24)的制备：采用23v和LEE37的合成方法，以化合物8和13b为原料，得到白色固体的24，产率为50％。 1H NMR(400MHz,DMSO-d 6)δ10.34(s,1H),10.08(s,1H),7.74(d,J＝10.2Hz,2H),7.64(d,J＝8.7Hz,2H),2.29(t,J＝7.4Hz,2H),1.55(t,J＝7.0Hz,2H),1.48(q,J＝6.9Hz,4H),1.40(s,4H),1.28(d,J＝3.7Hz,13H),0.84(t,J＝7.2Hz,3H).ESI-MS m/z:461.12[M+H] +.
实施例15：
Figure PCTCN2022109469-appb-000081
(E)-N-(4-(2-丙基肼-1-羰基)苄基)-3-(吡啶-3-基)丙烯酰胺(LEE12)的制备：采用LEE1的合成方法，以化合物13b和(E)-3-(吡啶-3-基)丙烯酸为原料，得到白色固体状的LEE12，产率为52％。 1H NMR(600MHz,DMSO-d 6)δ9.98(s,1H),8.88-8.68(m,2H),8.56(dd,J＝4.8,1.6Hz,1H),8.00(dt,J＝8.0,2.0Hz,1H),7.85-7.76(m,2H),7.53(d,J＝15.9Hz,1H),7.50-7.42(m,1H),7.37(d,J＝8.3Hz,2H),6.82(d,J＝15.9Hz,1H),5.11(s,1H),4.46(d,J＝6.0Hz,2H),2.75(t,J＝7.1Hz,2H),1.47(h,J＝7.4Hz,2H),0.91(t,J＝7.5Hz,3H). 13C NMR(101MHz,DMSO-d 6)δ165.57,165.14,150.67,149.64,143.10,136.30,134.46,132.34,131.12,127.61,124.45,124.39,53.58,42.55,21.33,12.13.ESI-MS m/z:339.08[M+H] +。
实施例16：
Figure PCTCN2022109469-appb-000082
N-(4-(2-丙基肼-1-羰基)苄基)-1H-吡咯并[3，2-c]吡啶-2-甲酰胺(LEE18)的制备：将23c(0.22g，0.5mmol)溶于15mL二氯甲烷中，加入三氟乙酸(TFA，0.11g，1.0mmol)，室温下反应过夜。反应完全后，加入三乙胺(0.1g，1.0mmol)将pH调至8。然后用盐水洗涤溶液，并用Na 2SO 4干燥。过滤后蒸干溶剂，通过快速色谱纯化，得到化合物LEE18，白色固体粉末(0.14g，85％)。 1H NMR(400MHz,DMSO-d 6)δ9.42(t,J＝6.1Hz,1H),9.24(s,1H),8.30(d,J＝6.5Hz,1H),7.73-7.68(m,2H),7.67-7.63(m,1H),7.52(d,J＝0.9Hz,1H),7.35-7.28(m,2H),4.49(d,J＝6.0Hz,2H),2.64(t,J＝7.1Hz,2H),1.35(h,J＝7.3Hz,2H),0.79(t,J＝7.4Hz,3H). 13C NMR(101MHz,DMSO-d 6)δ165.56,160.21,142.79,141.87,140.84,136.81,135.09,132.44,127.68,127.57,124.42,109.75,104.54,53.55,42.64,21.28,12.12.ESI-MS m/z:351.86[M+H] +。
实施例17：
Figure PCTCN2022109469-appb-000083
N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺(LEE16)的制备：采用LEE18的合成方法，以化合物23a和三氟乙酸为原料，得到LEE16，为白色固体，产率为79％。 1H NMR(400MHz,DMSO-d 6)δ9.90(d,J＝6.0Hz,1H),9.27(t,J＝6.2Hz,1H),7.75-7.68(m,3H),7.59(dd,J＝8.4,1.1Hz,1H),7.51(d,J＝1.1Hz,1H),7.40(ddd,J＝8.4,7.2,1.4Hz,1H),7.33(d,J＝8.1Hz,2H),7.30-7.24(m,1H),5.00(q,J＝6.0Hz,1H),4.47(t,J＝6.3Hz,2H),2.67(q,J＝6.8Hz,2H),1.39(h,J＝7.3Hz,2H),0.83(t,J＝7.4Hz,3H). 13C NMR(101MHz,DMSO-d 6)δ165.60,158.72,154.75,149.49,143.00,132.36,127.64,127.60,127.35,124.20,123.26,112.27,110.12,53.57,42.42,21.32,12.12.ESI-MS m/z:352.07[M+H] +。
实施例18：
Figure PCTCN2022109469-appb-000084
N-(4-(2-丙基肼-1-羰基)苄基)苯并[b]噻吩-2-甲酰胺(LEE17)的制备：采用LEE18的合成方法，以化合物23b和三氟乙酸为原料，得到LEE17，为白色固体，产率为83％。 1H NMR(400MHz,DMSO-d 6)δ9.95-9.87(m,1H),9.30(t,J＝6.0Hz,1H),8.08(s,1H),7.98-7.93(m,1H),7.90-7.85(m,1H),7.76-7.70(m,2H),7.41-7.30(m,4H),5.01(s,1H),4.47(d,J＝5.9Hz, 2H),2.73-2.60(m,2H),1.39(h,J＝7.3Hz,2H),0.83(t,J＝7.4Hz,3H). 13C NMR(101MHz,DMSO-d 6)δ165.59,162.10,143.03,140.72,140.17,139.64,132.41,127.64,127.60,126.72,125.69,125.46,125.41,123.29,53.58,42.95,21.32,12.13.ESI-MS m/z:367.96[M+H] +。
实施例19：
Figure PCTCN2022109469-appb-000085
N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-甲酰胺(LEE15)的制备：采用LEE12的合成方法，以化合物15a和1H-吲哚-2-羧酸为原料，缩合，脱保护后得到LEE15，为白色固体，产率为56％。 1H NMR(400MHz,DMSO-d 6)δ11.54(s,1H),9.89(d,J＝5.6Hz,1H),9.01(t,J＝6.1Hz,1H),7.75-7.70(m,2H),7.55(dt,J＝7.9,1.0Hz,1H),7.40-7.30(m,3H),7.15-7.08(m,2H),6.97(ddd,J＝8.0,6.9,1.0Hz,1H),5.05-4.94(m,1H),4.48(d,J＝6.0Hz,2H),2.67(td,J＝7.1,5.7Hz,2H),1.39(h,J＝7.3Hz,2H),0.83(t,J＝7.4Hz,3H). 13C NMR(101MHz,DMSO-d 6)δ165.60,161.70,143.45,136.99,132.29,131.98,127.60,127.57,127.44,123.83,121.99,120.22,112.79,103.16,53.58,42.41,21.32,12.13.ESI-MS m/z:350.86[M+H] +。
实施例21：
Figure PCTCN2022109469-appb-000086
N-(4-(2-丙基肼-1-羰基)苄基)-1H-吡咯并[2,3-c]吡啶-2-羧酰胺(LEE21)的制备：采用LEE18的合成方法，以化合物23e和三氟乙酸为原料，得到LEE21，为白色固体，产率为82％。 1H NMR(400MHz,DMSO-d 6)δ12.06(s,1H),9.90(s,1H),9.25(t,J＝6.1Hz,1H),8.76-8.71(m,1H),8.07(d,J＝5.6Hz,1H),7.78-7.68(m,2H),7.55(dd,J＝5.5,1.2Hz,1H),7.38-7.30(m,2H),7.15(s,1H),5.04(s,1H),4.51(d,J＝6.0Hz,2H),2.67(t,J＝7.1Hz,2H),1.39(h,J＝7.3Hz,2H),0.84(t,J＝7.4Hz,4H).13C NMR(101MHz,DMSO-d 6)δ165.57,161.10,143.05,138.53,136.28,135.24,133.77,132.39,131.72,127.64,127.50,116.28,101.96,53.57,42.55,21.32,12.13.ESI-MS m/z:352.06[M+H] +。
实施例22：
Figure PCTCN2022109469-appb-000087
(E)-N-(4-(2-丙基肼-1-羰基)苄基)-3-(吡啶-2-基)丙烯酰胺(LEE13)的制备：采用LEE18的合成方法，以化合物15a和3-(2-吡啶基)丙烯酸为原料，得到LEE13，为白色固体，产率57％。 1H NMR(400MHz,DMSO-d 6)δ9.96(s,1H),8.82(t,J＝6.1Hz,1H),8.58(d,J＝5.2Hz,2H),7.76(d,J＝7.9Hz,2H),7.58–7.25(m,6H),6.88(d,J＝15.9Hz,1H),5.06(s,1H),4.42(d,J＝6.0Hz,2H),2.70(t,J＝7.5Hz,2H),1.42(h,J＝7.1Hz,2H),0.86(t,J＝7.5Hz,3H). 13C NMR(101MHz,DMSO-d 6)δ165.57,165.31,165.13,153.46,150.67,150.33,149.64,143.13,143.10,139.02,137.66,136.29,134.46,132.34,131.12,127.61,127.53,125.95,124.74,124.53,124.45,124.39,53.58,42.55,42.52,21.32,12.13.ESI-MS m/z:339.06[M+H] +。
实施例23：
Figure PCTCN2022109469-appb-000088
(E)-N-(4-(2-丙基肼-1-羰基)苄基)-3-(吡啶-4-基)丙烯酰胺(LEE14)的制备：采用LEE18的合成方法，以化合物15a和3-(4-吡啶基)丙烯酸为原料，得到LEE14，为白色固体，产率55％。 1H NMR(400MHz,DMSO-d 6)δ9.98(s,1H),8.81-8.69(m,2H),8.56(dd,J＝4.8,1.6Hz,1H),8.00(dt,J＝8.0,2.0Hz,1H),7.85-7.76(m,2H),7.53(d,J＝15.9Hz,1H),7.45(dd,J＝8.0,4.8Hz,1H),7.40-7.32(m,2H),6.82(d,J＝15.9Hz,1H),5.09(s,1H),4.46(d,J＝5.9Hz,2H),2.75(t,J＝7.1Hz,2H),1.47(h,J＝7.3Hz,2H),0.91(t,J＝7.4Hz,3H). 13C NMR(101MHz,DMSO-d 6)δ165.57,165.14,150.67,149.64,143.10,136.30,134.46,132.34,131.12,127.61,124.45,124.39,53.58,42.55,21.33,12.13.ESI-MS m/z:339.05[M+H] +。
实施例24：
N-(4-(2-丙基肼-1-羰基)苄基)噻吩并[3,2-c]吡啶-2-甲酰胺(LEE19)。采用LEE18的合成方法，以化合物15a和噻吩并[3,2-c]吡啶-2-羧酸为原料，得到LEE19，为白色固体，产率54％。ESI-MS m/z:368.92[M+H] +。
实施例25：
Figure PCTCN2022109469-appb-000089
N-(4-(2-丙基肼-1-羰基)苄基)呋喃[3,2-c]吡啶-2-甲酰胺(LEE20)的制备：采用LEE18的合成方法，以化合物23d和三氟乙酸为原料，得到LEE20，为白色固体，产率为78％。 1H NMR(400MHz,DMSO-d 6)δ9.98(s,1H),9.49(t,J＝6.2Hz,1H),9.05(s,1H),8.55(d,J＝5.8Hz,1H),7.80-7.63(m,4H),7.36(d,J＝7.9Hz,2H),4.49(d,J＝6.1Hz,2H),2.70(t,J＝7.2Hz,2H),1.42(h,J＝7.4Hz,2H),0.86(t,J＝7.4Hz,3H)； 13C NMR(101MHz,DMSO-d 6)δ165.64,158.98,158.21,150.06,146.84,146.48,142.86,132.38,127.68,125.05,108.53,108.17,53.58,42.53,21.33,12.19.ESI-MS m/z:352.88[M+H] +。
实施例26：
Figure PCTCN2022109469-appb-000090
5-溴-N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-甲酰胺(LEE26)的制备：采用LEE18的合成方法，以化合物23j和三氟乙酸为原料，得到LEE26，为白色固体，收率84％。 1H NMR(500MHz,DMSO-d 6)δ11.88(s,1H),9.98(s,1H),9.27(d,J＝6.9Hz,1H),7.84(d,J＝1.9Hz,1H),7.81-7.76(m,2H),7.42-7.36(m,3H),7.31-7.26(m,1H),7.18(s,1H),5.06(s,1H),4.53(d,J＝6.0Hz,2H),2.73(t,J＝7.1Hz,2H),1.50-1.39(m,2H),0.89(t,J＝7.4Hz,4H).
实施例27：
Figure PCTCN2022109469-appb-000091
5-氟-N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-羧酰胺(LEE28)的制备：采用LEE18的合成方法，以化合物23l和三氟乙酸为原料，得到LEE28，为白色固体，收率85％。 1H NMR(500MHz,DMSO-d 6)δ11.78(s,1H),9.99(s,1H),9.26(t,J＝6.1Hz,1H),7.79(d,J＝8.3Hz,2H),7.45-7.36(m,4H),7.19(s,1H),7.07-6.99(m,1H),5.06(s,1H),4.53(d,J＝6.1Hz,2H),2.73(t,J＝7.1Hz,2H),1.49-1.39(m,2H),0.89(t,J＝7.4Hz,4H).
实施例28：
Figure PCTCN2022109469-appb-000092
6-(二甲基氨基)-N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-甲酰胺(LEE27)的制备：采用LEE18的合成方法，以化合物23k和三氟乙酸为原料，得到LEE27，为白色固体，收率82％。
实施例29：
Figure PCTCN2022109469-appb-000093
5-氯-N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-甲酰胺(LEE29)的制备：采用LEE18的合成方法，以化合物23m和三氟乙酸为原料，得到LEE29，为白色固体，收率80％。 1H NMR(500MHz,DMSO-d 6)δ11.87(s,1H),9.98(s,1H),9.26(t,J＝6.1Hz,1H),7.79(d,J＝8.3Hz,2H),7.69(d,J＝2.1Hz,1H),7.43(d,J＝8.7Hz,1H),7.39(d,J＝8.3Hz,2H),7.21-7.15(m,2H),5.06(s,1H),4.53(d,J＝6.1Hz,2H),2.73(t,J＝7.1Hz,2H),1.50-1.39(m,2H),0.89(t,J＝7.4Hz,4H).
实施例30：
Figure PCTCN2022109469-appb-000094
5-甲氧基-N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺(LEE30)的制备：采用LEE18的合成方法，以化合物23n和三氟乙酸为原料，得到LEE30，为白色固体，收率85％。 1H NMR(500MHz,DMSO-d 6)δ9.98(s,1H),9.31(t,J＝6.2Hz,1H),7.81-7.75(m,2H),7.57-7.52(m,1H),7.51(d,J＝1.0Hz,1H),7.42-7.34(m,2H),7.26(d,J＝2.6Hz,1H),7.05(dd,J＝9.0,2.7Hz,1H),4.50(d,J＝6.1Hz,2H),3.79(s,3H),2.73(t,J＝7.1Hz,2H),1.51-1.40(m,2H),0.89(t,J＝7.4Hz,3H).
实施例31：
Figure PCTCN2022109469-appb-000095
5-氟-N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺(LEE31)的制备：采用LEE18的合成方法，以化合物23o和三氟乙酸为原料，得到LEE31，为白色固体，收率86％。 1H NMR(500MHz,DMSO-d 6)δ9.96(d,J＝6.2Hz,1H),9.40(t,J＝6.1Hz,1H),7.81-7.74(m,2H),7.72-7.65(m,1H),7.62-7.55(m,2H),7.41-7.36(m,2H),7.35-7.28(m,1H),5.07(s,1H),4.51(d,J＝6.1Hz,2H),2.73(t,J＝7.1Hz,2H),1.45(h,J＝7.3Hz,2H),0.89(t,J＝7.5Hz,3H).
实施例32：
Figure PCTCN2022109469-appb-000096
5-溴-N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺(LEE32)的制备：采用LEE18的合成方法，以化合物23p和三氟乙酸为原料，得到LEE32，为白色固体，收率84％。 1H NMR(500MHz,DMSO-d 6)δ9.98(s,1H),9.45(t,J＝6.2Hz,1H),8.01(d,J＝2.0Hz,1H),7.81-7.75(m,2H),7.64(d,J＝8.8Hz,1H),7.60(d,J＝2.0Hz,1H),7.58(d,J＝0.9Hz,2H),7.39(d,J＝8.2Hz,2H),5.07(s,1H),4.51(d,J＝6.1Hz,2H),2.73(t,J＝7.1Hz,2H),1.53-1.35(m,2H),0.89(t,J＝7.4Hz,3H).
实施例33：
Figure PCTCN2022109469-appb-000097
5-甲氧基-N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-甲酰胺(LEE33)的制备：采用LEE18的合成方法，以化合物23q和三氟乙酸为原料，得到LEE33，为白色固体，收率82％。 1H NMR(500MHz,DMSO-d 6)δ11.47(s,1H),9.98(s,1H),9.07(t,J＝6.1Hz,1H),7.81-7.76(m,2H),7.42-7.36(m,2H),7.31(d,J＝8.9Hz,1H),7.12-7.05(m,2H),6.83(dd,J＝8.9,2.5Hz,1H),5.09(s,1H),4.53(d,J＝6.1Hz,2H),3.75(s,3H),2.73(t,J＝7.1Hz,2H),1.50-1.40(m,2H),0.89(t,J＝7.4Hz,3H)。
实施例34：
Figure PCTCN2022109469-appb-000098
5-氯-N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺(LEE34)的制备：采用LEE18的合成方法，以化合物23r和三氟乙酸为原料，得到LEE34，为白色固体，收率86％。 1H NMR(500MHz,DMSO-d 6)δ9.99(d,J＝4.1Hz,1H),9.49(t,J＝6.1Hz,1H),7.87(d,J＝2.2Hz,1H),7.81-7.75(m,2H),7.69(d,J＝8.8Hz,1H),7.60(d,J＝1.0Hz,1H),7.48(dd,J＝8.8,2.2Hz,1H),7.42-7.36(m,2H),5.09(s,1H),4.51(d,J＝6.1Hz,2H),2.73(t,J＝7.1Hz,2H),1.49-1.39(m,3H),0.89(t,J＝7.4Hz,4H).
实施例35：
Figure PCTCN2022109469-appb-000099
5-(丙-2-炔-1-基氧基)-N-(4-(2-丙肼-1-羰基)苄基)苯并呋喃-2-甲酰胺(LEE35)的制备：采用LEE18的合成方法，以化合物23s和三氟乙酸为原料，得到LEE35，为白色固体，收率83％。 1H NMR(500MHz,DMSO-d 6)δ9.97(s,1H),9.31(t,J＝6.2Hz,1H),7.81-7.74(m,2H),7.61-7.55(m,1H),7.52(d,J＝0.9Hz,1H),7.43-7.36(m,2H),7.34(d,J＝2.6Hz,1H),7.13-7.07(m,1H),5.08(s,1H),4.83(d,J＝2.4Hz,2H),4.51(d,J＝6.1Hz,2H),3.56(t,J＝2.4Hz,1H),2.73(t,J＝7.1Hz,2H),1.45(h,J＝7.4Hz,2H),0.89(t,J＝7.4Hz,4H).
实施例36：
Figure PCTCN2022109469-appb-000100
5-羟基-N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺(LEE36)的制备：采用LEE18的合成方法，以化合物23u和三氟乙酸为原料，得到LEE36，为白色固体，收率79％。 1H NMR(500MHz,DMSO-d 6)δ9.96(s,1H),9.39(s,1H),9.24(t,J＝6.2Hz,1H),7.81-7.74(m,2H),7.46-7.35(m,4H),7.02(d,J＝2.5Hz,1H),6.90(dd,J＝8.9,2.5Hz,1H),4.49(d,J＝6.2Hz,2H),2.73(t,J＝7.1Hz,2H),1.50-1.39(m,2H),0.89(t,J＝7.4Hz,3H).
实施例37：
Figure PCTCN2022109469-appb-000101
(E)-N-(4-(2-丙基肼-1-羰基)苄基)-8-(3-(吡啶-3-基)丙烯酰胺基)辛酰胺(LEE37)的制备：采用LEE18的合成方法，以化合物23v和三氟乙酸为原料，得到LEE37，为白色固体，产率为78％。 1H NMR(400MHz,DMSO-d 6)δ9.93(s,1H),8.71(s,1H),8.50(d,J＝4.7Hz,1H),8.31(t,J＝6.0Hz,1H),8.12(t,J＝5.7Hz,1H),7.93(dt,J＝8.0,2.0Hz,1H),7.73(d,J＝8.0Hz,2H),7.45-7.36(m,2H),7.25(d,J＝8.0Hz,2H),6.69(d,J＝15.9Hz,1H),5.06(s,1H),4.25(d,J＝5.9Hz,2H),3.13(q,J＝6.6Hz,2H),2.70(t,J＝7.1Hz,2H),2.10(t,J＝7.4Hz,2H),1.55-1.35(m,6H),1.31-1.15(m,7H),0.86(t,J＝7.4Hz,3H). 13C NMR(126MHz,DMSO-d 6)δ172.69,165.56,164.83,150.49,149.51,143.60,135.52,134.34,132.12,131.22,127.49,127.36,124.81,124.41,53.55,42.16,39.17,35.76,29.53,29.09,28.94,26.84,25.70,21.30,12.12.ESI-MS m/z:479.96[M+H] +。
实施例38：
Figure PCTCN2022109469-appb-000102
(E)-N-(4-(2-丙基肼-1-羰基)苄基)-7-(3-(吡啶-3-基)丙烯酰胺基)庚酰胺(LEE39)的制备：采用LL289的合成方法，以化合物15a和2d为原料，得到为白色固体的YKR-31，产率为58％。 1H NMR(400MHz,DMSO-d 6)δ9.93(s,1H),8.51(dd,J＝4.7,1.6Hz,1H),8.32(t,J＝6.1Hz,1H),8.13(t,J＝5.6Hz,1H),7.73(d,J＝8.0Hz,2H),7.46-7.36(m,2H),7.26(d,J＝8.0Hz,2H),6.69(d,J＝15.9Hz,1H),5.03(s,1H),4.25(d,J＝5.8Hz,2H),3.13(q,J＝6.6Hz,2H),7.97-7.90(m,1H),2.70(t,J＝7.0Hz,2H),2.11(t,J＝7.4Hz,2H),1.53-1.38(m,6H),1.24(dq,J＝9.4,5.7,5.0Hz,4H),0.86(t,J＝7.4Hz,3H),8.75-8.67(m,1H). 13C NMR(126MHz,DMSO-d 6)δ172.67,165.57,164.84,153.46,150.50,149.51,143.60,135.53,135.52,134.34,132.12,131.22,127.98,127.50,127.42,127.35,124.81,124.41,53.56,42.19,42.16,39.15,35.75,29.45,28.86,26.69,25.89,25.70,21.30,12.12,11.10.ESI-MS m/z:465.89[M+H] +。
实施例39：
Figure PCTCN2022109469-appb-000103
(E)-N-(4-(2-丙基肼-1-羰基)苯基)-7-(3-(吡啶-3-基)丙烯酰胺基)庚酰胺(LEE40)的制备：采用LEE18的合成方法，以化合物23x和三氟乙酸为原料，得到LEE40，为白色固体，产率为65％。 1H NMR(400MHz,DMSO-d 6)δ10.06(s,1H),9.84(d,J＝5.5Hz,1H),8.71(d,J＝2.2Hz,1H),8.50(dd,J＝4.8,1.6Hz,1H),8.13(t,J＝5.7Hz,1H),7.93(dt,J＝8.1,2.0Hz,1H),7.72(d,J＝8.6Hz,2H),7.61(d,J＝8.6Hz,2H),7.45-7.35(m,2H),6.69(d,J＝15.9Hz,1H),5.00(d,J＝5.5Hz,1H),3.14(q,J＝6.5Hz,2H),2.69(q,J＝6.4Hz,2H),2.29(t,J＝7.4Hz,2H),1.57(d,J＝7.3Hz,2H),1.43(p,J＝7.3Hz,4H),1.31-1.26(m,4H),0.87(t,J＝7.4Hz,3H). 13C NMR(126MHz,DMSO-d 6)δ172.09,165.33,164.83,150.50,149.51,142.38,135.51,134.34,131.22,128.82,128.28,127.85,124.81,124.41,118.63,53.62,39.14,36.87,29.42,28.84,26.71,25.41,21.31,12.12,11.14.ESI-MS m/z:451.97[M+H] +。
实施例40：
Figure PCTCN2022109469-appb-000104
(E)-N-(4-(2-丙基肼-1-羰基)苯基)-8-(3-(吡啶-3-基)丙烯酰胺基)酰胺(LEE38)的制备：采用LEE18的合成方法，以化合物23w和三氟乙酸为原料，得到LEE38，为白色固体，产率为67％。 1H NMR(400MHz,DMSO-d 6)δ10.04(s,1H),9.85(s,1H),8.71(d,J＝2.3Hz,1H),8.50(dd,J＝4.7,1.6Hz,1H),8.11(t,J＝5.6Hz,1H),7.93(dt,J＝8.0,2.0Hz,1H),7.72(d,J＝8.7Hz,2H),7.62(s,2H),7.45-7.35(m,2H),6.68(d,J＝15.9Hz,1H),3.13(q,J＝6.6Hz,2H),2.69(t,J＝7.1Hz,2H),2.29(t,J＝7.4Hz,2H),1.56(p,J＝6.8Hz,2H),1.42(h,J＝7.3Hz,4H),1.27(s,6H),0.87(t,J＝7.4Hz,3H). 13C NMR(126MHz,DMSO-d 6)δ172.10,165.34,164.82,150.49,149.50,142.39,135.51,134.34,131.22,128.29,127.83,124.81,124.41,118.62,53.60,39.15,36.90,29.53,29.08,28.98,26.82,25.42,21.29,12.12.ESI-MS m/z:465.97[M+H] +。
实施例41：
Figure PCTCN2022109469-appb-000105
N-(4-(2-丙基肼-1-羰基)苯基)-8-(4-(吡啶-3-基)-1H-1,2,3-三唑-1-基)辛酰胺(LEE41)的制备：采用LEE18的合成方法，以化合物24和三氟乙酸为原料，得到白色固体，产率为67％。
采用LEE11的合成方法，以上一步的产物和3-乙炔基吡啶为原料，得到白色固体的LEE41，产率为80％。 1H NMR(400MHz,DMSO-d 6)δ10.08(s,1H),9.93(s,1H),9.01(d,J＝2.2Hz,1H),8.68(s,1H),8.49(d,J＝4.7Hz,1H),8.16(dd,J＝8.0,2.0Hz,1H),7.72(d,J＝8.5Hz,2H),7.61(d,J＝8.5Hz,2H),7.44(dd,J＝8.0,4.8Hz,1H),4.38(t,J＝7.1Hz,2H),2.71(t,J＝7.1Hz,2H),2.28(t,J＝7.3Hz,2H),1.83(p,J＝7.2Hz,2H),1.54(p,J＝7.3Hz,2H),1.43(h,J＝7.3Hz,2H),1.35-1.16(m,8H),0.87(t,J＝7.4Hz,3H). 13C NMR(126MHz,DMSO-d 6)δ172.10,165.34,149.24,146.77,143.92,142.46,132.82,128.33,127.67,127.27,124.47,122.41,118.62,53.53,50.07,36.84,30.01,28.91,28.61,26.17,25.34,21.15,12.08.ESI-MS m/z:464.03[M+H] +。
实施例42：
Figure PCTCN2022109469-appb-000106
(E)-4-((3-(吡啶-3-yl)丙烯酰胺)甲基)苯甲酸甲酯(LEE43)的制备：采用1a的合成方法，得到LEE43，为白色固体，收率51％。 1H NMR(400MHz,DMSO-d 6)δ8.80-8.78(m,2H),8.57(dd,J＝1.1Hz,J＝3.1Hz,1H),8.03(td,J＝1.1Hz,J＝5.4Hz,1H),7.95(td,J＝1.3Hz,J＝5.5Hz,2H),7.55(d,J＝10.6Hz,1H),7.47-7.43(m,3H),6.84(d,J＝10.6Hz,1H),4.51(d,J＝4.0Hz,2H),3.85(s,3H)。
实施例43：
Figure PCTCN2022109469-appb-000107
4-((1H-吡咯[3,2-c]吡啶-2-羰酰胺)甲基)苯甲酸甲酯(LEE44)的制备：采用1a的合成方法，得到LEE44，为白色固体，收率46％。1H NMR(400MHz,DMSO-d 6)δ12.94(s,1H),9.65(t,J＝4.0Hz,1H),9.31(s,1H),8.39(d,J＝4.3Hz,1H),7.96(d,J＝5.6Hz,2H),7.74(d,J＝4.2Hz, 1H),7.64(s,1H),7.51(d,J＝5.5Hz,2H),4.64(d,J＝4.0Hz,2H),3.85(s,3H)。
实施例44：化合物对HDAC1，2，3的抑制活性。
实验材料：HDAC缓冲液:15mM Tris-HCl(PH 8.0)，250μM EDTA，250mM NaCl，10％甘油。胰蛋白酶终止液：10mg/ml胰酶，50mM Tris-HCL(pH8.0),100mM NaCl，2μM TSA。底物：HDAC1，HDAC2和HDAC3的专属底物二甲基亚砜溶解配成30mM的储液，用HDAC缓冲液稀释至300μM，使得二甲基亚砜含量为1％左右。酶液：HDAC1，HDAC2和HDAC3按1:20用HDAC缓冲液稀释。
实验步骤：
a)100％溶液的配制：50μL HDAC缓冲液与10μL酶液混合，5min后加入40μL底物在37℃下反应30min，然后加入100μL胰蛋白酶终止液终止上述反应，并在37℃下反应20min，于390nm/460nm测定荧光强度，即得100％吸收。用AMC作为标准品做标准曲线，计算酶活。
b)空白溶液的配制：60μL HDAC buffer加入40μL底物后在37℃下反应30min，然后加入100μL胰蛋白酶终止液，并在37℃下反应20min，于390nm/460nm测定荧光强度，即得空白吸收。
6.药物抑制HDAC酶活的测定步骤：50μL含有药物的HDAC buffer与10μL酶液混合预先孵育5min，加入40μL底物后，在37℃下反应30min，然后加入100μL胰蛋白酶终止液终止上述反应，并在37℃下反应20min，于390nm/460nm测定荧光强度。
Figure PCTCN2022109469-appb-000108
最后将化合物的抑制率(％)和其相应浓度进行S曲线拟合，计算出IC 50值。
本发明所述结构通式(I、II、III或IV)所示的部分化合物对HDAC1，HDAC2，HDAC3的抑制活性结果见下面表1：
表1、部分化合物对HDAC1、2、3的抑制IC 50值。
Figure PCTCN2022109469-appb-000109
Figure PCTCN2022109469-appb-000110
Figure PCTCN2022109469-appb-000111
a表中数据均来自于三次独立的实验，数值为平均值，标准偏差<10％；ND：未检测。
实验结果表明，表格中化合物绝大多数对HDAC1/2/3均具有纳摩尔级的抑制IC 50值，对HDAC1和HDAC3的活性普遍高于HDAC2。活性普遍高于阳性对照药SAHA和MS275。
实施例45：化合物对NAMPT的抑制活性。
表2、部分化合物对NAMPT的抑制IC 50值。
Figure PCTCN2022109469-appb-000112
Figure PCTCN2022109469-appb-000113
a表中数据均来自于三次独立的实验，数值为平均值，标准偏差<10％。
实验结果表明，LEE12、LEE18、LEE7、LEE43、LEE44均表现出微摩尔级的NAMPT的抑制活性，上述化合物相较于对比例1和对比例2中的化合物取得了预料不到的技术效果。
实施例46：化合物对肿瘤细胞半数生长抑制浓度(GI 50)和半数致死浓度(LC 50)
半数生长抑制浓度(GI 50)和半数致死浓度(LC 50)由NCI方法测定。白血病细胞株MV4-11、HL60、PL21、KASUMI-1、MONO-MAC-1、NB-4培养于含有10％胎牛血清的IMDM培养液中，以10000个/100uL的密度接种于96孔细胞培养板中，培养过夜。选定6个孔为Tz孔，加入0.125mg/mL的Cell Titer-Blue染料，培养4h后于560nM/590nM(激发波长/发射波长)处读取荧光强度。其余孔加入不同浓度的化合物，同时设置100％对照组，培养48h后加入0.125mg/mL的Cell Titer-Blue染料，4h后于560nM/590nM处读取荧光强度。加药组的荧光强度以Ti表示，100％对照组的荧光强度以C表示，加药之前的荧光强度以Tz表示。
如果Ti≥Tz，使用公式[(Ti-Tz)/(C-Tz)]×100
如果Ti<Tz，使用公式[(Ti-Tz)/Tz]×100
半数生长抑制浓度(GI 50)为[(Ti-Tz)/(C-Tz)]×100＝50的化合物浓度，半数致死浓度(LC 50)为[(Ti-Tz)/Tz]×100＝-50的浓度。部分化合物对白血病细胞株MV4-11、HL60、PL21、KASUMI-1、MONO-MAC-1、NB-4的GI 50和LC 50值见图1、表3、图2和表4所示。
表3、部分化合物对急性髓性白血病细胞MV4-11和HL60的GI 50和LC 50值。
Figure PCTCN2022109469-appb-000114
Figure PCTCN2022109469-appb-000115
注：KF866为文献报道的NAMPT抑制剂。
a表中数据均来自于三次独立的实验，数值为平均值，标准偏差<10％； bND：未检测。
实验结果表明，在wt-p53细胞株MV4-11中，所测试化合物均表现出纳摩尔级的GI 50值和LC 50值，说明它们不仅可以导致细胞增殖抑制，也可以导致细胞死亡，大多数化合物的活性显著强于对比例1化合物LP411(3b)的抑制活性。而在p53-null细胞株HL60中，所测试化合物均表现出纳摩尔级的GI 50值，说明它们具有良好的抗增值活性，而仅有LEE12、LEE14、LEE18、LEE7具有纳摩尔级的LC 50值，可以导致细胞死亡。说明同时抑制HDAC和NAMPT可能对p53-null的细胞株具有合成致死作用。
表4、部分化合物对白血病细胞系的GI 50和LC 50值。
Figure PCTCN2022109469-appb-000116
Figure PCTCN2022109469-appb-000117
a表中数据均来自于三次独立的实验，数值为平均值，标准偏差<10％。
图2和表4也表明了，HDAC和NAMPT双抑制剂LEE12和LEE18对p53-mutant以及p53-null的细胞株具有致死作用，绝大多数化合物相对于对比例1中化合物，取得了预料不到的技术效果。
实施例47：目标化合物LEE17体内抗结肠癌活性活性
5-FU:5-氟尿嘧啶，传统的抗肿瘤化疗药物，Oxaliplatin：奥沙利铂，第3代铂类抗癌药，为抗肿瘤化疗药物。
于裸鼠右肩皮下接种结肠癌细胞HCT116，每只100uL(细胞计数：1.8*10 8个/mL)，一周后开始分组给药，将荷瘤鼠分组，灌胃给药，分组情况如下：
待测组：化合物LL341，给药剂量8mg/kg/d口服，给药体积：每次每只0.2mL
Figure PCTCN2022109469-appb-000118
相对肿瘤体积(RTV)＝Vt/Vo
抗肿瘤活性的评价指标为相对肿瘤增殖率T/C(％)，
Figure PCTCN2022109469-appb-000119
表7、化合物LEE17对HCT116荷瘤模型体内研究数据结果
Figure PCTCN2022109469-appb-000120
注：5-FU：5-氟尿嘧啶，传统的抗肿瘤化疗药物；Oxaliplatin：奥沙利铂，第3代铂类抗癌药，为抗肿瘤化疗药物。
实验结果表明，在8mg/kg/d给药剂量下，化合物LEE17能显著抑制HCT116肿瘤的生长，抑瘤率达到了85.5％，明显高于阳性药5-FU+Oxaliplatin。实验结束，裸鼠体重并无明显变化，说明了LEE17在给药剂量下具有一定的安全性。
实施例48：目标化合物LEE15、LEE16体内抗白血病活性活性Panobinostat:帕比司他，上市的广谱HDAC抑制剂
于裸鼠右肩皮下接种急性髓性白血病细胞MV4-11，每只100uL(细胞计数：1.8*10 8个/mL)，一周后开始分组给药，将荷瘤鼠分组，灌胃给药，分组情况如下：
待测组：化合物LEE15、LEE16，给药剂/4mg/kg/d，给药体积：每次每只200uL/20g；
阳性对照组：阳性药panobinostat，给药剂量4mg/kg/d，腹腔注射。
空白对照组：给予相同体积的PBS。
每天给药一次，每隔3-4天测量肿瘤体积，取各组平均值，绘制肿瘤生长曲线(见图6),MV4-11荷瘤模型给药第23天处死裸鼠，解剖肿瘤及内脏，实验结束，称出瘤块的重量，并按照公式计算抑瘤率。测量肿瘤最大径(a)和最小径(b)，计算肿瘤体积(V)：V＝ab 2/2，并计算相对肿瘤增殖率T/C(％)。
Figure PCTCN2022109469-appb-000121
相对肿瘤体积(RTV)＝Vt/Vo
抗肿瘤活性的评价指标为相对肿瘤增殖率T/C(％)，
Figure PCTCN2022109469-appb-000122
表8、化合物LEE15、LEE16对MV4-11荷瘤模型体内研究数据结果
Figure PCTCN2022109469-appb-000123
注：Panobinostat，阳性对照药，上市的广谱HDAC抑制剂。
实验结果表明，在4mg/kg的给药剂量下，LEE15和LEE16具有显著的体内抗急性髓性白血病作用，肿瘤抑制率为81.3％和78.4％。体内抗肿瘤活性明显高于阳性对照药panobinostat。
实施例49：目标化合物LEE15、LEE16、LEE17的药代动力学性质
LEE15、LEE16、LEE17溶解于40％PEG300和60％H2O。3只老鼠每组，分别以口服灌胃(po)20mg/kg和静脉注射(iv)5mg/kg的方式单次给药。给药结束后分别在0.083、0.25、0.5、1、2、4、6、8和24h取血，制备样品后分别测定t 1/2、C 0、AUC、Vss、CLp、MRT、C max、t max、F％等参数。
Po administration
Figure PCTCN2022109469-appb-000124
由上述代谢数据结果可知：化合物LEE16、LEE17、LEE15的代谢效果明显优于对比例2中的化合物的代谢效果，上述化合物LEE16、LEE17、LEE15在代谢上具有预料不到的技术效果。
以上所述仅为本申请的优选实施例而已，并不用于限制本申请，对于本领域的技术人员来说，本申请可以有各种更改和变化。凡在本申请的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本申请的保护范围之内。

Claims (14)

  1. 一种具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，其特征在于，所述的多靶点HDAC化合物具有如通式I，
    环E-B-L-C(O)-(NH)r-R(通式I)
    其中环E选自
    Figure PCTCN2022109469-appb-100001
    r＝1、2，R选自R 1、
    Figure PCTCN2022109469-appb-100002
    所述R 1选自H、C 1- 4烷基、C 3-5环烷基或C 1-2烷基取代的C 3-5环烷基；
    A环选自一个或多个取代或未取代的5元芳环或芳杂环，所述芳杂环含有1-2个N、O或S的杂原子；所述的取代基为H、卤素、C 1-2烷基、卤代甲基、OH、OCH 3、O(CH 2) nCH 3、环丙基氧基、OC(CH 3) 3、OCH(CH 3) 2、NH 2、N(CH 3) 2、NH(CH 2) nCH 3、CN、N 3等，其中n为0-9；
    X 1选自CR 4或N；
    X 2选自CR 5或N；
    X 3选自CR 6或N；
    X 4选自CR 7或N；
    X 5、X 6或X 7独立地选自CH或N；
    R 2和R 3各自独立地选自H、卤素、CH 3、OCH 3；
    R 4、R 5、R 6和R 7选自H、卤素、C 1-2烷基、卤代甲基、OH、OCH 3、O(CH 2) nCH 3、OC(CH 3) 3、OCH(CH 3) 2、环丙基氧基、5-6元烷氧基、NH 2、N(CH 3) 2、NH(CH 2) nCH 3、CN、N 3，其中n为0-9；
    B选自
    Figure PCTCN2022109469-appb-100003
    Figure PCTCN2022109469-appb-100004
    表示与A环相连的键；
    Figure PCTCN2022109469-appb-100005
    表示与L相连的键；
    L选自由C 1-14烷基、C 1-14烷氧基、C 2-14烯基、C 2-14炔基、C 3-10环烷基、C 6-10芳基或苄基组成的组。
  2. 根据权利要求1所述一种具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药：其特征在于，所述通式I化合物进一步具有如通式Ⅱ、通式Ⅲ、通式Ⅳ、通式Ⅴ所示的结构，
    Figure PCTCN2022109469-appb-100006
  3. 根据权利要求1-2任一项所述一种具有多靶点抑制活性的HDAC化合物及其药学上可其中，
    A环选自如下环系：
    Figure PCTCN2022109469-appb-100007
    其中，
    G选自CH 2、NH、N(CH 2) nCH 3、O或S，其中n为0-9；
    Figure PCTCN2022109469-appb-100008
    表示与相邻并环相连的键；
    Figure PCTCN2022109469-appb-100009
    表示与B相连的键。
  4. 根据权利要求1-2任一项所述一种具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，其中X 1、X 2、X 3或X 4为N，并且其中1-2个同时为N；。
  5. 根据权利要求1-2任一项所述一种具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，其中X 5为N。
  6. 根据权利要求1-2任一项所述一种具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，X 6或X 7为N。
  7. 根据权利要求1-2任一项所述一种具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，R 4、R 5、R 6或R 7各自独立选自H、卤素、(C 1-2)烷基、卤代甲基、OH、OCH 3、O(CH 2) nCH 3、环丙基氧基、OC(CH 3) 3、OCH(CH 3) 2、5-6元烷氧基、NH 2、N(CH 3) 2、NH(CH 2) nCH 3、CN、N 3，其中n为0-9。
  8. 根据权利要求1-7任一项所述一种具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，其中L选自直链C 1-14烷基或L选自直链C 1- 14烷氧基例如-(CH 2CH 2O) m-,-(CH 2CH 2O) m-C 2H 4-,-(CH 2CH 2O) m-CH 2-,-(CH 2CH 2O) n-,-C 2H 4-(CH 2CH 2O) n-,-CH 2-(OCH 2CH 2) n-,-(CH 2) p-O-(CH 2) q-；其中m、n、p或q独立选自1、2、3或4；或L为C 3-10环烷基、C 6-10芳基或苄基时，所述环烷基、芳基或苄基可被C 1-9烷基、C 1 -9烷氧基、(C 1-9烷基)-(C＝O)NH取代；或L为C 3-10环烷基时，所述环烷基可含有杂原子，如N、O或S；或L为芳基或苄基时，所述芳基或苄基可为芳杂环，如N、O或S。
  9. 根据权利要求1-2任一项所述一种具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，其中化合物选自：
    (E)-N-(3-氧代-3-(2-丙基肼基)丙基)-3-(吡啶-3-基)丙烯酰胺
    (E)-N-(5-氧代-5-(2-丙基肼基)戊基)-3-(吡啶-3-基)丙烯酰胺
    (E)-N-(7-氧代-7-(2-丙基肼基)庚基)-3-(吡啶-3-基)丙烯酰胺
    (E)-N-(8-氧代-8-(2-丙基肼基)辛基)-3-(吡啶-3-基)丙烯酰胺
    1-(7-氧代-7-(2-丙基肼基)庚基)-3-(吡啶-3-基甲基)脲N-(7-氧代-7-(2-丙基肼基)庚基)-3H-吡咯并[3,2-c]吡啶-2-甲酰胺
    8-叠氮-N'-丙辛烷酰肼
    (E)N-(2-氨基苯基)-8-(3-(吡啶-3-基)丙烯酰胺)辛酰胺
    N-(2-氨基苯基)-8-叠氮辛酰胺
    正丙基-8-(4-(吡啶-3-基)-1H-1,2,3-三唑-1-基)辛烷酰肼
    N-(2-氨基苯基)-8-(4-(吡啶-3-基)-1H-1,2,3-三唑-1-基)辛酰胺
    (E)-N-(4-(2-丙基肼-1-羰基)苄基)-3-(吡啶-3-基)丙烯酰胺
    N-(4-(2-丙基肼-1-羰基)苄基)-1H-吡咯并[3，2-c]吡啶-2-甲酰胺
    N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺
    N-(4-(2-丙基肼-1-羰基)苄基)苯并[b]噻吩-2-甲酰胺
    N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-甲酰胺
    N-(4-(2-丙基肼-1-羰基)苄基)-1H-吡咯并[2,3-c]吡啶-2-羧酰胺
    (E)-N-(4-(2-丙基肼-1-羰基)苄基)-3-(吡啶-2-基)丙烯酰胺
    (E)-N-(4-(2-丙基肼-1-羰基)苄基)-3-(吡啶-4-基)丙烯酰胺
    N-(4-(2-丙基肼-1-羰基)苄基)噻吩并[3,2-c]吡啶-2-甲酰胺
    N-(4-(2-丙基肼-1-羰基)苄基)呋喃[3,2-c]吡啶-2-甲酰胺
    5-溴-N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-甲酰胺
    5-氟-N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-甲酰胺
    6-(二甲基氨基)-N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-甲酰胺
    5-氯-N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-甲酰胺
    5-甲氧基-N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺
    5-氟-N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺
    5-溴-N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺
    5-甲氧基-N-(4-(2-丙基肼-1-羰基)苄基)-1H-吲哚-2-甲酰胺
    5-氯-N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺
    5-(丙-2-炔-1-基氧基)-N-(4-(2-丙肼-1-羰基)苄基)苯并呋喃-2-甲酰胺
    5-羟基-N-(4-(2-丙基肼-1-羰基)苄基)苯并呋喃-2-甲酰胺
    (E)-N-(4-(2-丙基肼-1-羰基)苄基)-8-(3-(吡啶-3-基)丙烯酰胺基)辛酰胺
    (E)-N-(4-(2-丙基肼-1-羰基)苄基)-7-(3-(吡啶-3-基)丙烯酰胺基)庚酰胺
    (E)-N-(4-(2-丙基肼-1-羰基)苯基)-7-(3-(吡啶-3-基)丙烯酰胺基)庚酰胺
    (E)-N-(4-(2-丙基肼-1-羰基)苯基)-8-(3-(吡啶-3-基)丙烯酰胺基)辛酰胺
    N-(4-(2-丙基肼-1-羰基)苯基)-8-(4-(吡啶-3-基)-1H-1,2,3-三唑-1-基)辛酰胺
    (E)-4-((3-(吡啶-3-yl)丙烯酰胺)甲基)苯甲酸甲酯
    4-((1H-吡咯[3,2-c]吡啶-2-羰酰胺)甲基)苯甲酸甲酯
  10. 根据权利要求1-9中所述一种具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药，其中药学上可接受的盐包括无机酸盐如硫酸、硝酸、氢溴酸、磷酸、盐酸、硼酸、氨基磺酸等；或有机酸如乙酸、丙酸、丁酸、樟脑酸、癸酸、己酸、辛酸、碳酸、肉桂酸、羟基乙酸、三氟乙酸、己二酸、海藻酸、2-羟基丙酸、2-氧代丙酸、硬脂酸、乳酸、柠檬酸、草酸、丙二酸、琥珀酸、焦谷氨酸、抗坏血酸、天冬氨酸、苯乙酸、谷氨酸、苯甲酸、水杨酸、羟基马来酸、棕榈酸、肉桂酸、异丁酸、月桂酸、扁桃酸,、马来酸、富马酸、苹果酸、酒石酸、对氨基苯磺酸、2-乙酰氧基一苯甲酸、2- 羟基-1,2,3-丙三酸、辛二酸、葡糖酸、葡萄糖醛酸、谷氨酸、戊二酸、甲酸、反丁烯二酸、粘酸、龙胆酸、丙酮酸、水杨酸、甲磺酸、乙基磺酸、苯甲磺酸、对甲苯磺酸、环己基亚磺酸、羟乙基磺酸、乙烷二磺酸、4-(笏甲氧羰基氨基)丁酸、二氯乙酸、1，2-乙烷二磺酸、樟脑-10-磺酸、2，4-二羟基苯甲酸、α-酮戊二酸、1-羟基-2-萘甲酸、对乙酰氨基苯甲酸、2-羟基苯甲酸、4-氨基-2-羟基苯甲酸、全反式维甲酸、丙戊酸。
  11. 一种药物组合物，其包含根据权利要求1至9中任一项所述具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药和/或其他治疗剂和/或至少一种药学上可接受的载体，所述其他治疗效果的药物例如选自HDAC抑制剂。
  12. 根据权利要求11任一项所述的药物组合物，其特征在于，所述药物组合物为注射制剂、口服制剂，例如药物组合物的剂型为片剂、丸剂、散剂、锭剂、囊剂、扁囊剂、酏剂、混悬剂、乳剂、溶液剂、糖浆剂、气雾剂(作为固体或在液体介质中)、软膏剂、软的和硬的明胶胶囊剂、栓剂、无菌可注射溶液剂和无菌包装的散剂。
  13. 根据权利要求1-9任一项所述一种具有多靶点抑制活性的HDAC化合物及其药学上可接受的盐、水合物、氘代物、异构体、或前药或权利要求11-12任一项所述的药物组合物在制备预防或治疗与组蛋白去乙酰化酶活性异常表达或NAD合成相关疾病的药物中用途，其中所述与组蛋白去乙酰化酶活性异常表达或NAD合成相关疾病例如选自恶性肿瘤、神经退行性疾病、病毒感染、艾滋病、炎症、疟疾、糖尿病、真菌感染、细菌感染，其中恶性肿瘤包括各类白血病、淋巴瘤、骨髓瘤、结直肠癌、黑色素瘤、胃癌、乳腺癌、卵巢癌、胰腺癌、肝癌、脑胶质瘤、脑内肿瘤、肾癌、前列腺癌、膀胱癌、肺癌、胰腺癌、卵巢癌、皮肤癌、上皮细胞癌、鼻咽癌、表皮细胞癌、宫颈癌、口腔癌、舌癌、人纤维肉瘤。
  14. 一种具有多靶点抑制活性的HDAC化合物的制备方法，其制备步骤包括：
    Figure PCTCN2022109469-appb-100010
    试剂与条件:(a)丙醛，甲醇，产率98％；氰基硼氢化钠，甲醇，浓盐酸，甲基橙，产率60％；(b)二碳酸二叔丁酯，三乙胺，乙醇，产率85％；(c)钯碳，氢气，甲醇，产率85％；(d)三氟乙酸酐，二氯甲烷，产率85％；(e)1-(3-二甲胺基丙基)-3-乙基碳二亚胺，1-羟基苯并三唑，三乙胺，二氯甲烷，产率55％；(f)碳酸钾，甲醇，产率70％；(g)O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸，三乙胺，二氯甲烷，产率55％；(h)三氟乙酸，二氯甲烷，三乙胺，产率85％。
PCT/CN2022/109469 2021-08-04 2022-08-01 靶向于hdac和nad合成的多靶点抑制剂及其用途 WO2023011416A1 (zh)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116496178A (zh) * 2023-04-27 2023-07-28 合肥工业大学 具有丁香酸和氨基酸结构片段的化合物及其合成与用途

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004058234A2 (en) * 2002-12-27 2004-07-15 Schering Aktiengesellschaft Pharmaceutical combinations of phthalazine vegf inhibitors and benzamide hdac inhibitors
CN103880736A (zh) * 2014-04-04 2014-06-25 深圳微芯生物科技有限责任公司 一种e构型苯甲酰胺类化合物及其药用制剂与应用
CN106916101A (zh) * 2017-02-15 2017-07-04 聚缘（上海）生物科技有限公司 Nampt/hdac双靶点抑制剂及其制备方法
WO2018071740A1 (en) * 2016-10-13 2018-04-19 Musc Foundation For Research Development Histone deacetylase inhibitors and uses thereof
CN108558743A (zh) * 2018-06-08 2018-09-21 湖南华腾制药有限公司 一种氘代苯甲酰胺衍生物、制备方法及其应用
CN113831338A (zh) * 2021-09-20 2021-12-24 中国海洋大学 一种组蛋白去乙酰化酶抑制剂及其制备方法和应用

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004058234A2 (en) * 2002-12-27 2004-07-15 Schering Aktiengesellschaft Pharmaceutical combinations of phthalazine vegf inhibitors and benzamide hdac inhibitors
CN103880736A (zh) * 2014-04-04 2014-06-25 深圳微芯生物科技有限责任公司 一种e构型苯甲酰胺类化合物及其药用制剂与应用
WO2018071740A1 (en) * 2016-10-13 2018-04-19 Musc Foundation For Research Development Histone deacetylase inhibitors and uses thereof
CN106916101A (zh) * 2017-02-15 2017-07-04 聚缘（上海）生物科技有限公司 Nampt/hdac双靶点抑制剂及其制备方法
CN108558743A (zh) * 2018-06-08 2018-09-21 湖南华腾制药有限公司 一种氘代苯甲酰胺衍生物、制备方法及其应用
CN113831338A (zh) * 2021-09-20 2021-12-24 中国海洋大学 一种组蛋白去乙酰化酶抑制剂及其制备方法和应用

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BAILEY, M. D. ET AL.: "Novel Azapeptide Inhibitors of Hepatitis C Virus Serine Protease.", JOURNAL OF MEDICINAL CHEMISTRY, vol. 47, no. 15, 17 June 2004 (2004-06-17), XP055029374, ISSN: 0022-2623, DOI: 10.1021/jm049864b *
JIANG YUQI, XU JIE, YUE KAIRUI, HUANG CHAO, QIN MENGTING, CHI DONGYU, YU QIXIN, ZHU YUE, HOU XIAOHAN, XU TONGQIANG, LI MIN, CHOU C: "Potent Hydrazide-Based HDAC Inhibitors with a Superior Pharmacokinetic Profile for Efficient Treatment of Acute Myeloid Leukemia In Vivo", JOURNAL OF MEDICINAL CHEMISTRY, AMERICAN CHEMICAL SOCIETY, US, vol. 65, no. 1, 13 January 2022 (2022-01-13), US , pages 285 - 302, XP093032384, ISSN: 0022-2623, DOI: 10.1021/acs.jmedchem.1c01472 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116496178A (zh) * 2023-04-27 2023-07-28 合肥工业大学 具有丁香酸和氨基酸结构片段的化合物及其合成与用途

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