Multi-target inhibitors targeting HDAC and NAD synthesis and uses thereof

Abstract

The present invention provides a compound targeting HDAC and NAD synthesis and a pharmaceutically acceptable salt, hydrate, deuterated product, isomer, or prodrug thereof, as well as preparation and application thereof. Specifically, a compound shown in the general structural formula (I) and a pharmaceutically acceptable salt, hydrate, deuterated product, isomer, or prodrug thereof are provided. The compound of the general structural formula (I) is a multi-target inhibitor, targeting HDAC and NAD targets, and exhibiting significant HDAC inhibitory activity, while representative compounds exhibit certain NAD inhibitory activity.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology, and in particular relates to a multi-target inhibitor targeting HDAC and NAD synthesis, a pharmaceutically acceptable salt, solvate and prodrug thereof, and applications thereof. Background of the Invention

[0002] The focus of traditional drug research is to find molecules with high affinity and high selectivity for a single target. However, the occurrence and development of tumors depend on multiple receptors or signaling pathways, which makes anti-tumor drugs acting on a single target unable to completely kill tumor cells or produce drug resistance due to compensatory resistance. In order to overcome these limitations, multi-target anti-tumor drug design has been considered an effective strategy and has attracted widespread attention in drug development. Ideally, multi-target drugs can simultaneously regulate a network of disease-related targets and produce synergistic effects (Proc Natl Acad Sci US A. 2019, 116, 7129-7136). Compared with combination therapy, multi-target drugs can avoid drug-drug interactions, reduce toxic side effects, and improve patient compliance.

[0003] Histone deacetylase (HDAC) is a family of enzymes involved in the regulation of many cellular processes, including cell proliferation, apoptosis, and cytoskeletal assembly. HDAC affects cell function by regulating the acetylation levels of histones and non-histones. This process involves a mutual balance between histone acetyltransferases (HATs) and HDACs, both of which are involved in the post-translational modification of histones (Cancer Chemoth Pharm. 2001, 48, 20-26). HATs and HDACs have opposite effects on acetylation and deacetylation of highly conserved lysine residues on the N-terminal tails of histones, thereby altering the assembly and transcriptional activity of chromatin. HDACs are also involved in regulating the acetylation of many non-histone proteins, such as α-tubulin and tumor suppressor p53 (Pharmacol Res. 2021, 163, 105274; J Invest Dermatol. 2020, 140, 2009-2022). These results, together with reports of aberrant HDAC activity in many tumor types, suggest that HDAC inhibition represents a viable anticancer strategy.

[0004] Given the important role of HDAC in tumorigenesis, HDAC has synergistic anti-tumor effects with a variety of tumor targets (such as tubulin and heat shock protein 90 (Hsp90), which has led to the extensive study of multi-target molecules designed based on the inhibition of HDAC (Eur J Med Chem, 2020, 208, 112831). Nicotinamide adenine dinucleotide (NAD+ ) is the most important coenzyme and core metabolite in organisms. It is widely involved in the redox reaction of energy metabolism. Due to the rapid proliferation of tumor cells, they have higher energy demand. + The biosynthesis of NAD is also often upregulated (Nat Rev Cancer. 2012, 12, 741-752). Nicotinamide phosphoribosyltransferase (NAMPT) is one of the most representative metabolic targets. NAMPT catalyzes nicotinamide (NAM) to generate nicotinamide 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 in the NAD production pathway and plays a vital role in cell physiological activities. Studies have shown that targeting NAD synthesis has important anti-tumor effects. 1) Tumor cells have higher NAD consumption and metabolic rates than normal cells, and they are more susceptible to NAMPT inhibitors; 2) NAD is an essential coenzyme involved in the synthesis of a variety of essential substances in tumor cells. NAD can significantly reduce the content of reactive oxygen species (ROS) in the environment to protect tumor cells; 3) NAMPT plays a vital role in angiogenesis and induces the production of vascular endothelial growth factor. Currently, two NAMPT inhibitors targeting NAD synthesis, FK866 and CHS-828, have entered clinical research (Cancer Res. 2003, 63, 7436-7442; Cancer Res. 1999, 59, 5751-5757).

[0005] Targeting HDAC and NAD synthesis has a synergistic anti-tumor effect. Certain specific genotypes, such as p53-deficient or mutated tumor cells, develop primary resistance to HDAC inhibitors, and combined use with NAD synthesis blocking drugs may produce synergistic lethality (Synthetic Lethality) on these cells, thereby achieving better anti-tumor effects. Therefore, the design of multi-target inhibitors targeting HDAC and NAD synthesis is of great significance for tumor treatment. In addition, analysis of the pharmacophores of NAMPT and HDAC inhibitors showed that the two have similar structural characteristics, providing a basis for the design of dual inhibitors. Summary of the invention

[0006] The present application particularly provides an HDAC compound having multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterated substance, isomer, or prodrug thereof, characterized in that the multi-target HDAC compound has a general formula I,

[0007] CycloEBLC(O)-(NH)rR (Formula I)

[0008] wherein ring E is selected from r=1, 2, R is selected from H, C 1-4 Alkyl, C 3-5 Cycloalkyl or C 1-2 Alkyl substituted C 3-5 Cycloalkyl,

[0009] Further, the compound of formula I is further selected from the compounds of formula II, formula III, formula IV, and formula V:

[0010]

[0011] Formula Ⅱ

[0012]

[0013] Formula III

[0014]

[0015] Formula IV

[0016]

[0017] Formula V

[0018] or a pharmaceutically acceptable salt, hydrate, deuterated substance or prodrug thereof, wherein:

[0019] Ring A is selected from

[0020] G is selected from CH2, NH, N(CH2) n CH3, O or S, wherein n is 0-9;

[0021] represents the bond connecting the adjacent ring;

[0022] --- indicates the bond connected to B;

[0023] X 1 Selected from CR 4 or N;

[0024] X 2 Selected from CR 5 or N;

[0025] X 3 Selected from CR 6 or N;

[0026] X 4 Selected from CR 7 or N;

[0027] X 5 , X6 or X 7 are independently selected from CH or N;

[0028] R 1 Selected from H, C1-C4 alkyl, C3-C5 cycloalkyl or C1-C2 alkyl substituted C3-C5 cycloalkyl;

[0029] R 2 and R 3 Each is independently selected from H, halogen, CH3, OCH3;

[0030] B is selected from

[0031] represents the bond to the A ring;

[0032] --- indicates the bond connected to L;

[0033] L chooses from C 1-14 Alkyl, C 1-14 Alkoxy, C 2-14 Alkenyl, C 2-14 Alkynyl, C 3-10 Cycloalkyl, C 1-9 Alkyl substituted C 3-10 Cycloalkyl, C 1-9 Alkoxy substituted C 3-10 Cycloalkyl, C 6-10 Aryl, C 1-9 Alkyl substituted C 6-10 Aryl, C 1-9 Alkoxy substituted C 6-10 Aryl, (C 1-9 alkyl))-(C=O)NH substituted aryl, benzyl or (C 1-8 alkyl)-(C=O)NH substituted benzyl;

[0034] R 4 , R 5 , R 6 , R 7 are independently selected from H, halogen, (C 1-2 )alkyl, halomethyl, OH, OCH3, O(CH2) n CH3, cyclopropyloxy, OC(CH3)3, OCH(CH3)2, 5-6 membered alkoxy, NH2, N(CH3)2, NH(CH2) n CH3, CN, N3, etc., wherein n is 0-9.

[0035] The present invention preferably defines compounds as shown in general formula I to general formula V and pharmaceutically acceptable salts, hydrates, deuterated substances, isomers, or prodrugs thereof,

[0036] in,

[0037] Ring A is selected from the following ring systems:

[0038] G is selected from NH, O or S;

[0039] represents the bond connecting the adjacent ring;

[0040] --- indicates the bond connected to B;

[0041] X 1 Selected from CR 4 or N;

[0042] X 2 Selected from CR 5 or N;

[0043] X 3 Selected from CR 6 or N;

[0044] X 4 Selected from CR 7 or N;

[0045] R 4 , R 5 , R 6 , R 7 All are H;

[0046] R 1 Selected from H, C 1-4 Alkyl, C 3-5 Cycloalkyl or C 1-2 Alkyl substituted C 3-5 Cycloalkyl;

[0047] B is selected from the following structures

[0048] represents the bond to the A ring;

[0049] --- indicates the bond connected to L;

[0050] L is selected from C 1-14 Alkyl, C 1-14 Alkoxy, C 2-14 Alkenyl, C 2-14 Alkynyl, C 1-9 Alkyl substituted C 3-10 Cycloalkyl, C 1-9 Alkoxy substituted C 3-10 Cycloalkyl, C 6-10 Aryl, C 1-9 Alkyl substituted C 6-10 Aryl, C1-9 Alkoxy substituted C 6-10 Aryl, (C 1-9 alkyl))-(C=O)NH substituted aryl, benzyl or (C 1-8 alkyl)-(C=O)NH substituted benzyl;

[0051] The present invention preferably defines a compound as shown in Formula III and a pharmaceutically acceptable salt, hydrate, deuterated substance, isomer, or prodrug thereof,

[0052] in,

[0053] X 1 Selected from CR 4 or N;

[0054] X 2 Selected from CR 5 or N;

[0055] X 3 Selected from CR 6 or N;

[0056] X 4 Selected from CR 7 or N;

[0057] X 5 is selected from CH or N;

[0058] R 4 , R 5 , R 6 and R 7 All are H;

[0059] R 1 Selected from H, C 1-4 Alkyl, C 3-5 Cycloalkyl or C 1-2 Alkyl substituted C 3-5 Cycloalkyl;

[0060] B is selected from the following structures

[0061] represents the bond to the A ring;

[0062] --- indicates the bond connected to L;

[0063] L is selected from C 1-14 Alkyl, C 1-14 Alkoxy, C 2-14 Alkenyl, C 2-14 Alkynyl, C 1-9 Alkyl substituted C 3-10 Cycloalkyl, C 1-9 Alkoxy substituted C 3-10 Cycloalkyl, C6-10 Aryl, C 1-9 Alkyl substituted C 6-10 Aryl, C 1-9 Alkoxy substituted C 6-10 Aryl, (C 1-9 alkyl)-(C=O)NH substituted aryl, benzyl or (C 1-8 alkyl)-(C=O)NH substituted benzyl;

[0064] The present invention preferably defines a compound as shown in formula IV and a pharmaceutically acceptable salt, hydrate, deuterated substance, isomer, or prodrug thereof,

[0065] in,

[0066] Ring A is selected from the following ring systems:

[0067] G is selected from NH, O or S;

[0068] represents the bond connecting the adjacent ring;

[0069] --- indicates the bond connected to B;

[0070] X 1 Selected from CR 4 or N;

[0071] X 2 Selected from CR 5 or N;

[0072] X 3 Selected from CR 6 or N;

[0073] X 4 Selected from CR 7 or N;

[0074] X 6 or X 7 are independently selected from CH or N;

[0075] R 4 , R 5 , R 6 , R 7 All are H;

[0076] R 1 Selected from H, C 1-4 Alkyl, C 3-5 Cycloalkyl or C 1-2 Alkyl substituted C 3-5 Cycloalkyl;

[0077] R 2 and R 3 All are H;

[0078] B is selected from the following structures

[0079] represents the bond to the A ring;

[0080] --- indicates the bond connected to L;

[0081] L is selected from C 1-14 Alkyl, C 1-14 Alkoxy, C 2-14 Alkenyl, C 2-14 Alkynyl, C 1-9 Alkyl substituted C 3-10 Cycloalkyl, C 1-9 Alkoxy substituted C 3-10 Cycloalkyl, C 6-10 Aryl, C 1-9 Alkyl substituted C 6-10 Aryl, C 1-9 Alkoxy substituted C 6-10 Aryl, (C 1-9 alkyl)-(C=O)NH substituted aryl, benzyl or (C 1-8 alkyl)-(C=O)NH substituted benzyl;

[0082] The present invention preferably defines a compound as shown in Formula V and a pharmaceutically acceptable salt, hydrate, deuterated substance, isomer, or prodrug thereof,

[0083] in,

[0084] X 1 Selected from CR 4 or N;

[0085] X 2 Selected from CR 5 or N;

[0086] X 3 Selected from CR 6 or N;

[0087] X 4 Selected from CR 7 or N;

[0088] X 5 , X 6 or X 7 are independently selected from CH or N

[0089] R 4 , R 5 , R 6 and R 7 All are H;

[0090] R1 Selected from H, C 1-4 Alkyl, C 3-5 Cycloalkyl or C 1-2 Alkyl substituted C 3-5 Cycloalkyl;

[0091] B is selected from the following structures

[0092] represents the bond to the A ring;

[0093] --- indicates the bond connected to L;

[0094] L is selected from C 1-14 Alkyl, C 1-14 Alkoxy, C 2-14 Alkenyl, C 2-14 Alkynyl, C 1-9 Alkyl substituted C 3-10 Cycloalkyl, C 1-9 Alkoxy substituted C 3-10 Cycloalkyl, C 6-10 Aryl, C 1-9 Alkyl substituted C 6-10 Aryl, C 1-9 Alkoxy substituted C 6-10 Aryl, (C 1-9 alkyl)-(C=O)NH substituted aryl, benzyl or (C 1-8 alkyl)-(C=O)NH substituted benzyl;

[0095] Unless otherwise defined, the compounds and salts provided herein may also contain all isotopes of atoms present in the intermediates or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers.

[0096] It will be appreciated by those skilled in the art that the described methods are not exclusive means by which the compounds provided herein can be synthesized, and that a wide range of synthetic organic reactions can be obtained that are potentially useful for synthesizing the compounds provided herein. Those skilled in the art know how to select and implement appropriate synthetic routes. Suitable synthetic methods for starting materials, intermediates, and products can be determined by reference to the literature including, for example, the following references: Progress in Heterocyclic Chemistry, Volumes 1-107 (Elsevier, 1963-2012); Journal of Heterocyclic Chemistry, Volumes 1-49 (Journal of Heterocyclic Chemistry, 1964-2012); Carreira et al. (eds.), Synthetic Science, Volumes 1-48 (2001-2010); Katritzky et al. (eds.), Comprehensive Organic Functional Group Transformations (Pergamon Press, 1996); Katritzky et al. (eds.); Comprehensive Organic Functional Group Transformations II (Elsevier, 2nd ed., 2004); Katritzky et al. (eds.), Comprehensive Heterocyclic Chemistry (Pergamon Press, 1984); Smith et al., Advanced Organic Chemistry: Reactions, Mechanisms and Structure, 6th ed. (Wiley, 2007); Trost et al. (eds.), Comprehensive Organic Synthesis (Pergamon Press, 1991).

[0097] The preparation of the compounds described herein may involve the protection and deprotection of various chemical groups. The need for protection and deprotection and the selection of appropriate protecting groups may be easily determined by those skilled in the art. The chemistry of protecting groups can be found in, for example, TW Greene and PGM Wuts, Protecting Groups in Organic Synthesis, 3rd Edition, Wiley & Sons, Inc., New York (1999).

[0098] The reaction can be monitored by any suitable method known in the art. For example, 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). 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.

[0099] As used herein, the phrase "optionally substituted" means unsubstituted or substituted. As used herein, the term "substituted" means that a hydrogen atom is removed and replaced by a substituent. It should be understood that substitution on a given atom is subject to valence restrictions.

[0100] Throughout all definitions, the term “C n-m " denotes a range including endpoints, wherein n and m are integers and represent the number of carbons. Examples include C 1-14 and C 2-14 wait.

[0101] As used herein, the term “C n-m "Alkyl" refers to a saturated hydrocarbon group that can be straight or branched and has n to m carbons. Examples of 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. In some embodiments, the alkyl group contains 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.

[0102] As used herein, the term “C n-m "Alkoxy" refers to a radical of the formula -O-alkyl, wherein the alkyl has n to m carbons. Examples of alkoxy include methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), and tert-butoxy, among others. In some embodiments, the alkyl has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0103] As used herein, "halogen" refers to F, Cl, Br, or I. In some embodiments, halogen is F, Cl, or Br. In some embodiments, halogen is F. In some embodiments, halogen is Cl. In some embodiments, halogen is Br. In some embodiments, halogen is I.

[0104] As used herein, the term “C n-m "Haloalkyl" refers to an alkyl group having from 1 halogen atom to 2s+1 halogen atoms, which may be the same or different, wherein "s" is the number of carbon atoms in the alkyl group, wherein the alkyl group has n to m carbon atoms. In some embodiments, the haloalkyl group is only fluorinated (e.g., C 1-6 In some embodiments, 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.

[0105] As used herein, the term "aryl" refers to an aromatic hydrocarbon group, which may be monocyclic or polycyclic (eg, having 2 fused rings). n-m "Aryl" refers to an aromatic group having n to m ring carbon atoms. Aryl groups include, for example, phenyl, naphthyl, and the like. In some embodiments, the aryl group has 6 to 10 carbon atoms. In some embodiments, the aryl group is a substituted or unsubstituted phenyl group.

[0106] As used herein, "cycloalkyl" refers to a non-aromatic cyclic hydrocarbon including cyclized alkyl and / or alkenyl groups. Cycloalkyl groups may include monocyclic or polycyclic (e.g., having 2 fused rings) groups. Cycloalkyl groups may have 3, 4, 5, or 6 ring carbon atoms (i.e., C 3-6 Cycloalkyl). The ring-forming carbon atoms of the cycloalkyl may be optionally substituted by oxo or sulfido (e.g., C(═O) or C(═S)). Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, or cyclohexadienyl. In some embodiments, the cycloalkyl is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. In some embodiments, the cycloalkyl has 3-6 ring-forming carbon atoms (i.e., C3-6 cycloalkyl).

[0107] As used herein, "heterocycloalkyl" refers to a non-aromatic monocyclic or polycyclic heterocycle having one or more ring-forming heteroatoms selected from O, N or S. Included in heterocycloalkyl are monocyclic 4-, 5- and 6-membered heterocycloalkyls. Examples of heterocycloalkyls include pyrrolidin-2-one, 1,3-isoxazolidin-2-one, pyranyl, tetrahydropyranyl, oxetanyl, azetidinyl, morpholino, thiomorpholino, piperazinyl, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl, pyrrolidinyl, isoxazolidinyl, isothiazolidinyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, imidazolidinyl and azaalkyl etc. The ring-forming carbon atoms and heteroatoms of heterocycloalkyl can be optionally substituted by oxy (=O). Heterocycloalkyl can be connected by ring-forming carbon atoms or ring-forming heteroatoms. In some embodiments, heterocycloalkyl contains 0 to 3 double bonds.

[0108] As used herein, the term "compound" is intended to include all stereoisomers, geometric isomers, tautomers, and isotopes of the depicted structure. Unless otherwise indicated, a compound identified herein by name or structure as one particular tautomeric form is intended to include the other tautomeric forms.

[0109] Compounds provided herein also include tautomeric forms. Tautomeric forms are caused by the exchange of a single bond with an adjacent double bond and the accompanying proton migration. Tautomeric forms include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of prototropic tautomers include keto-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which protons can occupy more than two positions of a heterocyclic system, for example, 1H- and 3H-imidazoles, 1H-, 2H- and 4H-1,2,4-triazoles, 1H- and 2H-isoindoles, and 1H- and 2H-pyrazoles. Tautomeric forms can be in equilibrium or stereologically locked into a form by appropriate substitution.

[0110] All compounds and pharmaceutically acceptable salts thereof may be found together with other substances such as water and solvents (eg, hydrates and solvates), or may be isolated.

[0111] In some embodiments, preparation of the compounds may involve the addition of acids or bases to effect, for example, catalysis of a desired reaction or the formation of salt forms such as acid addition salts.

[0112] Examples of acids can be inorganic or organic acids and include, but are not limited to, strong acids 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, enanthic acid, caprylic acid, nonanoic acid, and capric acid.

[0113] Examples of bases include lithium hydroxide, sodium hydroxide, potassium hydroxide, lithium carbonate, sodium carbonate, potassium carbonate, and sodium bicarbonate. Some examples of strong bases include, but are not limited to, hydroxides, alkoxides, metal amides, metal hydrides, metal dialkylamides, and arylamines, wherein alkoxides include lithium, sodium, and potassium salts of methyl, ethyl, and tert-butyl oxides; metal amides include sodium amide, potassium amide, and lithium amide; metal hydrides include sodium hydride, potassium hydride, and lithium hydride; and metal dialkylamides include lithium, sodium, and potassium salts of amides substituted with methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, trimethylsilyl, and cyclohexyl.

[0114] In some embodiments, the compound and salt provided herein are substantially separated.By "substantially separated" it is meant that the compound is separated at least partially or substantially from its formed or detected environment.Partial separation can include, for example, a composition rich in the compound provided herein.Substantially separating can include a composition 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% of the compound or its salt provided herein by weight.The method for separating compounds and their salts is conventional in the art.

[0115] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0116] The present application also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds, wherein the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; and alkali metal or organic salts of acidic residues such as carboxylic acids, etc. The pharmaceutically acceptable salts of the present application include, for example, 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, aminosulfonic 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, hydroxymaleic acid, palmitic acid, cinnamic acid, isobutyric acid, lauric acid, mandelic acid, maleic acid, fumaric acid, malic acid The pharmaceutically acceptable salts of the present invention can be substituted with 1,2-dihydroxy-1,2,3-tripropanecarboxylic acid, 1,2-dihydroxy-2,4-dihydroxybenzoic acid, α-ketoglutaric acid, 1,2-dihydroxy-2,4-dihydroxybenzo ... Generally, 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; generally, nonaqueous media such as ether, ethyl acetate, alcohols (e.g., methanol, ethanol, isopropanol or butanol), or acetonitrile (MeCN) are preferred.

[0117] In some embodiments, the disease is cancer. In some embodiments, cancer is selected from the group consisting of colorectal cancer, endometrial cancer, brain cancer (e.g., glioblastoma multiforme), melanoma, gastric cancer, breast cancer, ovarian cancer, pancreatic cancer, liver cancer, glioma, brain tumor, kidney cancer, prostate cancer, bladder cancer, lung cancer, pancreatic cancer, ovarian cancer, skin cancer, epithelial cell cancer, nasopharyngeal cancer, epidermal cell cancer, cervical cancer, oral cancer, tongue cancer, human fibrosarcoma, multiple myeloma and blood cancer. In some embodiments, cancer includes solid tumors. In some embodiments, cancer is selected from the group consisting of glioma, glioblastoma, non-small cell lung cancer and blood cancer.

[0118] In some embodiments, cancer is a blood cancer. In some embodiments, the blood cancer is selected from the group consisting of leukemia and lymphoma. In some embodiments, 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, activated B-cell like (ABC) diffuse large B cell lymphoma, and germinal center B cell (GCB) diffuse large B cell lymphoma.

[0119] The compounds of the present invention can be used alone or in combination with other therapeutic agents for treating the diseases or conditions of the present invention. The compounds of the present invention are combined with other anti-tumor drugs. The anti-tumor drugs include, but are not limited to, cyclophosphamide, nitrogen mustard, melphalan, leuprorelin, 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.

[0120] When used as a drug, the compounds and salts provided herein can be administered in the form of a pharmaceutical composition. These compositions can be prepared as described herein or elsewhere, and can be administered by a variety of routes, depending on whether local treatment or systemic treatment is desired and on the area to be treated. Administration can be topical (including transdermal, epidermal, ocular, and mucosal administration including intranasal delivery, vaginal delivery, and rectal delivery), pulmonary (e.g., by inhalation or insufflation of a powder or aerosol, including by means of a nebulizer; intratracheal or intranasal), oral, or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular injection or infusion; or intracranial (e.g., intrathecal or intraventricular administration). Parenteral administration can be in the form of a single bolus dose, or can be, for example, by a continuous infusion pump.

[0121] In some embodiments, 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.

[0122] Pharmaceutical compositions and preparations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquid preparations and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, and thickeners and the like may be necessary or desirable.

[0123] Also provided are pharmaceutical compositions comprising compounds provided herein as active ingredients or pharmaceutically acceptable salts thereof in combination with one or more pharmaceutically acceptable carriers (e.g., excipients). In the preparation of the compositions provided herein, the active ingredient is usually mixed with an excipient, diluted by the excipient or encapsulated in such a carrier in the form of, for example, a capsule, a sachet, paper or other container. When an excipient is used as a diluent, it can be a solid, semisolid or liquid material, which acts as a solvent, carrier or medium for the active ingredient. Therefore, the composition can be in the form of tablets, pills, powders, lozenges, capsules, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solids or in liquid media), ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.

[0124] Some examples of suitable excipients include without limitation lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The formulation may additionally include without limitation lubricants such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifiers and suspending agents; preservatives such as methyl and propyl hydroxybenzoates; sweeteners; flavoring agents; or combinations thereof.

[0125] The active ingredient can be effective over a wide dosage range and is generally administered in a pharmaceutically effective amount. However, it will be understood that the amount of compound actually administered will generally be determined by a physician based on relevant circumstances, including the condition to be treated, the selected route of administration, the actual compound administered, the age, weight and response of the individual subject, and the severity of the subject's symptoms, etc.

[0126] The beneficial effect of the present invention is that the drug design based on HDAC multi-targets provides new chemical entities for the treatment of various cancers, thereby transforming the current cancer treatment methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0127] Figure 1 , concentration-response curves of some compounds on acute myeloid leukemia cells MV4-11 and HL60.

[0128] Figure 2 , concentration-response curves of some compounds on acute myeloid leukemia cells PL21, KASUMI-1, MONO-MAC-1, and NB-4.

[0129] Figure 3 Concentration-response curves of CZ411, FK866 and their mixture on acute myeloid leukemia cells MV4-11 and HL60.

[0130] Figure 4 , NMN has a reversal effect on cell death caused by LEE18 and LEE12. NMN is nicotinamide mononucleotide.

[0131] Figure 5 , HCT116 tumor growth curve and tumor image. 5-FU: 5-fluorouracil, a traditional anti-tumor chemotherapy drug; Oxaliplatin: Oxaliplatin, a third-generation platinum anticancer drug, is an anti-tumor chemotherapy drug. 5-FU+Oxaliplatin is a positive control.

[0132] Figure 6 , tumor growth curve. Panobinostat is a marketed broad-spectrum HDAC inhibitor and serves as a positive control drug. DETAILED DESCRIPTION

[0133] The present invention will be described in more detail by way of specific examples. The following examples are provided for illustrative purposes and are not intended to limit the present invention in any way. Those skilled in the art will readily recognize that various non-critical parameters that can be changed or modified to obtain substantially the same result.

[0134] General Materials and Methods

[0135] All reactions insensitive to air and moisture were carried out under ambient atmosphere and with magnetic stirring. Tetrahydrofuran was distilled from dark purple sodium benzophenone ketyl. Dry DMF, dry DMSO. Dry acetonitrile, dry dichloromethane, dry toluene, and dry dioxane were purchased from Anaiji Chemical. All operations sensitive to air and moisture were carried out under nitrogen atmosphere using dried glassware.

[0136] Thin layer chromatography (TLC) was performed on EMD TLC plates pre-coated with 250 μm thick silica gel 60 F254 plates and visualized by fluorescence quenching under UV light and KMnO4 staining.

[0137] All deuterated solvents were purchased from Beijing Bailingwei. NMR spectra were recorded on the following instruments: 1 H and 13 C acquisition, JEOL 400 spectrometer operating at 400 MHz. The solvent resonance was used as the internal standard ( 1 H: CDCl3, δ7.26; DMSO-d6, δ2.50), ( 13 C: CDCl3, δ 77.16; DMSO-d6, δ 39.52), chemical shifts are reported in ppm. Data are reported as follows: s is singlet, d is doublet, t is triplet, q is quartet, m is multiplet; coupling constants in Hz; integration; unless otherwise noted, carbon signals are singlets.

[0138] The examples and preparations provided below further illustrate and illustrate the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparations does not limit the scope of the present invention in any way.

[0139] The following synthetic routes describe the preparation of compounds of Formula II, III, IV or V of the present invention, all starting materials being prepared by the methods described in these routes, by methods well known to those skilled in the art of organic chemistry or commercially available. All final compounds of the present invention are prepared by the methods described in these routes or by methods analogous thereto, which are well known to those skilled in the art of organic chemistry. All variables used in these routes are as defined below or as defined in the claims.

[0140] The preparation of the intermediate compound of formula II, III, IV or V in the present invention is shown in route 1 and route 2, and each substituent is as defined in the summary of the invention.

[0141] Synthesis route 1:

[0142]

[0143] Reagents and conditions: (a) different amine, TBTU, TEA, DCM, yield 55%; (b) CH3OH / H2O, 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%.

[0144] Synthesis route 2:

[0145]

[0146] Reagents and conditions: (a) propionaldehyde, MeOH, yield 98%; NaBH3CN, MeOH, concentrated hydrochloric acid, methyl orange, yield 60%; (b) (Boc)2O, triethylamine, EtOH, yield 85%; (c) Pd / C, H2, MeOH, yield 85%; (d) trifluoroaceticanhydride, DCM, yield 85%; (e) EDCI, HOBt, TEA, DCM, yield 55%; (f) Na2CO3, MeOH, yield 70%.

[0147] The preparation of the compound of formula II, III, IV or V of the present invention is shown in Scheme 3, Scheme 4, Scheme 5 and Scheme 6, and each substituent is as defined in the Summary of the Invention.

[0148] Route 3:

[0149]

[0150] Reagents and conditions: (a) TBTU, TEA, DCM, yield 55%; (b) TFA, DCM, triethylamine, yield 85%.

[0151] Route 4:

[0152]

[0153] Reagents and conditions: (a) TBTU, TEA, DCM, yield 55%.

[0154] Route 5:

[0155]

[0156] Reagents and conditions: (a) 3-ethynylpyridine, sodium ascorbate, CuSO4, THF, H2O, yield 85%.

[0157] Route 6:

[0158]

[0159] Reagents and conditions: (a) TBTU, TEA, DCM, yield 55%; (b) EDCI, HOBt, triethylamine, DCM, yield 50%; (c) TFA, DCM, triethylamine, yield 85%; (d) 3-ethynylpyridine, sodiumascorbate, CuSO4, THF, H2O, yield 85%.

[0160] Specific implementation plan:

[0161] The present invention is described in detail below through specific examples, but the use and purpose of these exemplary implementation modes are only used to illustrate the present invention, and do not constitute any form of limitation on the actual protection scope of the present invention, nor limit the protection scope of the present invention to them.

[0162] Preparation of the compound in route 1:

[0163] Embodiment 1:

[0164]

[0165] Preparation of (E)-3-(3-(pyridin-3-yl)acrylamido)propionic acid methyl ester (1a): 3-(pyridin-3-yl)acrylic acid (0.44 g, 3 mmol) was dissolved in 20 mL of dichloromethane, and 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroboric acid (TBTU, 1.05 g, 3.6 mmol) and TEA (0.6 mL, 4.5 mmol) were added under ice bath. After 30 minutes, 3-aminopropionic acid methyl ester hydrochloride (0.46 g, 3.3 mmol) was added, followed by 0.6 mL of TEA, and the reaction was allowed to react overnight. The mixture was washed with saturated NaHCO3 (2×30 mL) and saturated brine (2×30 mL), and dried over MgSO4. After evaporating the solvent, the mixture was purified by flash chromatography to give compound 1a as a white solid powder (0.35 g, 51%). 1 H NMR (600MHz, DMSO-d6) 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] + .

[0166]

[0167] Preparation of (E)-5-(3-(pyridin-3-yl)acrylamido)pentanoic acid methyl ester (1b): Using the synthetic method of 1a, 3-(pyridin-3-yl)acrylic acid and 5-aminopentanoic acid methyl ester hydrochloride were used as raw materials to obtain a white solid 1b with a yield of 55%. 1 HNMR(600MHz,DMSO-d6)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]+ .

[0168]

[0169] Preparation of (E)-7-(3-(pyridin-3-yl)acrylamido)heptanoic acid methyl ester (1c): Using the synthetic method of 1a, 3-(pyridin-3-yl)acrylic acid and 7-aminoheptanoic acid methyl ester hydrochloride were used as raw materials to obtain a white solid 1c with a yield of 53%. 1 HNMR(600MHz,DMSO-d6)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=1 5.9Hz,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] + .

[0170]

[0171] Preparation of (E)-7-(3-(pyridin-3-yl)acrylamido)octanoic acid methyl ester (1d): Using the synthetic method of 1a, 3-(pyridin-3-yl)acrylic acid and 8-aminooctanoic acid methyl ester hydrochloride were used as raw materials to obtain a white solid 1d with a yield of 53%. 1 HNMR(600MHz,DMSO-d6)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).

[0172]

[0173] Preparation of methyl 7-(3H-pyrrolo[3,2-c]pyridine-2-carboxamide)heptanoate (6): Using the synthetic method of 1a, 1H-pyrrolo[3,2-c]pyridine-2-carboxylic acid and methyl 7-aminoheptanoate hydrochloride were used as raw materials to obtain white solid 6 with a yield of 55%. 1 H NMR(600MHz,DMSO-d6)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] + .

[0174]

[0175] Preparation of (E)-3-(3-(pyridin-3-yl)acrylamido)propanoic acid (2a): Compound 1a (0.35 g, 1.5 mmol) was dissolved in 5 mL of methanol, and then 2 mL of 3 M aqueous KOH was added. The mixture was refluxed at 85 °C for 2 h. After the reaction was completed, MeOH was evaporated under vacuum. The residue was acidified to pH 5-6 with 1 N HCl and then filtered. The corresponding acidic 2a was precipitated as a white solid (0.28 g, 85%). The material was used directly in the next step without further purification. ESI-MS m / z: 219.15 [M–H] - .

[0176]

[0177] Preparation of (E)-5-(3-(pyridin-3-yl)acrylamido)pentanoic acid (2b): Using the synthetic method of 2a, 2b was obtained as a white solid with a yield of 88%. 1H NMR (400MHz, DMSO-d6) 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] - .

[0178]

[0179] Preparation of (E)-7-(3-(pyridin-3-yl)acrylamido)heptanoic acid (2c): The synthetic method of 2a was used with compound LL433 as raw material to obtain white solid 2c with a yield of 55%. 1 H NMR(400MHz,DMSO-d6)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] - .

[0180]

[0181] Preparation of (E)-8-(3-(pyridin-3-yl)acrylamido)octanoic acid (2d): The synthetic method of 2a was used with compound 1d as raw material to obtain white solid 2d with a yield of 87%. 1H NMR(400MHz,DMSO-d6)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] - .

[0182]

[0183] Preparation of 7-(3-(pyridin-3-ylmethyl)ureido)heptanoic acid (5): Using the synthetic method of 2a and compound 4 as the raw material, a white solid 5 was obtained with a yield of 85%. 1 H NMR (400MHz, DMSO-d6) 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] - .

[0184]

[0185] 7-(1H-Pyrrolo[3,2-c]pyridine-2-carboxamide)heptanoic acid (7). Using the synthetic method of 2a, compound 6 was used as the raw material to obtain white solid 7 with a yield of 85%. ESI-MS m / z: 288.21 [M–H] - .

[0186]

[0187] Preparation of 4-nitrophenyl (pyridin-3-ylmethyl) carbamate (3): 3-pyridinemethylamine (0.22 g, 2 mmol) and triethylamine (0.43 g, 2 mmol) were dissolved in DCM, and then 4-nitrophenyl chloroformate (0.61 g, 3 mmol) was added and stirred at room temperature for 5 hours. After the reaction was completed, it was washed with brine 3 times and dried over Na2SO4. The solvent was evaporated to give compound 3 as a white solid powder (0.07 g, 70% yield). 1 H NMR (400MHz, DMSO-d6) 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] + .

[0188]

[0189] Preparation of methyl 7-(3-(pyridin-3-ylmethyl)ureido)heptanoate (4): To a solution of methyl 7-aminoheptanoate hydrochloride (0.19 g, 1 mmol) in dichloromethane, triethylamine (0.12 g, 1.2 mmol) and compound 3 (0.33 g, 1.2 mmol) were added at 0°C. The reaction was allowed to proceed for 2 hours at room temperature. After the reaction was completed, the mixture was washed 3 times with brine and dried over Na2SO4. After evaporating the solvent, the mixture was purified by flash chromatography to give compound 4 (0.2 g, 70% yield) as a white solid powder. 1 H NMR (400MHz, DMSO-d6) δ8.42(s,1H),8.39(d,J=4.8Hz,1H),7.60(d,J=7.8Hz,1 H),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] + .

[0190]

[0191] Preparation of 8-azidooctanoic acid (8): 8-bromooctanoic acid (0.33 g, 1.5 mmol) was dissolved in DMF, and then NaN3 (0.15 g, 2.25 mmol) was added and reacted at 80°C overnight. 25 mL of DCM was added to the solution, and the solvent was washed with water 5 times, dried over Na2SO4, and the solvent was evaporated to obtain compound 8 as a colorless oil (0.27 g, 95% yield). 1 H NMR(400MHz, DMSO-d6)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] - .

[0192] Preparation of compounds in route 2:

[0193] Embodiment 2:

[0194]

[0195] Preparation of 2-propylhydrazine-1-carboxylic acid benzyl ester (9): Benzyl hydrazinecarboxylate (1.66 g, 10 mmol) was dissolved in 50 mL of methanol, and then propionaldehyde (0.61 g, 10.5 mmol) was added and reacted at room temperature for 2 hours. After the reaction was complete, methanol was removed. The resulting solid was dissolved in 30 mL of methanol, and then NaBH3CN (1.2 g, 20 mmol) and 2 drops of concentrated HCl / MeOH (v:v=1:1) solution were added and reacted overnight. After the reaction was complete, the solvent was evaporated and the crude product was purified by flash chromatography to obtain compound 9 as a white solid powder (1.2 g, 60%). 1 H NMR(400MHz,DMSO-d6)δ8.54(s,1H),7.36-7.23(m,5H),4.99(s,2H),4.46(s,1 H),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] + .

[0196]

[0197] Preparation of 2-benzyl 1-(tert-butyl)-1-propylhydrazine-1,2-dicarboxylate (10): 9 (1 g, 5 mmol) was dissolved in 50 mL of anhydrous dichloromethane, and triethylamine (1.5 g, 15 mmol) and (Boc)2O (0.22 g, 10 mmol) were added. After reacting at room temperature for 2 hours, the solution was washed with 1 M citric acid aqueous solution (3×100 mL) and brine (3×100 mL), then dried over MgSO4, and the solvent was evaporated to obtain compound 10 as a white solid (1.3 g, 85%). 1 H NMR (400MHz, DMSO-d6) 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] + .

[0198]

[0199] Preparation of 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. The mixture was then reacted in hydrogen at room temperature 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%). 1 H NMR (400MHz, DMSO-d6) 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] + .

[0200]

[0201] Preparation of 4-((2,2,2-trifluoroacetylamino)methyl)benzoic acid (13a): 10 mL of trifluoroacetic anhydride was slowly added dropwise to 4-(aminomethyl)benzoic acid (3.0 g, 20 mmol) in an ice bath, and the mixture was reacted at room temperature for 2 hours. After the reaction was complete, 100 mL of ice water was added to quench the reaction, and then the mixture was filtered and the filter cake was dried to obtain 13a (4.43 g, 95%). 1H NMR (400MHz, DMSO-d6) 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] - .

[0202]

[0203] Preparation of 4-((2,2,2-trifluoroacetylamino)methyl)benzoic acid (13b): The synthetic method of 13a was used to obtain 13b as a white solid with 4-aminobenzoic acid and trifluoroacetic anhydride as raw materials in a yield of 98%. 1 H NMR (400MHz DMSO-d6) 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] - .

[0204]

[0205] Preparation of tert-butyl 1-propyl-2-(4-((2,2,2-trifluoroacetylamino)methyl)benzoyl)hydrazine-1-carboxylate (14a): Compound 13a (0.7 g, 3 mmol) was dissolved in 20 mL of dichloromethane, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI·HCl, 0.7 g, 3.6 mmol), 1-hydroxybenzotriazole (HOBt, 0.44 g, 3.6 mmol) and triethylamine (0.6 mL, 4.5 mmol) were added under ice bath. After 30 minutes, compound 11 (0.57 g, 3.3 mmol) was added and the reaction was allowed to proceed overnight. The reaction solution was washed with saturated NaHCO3 (2×30 mL) and brine (2×30 mL), and the organic phases were combined and dried over anhydrous magnesium sulfate. After evaporating the solvent, it was purified by flash chromatography to obtain a white solid powder 14a (0.66 g, 55%). 1 H NMR (400MHz, DMSO-d6) 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]+ .

[0206]

[0207] Preparation of tert-butyl 1-propyl-2-(4-(2,2,2-trifluoroacetylamino)benzoyl)hydrazine-1-carboxylate (14b): Using the synthetic method of 14a and compounds 13b and 11 as raw materials, a white solid 14b was obtained in a yield of 50%. 1 H NMR(400MHz,DMSO-d6)δ11.46(s,1H),10.49(s,1H),7.85(dd,J=14.7,8.6Hz,2H),7.7 5(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] + .

[0208]

[0209] Preparation of tert-butyl 2-(4-(aminomethyl)benzoyl)-1-propylhydrazine-1-carboxylate (15a): Compound 14a (1.2 g, 3 mmol) was dissolved in 20 mL of methanol / water (v:v=1:1), and then K2CO3 (1.24 g, 9 mmol) was added and reacted overnight at room temperature. The methanol was evaporated to dryness and then extracted with ethyl acetate (2×20 mL). The organic phases were combined and washed with brine (2×30 mL) and dried over anhydrous Na2SO4. After filtration, the solvent was evaporated to dryness to obtain 15a (0.74 g, 80%). 1 H NMR (400MHz, DMSO-d6) δ10.42(s,1H),7.74(dd,J=16.8,7.9Hz,2H),7.39(d,J=8.0Hz,2 H),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] + .

[0210]

[0211] Preparation of tert-butyl 2-(4-aminobenzoyl)-1-propylhydrazine-1-carboxylate (15b): The synthetic method of 15a was used with compound 14b as raw material to obtain white solid 15b in a yield of 82%. 1H NMR (400MHz, DMSO-d6) 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] + .

[0212] Preparation of the compound in route 3

[0213] Embodiment 3:

[0214]

[0215] Preparation of (E)-N-(3-oxo-3-(2-propylhydrazine)propyl)-3-(pyridin-3-yl)acrylamide (LEE1): Compound 2b (0.53 g, 3 mmol) was dissolved in 15 mL of dichloromethane, and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU, 1.05 g, 3.6 mmol) and triethylamine (0.6 mL, 4.5 mmol) were added under ice bath. After 30 minutes of reaction, compound 11 (0.57 g, 3.3 mmol) was added and reacted at room temperature overnight. The reaction solution was washed with saturated NaHCO3 (2×30 mL) and saturated brine (2×30 mL), and dried with MgSO4. After filtration, the solvent was evaporated and purified by flash chromatography to obtain a white solid powder (0.35 g, 54%).

[0216] The product of the previous step (0.30 g, 0.8 mmol) was dissolved in 15 mL of dichloromethane, and then trifluoroacetic acid (TFA, 0.11 g, 1.0 mmol) was added and reacted overnight. After the reaction was complete, TEA (0.10 g, 1.0 mmol) was added to adjust the pH to 8. The solution was washed with brine and dried over Na2SO4. After filtration, the solvent was evaporated and then purified by flash chromatography to obtain a white solid powder compound LEE1 (0.18 g, 85%). 1H NMR (400MHz, DMSO-d6) 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). 13 CNMR(101MHz,DMSO-d6)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] + .

[0217] Embodiment 4:

[0218]

[0219] Preparation of (E)-N-(5-oxo-5-(2-propylhydrazine)pentyl)-3-(pyridin-3-yl)acrylamide (LEE2): Using the synthetic method of LEE1, compound 11 and compound 2b were used as raw materials to obtain white solid LEE2 with a yield of 55%. 1 HNMR(400MHz,DMSO-d6)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),7.49-7.3 8(m,2H),6.73(d,J=15.9Hz,1H),4.77(d,J=5.9Hz,1H),3.18(q,J=6.5Hz,2H),2.6 1(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-d6)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] + .

[0220] Embodiment 5:

[0221]

[0222] Preparation of (E)-N-(7-oxo-7-(2-propylhydrazine)heptyl)-3-(pyridin-3-yl)acrylamide (LEE3): Using the synthetic method of LEE1, compounds 11 and 2c were used as raw materials to obtain white solid LEE3 with a yield of 57%. 1 H NMR (400MHz, DMSO-d6) δ9.21(d,J=5.7Hz,1H),8.75(d,J=2.2Hz,1H),8.55(dd,J=4.8,1.6H z,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). 13 C NMR(101MHz,DMSO-d6)δ171.36,164.84,150.51,149.53,135.52,134.35,131.25,12 4.84,124.42,53.50,39.15,33.91,29.46,28.74,26.65,25.63,21.22,12.04.ESI-MS m / z:332.04[M+H] + .

[0223] Embodiment 6:

[0224]

[0225] Preparation of (E)-N-(8-oxo-8-(2-propylhydrazine)octyl)-3-(pyridin-3-yl)acrylamide (LEE4): Using the synthetic method of LEE1, compounds 11 and 2d were used as raw materials to obtain white solid LEE4 with a yield of 60%. 1 H NMR (400MHz, DMSO-d6) δ10.04(s,1H),9.85(s,1H),8.71(d,J=2.3Hz,1H),8.50(dd,J=4.7,1.6H z,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). 13 C NMR (126MHz, DMSO-d6) δ171.55,164.84,150.41,149.39,135.50,134.48,131.24,124.8 1,124.47,52.86,39.14,33.65,29.50,28.87,28.83,26.79,25.38,20.05,11.73.ESI-MS m / z:346.97[M+H] + .

[0226] Embodiment 7:

[0227]

[0228] Preparation of 1-(7-oxo-7-(2-propylhydrazine)heptyl)-3-(pyridin-3-ylmethyl)urea (LEE5): Using the synthetic method of LEE1 and compounds 11 and 5 as raw materials, white solid LEE5 was obtained with a yield of 61%. 1H NMR (400MHz, DMSO-d6) 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); 13 C NMR(126MHz,DMSO-d6)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] + .

[0229] Embodiment 8:

[0230]

[0231] Preparation of N-(7-oxo-7-(2-propylhydrazine)heptyl)-3H-pyrrolo[3,2-c]pyridine-2-carboxamide (LEE7): Using the synthetic method of LEE1, compounds 11 and 7 were used as raw materials to obtain white solid LEE7 with a yield of 56%. 1 H NMR (400MHz, DMSO-d6) 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). 13CNMR(101MHz,DMSO-d6)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] + .

[0232] Embodiment 9:

[0233]

[0234] Preparation of 8-azido-N'-propyloctane hydrazide (LEE6): Using the synthetic method of 1a, compounds 8 and 11 were used as raw materials to obtain colorless oily LEE6 with a yield of 56%. ESI-MS m / z: 241.98 [M+H] + .

[0235] Preparation of the compound in route 4

[0236] Embodiment 10:

[0237]

[0238] Preparation of (E)-N-(2-aminophenyl)-8-(3-(pyridin-3-yl)acrylamide)octanamide (LEE8): Using the synthetic method of 1a, compound 2d and 1,2-diaminobenzene were used as raw materials to obtain white solid LEE8 with a yield of 53%. 1 H NMR (400MHz, DMSO-d6) 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-d6) 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] + .

[0239] Embodiment 11:

[0240]

[0241] Preparation of N-(2-aminophenyl)-8-azidooctanamide (20): Using the synthetic method of 1a, compound 8 and 1,2-diaminobenzene were used as raw materials to obtain white solid 20 in a yield of 60%. 1 H NMR (400MHz, DMSO-d6) 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] + .

[0242] Preparation of the compound in route 5

[0243] Embodiment 12:

[0244]

[0245] Preparation of n-propyl-8-(4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)octanylhydrazide (LEE10): Compound 19 (60.2 mg, 0.25 mmol) and 3-ethynylpyridine (26 mg, 0.25 mmol) were dissolved in a solution of THF / H2O (v:v=2:1), sodium ascorbate (50 mg, 0.25 mmol) and copper sulfate (4 mg, 0.025 mmol) were added, and the mixture was reacted overnight at room temperature. After the reaction was complete, the reaction solution was washed 3 times with water and dried over Na2SO4. After filtration, the solvent was evaporated to obtain compound LEE10 as a white solid powder (0.07 g, 84%). 1H NMR (400MHz, DMSO-d6) 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). 13 C NMR(126MHz,DMSO-d6)δ171.33,149.26,146.79,143.92,132.79,127.26,124.46,12 2.39,53.44,50.06,33.84,30.00,28.76,28.50,26.17,25.52,21.16,12.01.ESI-MS m / z:345.06[M+H] + .

[0246] Embodiment 13:

[0247]

[0248] Preparation of N-(2-aminophenyl)-8-(4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)octanamide (LEE11): Using the synthetic method of LEE10, compound 20 and 3-ethynylpyridine were used as raw materials to obtain compound LEE11 as a white solid with a yield of 76%. 1H NMR(400MHz,DMSO-d6)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). 13 C NMR (126MHz, DMSO-d6) 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] + .

[0249] Preparation of the compound in route 6

[0250] Embodiment 14:

[0251]

[0252] Preparation of tert-butyl 2-(4-((3H-pyrrolo[3,2-c]pyridine-2-carboxamide)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23c): 1H-pyrrolo[3,2-c]pyridine-2-carboxylic acid (0.48 g, 3 mmol) was dissolved in 15 mL of dichloromethane, and O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU, 1.05 g, 3.6 mmol) and triethylamine (0.6 mL, 4.5 mmol) were added to the reaction solution under ice bath. After 30 minutes, compound 15a (1.0 g, 3.3 mmol) was added, and then the reaction was allowed to react overnight. The reaction solution was washed with saturated NaHCO3 (2×30 mL) and brine (2×30 mL), and dried over MgSO4. After filtration, the solvent was evaporated and purified by flash chromatography to give a white solid powder (0.7 g, 54%). 1H NMR(400MHz,DMSO-d6)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] + .

[0253] Preparation of tert-butyl 2-(4-(((1H-indole-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23 g): Using the synthetic method of 23c, 2-indolecarboxylic acid and compound 15a were used as raw materials to obtain a white solid 23 g with a yield of 56%. 1 H NMR (400MHz, DMSO-d6) 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=5.9 Hz,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] + .

[0254]

[0255] Preparation of tert-butyl 2-(4-((benzofuran-2-carboxamide)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23a): Using the synthetic method of 23c, 2-benzofurancarboxylic acid and compound 15a were used as raw materials to obtain a white solid 23a in a yield of 56%. 1H NMR (400MHz, DMSO-d6) 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] + .

[0256]

[0257] Preparation of tert-butyl 2-(4-((Benzothiophene-2-carboxamide)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23b): Using the synthetic method of 23c, benzothiophene-2-carboxylic acid and compound 15a were used as raw materials to obtain a white solid 23b in a yield of 53%. 1 H NMR (400MHz, DMSO-d6) 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] + .

[0258]

[0259] Preparation of tert-butyl 2-(4-((3H-pyrrolo[2,3-c]pyridine-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23e): Using the synthetic method of 23c, 1H-pyrrolo[2,3-c]pyridine-2-carboxylic acid and compound 15a were used as raw materials to obtain a white solid 23e with a yield of 49%. 1H NMR (400MHz, DMSO-d6) 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] + .

[0260]

[0261] Preparation of tert-butyl 2-(4-(((furan[3,2-c]pyridine-2-carboxamido)methyl)benzoyl)-1-propylhydroxyazine-1-carboxylate (23d): Using the synthetic method of 23c, benzo[b]thiophene-2-carboxylic acid and compound 15a were used as raw materials to obtain 23d as a white solid in 53% yield. 1 H NMR (400MHz, DMSO-d6) 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] + .

[0262]

[0263] Preparation of tert-butyl 2-(4-((5-bromo-1H-indole-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23j): Using the synthetic method of 23c, 5-bromoindole-2-carboxylic acid and compound 15a were used as raw materials to obtain 23j as a white solid with a yield of 58%.

[0264]

[0265] Preparation of tert-butyl 2-(4-(((5-fluoro-1H-indole-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23l): Using the synthetic method of 23c, 5-fluoroindole-2-carboxylic acid and compound 15a were used as raw materials to obtain 23l as a white solid with a yield of 53%.

[0266]

[0267] Preparation of tert-butyl 2-(4-((6-(dimethylamino)-1H-indole-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23k): Using the synthetic method of 23c, 6-(dimethylamino)-1H-indole-2-carboxylic acid and compound 15a were used as raw materials to obtain 23k as a white solid with a yield of 50%.

[0268]

[0269] Preparation of tert-butyl 2-(4-((5-chloro-1H-indole-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23m): Using the synthetic method of 23c, 5-chloroindole-2-carboxylic acid and compound 15a were used as raw materials to obtain 23m as a white solid with a yield of 51%.

[0270]

[0271] Preparation of tert-butyl 2-(4-(((5-methoxybenzofuran-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23n): Using the synthetic method of 23c, 5-methoxybenzofuran-2-carboxylic acid and compound 15a were used as raw materials to obtain 23n as a white solid with a yield of 47%.

[0272]

[0273] Preparation of tert-butyl 2-(4-((5-fluorobenzofuran-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23o): Using the synthetic method of 23c, 5-fluorobenzofuran-2-carboxylic acid and compound 15a were used as raw materials to obtain 23o as a white solid with a yield of 59%.

[0274]

[0275] Preparation of tert-butyl 2-(4-((5-bromobenzofuran-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23p): Using the synthetic method of 23c, 5-bromobenzofuran-2-carboxylic acid and compound 15a were used as raw materials to obtain 23p as a white solid with a yield of 52%.

[0276]

[0277] Preparation of tert-butyl 2-(4-(((5-methoxy-1H-indole-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23q): Using the synthetic method of 23c, 5-methoxyindole-2-carboxylic acid and compound 15a were used as raw materials to obtain 23q as a white solid with a yield of 49%.

[0278]

[0279] Preparation of tert-butyl 2-(4-((5-chlorobenzofuran-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23r): Using the synthetic method of 23c, 5-chlorobenzofuran-2-carboxylic acid and compound 15a were used as raw materials to obtain 23r as a white solid with a yield of 46%.

[0280]

[0281] Preparation of tert-butyl 2-(4-(((5-(prop-2-yn-1-yloxy)benzofuran-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23s): Using the synthetic method of 23c, 5-(prop-2-yn-1-yloxy)benzofuran-2-carboxylic acid and compound 15a were used as raw materials to give 23s as a white solid with a yield of 50%.

[0282]

[0283] Preparation of tert-butyl 2-(4-(((5-hydroxybenzofuran-2-carboxamido)methyl)benzoyl)-1-propylhydrazine-1-carboxylate (23u): Using the synthetic method of 23c, 5-hydroxybenzofuran-2-carboxylic acid and compound 15a were used as raw materials to obtain 23u as a white solid with a yield of 58%.

[0284]

[0285] Preparation of (E)-1-propyl-2-(4-(((8-(3-(pyridin-3-yl)acrylamido)octanamido)methyl)benzoyl)hydrazine-1-carboxylate (23v): Compound 2d (0.29 g, 1 mmol) was dissolved in 15 mL of dichloromethane, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 0.23 g, 1.2 mmol) and 1-hydroxybenzotriazole (HOBt at 0°C, 0.18 g, 1.2 mmol) were added under ice bath. After 30 minutes, 15a (0.37 mg, 1.2 mmol) and triethylamine (0.17 mL, 1.2 mmol) were added and reacted at room temperature overnight. The reaction solution was washed with brine (2×30 mL) and dried over MgSO4. After filtration, the solvent was evaporated and purified by flash chromatography to give compound 23v as a white solid powder (0.3 g, 51%). 1 H NMR (400MHz, DMSO-d6) 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),7.4 5-7.36(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] + .

[0286]

[0287] Preparation of (E)-1-propyl-2-(4-(7-(3-(pyridin-3-yl)acrylamide)heptylamide)benzoyl)tert-butylhydrazine (carboxylic acid) (23x): Using the synthetic method of 23v and compounds 2c and 15b as raw materials, 23x was obtained as a white solid with a yield of 53%. 1H NMR (400MHz, DMSO-d6) δ10.36(d,J=3.6Hz,1H),10.12(s,1H),8.72(d,J=2.2Hz,1H),8.52(d d,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),1.4 5(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] + .

[0288]

[0289] Preparation of (E)-1-propyl-2-(4-(8-(3-(pyridin-3-yl)acrylamido)octanamido)benzoyl)hydrazine tert-butyl-1-carboxylate (23w): Using the synthetic method of 23v and compounds 2d and 15b as raw materials, 23w was obtained as a white solid with a yield of 51%. 1 H NMR(400MHz,DMSO-d6)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),7.42-7.3 7(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] + .

[0290]

[0291] Preparation of tert-butyl 2-(4-(8-azidooctaamino)benzoyl)-1-propylhydrazine-1-carboxylate (24): Using the synthetic method of 23v and LEE37, compounds 8 and 13b were used as raw materials to obtain 24 as a white solid with a yield of 50%.1 H NMR (400MHz, DMSO-d6) 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] + .

[0292] Embodiment 15:

[0293]

[0294] Preparation of (E)-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-3-(pyridin-3-yl)acrylamide (LEE12): Using the synthesis method of LEE1, compound 13b and (E)-3-(pyridin-3-yl)acrylic acid were used as raw materials to obtain LEE12 as a white solid with a yield of 52%. 1 H NMR (600MHz, DMSO-d6) δ9.98 (s, 1H), 8.88-8.68 (m, 2H), 8.56 (dd, J = 4.8, 1.6Hz, 1 H),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). 13 CNMR(101MHz,DMSO-d6)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] + .

[0295] Embodiment 16:

[0296]

[0297] Preparation of N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-1H-pyrrolo[3,2-c]pyridine-2-carboxamide (LEE18): 23c (0.22 g, 0.5 mmol) was dissolved in 15 mL of dichloromethane, trifluoroacetic acid (TFA, 0.11 g, 1.0 mmol) was added, and the reaction was allowed to proceed overnight at room temperature. After the reaction was complete, triethylamine (0.1 g, 1.0 mmol) was added to adjust the pH to 8. The solution was then washed with brine and dried over Na2SO4. After filtration, the solvent was evaporated and purified by flash chromatography to give compound LEE18 as a white solid powder (0.14 g, 85%). 1 H NMR (400MHz, DMSO-d6) 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). 13 C NMR(101MHz,DMSO-d6)δ165.56,160.21,142.79,141.87,140.84,136.81,135.09,1 32.44,127.68,127.57,124.42,109.75,104.54,53.55,42.64,21.28,12.12.ESI-MS m / z:351.86[M+H] + .

[0298] Embodiment 17:

[0299]

[0300] Preparation of N-(4-(2-propylhydrazine-1-carbonyl)benzyl)benzofuran-2-carboxamide (LEE16): Using the synthetic method of LEE18, compound 23a and trifluoroacetic acid were used as raw materials to obtain LEE16 as a white solid with a yield of 79%. 1H NMR(400MHz,DMSO-d6)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). 13 C NMR(101MHz,DMSO-d6)δ165.60,158.72,154.75,149.49,143.00,132.36,127.64,1 27.60,127.35,124.20,123.26,112.27,110.12,53.57,42.42,21.32,12.12.ESI-MS m / z:352.07[M+H] + .

[0301] Embodiment 18:

[0302]

[0303] Preparation of N-(4-(2-propylhydrazine-1-carbonyl)benzyl)benzo[b]thiophene-2-carboxamide (LEE17): Using the synthetic method of LEE18, compound 23b and trifluoroacetic acid were used as raw materials to obtain LEE17 as a white solid with a yield of 83%. 1 H NMR (400MHz, DMSO-d6) 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-d6)δ165.59,162.10,143.03,140.72,140.17,139.64,132.41,127.6 4,127.60,126.72,125.69,125.46,125.41,123.29,53.58,42.95,21.32,12.13.ESI-MS m / z:367.96[M+H] + .

[0304] Embodiment 19:

[0305]

[0306] Preparation of N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-1H-indole-2-carboxamide (LEE15): The synthetic method of LEE12 was adopted, with compound 15a and 1H-indole-2-carboxylic acid as raw materials, condensation and deprotection to obtain LEE15 as a white solid with a yield of 56%. 1 H NMR (400MHz, DMSO-d6) δ11.54(s,1H),9.89(d,J=5.6Hz,1H),9.01(t,J=6.1Hz,1 H),7.75-7.70(m,2H),7.55(dt,J=7.9,1.0Hz,1H),7.40-7.30(m,3H),7.15-7.0 8(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). 13 C NMR(101MHz,DMSO-d6)δ165.60,161.70,143.45,136.99,132.29,131.98,127.60,127.5 7,127.44,123.83,121.99,120.22,112.79,103.16,53.58,42.41,21.32,12.13.ESI-MS m / z:350.86[M+H] + .

[0307] Embodiment 21:

[0308]

[0309] Preparation of N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-1H-pyrrolo[2,3-c]pyridine-2-carboxamide (LEE21): Using the synthetic method of LEE18, compound 23e and trifluoroacetic acid were used as raw materials to obtain LEE21 as a white solid with a yield of 82%. 1 H NMR(400MHz,DMSO-d6)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-d6)δ165.57,161.10,143.05,138.53,136.28,135.24,133.77,1 32.39,131.72,127.64,127.50,116.28,101.96,53.57,42.55,21.32,12.13.ESI-MS m / z:352.06[M+H] + .

[0310] Embodiment 22:

[0311]

[0312] Preparation of (E)-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-3-(pyridin-2-yl)acrylamide (LEE13): Using the synthesis method of LEE18, compound 15a and 3-(2-pyridinyl)acrylic acid were used as raw materials to obtain LEE13 as a white solid with a yield of 57%. 1 H NMR(400MHz, DMSO-d6)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-d6)δ165.57,165.31,165.13,153.46,150.67,150.33,149.64,143.13,143.10,139.02,137.66,136.29,1 34.46,132.34,131.12,127.61,127.53,125.95,124.74,124.53,124.45,124.39,53.58,42.55,42.52,21.32,12.13.ESI-MS m / z:339.06[M+H] + .

[0313] Embodiment 23:

[0314]

[0315] Preparation of (E)-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-3-(pyridin-4-yl)acrylamide (LEE14): Using the synthesis method of LEE18, compound 15a and 3-(4-pyridinyl)acrylic acid were used as raw materials to obtain LEE14 as a white solid with a yield of 55%. 1 H NMR (400MHz, DMSO-d6) 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). 13 CNMR(101MHz,DMSO-d6)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] + .

[0316] Embodiment 24:

[0317] N-(4-(2-propylhydrazine-1-carbonyl)benzyl)thieno[3,2-c]pyridine-2-carboxamide (LEE19). Using the synthetic method of LEE18, compound 15a and thieno[3,2-c]pyridine-2-carboxylic acid were used as raw materials to obtain LEE19 as a white solid with a yield of 54%. ESI-MS m / z: 368.92[M+H] + .

[0318] Embodiment 25:

[0319]

[0320] Preparation of N-(4-(2-propylhydrazine-1-carbonyl)benzyl)furan[3,2-c]pyridine-2-carboxamide (LEE20): Using the synthetic method of LEE18, compound 23d and trifluoroacetic acid were used as raw materials to obtain LEE20 as a white solid with a yield of 78%. 1 H NMR(400MHz,DMSO-d6)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); 13 C NMR(101MHz,DMSO-d6)δ165.64,158.98,158.21,150.06,146.84,146.48,142.8 6,132.38,127.68,125.05,108.53,108.17,53.58,42.53,21.33,12.19.ESI-MS m / z:352.88[M+H] + .

[0321] Embodiment 26:

[0322]

[0323] Preparation of 5-bromo-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-1H-indole-2-carboxamide (LEE26): Using the synthetic method of LEE18, compound 23j and trifluoroacetic acid were used as raw materials to obtain LEE26 as a white solid with a yield of 84%. 1HNMR(500MHz,DMSO-d6)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).

[0324] Embodiment 27:

[0325]

[0326] Preparation of 5-fluoro-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-1H-indole-2-carboxamide (LEE28): Using the synthetic method of LEE18, compound 231 and trifluoroacetic acid were used as raw materials to obtain LEE28 as a white solid with a yield of 85%. 1 HNMR(500MHz,DMSO-d6)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).

[0327] Embodiment 28:

[0328]

[0329] Preparation of 6-(dimethylamino)-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-1H-indole-2-carboxamide (LEE27): Using the synthetic method of LEE18, compound 23k and trifluoroacetic acid were used as raw materials to obtain LEE27 as a white solid with a yield of 82%.

[0330] Embodiment 29:

[0331]

[0332] Preparation of 5-chloro-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-1H-indole-2-carboxamide (LEE29): Using the synthetic method of LEE18, compound 23m and trifluoroacetic acid were used as raw materials to obtain LEE29 as a white solid with a yield of 80%. 1 HNMR (500MHz, DMSO-d6) 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=6.1H z,2H),2.73(t,J=7.1Hz,2H),1.50-1.39(m,2H),0.89(t,J=7.4Hz,4H).

[0333] Embodiment 30:

[0334]

[0335] Preparation of 5-methoxy-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)benzofuran-2-carboxamide (LEE30): Using the synthetic method of LEE18, compound 23n and trifluoroacetic acid were used as raw materials to obtain LEE30 as a white solid with a yield of 85%. 1 H NMR(500MHz, DMSO-d6)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).

[0336] Embodiment 31:

[0337]

[0338] Preparation of 5-fluoro-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)benzofuran-2-carboxamide (LEE31): Using the synthetic method of LEE18, compound 23o and trifluoroacetic acid were used as raw materials to obtain LEE31 as a white solid with a yield of 86%.1 HNMR(500MHz,DMSO-d6)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).

[0339] Embodiment 32:

[0340]

[0341] Preparation of 5-bromo-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)benzofuran-2-carboxamide (LEE32): Using the synthetic method of LEE18, compound 23p and trifluoroacetic acid were used as raw materials to obtain LEE32 as a white solid with a yield of 84%. 1 HNMR(500MHz,DMSO-d6)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).

[0342] Embodiment 33:

[0343]

[0344] Preparation of 5-methoxy-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-1H-indole-2-carboxamide (LEE33): Using the synthetic method of LEE18, compound 23q and trifluoroacetic acid were used as raw materials to obtain LEE33 as a white solid with a yield of 82%. 1H NMR(500MHz,DMSO-d6)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),3.7 5(s,3H),2.73(t,J=7.1Hz,2H),1.50-1.40(m,2H),0.89(t,J=7.4Hz,3H).

[0345] Embodiment 34:

[0346]

[0347] Preparation of 5-chloro-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)benzofuran-2-carboxamide (LEE34): Using the synthetic method of LEE18, compound 23r and trifluoroacetic acid were used as raw materials to obtain LEE34 as a white solid with a yield of 86%. 1 HNMR(500MHz,DMSO-d6)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).

[0348] Embodiment 35:

[0349]

[0350] Preparation of 5-(prop-2-yn-1-yloxy)-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)benzofuran-2-carboxamide (LEE35): Using the synthetic method of LEE18, compound 23s and trifluoroacetic acid were used as raw materials to obtain LEE35 as a white solid with a yield of 83%. 1H NMR (500MHz, DMSO-d6) δ9.97 (s, 1H), 9.31 (t, J = 6.2Hz, 1H), 7.81-7.74 (m, 2H), 7.6 1-7.55(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).

[0351] Embodiment 36:

[0352]

[0353] Preparation of 5-hydroxy-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)benzofuran-2-carboxamide (LEE36): Using the synthetic method of LEE18, compound 23u and trifluoroacetic acid were used as raw materials to obtain LEE36 as a white solid with a yield of 79%. 1 HNMR (500MHz, DMSO-d6) 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).

[0354] Embodiment 37:

[0355]

[0356] Preparation of (E)-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-8-(3-(pyridin-3-yl)acrylamido)octanamide (LEE37): Using the synthesis method of LEE18, compound 23v and trifluoroacetic acid were used as raw materials to obtain LEE37 as a white solid with a yield of 78%. 1H NMR (400MHz, DMSO-d6) δ9.93 (s, 1H), 8.71 (s, 1H), 8.50 (d, J = 4.7Hz, 1H), 8.31 (t, J = 6.0Hz, 1H), 8.1 2(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). 13 C NMR (126MHz, DMSO-d6) δ172.69,165.56,164.83,150.49,149.51,143.60,135.52,134.34,132.12,131.22,127.4 9,127.36,124.81,124.41,53.55,42.16,39.17,35.76,29.53,29.09,28.94,26.84,25.70,21.30,12.12.ESI-MS m / z:479.96[M+H] + .

[0357] Embodiment 38:

[0358]

[0359] Preparation of (E)-N-(4-(2-propylhydrazine-1-carbonyl)benzyl)-7-(3-(pyridin-3-yl)acrylamido)heptylamide (LEE39): Using the synthesis method of LL289 and compounds 15a and 2d as raw materials, YKR-31 was obtained as a white solid with a yield of 58%. 1H NMR (400MHz, DMSO-d6) δ9.93(s,1H),8.51(dd,J=4.7,1.6Hz,1H),8.32(t,J=6.1Hz,1H),8.13(t,J=5.6H z,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). 13 CNMR(126MHz,DMSO-d6)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] + .

[0360] Embodiment 39:

[0361]

[0362] Preparation of (E)-N-(4-(2-propylhydrazine-1-carbonyl)phenyl)-7-(3-(pyridin-3-yl)acrylamido)heptylamide (LEE40): Using the synthesis method of LEE18, compound 23x and trifluoroacetic acid were used as raw materials to obtain LEE40 as a white solid with a yield of 65%. 1H NMR (400MHz, DMSO-d6) 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). 13 C NMR (126MHz, DMSO-d6) δ172.09,165.33,164.83,150.50,149.51,142.38,135.51,134.34,131.22,128.82,128.2 8,127.85,124.81,124.41,118.63,53.62,39.14,36.87,29.42,28.84,26.71,25.41,21.31,12.12,11.14.ESI-MS m / z:451.97[M+H] + .

[0363] Embodiment 40:

[0364]

[0365] Preparation of (E)-N-(4-(2-propylhydrazine-1-carbonyl)phenyl)-8-(3-(pyridin-3-yl)acrylamide)amide (LEE38): Using the synthesis method of LEE18, compound 23w and trifluoroacetic acid were used as raw materials to obtain LEE38 as a white solid with a yield of 67%. 1H NMR (400MHz, DMSO-d6) δ10.04(s,1H),9.85(s,1H),8.71(d,J=2.3Hz,1H),8.50(dd,J=4.7,1.6H z,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). 13 C NMR (126MHz, DMSO-d6) δ172.10,165.34,164.82,150.49,149.50,142.39,135.51,134.34,131.22,128.29,12 7.83,124.81,124.41,118.62,53.60,39.15,36.90,29.53,29.08,28.98,26.82,25.42,21.29,12.12.ESI-MS m / z:465.97[M+H] + .

[0366] Embodiment 41:

[0367]

[0368] Preparation of N-(4-(2-propylhydrazine-1-carbonyl)phenyl)-8-(4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)octanamide (LEE41): Using the synthetic method of LEE18, compound 24 and trifluoroacetic acid were used as raw materials to obtain a white solid with a yield of 67%.

[0369] The synthetic method of LEE11 was adopted, and the product of the previous step and 3-ethynylpyridine were used as raw materials to obtain LEE41 as a white solid with a yield of 80%. 1H NMR (400MHz, DMSO-d6) 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). 13 C NMR (126MHz, DMSO-d6) δ172.10,165.34,149.24,146.77,143.92,142.46,132.82,128.33,127.67,127.2 7,124.47,122.41,118.62,53.53,50.07,36.84,30.01,28.91,28.61,26.17,25.34,21.15,12.08.ESI-MS m / z:464.03[M+H] + .

[0370] Embodiment 42:

[0371]

[0372] Preparation of methyl (E)-4-((3-(pyridine-3-yl)acrylamide)methyl)benzoate (LEE43): Using the synthetic method of 1a, LEE43 was obtained as a white solid with a yield of 51%. 1 H NMR (400MHz, DMSO-d6) 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).

[0373] Embodiment 43:

[0374]

[0375] Preparation of methyl 4-((1H-pyrrolo[3,2-c]pyridine-2-carbonylamide)methyl)benzoate (LEE44): LEE44 was obtained as a white solid using the synthetic method of 1a in a yield of 46%. 1H NMR (400 MHz, DMSO-d6) δ 12.94 (s, 1H), 9.65 (t, J = 4.0 Hz, 1H), 9.31 (s, 1H), 8.39 (d, J = 4.3 Hz, 1H), 7.96 (d, J = 5.6 Hz, 2H), 7.74 (d, J = 4.2 Hz, 1H), 7.64 (s, 1H), 7.51 (d, J = 5.5 Hz, 2H), 4.64 (d, J = 4.0 Hz, 2H), 3.85 (s, 3H).

[0376] Example 44: Inhibitory activity of compounds against HDAC1, 2, and 3.

[0377] Experimental materials: HDAC buffer: 15mM Tris-HCl (PH 8.0), 250μM EDTA, 250mM NaCl, 10% glycerol. Trypsin stop solution: 10mg / ml trypsin, 50mM Tris-HCL (pH8.0), 100mM NaCl, 2μM TSA. Substrate: Dissolve the exclusive substrate dimethyl sulfoxide of HDAC1, HDAC2 and HDAC3 into a 30mM stock solution, dilute to 300μM with HDAC buffer, so that the dimethyl sulfoxide content is about 1%. Enzyme solution: HDAC1, HDAC2 and HDAC3 are diluted with HDAC buffer at a ratio of 1:20.

[0378] Experimental steps:

[0379] a) Preparation of 100% solution: 50 μL HDAC buffer was mixed with 10 μL enzyme solution, and 40 μL substrate was added after 5 minutes to react at 37°C for 30 minutes, and then 100 μL trypsin stop solution was added to terminate the above reaction, and the reaction was continued at 37°C for 20 minutes. The fluorescence intensity was measured at 390nm / 460nm, and the 100% absorption was obtained. AMC was used as a standard to make a standard curve and calculate the enzyme activity.

[0380] b) Preparation of blank solution: Add 40 μL substrate to 60 μL HDAC buffer and react at 37°C for 30 min, then add 100 μL trypsin stop solution and react at 37°C for 20 min. Measure the fluorescence intensity at 390 nm / 460 nm to obtain the blank absorption.

[0381] 6. Determination of drug inhibition of HDAC enzyme activity: 50 μL of HDAC buffer containing drugs was mixed with 10 μL of enzyme solution and pre-incubated for 5 minutes. After adding 40 μL of substrate, the reaction was carried out at 37°C for 30 minutes. Then, 100 μL of trypsin stop solution was added to terminate the above reaction, and the reaction was carried out at 37°C for 20 minutes. The fluorescence intensity was measured at 390nm / 460nm.

[0382]

[0383] Finally, the inhibition rate (%) of the compound and its corresponding concentration were fitted with an S curve to calculate the IC 50 value.

[0384] The inhibitory activity results of some compounds represented by the general structural formula (I, II, III or IV) of the present invention on HDAC1, HDAC2 and HDAC3 are shown in Table 1 below:

[0385] Table 1. Inhibitory IC values ​​of some compounds on HDAC1, 2, and 3 50 value.

[0386]

[0387]

[0388]

[0389] a The data in the table are from three independent experiments, the values ​​are mean values, and the standard deviation is <10%; ND: not detected.

[0390] The experimental results show that most of the compounds in the table have nanomolar inhibitory IC against HDAC1 / 2 / 3. 50 The activity against HDAC1 and HDAC3 is generally higher than that against HDAC2. The activity is generally higher than that of the positive control drugs SAHA and MS275.

[0391] Example 45: Inhibitory activity of compounds on NAMPT.

[0392] Table 2. Inhibitory IC values ​​of some compounds on NAMPT 50 value.

[0393]

[0394]

[0395] a The data in the table are from three independent experiments, the values ​​are average values, and the standard deviation is <10%.

[0396] The experimental results show that LEE12, LEE18, LEE7, LEE43, and LEE44 all exhibit micromolar-level inhibitory activity against NAMPT, and the above compounds have achieved unexpected technical effects compared to the compounds in Comparative Example 1 and Comparative Example 2.

[0397] Example 46: The half-maximal growth inhibitory concentration (GI 50 ) and the half-lethal concentration (LC 50 ) of the compound against tumor cells

[0398] The half-maximal growth inhibitory concentration (GI 50 ) and the half-lethal concentration (LC 50 ) were determined by the NCI method. The 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 inoculated into 96-well cell culture plates at a density of 10,000 cells / 100 μL and cultured overnight. Six wells were selected as the Tz wells, and 0.125 mg / mL of Cell Titer-Blue dye was added. After 4 hours of culture, the fluorescence intensity was read at 560 nM / 590 nM (excitation wavelength / emission wavelength). The remaining wells were added with different concentrations of the compound, and a 100% control group was set up. After 48 hours of culture, 0.125 mg / mL of Cell Titer-Blue dye was added, and the fluorescence intensity was read at 560 nM / 590 nM after 4 hours. The fluorescence intensity of the drug-added group was represented by Ti, the fluorescence intensity of the 100% control group was represented by C, and the fluorescence intensity before adding the drug was represented by Tz.

[0399] If Ti ≥ Tz, use the formula [(Ti - Tz) / (C - Tz)] × 100

[0400] If Ti < Tz, use the formula [(Ti - Tz) / Tz] × 100

[0401] The half-maximal growth inhibitory concentration (GI 50 ) is the compound concentration at which [(Ti - Tz) / (C - Tz)] × 100 = 50, and the half-lethal concentration (LC 50 ) is the concentration at which [(Ti - Tz) / Tz] × 100 = -50. The GI 50 and LC 50 values of some compounds against the 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.

[0402] Table 3. The GI of some compounds against acute myeloid leukemia cells MV4-11 and HL6050 and LC 50 value.

[0403]

[0404]

[0405] Note: KF866 is a NAMPT inhibitor reported in the literature.

[0406] a The data in the table are from three independent experiments, the values ​​are mean values, and the standard deviation is <10%; b ND: Not detected.

[0407] The experimental results showed that in the wt-p53 cell line MV4-11, all the tested compounds showed nanomolar GI 50 Value and LC 50 The values ​​indicate that they can not only inhibit cell proliferation but also cause cell death. The activity of most compounds is significantly stronger than that of the compound LP411 (3b) in Comparative Example 1. In the p53-null cell line HL60, all tested compounds showed nanomolar GI 50 values, indicating that they have good antiproliferative activity, while only LEE12, LEE14, LEE18, and LEE7 have nanomolar LC 50 This suggests that simultaneous inhibition of HDAC and NAMPT may have a synthetic lethal effect on p53-null cell lines.

[0408] Table 4. GI of some compounds on leukemia cell lines 50 and LC 50 value.

[0409]

[0410]

[0411] a The data in the table are from three independent experiments, the values ​​are average values, and the standard deviation is <10%.

[0412] Figure 2 Table 4 also shows that the HDAC and NAMPT dual inhibitors LEE12 and LEE18 have lethal effects on p53-mutant and p53-null cell lines. Most of the compounds have achieved unexpected technical effects compared with the compounds in Comparative Example 1.

[0413] Example 47: In vivo anti-colon cancer activity of target compound LEE17

[0414] 5-FU: 5-fluorouracil, a traditional anti-tumor chemotherapy drug, Oxaliplatin: Oxaliplatin, a third-generation platinum anticancer drug, is an anti-tumor chemotherapy drug.

[0415] Colon cancer cells HCT116 were inoculated subcutaneously on the right shoulder of nude mice, 100uL per mouse (cell count: 1.8*10 8 / mL), and one week later, the tumor-bearing mice were divided into groups and given the drug by gavage. The grouping was as follows:

[0416] Test group: Compound LL341, oral administration dose 8 mg / kg / d, administration volume: 0.2 mL per mouse each time

[0417]

[0418] Relative tumor volume (RTV) = Vt / Vo

[0419] The evaluation index of antitumor activity is the relative tumor proliferation rate T / C (%),

[0420]

[0421] Table 5. In vivo study data results of compound LEE17 on HCT116 tumor model

[0422]

[0423] Note: 5-FU: 5-fluorouracil, a traditional anti-tumor chemotherapy drug; Oxaliplatin: Oxaliplatin, a third-generation platinum anticancer drug, is an anti-tumor chemotherapy drug.

[0424] The experimental results showed that at a dose of 8 mg / kg / d, compound LEE17 could significantly inhibit the growth of HCT116 tumors, with an inhibition rate of 85.5%, significantly higher than the positive drug 5-FU+Oxaliplatin. At the end of the experiment, there was no significant change in the body weight of nude mice, indicating that LEE17 has a certain safety at the dose.

[0425] Example 48: In vivo anti-leukemia activity of target compound LEE12 Panobinostat: Panobinostat, a marketed broad-spectrum HDAC inhibitor

[0426] The acute myeloid leukemia cells MV4-11 were inoculated subcutaneously on the right shoulder of nude mice, 100uL per mouse (cell count: 1.8*10 8 / mL), and one week later, the tumor-bearing mice were divided into groups and given the drug by gavage. The grouping was as follows:

[0427] Test group: compounds LEE15 and LEE16, dosage / 4 mg / kg / d, administration volume: 200uL / 20g per mouse each time;

[0428] Positive control group: positive drug panobinostat, dosed at 4 mg / kg / d, intraperitoneal injection.

[0429] Blank control group: the same volume of PBS was given.

[0430] The drug was given once a day, and the tumor volume was measured every 3-4 days. The average value of each group was taken and the tumor growth curve was drawn (see Figure 6 ), MV4-11 tumor-bearing model was sacrificed on the 23rd day of administration, and the tumor and internal organs were dissected. At the end of the experiment, the tumor mass was weighed and the tumor inhibition rate was calculated according to the formula. The maximum diameter (a) and minimum diameter (b) of the tumor were measured, and the tumor volume (V) was calculated: V = ab 2 / 2, and the relative tumor proliferation rate T / C (%) was calculated.

[0431]

[0432] Relative tumor volume (RTV) = Vt / Vo

[0433] The evaluation index of antitumor activity is the relative tumor proliferation rate T / C (%),

[0434]

[0435] Table 6. In vivo study data results of compound LEE12 on MV4-11 tumor model

[0436]

[0437] Note: Panobinostat, positive control drug, is a marketed broad-spectrum HDAC inhibitor.

[0438] The experimental results showed that at a dose of 5 mg / kg, LEE15 and LEE16 had significant in vivo anti-acute myeloid leukemia effects, with tumor inhibition rates of 81.3% and 78.4%. The in vivo anti-tumor activity was significantly higher than that of the positive control drug panobinostat.

[0439] Example 49: Pharmacokinetic properties of target compounds LEE15, LEE16, and LEE17

[0440] LEE15, LEE16, and LEE17 were dissolved in 40% PEG300 and 60% H2O. Three mice in each group were given a single dose of 20 mg / kg orally (po) and 5 mg / kg intravenously (iv). Blood was collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h after administration, and the samples were prepared and measured for t 1 / 2 ,C0,AUC,Vss,CLp,MRT,C max ,t max , F% and other parameters.

[0441] Po administration

[0442] Table 7. Pharmacokinetic properties of target compounds LEE15, LEE16, and LEE17

[0443]

[0444] From the above metabolic data results, it can be seen that the metabolic effects of compounds LEE16, LEE17, and LEE15 are significantly better than those of the compounds in Comparative Example 2, and the above compounds LEE16, LEE17, and LEE15 have unexpected technical effects in metabolism.

[0445] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. An HDAC compound having multi-target inhibitory activity and a pharmaceutically acceptable salt thereof, characterized in that: The structure of the HDAC compound with multi-target inhibitory activity is as follows: ， ， ， ， ， ， ， ， ， 。 2. An HDAC compound having multi-target inhibitory activity and a pharmaceutically acceptable salt thereof according to claim 1, wherein the pharmaceutically acceptable salt comprises an inorganic acid salt and an organic acid salt.

3. An HDAC compound having multi-target inhibitory activity according to any one of claims 1 to 2 and a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt comprises an inorganic acid salt selected from the group consisting of salts formed with sulfuric acid, carbonic acid, nitric acid, hydrobromic acid, phosphoric acid, hydrochloric acid, boric acid, and aminosulfonic acid; or an organic acid salt selected from the group consisting of salts formed with acetic acid, propionic acid, butyric acid, camphoric acid, capric acid, caproic acid, caprylic 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, hydroxymaleic acid, palmitic acid, cinnamic acid, isobutyric acid, lauric acid, mandelic acid, Salts formed from maleic acid, fumaric acid, malic acid, tartaric acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, 2-hydroxy-1,2,3-propanetriol, suberic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, formic acid, fumaric acid, mucic acid, gentisic acid, pyruvic acid, salicylic acid, methanesulfonic acid, ethylsulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfinic acid, isethionic acid, ethanedisulfonic acid, 4-(w-methoxycarbonylamino)butyric acid, dichloroacetic acid, 1,2-ethanedisulfonic acid, camphor-10-sulfonic acid, 2,4-dihydroxybenzoic acid, α-ketoglutaric acid, 1-hydroxy-2-naphthoic acid, p-acetamidobenzoic acid, 2-hydroxybenzoic acid, 4-amino-2-hydroxybenzoic acid, all-trans retinoic acid, and valproic acid. 4 . A pharmaceutical composition comprising the HDAC compound having multi-target inhibitory activity according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof.

5. The pharmaceutical composition according to claim 4, characterized in that The pharmaceutical composition is in the form of tablets, pills, powders, lozenges, elixirs, suspensions, emulsions, solutions, syrups, aerosols, ointments, soft or hard gelatin capsules, suppositories or sterile injectable solutions.

6. Use of an HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 in the preparation of a medicament for preventing or treating diseases associated with abnormal expression of histone deacetylase activity and NAD synthesis, wherein the diseases are leukemia and colon cancer.

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