MULTI-TARGET INHIBITOR TARGETING HDAC AND NAD SYNTHESIS AND USE OF MULTI-TARGET INHIBITOR

Abstract

A compound targeting HDAC and NAD synthesis and a pharmaceutically acceptable salt, hydrate, deuterate, isomer or prodrug thereof, as well as preparation and application thereof. Specifically, a compound shown in structural general formula (I) and a pharmaceutically acceptable salt, hydrate, deuterate, isomer or prodrug thereof are provided. The compound of the structural general 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.

Ring E-B-L-C (O)(NH)r-R (General formula I)

Claims

1. An HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer or prodrug thereof, wherein the multi-target HDAC compound has a general formula I,
Ring E-B-L-C(O)(NH)r-R (General formula I) wherein, ring E is selected from ##STR00139## r is equal to 1 and 2, R is selected from R.sup.1, and ##STR00140## and the R.sup.1 is selected from H, C.sub.1-4 alkyl, C.sub.3-5 cycloalkyl or C.sub.1-2 alkyl substituted C.sub.3-5 cycloalkyl; ring A is selected from one or more substituted or unsubstituted 5-membered aromatic or heteroaromatic rings, the heteroaromatic ring contains 1 to 2 heteroatoms of N, O, or S; the substituents are H, halogen, C.sub.1-2 alkyl, halomethyl, OH, OCH.sub.3, O(CH.sub.2).sub.nCH.sub.3, cyclopropyloxy, OC(CH.sub.3).sub.3, OCH(CH.sub.3).sub.2, NH.sub.2, N(CH.sub.3).sub.2, NH(CH.sub.2).sub.nCH.sub.3, CN, N.sub.3, etc., wherein n is from 0 to 9; X.sup.1 is selected from CR.sup.4 or N; X.sup.2 is selected from CR.sup.5 or N; X.sup.3 is selected from CR.sup.6 or N; X.sup.4 is selected from CR.sup.7 or N; X.sup.5, X.sup.6 or X.sup.7 are independently selected from CH or N; R.sup.2 and R.sup.3 are each independently selected from H, halogen, CH.sub.3, and OCH.sub.3; R.sup.4, R.sup.5, R.sup.6 and R.sup.7 are selected from H, halogen, C.sub.1-2 alkyl, halomethyl, OH, OCH.sub.3, O(CH.sub.2).sub.nCH.sub.3, OC(CH.sub.3).sub.3, OCH(CH.sub.3).sub.2, cyclopropyloxy, 5 to 6-membered alkoxy, NH.sub.2, N(CH.sub.3).sub.2, NH(CH.sub.2).sub.nCH.sub.3, CN, and N.sub.3, wherein n is from 0 to 9; B is selected from ##STR00141## represents a bond connected to ring A; represents a bond connected to L; and L is selected from the group consisting of C.sub.1-14 alkyl, C.sub.1-14 alkoxy, C.sub.2-14 alkenyl, C.sub.2-14 alkynyl, C.sub.3-10 cycloalkyl, C.sub.6-10 aryl or benzyl.

2. The HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer, or prodrug thereof of claim 1, wherein the compound of general formula I further has a structure as shown in general formula II, general formula III, general formula IV, and general formula V, ##STR00142##

3. The HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer, or prodrug thereof of claim 1, wherein, ring A is selected from the following ring systems: ##STR00143## wherein, G is selected from CH.sub.2, NH, N(CH.sub.2).sub.nCH.sub.3, O or S, wherein n is from 0 to 9; represents a bond connected to an adjacent fused ring; and represents a bond connected to B.

4. The HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer, or prodrug thereof of claim 1, wherein X.sup.1, X.sup.2, X.sup.3 or X.sup.4 is N and 1-2 of X.sup.1, X.sup.2, X.sup.3 and X.sup.4 are N at the same time.

5. The HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer, or prodrug thereof of claim 1, wherein X.sup.5 is N.

6. The HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer, or prodrug thereof of claim 1, wherein X.sup.6 or X.sup.7 is N.

7. The HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer, or prodrug thereof of claim 1, wherein R.sup.4, R.sup.5, R.sup.6 or R.sup.7 are each independently selected from H, halogen, (C.sub.1-2) alkyl, halomethyl, OH, OCH.sub.3, O(CH.sub.2).sub.nCH.sub.3, cyclopropyloxy, OC(CH.sub.3).sub.3, OCH(CH.sub.3).sub.2, 5 to 6-membered alkoxy, NH.sub.2, N(CH.sub.3).sub.2, NH(CH.sub.2).sub.nCH.sub.3, CN, and N.sub.3, wherein n is from 0 to 9.

8. The HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer, or prodrug thereof of claim 1, wherein L is selected from linear C.sub.1-14 alkyl or L is selected from linear C.sub.1-14 alkoxy, wherein the C.sub.1-14 alkyl and C.sub.1-14 alkoxy are selected from (CH.sub.2CH.sub.2O).sub.m, (CH.sub.2CH.sub.2O).sub.mC.sub.2H.sub.4, (CH.sub.2CH.sub.2O).sub.mCH.sub.2, (CH.sub.2CH.sub.2O).sub.n, C.sub.2H.sub.4(CH.sub.2CH.sub.2O).sub.n, CH.sub.2(OCH.sub.2CH.sub.2)n-, and (CH.sub.2).sub.pO(CH.sub.2).sub.q; wherein m, n, p or q is independently selected from 1, 2, 3 or 4; or when L is C.sub.3-10 cycloalkyl, and C.sub.6-10 aryl or benzyl, the cycloalkyl, aryl or benzyl is substituted by C.sub.1-9 alkyl, C.sub.1-9 alkoxy, and (C.sub.1-9 alkyl)-(C?O)NH; or when L is C.sub.3-10 cycloalkyl, the cycloalkyl contains heteroatoms such as N, O or S; or when L is aryl or benzyl, the aryl or benzyl is an aromatic heterocycle, such as N, O or S.

9. The HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer, or prodrug thereof of claim 1, wherein the compound is selected from: (E)-N-(3-oxo-3-(2-propylhydrazino)propyl)-3-(pyridin-3-yl) acrylamide, (E)-N-(5-oxo-5-(2-propylhydrazino)pentyl)-3-(pyridin-3-yl) acrylamide, (E)-N-(7-oxo-7-(2-propylhydrazino)heptyl)-3-(pyridin-3-yl) acrylamide, (E)-N-(8-oxo-8-(2-propylhydrazino)octyl)-3-(pyridin-3-yl) acrylamide, 1-(7-oxo-7-(2-propylhydrazino)heptyl)-3-(pyridin-3-ylmethyl)urea, N-(7-oxo-7-(2-propylhydrazino)heptyl)-3H-pyrrolo[3,2-c]pyridine-2-carboxyamide 8-azido-N-propyloctanehydrazide, (E)-N-(2-aminophenyl)-8-(3-(pyridin-3-yl)acrylamide) octanamide N-(2-aminophenyl)-8-azidooctanamide, n-propyl-8-(4-(pyridin-3-yl)-1H-1,2,3-triazole-1-yl) octanehydrazide, N-(2-aminophenyl)-8-(4-(pyridin-3-yl)-1H-1,2,3-triazole-1-yl) octanamide, (E)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-3-(pyridin-3-yl) acrylamide, N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-pyrrolo[3,2-c]pyridine-2-carboxyamide, N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide, N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzo[b]thiophene-2-carboxyamide, N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide, N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-pyrrolo[2,3-c]pyridine-2-carboxyamide, (E)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-3-(pyridine-2-yl) acrylamide, (E)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-3-(pyridine-4-yl) acrylamide, N-(4-(2-propylhydrazino-1-carbonyl)benzyl)thiopheno[3,2-c]pyridine-2-carboxyamide, N-(4-(2-propylhydrazino-1-carbonyl)benzyl)furan[3,2-c]pyridine-2-carboxyamide, 5-bromo-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide, 5-fluoro-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide, 6-(dimethylamino)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide, 5-chloro-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide, 5-methoxy-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide, 5-fluoro-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide, 5-bromo-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide, 5-methoxy-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide, 5-chloro-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide, 5-(prop-2-yn-1-yloxy)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide, 5-hydroxy-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide, (E)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-8-(3-(pyridin-3-yl)acrylamido) octanamide, (E)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-7-(3-(pyridin-3-yl)acrylamido)heptamide, (E)-N-(4-(2-propylhydrazino-1-carbonyl)phenyl)-7-(3-(pyridin-3-yl)acrylamido) heptamide, (E)-N-(4-(2-propylhydrazino-1-carbonyl)phenyl)-8-(3-(pyridin-3-yl)acrylamido) octanamide, N-(4-(2-propylhydrazino-1-carbonyl)phenyl)-8-(4-(pyridin-3-yl)-1H-1,2,3-triazole-1-yl) octanamide, methyl (E)-4-((3-(pyridine-3-yl)acrylamide) methyl) benzoate, or methyl 4-((1H-pyrrolo [3,2-c] pyridine-2-carbonylamide) methyl) benzoate.

10. The HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer, or prodrug thereof of claim 1, wherein the pharmaceutically acceptable salt comprises inorganic acid salts selected from salts of sulfuric acid, nitric acid, hydrobromic acid, phosphoric acid, hydrochloric acid, boric acid, and sulphamic acid; or organic acid salts selected from salts of acetic acid, propionic acid, butyric acid, camphoric acid, decanoic acid, hexanoic acid, octanoic acid, carbonic acid, cinnamic acid, hydroxyacetic 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, tartaric acid, sulfanilic acid, 2-acetyloxybenzoic acid, 2-hydroxy-1,2,3-tricarballylic acid, octanedioic 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, benzene methanesulfonic acid, p-toluenesulfonic acid, cyclohexyl sulfinic acid, isethionic acid, ethanedisulfonic acid, 4-(fluorenemethoxycarbonylamino)butyric acid, dichloroacetic acid, 1,2-ethanedisulfonic acid, camphore-10 sulfonic acid, 2,4-dihydroxybenzoic acid, ?-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.

11. A pharmaceutical composition, comprising the HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer, or prodrug thereof of claim 1.

12. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is an injectable or oral preparation.

13. A process comprising preventing or treating a disease related to abnormal expression of histone deacetylase activity or NAD synthesis with the HDAC compound with multi-target inhibitory activity and a pharmaceutically acceptable salt, hydrate, deuterate, isomer, or prodrug thereof of claim 1, wherein the diseases related to abnormal expression of histone deacetylase activity or NAD synthesis are selected from malignant tumors, neurodegenerative diseases, viral infections, acquired immune deficiency syndrome, inflammation, malaria, diabetes, fungal infections, and bacterial infections, among which the malignant tumors comprise all kinds of leukemia, lymphoma, myeloma, colorectal cancer, melanoma, gastric cancer, breast cancer, ovarian cancer, pancreatic cancer, liver cancer, glioma, intracerebral 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, and human fibrosarcoma.

14. A preparation method of an HDAC compound with multi-target inhibitory activity, comprising the following steps: ##STR00144## ##STR00145## ##STR00146## wherein the reagents and conditions are: (a) propanal, methanol, yield 98%; sodium cyanoborohydride, methanol, concentrated hydrochloric acid, methyl orange, yield 60%; (b) di-tert-butyl dicarbonate, triethylamine, ethanol, yield 85%; (c) palladium carbon, hydrogen gas, methanol, yield 85%; (d) trifluoroacetic anhydride, dichloromethane, yield 85%; (e) 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, 1-hydroxybenzotriazole, triethylamine, dichloromethane, yield 55%; (f) potassium carbonate, methanol, yield 70%; (g) O-benzotriazol-N,N,N,N-tetramethylurea tetrafluoroborate, triethylamine, dichloromethane, yield 55%; (h) trifluoroacetic acid, dichloromethane, triethylamine, yield 85%.

15. The pharmaceutical composition of claim 12, wherein the injectable or oral dosage forms are selected from pills, powders, lozenges, capsules, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solid or in liquid media), ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0118] FIG. 1 shows concentration-response curves of some compounds on acute myelogenous leukemia cells MV4-11 and HL60.

[0119] FIG. 2 shows concentration-response curves of some compounds on acute myelogenous leukemia cells PL21, KASUMI-1, MONO-MAC-1 and NB-4.

[0120] FIG. 3 shows concentration-response curves of CZ411 and FK866, and their mixed liquors on acute myelogenous leukemia cells MV4-11 and HL60.

[0121] FIG. 4 shows that NMN has a reversal effect on cell death caused by LEE18 and LEE12. NMN is nicotinamide mononucleotide.

[0122] FIG. 5 shows HCT116 tumor growth curves and tumor images. 5-FU: 5-fluorouracil, a traditional anti-tumor chemotherapy drug; Oxaliplatin, a third-generation platinum anticancer drug, is an anti-tumor chemotherapy drug. 5-FU+Oxaliplatin is positive control.

[0123] FIG. 6 shows a tumor growth curve. Panobinostat is a broad-spectrum HDAC inhibitor marketed as a positive control drug.

DETAILED DESCRIPTION

[0124] The present disclosure will be described in more detail through specific embodiments. The following embodiments are provided for illustrative purposes and are not intended to limit the present disclosure in any way. Those skilled in the art will easily recognize a variety of non-critical parameters that can be changed or modified to achieve essentially the same results.

General Materials and Methods

[0125] All reactions that are insensitive to air and moisture are carried out in an ambient atmosphere and subjected to magnetic stirring. Tetrahydrofuran is distilled from deep purple sodium benzophenone ketyl. Dry DMF, dry DMSO, dry acetonitrile, dry dichloromethane, dry toluene, and dry dioxane are purchased from Energy Chemical. All operations sensitive to air and moisture are carried out using oven-dried glassware in a nitrogen atmosphere.

[0126] Thin layer chromatography (TLC) is carried out with an EMD TLC plate pre-coated with a 250 ?m thick silica gel 60F254 plate, and visualized through fluorescence quenching under UV light and KMnO.sub.4 staining.

[0127] All deuterated solvents are purchased from Beijing J&K Scientific. NMR spectra are recorded on the following instrument: JEOL 400 spectrometer running at 400 MHz for .sup.1H and .sup.13C acquisition. Using solvent resonance as the internal standard (.sup.1H: CDCl.sub.3, ? 7.26; DMSO-d.sub.6, ? 2.50), (.sup.13C: CDCl.sub.3, ? 77.16; DMSO-d.sub.6, ? 39.52), the chemical shifts are reported in ppm. The data are reported as follows: s is singlet, d is doublet, t is triplet, q is quartet, and m is multiplet; coupling constant in Hz; Integral; Unless otherwise noted, carbon signals are unimodal.

[0128] The compounds of the present disclosure and their preparation methods are further elucidated and illustrated by the examples and preparation examples provided below. It should be understood that the scope of the following examples and preparation examples does not limit the scope of the present disclosure in any way.

[0129] The following synthesis routes describe the preparation of compounds of formulas II, III, IV, or V in the present disclosure, and all raw materials are prepared by the methods described in these routes, and methods well-known by those of ordinary skill in the field of organic chemistry, or commercially available. All the final compounds of the present disclosure are prepared by the methods described in these routes or by methods similar therewith, which are well-known to those of ordinary skill in the field of organic chemistry. All variable factors applied in these routes are as defined below or as defined in the claims.

[0130] The preparation of intermediate compounds of formulas II, III, IV or V in the present disclosure is as shown in routes 1 and 2, and each substituent is as defined in the summary of the present disclosure.

Synthesis Route 1:

[0131] ##STR00012## ##STR00013## ##STR00014##

[0132] Reagents and conditions: (a) different amine, TBTU, TEA, DCM, yield 55%; (b) CH.sub.3OH/H.sub.2O, KOH, reflux, yield 80%; (c) 4-nitrophenyl chloroformate, TEA, DCM, yield 80%; (d) methyl 8-aminocaprylate hydrochlorid, TEA, DCM, yield 65%; (e) sodium azide, DMF, 80? C., yield 80%.

Synthetic Route 2:

[0133] ##STR00015## ##STR00016##

[0134] Reagents and conditions: (a) propionaldehyde, MeOH, yield 98%; NaBH.sub.3CN, MeOH, concentrated hydrochloric acid, methyl orange, yield 60%; (b) (Boc).sub.2O, triethylamine, EtOH, yield 85%; (c) Pd/C, H.sub.2, MeOH, yield 85%; (d) trifluoroacetic anhydride, DCM, yield 85%; (e) EDCI, HOBt, TEA, DCM, yield 55%; (f) Na.sub.2CO.sub.3, MeOH, yield 70%.

[0135] The preparation of compounds of formulas II, III, IV or V in the present disclosure is as shown in routes 3, 4, 5 and 6, and each substituent is as defined in the summary of the present disclosure.

Route 3:

[0136] ##STR00017##

[0137] Reagents and conditions: (a) TBTU, TEA, DCM, yield 55%; (b) TFA, DCM, triethylamine, yield 85%.

Route 4:

[0138] ##STR00018##

[0139] Reagents and conditions: (a) TBTU, TEA, DCM, yield 55%.

Route 5:

[0140] ##STR00019##

[0141] Reagents and conditions: (a) 3-ethynylpyridine, sodium ascorbate, CuSO.sub.4, THF. H.sub.2O, yield 85%.

Route 6:

[0142] ##STR00020## ##STR00021## ##STR00022## ##STR00023##

[0143] 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, sodium ascorbate, CuSO.sub.4, THF, H.sub.2O, yield 85%.

SPECIFIC EMBODIMENTS

[0144] Next, the present disclosure will be explained in detail through specific examples, but the use and purpose of these exemplary embodiments are only used for exemplifying the present disclosure and do not constitute any form of limitation on the actual scope of protection of the present disclosure, rather than limiting the scope of protection of the present disclosure thereto.

Preparation of Compounds in Route 1

Example 1

[0145] ##STR00024##

[0146] Preparation of methyl (E)-3-(3-(pyridin-3-yl)acrylamido) propionate (1a): 3-(pyridin-3-yl)acrylic acid (0.44 g, 3 mmol) was dissolved in 20 mL of dichloromethane, and 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylurea tetrafluoroborate (TBTU, 1.05 g, 3.6 mmol) and TEA (0.6 mL, 4.5 mmol) were added in an ice bath. After 30 minutes, methyl 3-aminopropionate hydrochloride (0.46 g, 3.3 mmol) was added, then 0.6 mL of TEA was added and the above materials reacted overnight. The reaction product was washed with saturated NaHCO.sub.3 (2?30 mL) and saturated brine (2?30 mL) and dried over MgSO.sub.4. After the solvent was completely evaporated, the product obtained after evaporation was purified by flash chromatography to obtain compound 1a as a white solid powder (0.35 g, 51%). .sup.1H NMR (600 MHz, DMSO-d.sub.6) ? 8.75 (d, J=2.3 Hz, 1H), 8.55 (dd, J=4.8, 1.6 Hz, 1H), 8.28 (t, J=5.7 Hz, 1H), 7.98 (dt, J=8.0, 2.0 Hz, 1H), 7.49-7.42 (m, 2H), 6.73 (d, J=15.9 Hz, 1H), 3.62 (s, 3H), 3.42 (td, J=6.8, 5.6 Hz, 2H), 2.55 (t, J=6.8 Hz, 2H). ESI-MS m/z: 234.87 [M+H].sup.+.

##STR00025##

[0147] Preparation of methyl (E)-5-(3-(pyridin-3-yl)acrylamido) pentanoate (1b): the synthesis method of 1a was used, and 3-(pyridin-3-yl)acrylic acid and methyl 5-aminopentanoate hydrochloride were used as raw material to obtain compound 1b as a white solid, with a yield of 55%. .sup.1H NMR (600 MHz, DMSO-d.sub.6) ? 8.75 (d, J=2.3 Hz, 1H), 8.55 (dd, J=4.8, 1.6 Hz, 1H), 8.18 (t, J=5.8 Hz, 1H), 7.98 (dt, J=7.9, 2.0 Hz, 1H), 7.48-7.42 (m, 2H), 6.72 (d, J=15.9 Hz, 1H), 3.59 (s, 3H), 3.22-3.16 (m, 2H), 2.34 (t, J=7.4 Hz, 2H), 1.60-1.52 (m, 2H), 1.52-1.43 (m, 2H). ESI-MS m/z: 248.97 [M+H].sup.+.

##STR00026##

[0148] Preparation of methyl (E)-7-(3-(pyridin-3-yl)acrylamido) heptanoate (1c): the synthesis method of 1a was used, and 3-(pyridin-3-yl)acrylic acid and methyl 7-aminoheptanoate hydrochloride were used as raw material to obtain compound 1c as a white solid, with a yield of 53%. .sup.1H NMR (600 MHz, DMSO-d.sub.6) ? 8.75 (d, J=2.3 Hz, 1H), 8.55 (dd, J=4.7, 1.6 Hz, 1H), 8.14 (t, J=5.7 Hz, 1H), 7.97 (dt, J=7.9, 2.0 Hz, 1H), 7.50-7.41 (m, 2H), 6.72 (d, J=15.9 Hz, 1H), 3.58 (s, 3H), 3.17 (td, J=7.0, 5.7 Hz, 2H), 2.34-2.26 (m, 2H), 1.53 (qd, J=7.4, 3.2 Hz, 2H), 1.45 (p, J=7.4 Hz, 2H), 1.30 (h, J=4.5, 3.5 Hz, 4H). ESI-MS m/z: 290.86 [M+H].sup.+.

##STR00027##

[0149] Preparation of methyl (E)-7-(3-(pyridin-3-yl)acrylamido) octanoate (1d): the synthesis method of 1a was used, and 3-(pyridin-3-yl)acrylic acid and methyl 8-aminooctanoate hydrochloride were used as raw material to obtain compound 1d as a white solid, with a yield of 53%. .sup.1H NMR (600 MHz, DMSO-d.sub.6) ? 8.75 (d, J=2.3 Hz, 1H), 8.55 (dd, J=4.7, 1.6 Hz, 1H), 8.14 (t, J=5.7 Hz, 1H), 7.97 (dt, J=7.9, 2.0 Hz, 1H), 7.47-7.42 (m, 2H), 6.72 (d, J=15.9 Hz, 1H), 3.58 (s, 3H), 3.20-3.14 (m, 2H), 2.30 (t, J=7.4 Hz, 2H), 1.56-1.50 (m, 2H), 1.49-1.40 (m, 2H), 1.31-1.24 (m, 7H).

##STR00028##

[0150] Preparation of methyl 7-(3H pyrrolo[3,2-c]pyridin-2-carboxamide) heptanate (6): the synthesis method of 1a was used, and 1H-pyrrolo[3,2-c]pyridin-2-carboxylic acid and methyl 7-aminoheptanoate hydrochloride were used as raw material to obtain compound 6 as a white solid, with a yield of 55%. .sup.1H NMR (600 MHz, DMSO-d.sub.6) ? 8.75 (d, J=2.3 Hz, 1H), 8.55 (dd, J=4.7, 1.6 Hz, 1H), 8.14 (t, J=5.7 Hz, 1H), 7.97 (dt, J=7.9, 2.0 Hz, 1H), 7.47-7.42 (m, 2H), 6.72 (d, J=15.9 Hz, 1H), 3.58 (s, 3H), 3.17 (td, J=7.0, 5.7 Hz, 2H), 2.30 (t, J=7.4 Hz, 2H), 1.52 (t, J=7.2 Hz, 2H), 1.45 (q, J=7.1 Hz, 2H), 1.31-1.24 (m, 7H). ESI-MS m/z: 304.89 [M+H].sup.+.

##STR00029##

[0151] Preparation of (E)-3-(3-(pyridin-3-yl)acrylamido) propionic acid (2a): compound 1a (0.35 g, 1.5 mmol) was dissolved in 5 mL of methanol, and then 2 mL of 3M KOH aqueous solution 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 with 1 N HCl to pH 5-6, and then filtered. The precipitate was corresponding acidic compound 2a as a white solid (0.28 g, 85%). The raw materials were directly used for the next step without further purification. ESI-MS m/z: 219.15 [M?H].sup.?.

##STR00030##

[0152] Preparation of (E)-5-(3-(pyridin-3-yl)acrylamido) pentanoic acid (2b): the synthesis method of 2a was used to obtain compound 2b as a white solid, with a yield of 88%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 8.39 (d, J=13.3 Hz, 2H), 7.59 (d, J=7.9 Hz, 1H), 7.29 (dd, J=8.1, 4.3 Hz, 1H), 6.38 (t, J=6.0 Hz, 1H), 5.98 (t, J=5.8 Hz, 1H), 4.17 (d, J=5.9 Hz, 2H), 2.97-2.92 (m, 2H), 2.14 (t, J=7.3 Hz, 2H), 1.45-1.24 (m, 8H). ESI-MS m/z: 278. 18 [M?H].sup.?.

##STR00031##

[0153] Preparation of (E)-7-(3-(pyridin-3-yl)acrylamido) heptanoic acid (2c): the synthesis method of 2a was used and compound LL433 was used as a raw material to obtain compound 2c as a white solid, with a yield of 55%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 8.97 (d, J=2.1 Hz, 1H), 8.73 (dd, J=5.4, 1.4 Hz, 1H), 8.46 (dt, J=8.2, 1.8 Hz, 1H), 8.29 (t, J=5.7 Hz, 1H), 7.85 (dd, J=8.1, 5.4 Hz, 1H), 7.50 (d, J=15.9 Hz, 1H), 6.85 (d, J=15.9 Hz, 1H), 3.14 (q, J=6.6 Hz, 2H), 2.26 (t, J=7.4 Hz, 2H), 1.45 (dt, J=28.5, 7.1 Hz, 4H), 1.25 (p, J=3.6 Hz, 4H). ESI-MS m/z: 275. 24 [M?H].sup.?.

##STR00032##

[0154] Preparation of (E)-8-(3-(pyridin-3-yl)acrylamido) octanoic acid (2d): the synthesis method of 2a was used and compound 1d was used as a raw material to obtain compound 2d as a white solid, with a yield of 87%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 8.97 (d, J=2.1 Hz, 1H), 8.73 (dd, J=5.4, 1.4 Hz, 1H), 8.46 (dt, J=8.2, 1.8 Hz, 1H), 8.29 (t, J=5.7 Hz, 1H), 7.85 (dd, J=8.1, 5.4 Hz, 1H), 7.50 (d, J=15.9 Hz, 1H), 6.85 (d, J=15.9 Hz, 1H), 3.14 (q, J=6.6 Hz, 2H), 2.26 (t, J=7.4 Hz, 2H), 1.45 (dt, J=28.5, 7.1 Hz, 4H), 1.25 (p, J=3.6 Hz, 4H). ESI-MS m/z: 275. 24 [M?H].sup.?.

##STR00033##

[0155] Preparation of 7-(3-(pyridin-3-ylmethyl) ureido) heptanoic acid (5): the synthesis method of 2a was used and compound 4 was used as a raw material to obtain compound 5 as a white solid, with a yield of 85%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 8.39 (d, J=13.3 Hz, 2H), 7.59 (d, J=7.9 Hz, 1H), 7.29 (dd, J=8.1, 4.3 Hz, 1H), 6.38 (t, J=6.0 Hz, 1H), 5.98 (t, J=5.8 Hz, 1H), 4.17 (d, J=5.9 Hz, 2H), 2.97-2.92 (m, 2H), 2.14 (t, J=7.3 Hz, 2H), 1.45-1.24 (m, 8H). ESI-MS m/z: 278. 18 [M?H].sup.?.

##STR00034##

[0156] 7-(1H-pyrrolo [3,2-c] pyridin-2-carboxamide) heptanoic acid (7). The synthesis method of 2a was used and compound 6 was used as a raw material to obtain compound 7 as a white solid, with a yield of 85%. ESI-MS m/z: 288. 21 [M?H].sup.?.

##STR00035##

[0157] Preparation of 4-nitrophenyl (pyridin-3-ylmethyl) carbamate (3): 3-pyridinemethanimine (0.22 g, 2 mmol) and triethylamine (0.43 g, 2 mmol) were dissolved in DCM, then 4-nitrophenyl chloroformate (0.61 g, 3 mmol) was added, and the above materials were stirred at room temperature for 5 hours. After the reaction was completed, the reaction product was washed with brine three times and dried over Na.sub.2SO.sub.4. The solvent was completely evaporated to obtain compound 3 as a white solid powder (0.07 g, 70% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 8.61 (t, J=6.1 Hz, 1H), 8.52 (s, 1H), 8.46 (d, J=4.8 Hz, 1H), 8.26-8.19 (m, 2H), 7.72 (d, J=8.0 Hz, 1H), 7.43-7.38 (m, 2H), 7.37-7.34 (m, 1H), 4.31 (d, J=6.0 Hz, 2H). ESI-MS m/z: 273.84 [M+H].sup.+.

##STR00036##

[0158] 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 were added triethylamine (0.12 g, 1.2 mmol) and compound 3 (0.33 g, 1.2 mmol) at 0? C. The above materials reacted at room temperature for 2 hours. After the reaction was completed, the reaction product was washed with brine three times and dried over Na.sub.2SO.sub.4. After the solvent was completely evaporated, the product obtained after evaporation was purified by flash chromatography to obtain compound 4 as a white solid powder (0.2 g, 70% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 8.42 (s, 1H), 8.39 (d, J=4.8 Hz, 1H), 7.60 (d, J=7.8 Hz, 1H), 7.29 (dd, J=7.8, 4.8 Hz, 1H), 6.31 (t, J=6.0 Hz, 1H), 5.92 (t, J=5.7 Hz, 1H), 4.17 (d, J=6.0 Hz, 2H), 3.54 (s, 3H), 2.94 (q, J=6.5 Hz, 2H), 2.24 (t, J=7.4 Hz, 2H), 1.47 (p, J=7.2 Hz, 2H), 1.31 (p, J=6.9 Hz, 2H), 1.21 (h, J=3.8, 3.0 Hz, 4H). ESI-MS m/z: 293.92 [M+H].sup.+.

##STR00037##

[0159] Preparation of 8-azido-octanoic acid (8): 8-bromooctanoic acid (0.33 g, 1.5 mmol) was dissolved in DMF, then NaN3 (0.15 g, 2.25 mmol) was added, and the above materials reacted overnight at 80? C. 25 mL of DCM was added to the solution, the solvent was washed with water 5 times and dried over Na2SO4. After the solvent was completely evaporated, compound 8 as a colorless oil (0.27 g, 95% yield) was obtained. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 2.15 (t, J=7.4 Hz, 2H), 1.46 (dp, J=14.4, 7.1 Hz, 4H), 1.28-1.19 (m, 6H). ESI-MS m/z: 184. 15 [M?H].sup.?.

Preparation of Compounds in Route 2

Example 2

[0160] ##STR00038##

[0161] Preparation of benzyl 2-propylhydrazin-1-carboxylate (9): benzyl hydrazinecarboxylate (1.66 g, 10 mmol) was dissolved in 50 mL of methanol, then propionaldehyde (0.61 g, 10.5 mmol) was added, and the above materials reacted at room temperature for 2 hours. After the reaction was completed, methanol was removed. The obtained solid was dissolved in 30 mL of methanol, then NaBH.sub.3CN (1.2 g, 20 mmol) and 2 drops of concentrated HCl/MeOH (v: v=1:1) solution were added, and the above materials reacted overnight. After the reaction was completed, the solvent was completely evaporated, and then a crude product was purified by flash chromatography to obtain compound 9 as a white solid powder (1.2 g, 60%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 8.54 (s, 1H), 7.36-7.23 (m, 5H), 4.99 (s, 2H), 4.46 (s, 1H), 2.61 (t, J=7.1 Hz, 2H), 1.32 (h, J=7.3 Hz, 2H), 0.81 (t, J=7.4 Hz, 3H). ESI-MS m/z: 108.92 [M+H].sup.+.

##STR00039##

[0162] Preparation of 2-benzyl 1-(tert-butyl) 1-propylhydrazine-1,2-dicarboxylate (10): the compound 9 (1 g, 5 mmol) was dissolved in 50 mL of anhydrous dichloromethane, and then triethylamine (1.5 g, 15 mmol) and (Boc).sub.2O (0.22 g, 10 mmol) were added. After reacting at room temperature for 2 hours, the solution was washed with a 1 M citric acid aqueous solution (3?100 mL and brine (3?100 mL) and then dried over MgSO.sub.4, and the solvent was completely evaporated to obtain compound 10 as a white solid (1.3 g, 85%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.45 (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=7.3 Hz, 3H). ESI-MS m/z: 308.86 [M+H].sup.+.

##STR00040##

[0163] Preparation of tert-butyl 1-propylhydrazin-1-carboxylate (11): compound 10 (0.6 g, 2 mmol) was dissolved in methanol and Pd/C (0.06 g) was added. Then the above materials reacted in hydrogen at room temperature for 4 hours. After the reaction was completed, Pd/C was removed by filtration, and the filtrate was completely evaporated to obtain compound 11 as a colorless oil (0.25 g, 72%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 4.37 (s, 1H), 3.16 (t, J=7.0 Hz, 2H), 1.45 (h, J=14.3, 7.2 Hz, 2H), 1.36 (s, 9H), 0.76 (t, J=7.4 Hz, 3H). ESI-MS m/z: 174.87 [M+H].sup.+.

##STR00041##

[0164] Preparation of 4-((2,2,2-trifluoroacetamido)methyl) benzoic acid (13a): 10 mL of trifluoroacetic anhydride was slowly added to 4-(aminomethyl) benzoic acid (3.0 g, 20 mmol) in an ice bath, and then reacted for 2 hours at room temperature after dropwise addition. After the reaction was completed, 100 mL of ice water was added to quench the reaction, then the reaction was filtered, and a filter cake was dried to obtain compound 13a (4.43 g, 95%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.05 (t, J=6.0 Hz, 1H), 7.89 (d, J=6.4 Hz, 2H), 7.35 (d, J=8.0 Hz, 2H), 4.43 (d, J=6.0 Hz, 2H). ESI-MS m/z: 246. 16 [M?H].sup.?.

##STR00042##

[0165] Preparation of 4-(2,2,2-trifluoroacetamido) methyl) benzoic acid (13b): the synthesis method of 13a was used, and 4-aminobenzoic acid and trifluoroacetic anhydride were used as raw materials to obtain compound 13b as a white solid, with a yield of 98%. .sup.1H NMR (400 MHz DMSO-d.sub.6) ? 12.94 (s, 1H), 10.07 (s, 1H), 7.94 (d, J=8.2 Hz, 2H), 7.40 (d, J=8.2 Hz, 2H), 4.47 (d, J=6.0 Hz, 2H). ESI-MS m/z: 232.12 [M?H].sup.?.

##STR00043##

[0166] Preparation of tert-butyl 1-propyl-2-(4-((2,2,2-trifluoroacetamido)methyl)benzoyl)hydrazine-1-carboxylate (14a): compound 13a (0.7 g, 3 mmol) was dissolved in 20 mL of dichloromethane, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI.Math.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 in an ice bath. After 30 minutes, compound 11 (0.57 g, 3.3 mmol) was added and the above materials reacted overnight. The reaction solution was washed with saturated NaHCO.sub.3 (2?30 mL) and brine (2?30 mL), and organic phases were combined and dried over anhydrous magnesium sulfate. After the solvent was completely evaporated, the obtained reaction product was purified by flash chromatography to obtain compound 14a as a white solid powder (0.66 g, 55% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.47 (s, 1H), 10.05 (t, J=6.1 Hz, 1H), 7.78 (dd, J=16.9, 7.8 Hz, 2H), 7.39-7.29 (m, 2H), 4.41 (d, J=5.9 Hz, 2H), 3.34 (s, 2H), 1.47-1.27 (m, 11H), 0.84 (t, J=7.1 Hz, 3H). ESI-MS m/z: 403.89 [M+H].sup.+.

##STR00044##

[0167] Preparation of tert-butyl 1-propyl-2-(4-(2,2,2-trifluoroacetamido) benzoyl) hydrazine-1-carboxylate (14b): the synthesis method of 14a was used and compounds 13b and 11 were used as raw materials to obtain compound 14b as a white solid, with a yield of 50%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 11.46 (s, 1H), 10.49 (s, 1H), 7.85 (dd, J=14.7, 8.6 Hz, 2H), 7.75 (d, J=8.4 Hz, 2H), 1.55-1.43 (m, 2H), 1.40-1.29 (m, 9H), 0.84 (t, J=6.9 Hz, 3H). ESI-MS m/z: 389.91 [M+H].sup.+.

##STR00045##

[0168] Preparation of tert-butyl 2-(4-(aminomethyl)benzoyl)-1-propylhydrazin-1-carboxylate (15a): compound 14a (1.2 g, 3 mmol) was dissolved in 20 mL of methanol/water (v: v=1:1) solution, then K.sub.2CO.sub.3 (1.24 g, 9 mmol) was added and the above materials reacted overnight at room temperature. After methanol was completely evaporated, the reaction solution was extracted with ethyl acetate (2?20 mL), organic phases were combined, washed with brine (2?30 mL) and dried over anhydrous Na.sub.2SO.sub.4. After filtration, the solvent was completely evaporated to obtain compound 15a (0.74 g, 80%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.42 (s, 1H), 7.74 (dd, J=16.8, 7.9 Hz, 2H), 7.39 (d, J=8.0 Hz, 2H), 3.72 (s, 2H), 1.47 (p, J=7.3 Hz, 3H), 1.39-1.28 (m, 9H), 0.83 (t, J=7.0 Hz, 4H). ESI-MS m/z: 307.94 [M+H].sup.+.

##STR00046##

[0169] Preparation of tert-butyl 2-(4-aminobenzoyl)-1-propylhydrazin-1-carboxylate (15b): the synthesis method of 15a was used and compound 14b was used as a raw material to obtain compound 15b as a white solid, with a yield of 82%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.00 (s, 1H), 10.00 (s, OH), 7.58-7.49 (m, 2H), 6.54-6.46 (m, 2H), 5.68 (d, J=4.7 Hz, 2H), 3.32-3.25 (m, 2H), 1.46 (h, J=7.5 Hz, 2H), 1.38-1.27 (m, 9H), 0.82 (t, J=7.1 Hz, 3H). ESI-MS m/z: 293.87 [M+H].sup.+.

Preparation of Compounds in Route 3

Example 3

[0170] ##STR00047##

[0171] Preparation of (E)-N-(3-oxo-3-(2-propylhydrazino) 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-tetramethylurea tetrafluoroborate (TBTU, 1.05 g, 3.6 mmol) and triethylamine (0.6 mL, 4.5 mmol) were added in an ice bath. After reacting for 30 minutes, compound 11 (0.57 g, 3.3 mmol) was added and the above materials reacted overnight at room temperature. The reaction solution was washed with saturated NaHCO.sub.3 (2?30 mL) and saturated brine (2?30 mL) and dried over MgSO.sub.4. After filtration, the solvent was completely evaporated, and then the obtained product was purified by flash chromatography to obtain a white solid powder (0.35 g, 54%).

[0172] The product (0.30 g, 0.8 mmol) of the previous step was dissolved in 15 mL of dichloromethane, then trifluoroacetic acid (TFA, 0.11 g, 1.0 mmol) was added and the above materials reacted overnight. After the reaction was completed, TEA (0.10 g, 1.0 mmol) was added to adjust the pH to 8. The solution was washed with brine and dried over Na.sub.2SO.sub.4. After filtration, the solvent was completely evaporated, and then the obtained product was then purified by flash chromatography to obtain compound LEE1 as a white solid powder (0.18 g, 85%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.32 (s, 1H), 8.75 (d, J=2.2 Hz, 1H), 8.55 (dd, J=4.7, 1.6 Hz, 1H), 8.21 (t, J=5.8 Hz, 1H), 7.97 (dt, J=8.0, 2.0 Hz, 1H), 7.50-7.40 (m, 2H), 6.74 (d, J=15.9 Hz, 1H), 4.79 (s, 1H), 3.39 (q, J=6.6 Hz, 2H), 2.60 (dt, J=18.8, 6.8 Hz, 2H), 2.27 (t, J=7.0 Hz, 2H), 1.38 (h, J=7.3 Hz, 2H), 0.85 (t, J=7.4 Hz, 3H). .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 169.49, 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].sup.+.

Example 4

[0173] ##STR00048##

[0174] Preparation of (E)-N-(5-oxo-5-(2-propylhydrazino)pentyl)-3-(pyridin-3-yl) acrylamide (LEE2): the synthesis method of LEE1 was used and compound 11 and compound 2b were used as raw materials to obtain compound LEE2 as a white solid, with a yield of 55%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.23 (d, J=5.6 Hz, 1H), 8.75 (d, J=2.2 Hz, 1H), 8.55 (dd, J=4.8, 1.6 Hz, 1H), 8.16 (q, J=5.8, 4.4 Hz, 1H), 7.97 (dt, J=8.0, 2.0 Hz, 1H), 7.49-7.38 (m, 2H), 6.73 (d, J=15.9 Hz, 1H), 4.77 (d, J=5.9 Hz, 1H), 3.18 (q, J=6.5 Hz, 2H), 2.61 (h, J=4.4 Hz, 2H), 2.04 (t, J=7.2 Hz, 2H), 1.60-1.32 (m, 6H), 0.86 (t, J=7.4 Hz, 3H)..sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 171.22, 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].sup.+.

Example 5

[0175] ##STR00049##

[0176] Preparation of (E)-N-(7-oxo-7-(2-propylhydrazino)heptyl)-3-(pyridin-3-yl) acrylamide (LEE3): the synthesis method of LEE1 was used and compounds 11 and 2c were used as raw materials to obtain compound LEE3 as a white solid, with a yield of 57%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.21 (d, J=5.7 Hz, 1H), 8.75 (d, J=2.2 Hz, 1H), 8.55 (dd, J=4.8, 1.6 Hz, 1H), 8.14 (t, J=5.6 Hz, 1H), 7.97 (dt, J=8.0, 2.0 Hz, 1H), 7.49-7.39 (m, 2H), 6.73 (d, J=15.9 Hz, 1H), 4.76 (d, J=6.2 Hz, 1H), 3.17 (q, J=6.6 Hz, 2H), 2.61 (td, J=6.9, 4.5 Hz, 2H), 2.01 (t, J=7.4 Hz, 2H), 1.57-1.34 (m, 6H), 1.28 (tq, J=14.3, 8.6, 8.0 Hz, 4H), 0.86 (t, J=7.4 Hz, 3H). .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 171.36, 164.84, 150.51, 149.53, 135.52, 134.35, 131.25, 124.84, 124.42, 53.50, 39.15, 33.91, 29.46, 28.74, 26.65, 25.63, 21.22, 12.04. ESI-MS m/z: 332.04 [M+H].sup.+.

Example 6

[0177] ##STR00050##

[0178] Preparation of (E)-N-(8-oxo-8-(2-propylhydrazino)octyl)-3-(pyridin-3-yl) acrylamide (LEE4): the synthesis method of LEE1 was used and compounds 11 and 2d were used as raw materials to obtain compound LEE4 as a white solid, with a yield of 60%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.04 (s, 1H), 9.85 (s, 1H), 8.71 (d, J=2.3 Hz, 1H), 8.50 (dd, J=4.7, 1.6 Hz, 1H), 8.11 (t, J=5.6 Hz, 1H), 7.93 (dt, J=8.0, 2.0 Hz, 1H), 7.72 (d, J=8.7 Hz, 2H), 7.62 (s, 2H), 7.45-7.35 (m, 2H), 6.68 (d, J=15.9 Hz, 1H), 3.13 (q, J=6.6 Hz, 2H), 2.69 (t, J=7.1 Hz, 2H), 2.29 (t, J=7.4 Hz, 2H), 1.56 (p, J=6.8 Hz, 2H), 1.42 (h, J=7.3 Hz, 4H), 1.27 (s, 6H), 0.87 (t, J=7.4 Hz, 3H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) ? 171.55, 164.84, 150.41, 149.39, 135.50, 134.48, 131.24, 124.81, 124.47, 52.86, 39.14, 33.65, 29.50, 28.87, 28.83, 26.79, 25.38, 20.05, 11.73. ESI-MS m/z: 346.97 [M+H].sup.+.

Example 7

[0179] ##STR00051##

[0180] Preparation of 1-(7-oxo-7-(2-propylhydrazino)heptyl)-3-(pyridin-3-ylmethyl) urea (LEE5): the synthesis method of LEE1 was used and compounds 11 and 5 were used as raw materials to obtain compound LEES as a white solid, with a yield of 61%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.19 (s, 1H), 8.41 (d, J=1.9 Hz, 1H), 8.39 (dd, J=4.8, 1.7 Hz, 1H), 7.59 (dt, J=7.9, 1.9 Hz, 1H), 7.29 (dd, J=7.8, 4.8, 0.9 Hz, 1H), 6.32 (t, J=6.1 Hz, 1H), 5.93 (t, J=5.7 Hz, 1H), 4.17 (d, J=5.9 Hz, 2H), 2.93 (q, J=6.6 Hz, 2H), 2.56 (t, J=7.1 Hz, 2H), 1.95 (t, J=7.3 Hz, 2H), 1.43 (t, J=7.2 Hz, 2H), 1.37-1.29 (m, 3H), 1.19 (s, 4H), 0.81 (t, J=7.4 Hz, 3H); .sup.13C NMR (126 MHz, DMSO-d.sub.6) ? 171.35, 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].sup.+.

Example 8

[0181] ##STR00052##

[0182] Preparation of N-(7-oxo-7-(2-propylhydrazino) heptyl)-3H-pyrrolo[3,2-c]pyridin-2-carboxamide (LEE7): the synthesis method of LEE1 was used and compounds 11 and 7 were used as raw materials to obtain compound LEE7 as a white solid, with a yield of 56%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.19 (d, J=7.9 Hz, 1H), 8.93 (s, 1H), 8.60 (t, J=5.7 Hz, 1H), 8.21 (d, J=5.8 Hz, 1H), 7.37 (d, J=5.8 Hz, 1H), 7.25 (s, 1H), 3.27 (s, 2H), 2.60 (t, J=7.1 Hz, 2H), 2.01 (t, J=7.3 Hz, 2H), 1.59-1.46 (m, 4H), 1.42-1.23 (m, 7H), 0.86 (t, J=7.4 Hz, 3H). .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 171.36, 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].sup.+.

Example 9

[0183] ##STR00053##

[0184] Preparation of 8-azido-N-propyloctanehydrazide (LEE6): the synthesis method of 1a was used, compounds 8 and 11 were used as raw materials to obtain compound LEE6 as colorless oil, with a yield of 56%. ESI-MS m/z: 241.98 [M+H].sup.+.

Preparation of Compounds in Route 4

Example 10

[0185] ##STR00054##

[0186] Preparation of (E)-N-(2-aminophenyl)-8-(3-(pyridin-3-yl)acrylamido) octanamide (LEE8): the synthesis method of 1a was used and compound 2d and 1,2-diaminobenzene were used as raw materials to obtain compound LEE8 as a white solid, with a yield of 53%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.05 (s, 1H), 8.71 (s, 1H), 8.51 (d, J=4.8 Hz, 1H), 8.12 (t, J=5.7 Hz, 1H), 7.93 (d, J=7.9 Hz, 1H), 7.47-7.34 (m, 2H), 7.11 (d, J=7.8 Hz, 1H), 6.85 (t, J=7.6 Hz, 1H), 6.74-6.63 (m, 2H), 6.50 (t, J=7.5 Hz, 1H), 4.82 (s, 1H), 3.14 (q, J=6.5 Hz, 2H), 2.27 (t, J=7.4 Hz, 2H), 1.56 (p, J=7.0 Hz, 2H), 1.43 (p, J=7.8 Hz, 2H), 1.28 (s, 7H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) ? 171.62, 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].sup.+.

Example 11

[0187] ##STR00055##

[0188] Preparation of N-(2-aminophenyl)-8-azidooctanamide (20): the synthesis method of 1a was used and compound 8 and 1,2-diaminobenzene were used as raw materials to obtain compound 20 as a white solid, with a yield of 60%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.05 (s, 1H), 7.10 (d, J=7.9 Hz, 1H), 6.85 (t, J=7.6 Hz, 1H), 6.67 (d, J=8.0 Hz, 1H), 6.50 (d, J=7.5 Hz, 1H), 4.75 (s, 2H), 2.27 (t, J=7.4 Hz, 2H), 1.52 (dt, J=20.1, 7.0 Hz, 4H), 1.28 (s, 8H). ESI-MS m/z: 375.06 [M+H].sup.+.

Preparation of Compounds in Route 5

Example 12

[0189] ##STR00056##

[0190] Preparation of n-propyl-8-(4-(pyridin-3-yl)-1H-1,2,3-triazole-1-yl) octanehydrazide (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), then sodium ascorbate (50 mg, 0.25 mmol) and copper sulfate (4 mg, 0.025 mmol) were added, and the above materials reacted overnight at room temperature. After the reaction was completed, the reaction solution was washed with water three times and dried over Na2SO4. After filtration, the solvent was completely evaporated to obtain compound LEE10 as a white solid powder (0.07 g, 84%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.19 (s, 1H), 9.01 (d, J=2.2 Hz, 1H), 8.68 (s, 1H), 8.50 (dd, J=4.8, 1.7 Hz, 1H), 8.17 (dt, J=7.9, 2.0 Hz, 1H), 7.44 (dd, J=8.0, 4.8 Hz, 1H), 4.37 (t, J=7.1 Hz, 2H), 2.55 (t, J=7.1 Hz, 1H), 1.95 (t, J=7.3 Hz, 1H), 1.82 (p, J=7.1 Hz, 2H), 1.43 (p, J=7.3 Hz, 2H), 1.35-1.16 (m, 8H), 0.80 (t, J=7.4 Hz, 2H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) ? 171.33, 149.26, 146.79, 143.92, 132.79, 127.26, 124.46, 122.39, 53.44, 50.06, 33.84, 30.00, 28.76, 28.50, 26.17, 25.52, 21.16, 12.01. ESI-MS m/z: 345.06 [M+H].sup.+.

Example 13

[0191] ##STR00057##

[0192] Preparation of N-(2-aminophenyl)-8-(4-(pyridin-3-yl)-1H-1,2,3-triazole-1-yl) octanamide (LEE11): the synthesis method of LEE10 was used, compound 20 and 3-ethynylpyridine were used as raw materials to obtain compound LEE11 as a white solid, with a yield of 76%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.08 (s, 1H), 9.01 (d, J=2.2 Hz, 1H), 8.68 (s, 1H), 8.50 (dd, J=4.9, 1.7 Hz, 1H), 8.17 (dt, J=7.9, 2.1 Hz, 1H), 7.44 (dd, J=7.9, 4.8 Hz, 1H), 7.10 (dd, J=7.9, 1.5 Hz, 1H), 6.84 (td, J=7.6, 1.6 Hz, 1H), 6.67 (dd, J=8.0, 1.5 Hz, 1H), 6.49 (td, J=7.6, 1.5 Hz, 1H), 4.90 (s, 1H), 4.39 (t, J=7.1 Hz, 2H), 2.27 (t, J=7.4 Hz, 2H), 1.84 (p, J=7.2 Hz, 2H), 1.54 (p, J=7.3 Hz, 2H), 1.40-1.20 (m, 6H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) ? 171.60, 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].sup.+.

Preparation of Compounds in Route 6

Example 14

[0193] ##STR00058##

[0194] Preparation of tert-butyl 2-(4-((3H-pyrrolo[3,2-c]pyridin-2-carboxamide)methyl) benzoyl)-1-propylhydrazine-1-carboxylate (23c): 1H-pyrrolo[3,2-c]pyridin-2-carboxylic acid (0.48 g, 3 mmol) was dissolved in 15 mL of dichloromethane, and O-benzotriazole-N,N,N,N-tetramethylurea tetrafluoroborate (TBTU, 1.05 g, 3.6 mmol) and triethylamine (0.6 mL, 4.5 mmol) were added to the reaction solution in an ice bath. After 30 minutes, compound 15a (1.0 g, 3.3 mmol) was added and then the above materials reacted overnight. The reaction solution was washed with saturated NaHCO.sub.3 (2?30 mL) and brine (2?30 mL) and dried over MgSO.sub.4. After filtration, the solvent was completely evaporated, and then the obtained product was purified by flash chromatography to obtain a white solid powder (0.7 g, 54%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.46 (d, J=5.5 Hz, 1H), 9.29 (t, J=6.1 Hz, 1H), 8.93 (s, 1H), 8.20 (d, J=5.8 Hz, 1H), 7.78 (dd, J=16.8, 7.9 Hz, 2H), 7.41 (d, J=8.1 Hz, 2H), 7.36 (d, J=5.9 Hz, 1H), 7.32 (d, J=4.0 Hz, 1H), 4.55 (d, J=5.9 Hz, 2H), 3.05 (q, J=7.3 Hz, 1H), 1.53-1.42 (m, 2H), 1.39-1.28 (m, 9H), 0.83 (t, J=7.1 Hz, 3H). ESI-MS m/z: 451.88 [M+H].sup.+.

[0195] Preparation of tert-butyl 2-(4-(((1H-indole-2-carboxyamido) methyl) benzoyl)-1-propylhydrazin-1-carboxylate (23g): the synthesis method of 23c was used, and 2-indolecarboxylic acid and compound 15a were used as raw materials to obtain compound 23g as a white solid, with a yield of 56%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 11.60 (s, 1H), 10.46 (d, J=7.5 Hz, 1H), 9.09 (t, J=6.0 Hz, 1H), 7.78 (dd, J=16.6, 7.8 Hz, 2H), 7.58 (d, J=8.0 Hz, 1H), 7.40 (dd, J=8.2, 3.6 Hz, 3H), 7.15 (d, J=2.6 Hz, 2H), 7.00 (t, J=7.5 Hz, 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, 6.7 Hz, 3H). ESI-MS m/z: 451.05 [M+H].sup.+.

##STR00059##

[0196] Preparation of tert-butyl 2-(4-((benzofuran-2-carboxyamido)methyl)benzoyl)-1-propylhydrazin-1-carboxylate (23a): the synthesis method of 23c was used, and 2-benzofurancarboxylic acid and compound 15a were used as raw materials to obtain compound 23a as a white solid, with a yield of 56%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.47 (d, J=5.4 Hz, 1H), 9.36 (t, J=6.2 Hz, 1H), 7.82-7.72 (m, 3H), 7.63 (d, J=8.3 Hz, 1H), 7.56 (s, 1H), 7.42 (t, J=7.2 Hz, 3H), 7.30 (t, J=7.5 Hz, 1H), 4.51 (d, J=5.9 Hz, 2H), 1.46 (q, J=7.1 Hz, 2H), 1.39-1.28 (m, 9H), 0.83 (t, J=7.0 Hz, 3H). ESI-MS m/z: 451.08 [M+H].sup.+.

##STR00060##

[0197] Preparation of tert-butyl 2-(4-((benzothiophene-2-carboxyamido) methyl) benzoyl)-1-propylhydrazin-1-carboxylate (23b): the synthesis method of 23c was used, and benzothiophene-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23b as a white solid, with a yield of 53%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.47 (d, J=5.9 Hz, 1H), 9.38 (t, J=6.0 Hz, 1H), 8.11 (s, 1H), 8.05-7.95 (m, 1H), 7.96-7.87 (m, 1H), 7.78 (dd, J=16.9, 7.9 Hz, 2H), 7.47-7.37 (m, 5H), 4.51 (d, J=5.8 Hz, 2H), 1.46 (q, J=7.1 Hz, 2H), 1.39-1.29 (m, 6H), 0.83 (t, J=7.3 Hz, 3H). ESI-MS m/z: 467.98 [M+H].sup.+.

##STR00061##

[0198] Preparation of tert-butyl 2-(4-((3H-pyrrolo[2,3-c]pyridin-2-carboxyamido)methyl)benzoyl)-1-propylhydrazin-1-carboxylate (23e): the synthesis method of 23c was used, and 1H-pyrrolo[2,3-c]pyridin-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23e as a white solid, with a yield of 49%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 12.11 (s, 1H), 10.46 (s, 1H), 9.32 (s, 1H), 8.76 (s, 1H), 8.10 (d, J=5.5 Hz, 1H), 7.85-7.72 (m, 2H), 7.59 (d, J=5.6 Hz, 1H), 7.41 (d, J=8.1 Hz, 2H), 7.19 (s, 1H), 4.55 (d, J=5.9 Hz, 2H), 1.46 (d, J=7.2 Hz, 2H), 1.28 (s, 9H), 0.82 (d, J=7.0 Hz, 3H). ESI-MS m/z: 452.07 [M+H].sup.+.

##STR00062##

[0199] Preparation of tert-butyl 2-(4-(((furan [3,2-c]pyridin-2-carboxyamido) methyl) benzoyl)-1-propylhydroxyzine-1-carboxylate (23d): the synthesis method of 23c was used, and benzo[b]thiophene-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23d as a white solid, with a yield of 53%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.46 (s, 1H), 9.49 (t, J=6.1 Hz, 1H), 9.05 (s, 1H), 8.55 (d, J=5.8 Hz, 1H), 7.83-7.70 (m, 3H), 7.68 (s, 1H), 7.40 (d, J=8.2 Hz, 2H), 4.51 (d, J=6.0 Hz, 2H), 1.52-1.42 (m, 2H), 1.39-1.28 (m, 9H), 0.83 (t, J=7.2 Hz, 3H). ESI-MS m/z: 453.08 [M+H].sup.+.

##STR00063##

[0200] Preparation of tert-butyl 2-(4-((5-bromo-1H-indolo-2-carboxyamido)methyl)benzoyl)-1-propylhydrazide-1-carboxylate (23j): the synthesis method of 23c was used, and 5-bromoindole-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23j as a white solid, with a yield of 58%.

##STR00064##

[0201] Preparation of tert-butyl 2-(4-(((5-fluoro-1H-indole-2-carboxyamido)methyl)benzoyl)-1-propylhydrazin-1-carboxylate (23l): the synthesis method of 23c was used, and 5-fluoroindole-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23l as a white solid, with a yield of 53%.

##STR00065##

[0202] Preparation of tert-butyl 2-(4-((6-(dimethylamino)-1H-indole-2-carboxyamido)methyl)benzoyl)-1-propylhydrazin-1-carboxylate (23k): the synthesis method of 23c was used, and 6-(dimethylamino)-1H-indole-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23k as a white solid, with a yield of 50%.

##STR00066##

[0203] Preparation of tert-butyl 2-(4-((5-chloro-1H-indole-2-carboxyamido)methyl)benzoyl)-1-propylhydrazin-1-carboxylate (23m): the synthesis method of 23c was used, and 5-chloroindole-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23m as a white solid, with a yield of 51%.

##STR00067##

[0204] Preparation of tert-butyl 2-(4-(((5-methoxybenzofuran-2-carboxyamido)methyl)benzoyl)-1-propylhydrazin-1-carboxylate (23n): the synthesis method of 23c was used, and 5-methoxybenzofuran-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23n as a white solid, with a yield of 47%.

##STR00068##

[0205] Preparation of tert-butyl 2-(4-(((5-fluorobenzofuran-2-carboxyamido)methyl)benzoyl)-1-propylhydrazin-1-carboxylate (23o): the synthesis method of 23c was used, and 5-fluorobenzofuran-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23o as a white solid, with a yield of 59%.

##STR00069##

[0206] Preparation of tert-butyl 2-(4-((5-bromobenzofuran-2-carboxyamido)methyl)benzoyl)-1-propylhydrazin-1-carboxylate (23p): the synthesis method of 23c was used, and 5-bromobenzofuran-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23p as a white solid, with a yield of 52%.

##STR00070##

[0207] Preparation of tert-butyl 2-(4-(((5-methoxy-1H-indole-2-carboxyamido) methyl) benzoyl)-1-propylhydrazin-1-carboxylate (23q): the synthesis method of 23c was used, and 5-methoxyindole-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23q as a white solid, with a yield of 49%.

##STR00071##

[0208] Preparation of tert-butyl 2-(4-((5-chlorobenzofuran-2-carboxyamido)methyl)benzoyl)-1-propylhydrazin-1-carboxylate (23r): the synthesis method of 23c was used, and 5-chlorobenzofuran-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23r as a white solid, with a yield of 46%.

##STR00072##

[0209] Preparation of tert-butyl 2-(4-(((5-(prop-2-yn-1-yloxy)benzofuran-2-carboxyamido) methyl)benzoyl)-1-propylhydrazin-1-carboxylate (23s): the synthesis method of 23c was used, and 5-(prop-2-yn-1-yloxy)benzofuran-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23s as a white solid, with a yield of 50%.

##STR00073##

[0210] Preparation of tert-butyl 2-(4-(((5-hydroxybenzofuran-2-carboxyamido)methyl)benzoyl)-1-propylhydrazin-1-carboxylate (23u): the synthesis method of 23c was used, and 5-hydroxybenzofuran-2-carboxylic acid and compound 15a were used as raw materials to obtain compound 23u as a white solid, with a yield of 58%.

##STR00074##

[0211] 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 in an ice bath. After 30 minutes, compound 15a (0.37 mg, 1.2 mmol) and triethylamine (0.17 mL, 1.2 mmol) were added and the above materials reacted overnight at room temperature. The reaction solution was washed with brine (2?30 mL) and dried over MgSO.sub.4. After filtration, the solvent was completely evaporated, and then the obtained product was purified by flash chromatography to obtain compound 23v as a white solid powder (0.3 g, 51%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.81 (s, 1H), 8.71 (d, J=2.2 Hz, 1H), 8.51 (dd, J=4.8, 1.6 Hz, 1H), 8.11 (t, J=5.7 Hz, 1H), 7.95 (dt, J=8.0, 1.9 Hz, 1H), 7.45-7.36 (m, 2H), 6.68 (d, J=15.9 Hz, 1H), 3.21 (s, 3H), 3.16-3.09 (m, 3H), 2.01 (t, J=7.3 Hz, 2H), 1.56-1.15 (m, 25H), 0.78 (t, J=7.4 Hz, 3H). ESI-MS m/z: 579.96 [M+H].sup.+.

##STR00075##

[0212] Preparation of (E)-1-propyl-2-(4-(7-(3-(pyridin-3-yl)acrylamido)heptanamido)benzoyl) tert-butylhydrazine (carboxylic acid) (23x): the synthesis method of 23v was used, and compounds 2c and 15b were used as raw materials to obtain compound 23x as a white solid, with a yield of 53%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.36 (d, J=3.6 Hz, 1H), 10.12 (s, 1H), 8.72 (d, J=2.2 Hz, 1H), 8.52 (dd, J=4.8, 1.6 Hz, 1H), 8.15 (t, J=5.7 Hz, 1H), 7.96 (dd, J=8.0, 2.0 Hz, 1H), 7.69-7.67 (m, 4H), 7.49 (d, J=1.3 Hz, 1H), 6.69 (d, J=15.9 Hz, 1H), 3.13 (t, J=6.5 Hz, 2H), 2.32-2.26 (m, 2H), 1.45 (d, J=7.2 Hz, 2H), 1.41-1.26 (m, 13H), 1.22 (t, J=6.2 Hz, 4H), 0.83 (t, J=6.1 Hz, 3H). ESI-MS m/z: 552.06 [M+H].sup.+.

##STR00076##

[0213] Preparation of (E)-1-propyl-2-(4-(8-(3-(pyridin-3-yl)acrylamido)octanamido)benzoyl)hydrazine tert-butyl-1-carboxylate (23w): the synthesis method of 23v was used, and compounds 2d and 15b were used as raw materials to obtain compound 23w as a white solid, with a yield of 51%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.34 (s, 1H), 10.09 (s, 1H), 8.71 (s, 1H), 8.50 (s, 1H), 8.11 (t, J=5.7 Hz, 1H), 7.93 (d, J=7.9 Hz, 1H), 7.74 (d, J=9.7 Hz, 2H), 7.65 (s, 2H), 7.42-7.37 (m, 3H), 6.68 (d, J=15.9 Hz, 1H), 3.13 (q, J=6.7 Hz, 3H), 2.30 (t, J=7.4 Hz, 2H), 1.56 (t, J=7.1 Hz, 2H), 1.52-1.34 (m, 8H), 1.33-1.16 (m, 13H), 0.84 (t, J=8.4 Hz, 3H). ESI-MS m/z: 566.04 [M+H].sup.+.

##STR00077##

[0214] Preparation of tert-butyl 2-(4-(8-azidooctaamino)benzoyl)-1-propylhydrazin-1-carboxylate (24): the synthesis methods of 23v and LEE37 were used and compound 8 and 13b were used as raw materials to obtain compound 24 as a white solid, with a yield of 50%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.34 (s, 1H), 10.08 (s, 1H), 7.74 (d, J=10.2 Hz, 2H), 7.64 (d, J=8.7 Hz, 2H), 2.29 (t, J=7.4 Hz, 2H), 1.55 (t, J=7.0 Hz, 2H), 1.48 (q, J=6.9 Hz, 4H), 1.40 (s, 4H), 1.28 (d, J=3.7 Hz, 13H), 0.84 (t, J=7.2 Hz, 3H). ESI-MS m/z: 461.12 [M+H].sup.+.

Example 15

[0215] ##STR00078##

[0216] Preparation of (E)-N-(4-(2-propylhydrazino-1-carbonyl) benzyl)-3-(pyridin-3-yl) acrylamide (LEE12): the synthesis method of LEE1 was used and compounds 13b and (E)-3-(pyridin-3-yl)acrylic acid were used as raw materials to obtain compound LEE12 as a white solid, with a yield of 52%. .sup.1H NMR (600 MHz, DMSO-d.sub.6) ? 9.98 (s, 1H), 8.88-8.68 (m, 2H), 8.56 (dd, J=4.8, 1.6 Hz, 1H), 8.00 (dt, J=8.0, 2.0 Hz, 1H), 7.85-7.76 (m, 2H), 7.53 (d, J=15.9 Hz, 1H), 7.50-7.42 (m, 1H), 7.37 (d, J=8.3 Hz, 2H), 6.82 (d, J=15.9 Hz, 1H), 5.11 (s, 1H), 4.46 (d, J=6.0 Hz, 2H), 2.75 (t, J=7.1 Hz, 2H), 1.47 (h, J=7.4 Hz, 2H), 0.91 (t, J=7.5 Hz, 3H). .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 165.57, 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].sup.+.

Example 16

[0217] ##STR00079##

[0218] Preparation of N-(4-(2-propylhydrazin-1-carbonyl)benzyl)-1H-pyrrolo[3,2-c]pyridin-2-carboxamide (LEE18): compound 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 above materials reacted overnight at room temperature. After the reaction was completed, triethylamine (0.1 g, 1.0 mmol) was added to adjust the pH to 8. Then the solution was washed with brine and dried over Na.sub.2SO.sub.4. After filtration, the solvent was completely evaporated, and then the obtained product was purified by flash chromatography to obtain compound LEE18 as a white solid powder (0.14 g, 85%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.42 (t, J=6.1 Hz, 1H), 9.24 (s, 1H), 8.30 (d, J=6.5 Hz, 1H), 7.73-7.68 (m, 2H), 7.67-7.63 (m, 1H), 7.52 (d, J=0.9 Hz, 1H), 7.35-7.28 (m, 2H), 4.49 (d, J=6.0 Hz, 2H), 2.64 (t, J=7.1 Hz, 2H), 1.35 (h, J=7.3 Hz, 2H), 0.79 (t, J=7.4 Hz, 3H). .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 165.56, 160.21, 142.79, 141.87, 140.84, 136.81, 135.09, 132.44, 127.68, 127.57, 124.42, 109.75, 104.54, 53.55, 42.64, 21.28, 12.12. ESI-MS m/z: 351.86 [M+H].sup.+.

Example 17

[0219] ##STR00080##

[0220] Preparation of N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide (LEE16): the synthesis method of LEE18 was used and compounds 23a and trifluoroacetic acid were used as raw materials to obtain compound LEE16 as a white solid, with a yield of 79%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.90 (d, J=6.0 Hz, 1H), 9.27 (t, J=6.2 Hz, 1H), 7.75-7.68 (m, 3H), 7.59 (dd, J=8.4, 1.1 Hz, 1H), 7.51 (d, J=1.1 Hz, 1H), 7.40 (ddd, J=8.4, 7.2, 1.4 Hz, 1H), 7.33 (d, J=8.1 Hz, 2H), 7.30-7.24 (m, 1H), 5.00 (q, J=6.0 Hz, 1H), 4.47 (t, J=6.3 Hz, 2H), 2.67 (q, J=6.8 Hz, 2H), 1.39 (h, J=7.3 Hz, 2H), 0.83 (t, J=7.4 Hz, 3H). .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 165.60, 158.72, 154.75, 149.49, 143.00, 132.36, 127.64, 127.60, 127.35, 124.20, 123.26, 112.27, 110.12, 53.57, 42.42, 21.32, 12.12. ESI-MS m/z: 352.07 [M+H].sup.+.

Example 18

[0221] ##STR00081##

[0222] Preparation of N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzo[b]thiophene-2-carboxyamide (LEE17): the synthesis method of LEE18 was used and compounds 23b and trifluoroacetic acid were used as raw materials to obtain compound LEE17 as a white solid, with a yield of 83%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.95-9.87 (m, 1H), 9.30 (t, J=6.0 Hz, 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=5.9 Hz, 2H), 2.73-2.60 (m, 2H), 1.39 (h, J=7.3 Hz, 2H), 0.83 (t, J=7.4 Hz, 3H). .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 165.59, 162.10, 143.03, 140.72, 140.17, 139.64, 132.41, 127.64, 127.60, 126.72, 125.69, 125.46, 125.41, 123.29, 53.58, 42.95, 21.32, 12.13. ESI-MS m/z: 367.96 [M+H].sup.+.

Example 19

[0223] ##STR00082##

[0224] Preparation of N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide (LEE15): the synthesis method of LEE12 was used and compounds 15a and 1H-indole-2-carboxylic acid were used as raw materials to carry out condensation and deprotection to obtain compound LEE15 as a white solid, with a yield of 56%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 11.54 (s, 1H), 9.89 (d, J=5.6 Hz, 1H), 9.01 (t, J=6.1 Hz, 1H), 7.75-7.70 (m, 2H), 7.55 (dt, J=7.9, 1.0 Hz, 1H), 7.40-7.30 (m, 3H), 7.15-7.08 (m, 2H), 6.97 (ddd, J=8.0, 6.9, 1.0 Hz, 1H), 5.05-4.94 (m, 1H), 4.48 (d, J=6.0 Hz, 2H), 2.67 (td, J=7.1, 5.7 Hz, 2H), 1.39 (h, J=7.3 Hz, 2H), 0.83 (t, J=7.4 Hz, 3H). .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 165.60, 161.70, 143.45, 136.99, 132.29, 131.98, 127.60, 127.57, 127.44, 123.83, 121.99, 120.22, 112.79, 103.16, 53.58, 42.41, 21.32, 12.13. ESI-MS m/z: 350.86 [M+H].sup.+.

Example 21

[0225] ##STR00083##

[0226] Preparation of N-(4-(2-propylhydrazino-1-carbonyl) benzyl)-1H-pyrrolo[2,3-c]pyridine-2-carboxamide (LEE21): the synthesis method of LEE18 was used and compounds 23e and trifluoroacetic acid were used as raw materials to obtain compound LEE21 as a white solid, with a yield of 82%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 12.06 (s, 1H), 9.90 (s, 1H), 9.25 (t, J=6.1 Hz, 1H), 8.76-8.71 (m, 1H), 8.07 (d, J=5.6 Hz, 1H), 7.78-7.68 (m, 2H), 7.55 (dd, J=5.5, 1.2 Hz, 1H), 7.38-7.30 (m, 2H), 7.15 (s, 1H), 5.04 (s, 1H), 4.51 (d, J=6.0 Hz, 2H), 2.67 (t, J=7.1 Hz, 2H), 1.39 (h, J=7.3 Hz, 2H), 0.84 (t, J=7.4 Hz, 4H). .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 165.57, 161.10, 143.05, 138.53, 136.28, 135.24, 133.77, 132.39, 131.72, 127.64, 127.50, 116.28, 101.96, 53.57, 42.55, 21.32, 12.13. ESI-MS m/z: 352.06 [M+H].sup.+.

Example 22

[0227] ##STR00084##

[0228] Preparation of (E)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-3-(pyridin-2-yl) acrylamide (LEE13): the synthesis method of LEE18 was used and compounds 15a and 3-(2-pyridyl) acrylic acid were used as raw materials to obtain compound LEE13 as a white solid, with a yield of 57%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.96 (s, 1H), 8.82 (t, J=6.1 Hz, 1H), 8.58 (d, J=5.2 Hz, 2H), 7.76 (d, J=7.9 Hz, 2H), 7.58-7.25 (m, 6H), 6.88 (d, J=15.9 Hz, 1H), 5.06 (s, 1H), 4.42 (d, J=6.0 Hz, 2H), 2.70 (t, J=7.5 Hz, 2H), 1.42 (h, J=7.1 Hz, 2H), 0.86 (t, J=7.5 Hz, 3H). .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 165.57, 165.31, 165.13, 153.46, 150.67, 150.33, 149.64, 143.13, 143.10, 139.02, 137.66, 136.29, 134.46, 132.34, 131.12, 127.61, 127.53, 125.95, 124.74, 124.53, 124.45, 124.39, 53.58, 42.55, 42.52, 21.32, 12.13. ESI-MS m/z: 339.06 [M+H].sup.+.

Example 23

[0229] ##STR00085##

[0230] Preparation of (E)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-3-(pyridin-4-yl)acrylamide (LEE14): the synthesis method of LEE18 was used and compounds 15a and 3-(4-pyridyl) acrylic acid were used as raw materials to obtain compound LEE14 as a white solid, with a yield of 55%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.98 (s, 1H), 8.81-8.69 (m, 2H), 8.56 (dd, J=4.8, 1.6 Hz, 1H), 8.00 (dt, J=8.0, 2.0 Hz, 1H), 7.85-7.76 (m, 2H), 7.53 (d, J=15.9 Hz, 1H), 7.45 (dd, J=8.0, 4.8 Hz, 1H), 7.40-7.32 (m, 2H), 6.82 (d, J=15.9 Hz, 1H), 5.09 (s, 1H), 4.46 (d, J=5.9 Hz, 2H), 2.75 (t, J=7.1 Hz, 2H), 1.47 (h, J=7.3 Hz, 2H), 0.91 (t, J=7.4 Hz, 3H). .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 165.57, 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].sup.+.

Example 24

[0231] N-(4-(2-propylhydrazin-1-carbonyl)benzyl)thiopheno[3,2-c]pyridin-2-carboxamide (LEE19). The synthesis method of LEE18 was used and compounds 15a and thiopheno[3,2-c]pyridine-2-carboxylic acid were used as raw materials to obtain compound LEE19 as a white solid, with a yield of 54%. ESI-MS m/z: 368.92 [M+H].sup.+.

Example 25

[0232] ##STR00086##

[0233] Preparation of N-(4-(2-propylhydrazino-1-carbonyl)benzyl)furan[3,2-c]pyridine-2-carboxyamide (LEE20): the synthesis method of LEE18 was used and compounds 23d and trifluoroacetic acid were used as raw materials to obtain compound LEE20 as a white solid, with a yield of 78%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.98 (s, 1H), 9.49 (t, J=6.2 Hz, 1H), 9.05 (s, 1H), 8.55 (d, J=5.8 Hz, 1H), 7.80-7.63 (m, 4H), 7.36 (d, J=7.9 Hz, 2H), 4.49 (d, J=6.1 Hz, 2H), 2.70 (t, J=7.2 Hz, 2H), 1.42 (h, J=7.4 Hz, 2H), 0.86 (t, J=7.4 Hz, 3H); .sup.13C NMR (101 MHz, DMSO-d.sub.6) ? 165.64, 158.98, 158.21, 150.06, 146.84, 146.48, 142.86, 132.38, 127.68, 125.05, 108.53, 108.17, 53.58, 42.53, 21.33, 12.19. ESI-MS m/z: 352.88 [M+H].sup.+.

Example 26

[0234] ##STR00087##

[0235] Preparation of 5-bromo-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide (LEE26): the synthesis method of LEE18 was used and compounds 23j and trifluoroacetic acid were used as raw materials to obtain compound LEE26 as a white solid, with a yield of 84%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) ? 11.88 (s, 1H), 9.98 (s, 1H), 9.27 (d, J=6.9 Hz, 1H), 7.84 (d, J=1.9 Hz, 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=6.0 Hz, 2H), 2.73 (t, J=7.1 Hz, 2H), 1.50-1.39 (m, 2H), 0.89 (t, J=7.4 Hz, 4H).

Example 27

[0236] ##STR00088##

[0237] Preparation of 5-fluoro-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide (LEE28): the synthesis method of LEE18 was used and compounds 23l and trifluoroacetic acid were used as raw materials to obtain compound LEE28 as a white solid, with a yield of 85%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) ? 11.78 (s, 1H), 9.99 (s, 1H), 9.26 (t, J=6.1 Hz, 1H), 7.79 (d, J=8.3 Hz, 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=6.1 Hz, 2H), 2.73 (t, J=7.1 Hz, 2H), 1.49-1.39 (m, 2H), 0.89 (t, J=7.4 Hz, 4H).

Example 28

[0238] ##STR00089##

[0239] Preparation of 6-(dimethylamino)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide (LEE27): the synthesis method of LEE18 was used and compounds 23k and trifluoroacetic acid were used as raw materials to obtain compound LEE27 as a white solid, with a yield of 82%.

Example 29

[0240] ##STR00090##

[0241] Preparation of 5-chloro-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide (LEE29): the synthesis method of LEE18 was used and compounds 23m and trifluoroacetic acid were used as raw materials to obtain compound LEE29 as a white solid, with a yield of 80%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) ? 11.87 (s, 1H), 9.98 (s, 1H), 9.26 (t, J=6.1 Hz, 1H), 7.79 (d, J=8.3 Hz, 2H), 7.69 (d, J=2.1 Hz, 1H), 7.43 (d, J=8.7 Hz, 1H), 7.39 (d, J=8.3 Hz, 2H), 7.21-7.15 (m, 2H), 5.06 (s, 1H), 4.53 (d, J=6.1 Hz, 2H), 2.73 (t, J=7.1 Hz, 2H), 1.50-1.39 (m, 2H), 0.89 (t, J=7.4 Hz, 4H).

Example 30

[0242] ##STR00091##

[0243] Preparation of 5-methoxy-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide (LEE30): the synthesis method of LEE18 was used and compounds 23n and trifluoroacetic acid were used as raw materials to obtain compound LEE30 as a white solid, with a yield of 85%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) ? 9.98 (s, 1H), 9.31 (t, J=6.2 Hz, 1H), 7.81-7.75 (m, 2H), 7.57-7.52 (m, 1H), 7.51 (d, J=1.0 Hz, 1H), 7.42-7.34 (m, 2H), 7.26 (d, J=2.6 Hz, 1H), 7.05 (dd, J=9.0, 2.7 Hz, 1H), 4.50 (d, J=6.1 Hz, 2H), 3.79 (s, 3H), 2.73 (t, J=7.1 Hz, 2H), 1.51-1.40 (m, 2H), 0.89 (t, J=7.4 Hz, 3H).

Example 31

[0244] ##STR00092##

[0245] Preparation of 5-fluoro-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide (LEE31): the synthesis method of LEE18 was used and compounds 23o and trifluoroacetic acid were used as raw materials to obtain compound LEE31 as a white solid, with a yield of 86%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) ? 9.96 (d, J=6.2 Hz, 1H), 9.40 (t, J=6.1 Hz, 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=6.1 Hz, 2H), 2.73 (t, J=7.1 Hz, 2H), 1.45 (h, J=7.3 Hz, 2H), 0.89 (t, J=7.5 Hz, 3H).

Example 32

[0246] ##STR00093##

[0247] Preparation of 5-bromo-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide (LEE32): the synthesis method of LEE18 was used and compounds 23p and trifluoroacetic acid were used as raw materials to obtain compound LEE32 as a white solid, with a yield of 84%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) ? 9.98 (s, 1H), 9.45 (t, J=6.2 Hz, 1H), 8.01 (d, J=2.0 Hz, 1H), 7.81-7.75 (m, 2H), 7.64 (d, J=8.8 Hz, 1H), 7.60 (d, J=2.0 Hz, 1H), 7.58 (d, J=0.9 Hz, 2H), 7.39 (d, J=8.2 Hz, 2H), 5.07 (s, 1H), 4.51 (d, J=6.1 Hz, 2H), 2.73 (t, J=7.1 Hz, 2H), 1.53-1.35 (m, 2H), 0.89 (t, J=7.4 Hz, 3H).

Example 33

[0248] ##STR00094##

[0249] Preparation of 5-methoxy-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-1H-indole-2-carboxyamide (LEE33): the synthesis method of LEE18 was used and compounds 23q and trifluoroacetic acid were used as raw materials to obtain compound LEE33 as a white solid, with a yield of 82%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) ? 11.47 (s, 1H), 9.98 (s, 1H), 9.07 (t, J=6.1 Hz, 1H), 7.81-7.76 (m, 2H), 7.42-7.36 (m, 2H), 7.31 (d, J=8.9 Hz, 1H), 7.12-7.05 (m, 2H), 6.83 (dd, J=8.9, 2.5 Hz, 1H), 5.09 (s, 1H), 4.53 (d, J=6.1 Hz, 2H), 3.75 (s, 3H), 2.73 (t, J=7.1 Hz, 2H), 1.50-1.40 (m, 2H), 0.89 (t, J=7.4 Hz, 3H).

Example 34

[0250] ##STR00095##

[0251] Preparation of 5-chloro-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide (LEE34): the synthesis method of LEE18 was used and compounds 23r and trifluoroacetic acid were used as raw materials to obtain compound LEE34 as a white solid, with a yield of 86%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) ? 9.99 (d, J=4.1 Hz, 1H), 9.49 (t, J=6.1 Hz, 1H), 7.87 (d, J=2.2 Hz, 1H), 7.81-7.75 (m, 2H), 7.69 (d, J=8.8 Hz, 1H), 7.60 (d, J=1.0 Hz, 1H), 7.48 (dd, J=8.8, 2.2 Hz, 1H), 7.42-7.36 (m, 2H), 5.09 (s, 1H), 4.51 (d, J=6.1 Hz, 2H), 2.73 (t, J=7.1 Hz, 2H), 1.49-1.39 (m, 3H), 0.89 (t, J=7.4 Hz, 4H).

Example 35

[0252] ##STR00096##

[0253] Preparation of 5-(prop-2-yn-1-yloxy)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide (LEE35): the synthesis method of LEE18 was used and compounds 23s and trifluoroacetic acid were used as raw materials to obtain compound LEE35 as a white solid, with a yield of 83%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) ? 9.97 (s, 1H), 9.31 (t, J=6.2 Hz, 1H), 7.81-7.74 (m, 2H), 7.61-7.55 (m, 1H), 7.52 (d, J=0.9 Hz, 1H), 7.43-7.36 (m, 2H), 7.34 (d, J=2.6 Hz, 1H), 7.13-7.07 (m, 1H), 5.08 (s, 1H), 4.83 (d, J=2.4 Hz, 2H), 4.51 (d, J=6.1 Hz, 2H), 3.56 (t, J=2.4 Hz, 1H), 2.73 (t, J=7.1 Hz, 2H), 1.45 (h, J=7.4 Hz, 2H), 0.89 (t, J=7.4 Hz, 4H).

Example 36

[0254] ##STR00097##

[0255] Preparation of 5-hydroxy-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)benzofuran-2-carboxyamide (LEE36): the synthesis method of LEE18 was used and compounds 23u and trifluoroacetic acid were used as raw materials to obtain compound LEE36 as a white solid, with a yield of 79%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) ? 9.96 (s, 1H), 9.39 (s, 1H), 9.24 (t, J=6.2 Hz, 1H), 7.81-7.74 (m, 2H), 7.46-7.35 (m, 4H), 7.02 (d, J=2.5 Hz, 1H), 6.90 (dd, J=8.9, 2.5 Hz, 1H), 4.49 (d, J=6.2 Hz, 2H), 2.73 (t, J=7.1 Hz, 2H), 1.50-1.39 (m, 2H), 0.89 (t, J=7.4 Hz, 3H).

Example 37

[0256] ##STR00098##

[0257] Preparation of (E)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-8-(3-(pyridine-3-yl)acrylamido) octanamide (LEE37): the synthesis method of LEE18 was used and compounds 23v and trifluoroacetic acid were used as raw materials to obtain compound LEE37 as a white solid, with a yield of 78%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.93 (s, 1H), 8.71 (s, 1H), 8.50 (d, J=4.7 Hz, 1H), 8.31 (t, J=6.0 Hz, 1H), 8.12 (t, J=5.7 Hz, 1H), 7.93 (dt, J=8.0, 2.0 Hz, 1H), 7.73 (d, J=8.0 Hz, 2H), 7.45-7.36 (m, 2H), 7.25 (d, J=8.0 Hz, 2H), 6.69 (d, J=15.9 Hz, 1H), 5.06 (s, 1H), 4.25 (d, J=5.9 Hz, 2H), 3.13 (q, J=6.6 Hz, 2H), 2.70 (t, J=7.1 Hz, 2H), 2.10 (t, J=7.4 Hz, 2H), 1.55-1.35 (m, 6H), 1.31-1.15 (m, 7H), 0.86 (t, J=7.4 Hz, 3H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) ? 172.69, 165.56, 164.83, 150.49, 149.51, 143.60, 135.52, 134.34, 132.12, 131.22, 127.49, 127.36, 124.81, 124.41, 53.55, 42.16, 39.17, 35.76, 29.53, 29.09, 28.94, 26.84, 25.70, 21.30, 12.12. ESI-MS m/z: 479.96 [M+H].sup.+.

Example 38

[0258] ##STR00099##

[0259] Preparation of (E)-N-(4-(2-propylhydrazino-1-carbonyl)benzyl)-7-(3-(pyridine-3-yl)acrylamido) heptanamide (LEE39): the synthesis method of LL289 was used and compounds 15a and 2d were used as raw materials to obtain compound YKR-31 as a white solid, with a yield of 58%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 9.93 (s, 1H), 8.51 (dd, J=4.7, 1.6 Hz, 1H), 8.32 (t, J=6.1 Hz, 1H), 8.13 (t, J=5.6 Hz, 1H), 7.73 (d, J=8.0 Hz, 2H), 7.46-7.36 (m, 2H), 7.26 (d, J=8.0 Hz, 2H), 6.69 (d, J=15.9 Hz, 1H), 5.03 (s, 1H), 4.25 (d, J=5.8 Hz, 2H), 3.13 (q, J=6.6 Hz, 2H), 7.97-7.90 (m, 1H), 2.70 (t, J=7.0 Hz, 2H), 2.11 (t, J=7.4 Hz, 2H), 1.53-1.38 (m, 6H), 1.24 (dq, J=9.4, 5.7, 5.0 Hz, 4H), 0.86 (t, J=7.4 Hz, 3H), 8.75-8.67 (m, 1H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) ? 172.67, 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].sup.+.

Example 39

[0260] ##STR00100##

[0261] Preparation of (E)-N-(4-(2-propylhydrazino-1-carbonyl)phenyl)-7-(3-(pyridine-3-yl)acrylamido) heptamide (LEE40): the synthesis method of LEE18 was used and compounds 23x and trifluoroacetic acid were used as raw materials to obtain compound LEE40 as a white solid, with a yield of 65%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.06 (s, 1H), 9.84 (d, J=5.5 Hz, 1H), 8.71 (d, J=2.2 Hz, 1H), 8.50 (dd, J=4.8, 1.6 Hz, 1H), 8.13 (t, J=5.7 Hz, 1H), 7.93 (dt, J=8.1, 2.0 Hz, 1H), 7.72 (d, J=8.6 Hz, 2H), 7.61 (d, J=8.6 Hz, 2H), 7.45-7.35 (m, 2H), 6.69 (d, J=15.9 Hz, 1H), 5.00 (d, J=5.5 Hz, 1H), 3.14 (q, J=6.5 Hz, 2H), 2.69 (q, J=6.4 Hz, 2H), 2.29 (t, J=7.4 Hz, 2H), 1.57 (d, J=7.3 Hz, 2H), 1.43 (p, J=7.3 Hz, 4H), 1.31-1.26 (m, 4H), 0.87 (t, J=7.4 Hz, 3H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) ? 172.09, 165.33, 164.83, 150.50, 149.51, 142.38, 135.51, 134.34, 131.22, 128.82, 128.28, 127.85, 124.81, 124.41, 118.63, 53.62, 39.14, 36.87, 29.42, 28.84, 26.71, 25.41, 21.31, 12.12, 11.14. ESI-MS m/z: 451.97 [M+H].sup.+.

Example 40

[0262] ##STR00101##

[0263] Preparation of (E)-N-(4-(2-propylhydrazino-1-carbonyl)phenyl)-8-(3-(pyridine-3-yl)acrylamido) amide (LEE38): the synthesis method of LEE18 was used and compounds 23w and trifluoroacetic acid were used as raw materials to obtain compound LEE38 as a white solid, with a yield of 67%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.04 (s, 1H), 9.85 (s, 1H), 8.71 (d, J=2.3 Hz, 1H), 8.50 (dd, J=4.7, 1.6 Hz, 1H), 8.11 (t, J=5.6 Hz, 1H), 7.93 (dt, J=8.0, 2.0 Hz, 1H), 7.72 (d, J=8.7 Hz, 2H), 7.62 (s, 2H), 7.45-7.35 (m, 2H), 6.68 (d, J=15.9 Hz, 1H), 3.13 (q, J=6.6 Hz, 2H), 2.69 (t, J=7.1 Hz, 2H), 2.29 (t, J=7.4 Hz, 2H), 1.56 (p, J=6.8 Hz, 2H), 1.42 (h, J=7.3 Hz, 4H), 1.27 (s, 6H), 0.87 (t, J=7.4 Hz, 3H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) ? 172.10, 165.34, 164.82, 150.49, 149.50, 142.39, 135.51, 134.34, 131.22, 128.29, 127.83, 124.81, 124.41, 118.62, 53.60, 39.15, 36.90, 29.53, 29.08, 28.98, 26.82, 25.42, 21.29, 12.12. ESI-MS m/z: 465.97 [M+H].sup.+.

Example 41

[0264] ##STR00102##

[0265] Preparation of N-(4-(2-propylhydrazino-1-carbonyl)phenyl)-8-(4-(pyridin-3-yl)-1H-1,2,3-triazole-1-yl) octanamide (LEE41): the synthesis method of LEE18 was used, compound 24 and trifluoroacetic acid were used as raw materials to obtain a white solid, with a yield of 67%.

[0266] The synthesis method of LEE11 was used, the product from the previous step and 3-ethynylpyridine were used as raw materials to obtain compound LEE41 as a white solid, with a yield of 80%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 10.08 (s, 1H), 9.93 (s, 1H), 9.01 (d, J=2.2 Hz, 1H), 8.68 (s, 1H), 8.49 (d, J=4.7 Hz, 1H), 8.16 (dd, J=8.0, 2.0 Hz, 1H), 7.72 (d, J=8.5 Hz, 2H), 7.61 (d, J=8.5 Hz, 2H), 7.44 (dd, J=8.0, 4.8 Hz, 1H), 4.38 (t, J=7.1 Hz, 2H), 2.71 (t, J=7.1 Hz, 2H), 2.28 (t, J=7.3 Hz, 2H), 1.83 (p, J=7.2 Hz, 2H), 1.54 (p, J=7.3 Hz, 2H), 1.43 (h, J=7.3 Hz, 2H), 1.35-1.16 (m, 8H), 0.87 (t, J=7.4 Hz, 3H). .sup.13C NMR (126 MHz, DMSO-d.sub.6) ? 172.10, 165.34, 149.24, 146.77, 143.92, 142.46, 132.82, 128.33, 127.67, 127.27, 124.47, 122.41, 118.62, 53.53, 50.07, 36.84, 30.01, 28.91, 28.61, 26.17, 25.34, 21.15, 12.08. ESI-MS m/z: 464.03 [M+H].sup.+.

Example 42

[0267] ##STR00103##

[0268] Preparation of methyl (E)-4-((3-(pyridine-3-yl)acrylamido)methyl) benzoate (LEE43): the synthesis method of 1a was used to obtain compound LEE43 as a white solid, with a yield of 51%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 8.80-8.78 (m, 2H), 8.57 (dd, J=1.1 Hz, J=3.1 Hz, 1H), 8.03 (td, J=1.1 Hz, J=5.4 Hz, 1H), 7.95 (td, J=1.3 Hz, J=5.5 Hz, 2H), 7.55 (d, J=10.6 Hz, 1H), 7.47-7.43 (m, 3H), 6.84 (d, J=10.6 Hz, 1H), 4.51 (d, J=4.0 Hz, 2H), 3.85 (s, 3H).

Example 43

[0269] ##STR00104##

[0270] Preparation of methyl 4-((1H-pyrrolo[3,2-c]pyridine-2-carbonylamide)methyl) benzoate (LEE44): the synthesis method of 1a was used to obtain compound LEE44 as a white solid, with a yield of 46%. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ? 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).

Example 44: Inhibitory Activity of Compounds on HDAC1, 2 and 3

[0271] Experimental materials: HDAC buffer: 15 mM Tris-HCl (pH 8.0), 250 ?M EDTA, 250 mM NaCl, 10% glycerin. Trypsin termination solution: 10 mg/ml pancreatin, 50 mM Tris-HCL (pH 8.0), 100 mM NaCl, 2 ?M TSA. Substrates: the dedicated substrates HDAC1, HDAC2 and HDAC3 were dissolved with dimethyl sulfoxide to be formulated into a 30 mM stock solution, the stock solution was diluted to 300 ?M with HDAC buffer, so that the content of dimethyl sulfoxide was around 1%. Enzyme solution: HDAC1, HDAC2 and HDAC3 were diluted with HDAC buffer in a ratio of 1:20.

Experimental Steps:

[0272] a) Formulation of 100% solution: 50 ?L of HDAC buffer was mixed with 10 ?L of enzyme solution, 40 ?L of substrate was added after 5 minutes and the above materials reacted at 37? C. for 30 minutes, then 100 ?L of trypsin termination 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 390 nm/460 nm to obtain 100% absorbence. AMC was used as the standard to create a standard curve and calculate enzyme activity.

[0273] b) Formulation of blank solution: 60 ?L of HDAC buffer was added with 40 ?L of substrate, and the above materials reacted at 37? C. for 30 minutes, then 100 ?L of trypsin termination solution was added and the reaction was carried out at 37? C. for 20 minutes, the fluorescence intensity was measured at 390 nm/460 nm to obtain blank absorbence.

[0274] 6. The determination steps for drug inhibition of HDAC enzyme activity: 50 ?L of HDAC buffer containing a drug was mixed with 10 ?L of enzyme solution and pre-incubated for 5 minutes, 40 ?L of substrate was added and then the above materials reacted at 37? C. for 30 minutes, then 100 ?L of trypsin termination solution was added to terminate the above reaction, and the reaction was carried out at 37? C. for 20 minutes, and the fluorescence intensity was measured at 390 nm/460 nm.

[00001]$\mathrm{Inhibitory}\mathrm{rate}\left(\%\right)=\frac{\begin{array}{c}100\%\mathrm{fluorescence}\mathrm{intensity}-\\ \mathrm{fluorescence}\mathrm{intensity}\mathrm{of}\mathrm{compound}\end{array}}{100\%\mathrm{fluorescence}\mathrm{intensity}-\mathrm{fluorescence}\mathrm{intensity}\mathrm{of}\mathrm{blank}}?100\%$

[0275] Finally, the inhibitory rate (%) of the compound was subjected to S-curve fitting with its corresponding concentration to calculate the IC.sub.50 value.

[0276] The inhibitory activity results of some compounds shown in structural general formulas (I, II, III or IV) of the present disclosure on HDAC1, HDAC2 and HDAC3 are shown in Table 1 below:

TABLE-US-00001 TABLE 1 Inhibitory IC.sub.50 values of some compounds on HDAC1, 2 and 3 IC.sub.50 (nM?) Structure HDAC1 HDAC2 HDAC3 LEE12 [00105] 25.10 140.8 3.12 LEE13 [00106] 13.20 49.17 1.82 LEE14 [00107] 12.71 88.48 2.74 LEE1 [00108] >10,000 13600 1556 LEE2 [00109] 689 2002 111.5 LEE3 [00110] 79.3 338.7 20.13 LEE18 [00111] 7.61 37.81 0.71 LEE20 [00112] 10.35 38.41 0.52 LEE16 [00113] 3.01 18.54 0.435 LEE15 [00114] 3.52 15.14 0.383 LEE17 [00115] 3.79 16.92 0.551 LEE19 [00116] 10.06 59.96 1.20 LEE7 [00117] 25.09 112.5 2.97 LEE21 [00118] 8.17 51.47 2.42 LEE5 [00119] 444 398 110.4 LEE37 [00120] 17.8 84.1 1.98 LEE8 [00121] 147.4 320.9 26.42 LEE39 [00122] 500.8 1176 84.31 LEE40 [00123] 53.87 122.7 12.37 LEE38 [00124] 620.1 539.1 97.37 LEE4 [00125] 241.0 505.8 76.68 LEE11 [00126] 56.06 187.7 8.78 LEE41 [00127] 18.02 78.67 1.08 LEE10 [00128] 57.37 150.0 5.43 Comparative example 1 LP411/3b [00129] 11.8 95.45 0.95 Comparative example 2 LP97/13a [00130] 5.17 49.5 0.28 SAHA 43.76 89.23 149.7 MS275 53.89 108.2 77.18 ?The data in the table are all from three independent experiments, and the values are mean values with a standard deviation of <10%; ND: not detected.

[0277] The experimental results show that the vast majority of compounds in the table have nanomolar level of inhibitory ICso values against HDAC1/2/3, and the activity against HDAC1 and HDAC3 is generally higher than that on HDAC2. The activity is generally higher than that of positive control drugs SAHA and MS275.

Example 45: Inhibitory Activity of cCmpounds on NAMPT

[0278]

TABLE-US-00002 TABLE 2 Inhibitory IC.sub.50 values of some compounds on NAMPT NAMPT IC.sup.50 Structure (nM?) LEE12 [00131] 6570 LEE18 [00132] 1618 LEE7 [00133] 4555 LEE19 [00134] >100,000 LEE43 [00135] 4675 LEE44 [00136] 6714 Comparative example 1 LP411/3b [00137] >20,000 Comparative example 2 LP97/13a [00138] >20,000 FK866 96.11 ?The data in the table are all from three independent experiments, and the values are mean values with a standard deviation of <10%.

[0279] The experimental results show that LEE12, LEE18, LEE7, LEE43 and LEE44 all exhibited micromolar level of inhibitory activity against NAMPT, and the above compounds achieved unexpected technical effects compared to compounds in comparative examples 1 and 2.

Example 46: Median Growth Inhibitory Concentration (GI.SUB.50.) and Median Lethal Concentration (LC.SUB.50.) of Compounds on Tumor Cells

[0280] The median growth inhibitory concentration (GI.sub.50) and median lethal concentration (LC.sub.50) were determined by NCI method. Leukemia cell strains MV4-11, HL60, PL21, KASUMI-1, MONO-MAC-1 and NB-4 were cultured in an Iscove's Modified Dulbecco Medium (IMDM) containing 10% fetal bovine serum, were inoculated into a 96-well cell culture plate at a density of 10,000 cells/100 ?L and cultured overnight. 6 wells were selected as Tz wells, 0.125 mg/mL Cell Titer-Blue dye was added, and the cells were cultured for 4 hours and then the fluorescence intensity at 560 nM/590 nM (excitation wavelength/emission wavelength) was read. Compounds of different concentrations were added to the remaining wells, and a 100% control group was set. After 48 hours of cultivation, 0.125 mg/mL Cell Titer-Blue dye was added and the fluorescence intensity at 560 nM/590 nM was read after 4 hours. The fluorescence intensity of the dosing group was represented by Ti, the fluorescence intensity of the 100% control group was represented by C, and the fluorescence intensity before dosing was represented by Tz.

[0281] If Ti?Tz, the formula [(Ti?Tz)/(C?Tz)]?100 was used.

[0282] If Ti<Tz, the formula [(Ti?Tz)/Tz]?100 was used.

[0283] The median growth inhibitory concentration (GI.sub.50) is a compound concentration of [(Ti?Tz)/(C?Tz)]?100=50, with a median lethal concentration (LC.sub.50) being a concentration of [(Ti?Tz)/Tz]?100=?50. The GI.sub.50 and LC.sub.50 values of some compounds on leukemia cell strains MV4-11, HL60, PL21, KASUMI-1, MONO-MAC-1 and NB-4 are shown in FIG. 1, Table 3, FIG. 2, and Table 4.

TABLE-US-00003 TABLE 3 GI.sub.50 and LC.sub.50 values of some compounds on acute myelogenous leukemia cells MV4-11 and HL60 MV4-11 HL60 MV4-11 HL60 Compound GI.sub.50 GI.sub.50 LC.sub.50 LC.sub.50 No. (nM.sup.a) (nMa) (nMa) (nMa) Comparative 65.99 120.9 68.80 >10,000 example 1 LP411/3b LEE12 38.31 42.15 218.5 590.7 LEE13 14.98 128.5 124.7 >10,000 LEE14 14.29 63.09 83.61 6058 LEE22 16.43 99.78 46.71 >10,000 LEE23 15.59 138.7 90.95 >10,000 LEE24 14.20 123.4 100.1 >10,000 LEE8 31.19 N.Db 455.7 N.Db LEE40 15.70 N.Db 349.2 N.Db LEE18 23.61 51.21 58.15 648.8 LEE21 25.41 81.49 57.95 >10,000 LEE7 64.70 158.0 163.7 724.1 LEE16 29.99 75.23 57.84 >10,000 LEE17 20.27 58.73 57.15 >10,000 LEE15 37.25 69.55 58.10 >10,000 LEE19 53.99 77.32 98.54 >10,000 LEE20 50.43 N.Db 109.1 >10,000 FK866 7.95 12.07 >10,000 >10,000 Note: KF866 is an NAMPT inhibitor reported in literature. .sup.aThe data in the table are all from three independent experiments, and the values are mean values with a standard deviation of <10%; .sup.bND: not detected.

[0284] The experimental results show that in the wt-p53 cell strain MV4-11, all the tested compounds exhibited nanomolar level of GI.sub.50 and LC.sub.50 values, indicating that they can cause not only cell proliferation inhibition but also cell death. The activity of most compounds was significantly stronger than that of compound LP411 (3b) of comparative example 1. While in the p53-null cell strain HL60, all the tested compounds exhibited nanomolar level of GI.sub.50 values, indicating that they have good anti-proliferative activity, while only LEE12, LEE14, LEE18 and LEE7 have nanomolar level of LC.sub.50 values, which could lead to cell death. It is indicated that inhibiting both HDAC and NAMPT may have a synthetic lethal effect on p53-null cell strains.

TABLE-US-00004 TABLE 4 GI.sub.50 and LC.sub.50 values of some compounds on leukemia cell lines MONO- MONO- PL21 PL21 KASUM KASUM MAC-1 MAC-1 NB4 NB4 Compound GI.sub.50 LC.sub.50 I-1 GI.sub.50 I-1 LC.sub.50 GI.sub.50 LC.sub.50 GI.sub.50 LC.sub.50 No. (nM.sup.a) (nM.sup.a) (nM.sup.a) (nM.sup.a) (nM.sup.a) (nM.sup.a) (nM.sup.a) (nM.sup.a) Comparative 522.8 >10,000 17.80 >10,000 16.26 >10,000 19.50 >10,000 example 1 LP411/3b LEE12 259.2 3816 13.46 285.3 73.73 9032 22.82 2296 LEE15 65.52 >10,000 17.49 8406 17.95 >10,000 31.87 >10,000 LEE16 34.28 >10,000 15.29 5362 25.32 >10,000 42.32 >10,000 LEE17 74.28 >10,000 25.32 7325 33.21 >10,000 51.17 >10,000 LEE18 109.7 2722 21.38 503.2 65.73 8682 16.47 1564 .sup.aThe data in the table are all from three independent experiments, and the values are mean values with a standard deviation of <10%.

[0285] FIG. 2 and Table 4 also indicate that the dual inhibitors of HDAC and NAMPT, i.e. LEE12 and LEE18, have lethal effects on p53-mutant and p53-null cell strains. The vast majority of compounds achieved unexpected technical effects compared to compounds in comparative example 1.

Example 47: In Vivo Anti-Colon Cancer Activity of Target Compound LEE17

[0286] 5-FU: 5-fluorouracil, a traditional anti-tumor chemotherapy drug; Oxaliplatin, a third-generation platinum anticancer drug, is an anti-tumor chemotherapy drug.

[0287] Colon cancer cell HCT116 was subcutaneously inoculated into the right shoulder of nude mice at 100 ?L per mouse (cell count: 1.8?10.sup.8 cells/mL). After one week, the mice were grouped and dosed, the tumor bearing mice were grouped and administered by gavage, and the grouping is as follows:

[0288] Test group: Compound LL341, administered orally at a dose of 8 mg/kg/d, with an administration volume of 0.2 mL per animal per dose

[00002]$\mathrm{Tumor}\mathrm{inhibitory}\mathrm{rate}\left(\%\right)=\frac{\begin{array}{c}\mathrm{average}\mathrm{tumor}\mathrm{weight}\mathrm{in}\mathrm{control}\mathrm{group}-\\ \mathrm{average}\mathrm{tumor}\mathrm{weight}\mathrm{in}\mathrm{treatment}\mathrm{group}\end{array}}{\mathrm{average}\mathrm{tumor}\mathrm{weight}\mathrm{of}\mathrm{control}\mathrm{group}}?100\%$

[0289] Relative tumor volume (RTV)=Vt/Vo

[0290] The evaluation index for anti-tumor activity is relative tumor proliferation rate T/C

[00003]$T/C\left(\%\right)=\frac{\mathrm{Treatment}\mathrm{group}\left(T\right)\mathrm{RTV}}{\mathrm{Negative}\mathrm{control}\mathrm{group}\left(C\right)\mathrm{RTV}}?100\%$

TABLE-US-00005 TABLE 7 In vivo study data results of compound LEE17 on HCT116 tumor bearing model Tumor Relative tumor Weight of nude inhibitory rate proliferation mice at the end of Compounds (TGI%) rate (T/C %) the experiment (g) LEE17 85.5 14.8 19.04 ? 1.31 5-FU + 23.5 52.3 19.77 ? 0.61 Oxaliplatin Blank control / / 21.39 ? 1.18 Note: 5-FU: 5-fluorouracil, a traditional anti-tumor chemotherapy drug; Oxaliplatin, a third-generation platinum anticancer drug, is an anti-tumor chemotherapy drug.

[0291] The experimental results show that at a dosage of 8 mg/kg/d, compound LEE17 can significantly inhibit the growth of HCT116 tumors, with an inhibitory rate of 85.5% that was significantly higher than that of positive drug 5-FU+ Oxaliplatin. At the end of the experiment, there was no significant change in the weight of the nude mice, indicating that LEE17 had a certain level of safety under the dosage of administration.

Example 48: In Vivo Anti-Leukemia Activity of Target Compounds LEE15 and LEE16 Panobinostat: Panobinostat is a Marketed Broad-Spectrum HDAC Inhibitor

[0292] Acute myeloid leukemia cell MV4-11 was subcutaneously inoculated into the right shoulder of nude mice at an amount of 100 ?L cells per mouse (cell count: 1.8?10.sup.8 cells/mL). After one week, the mice were grouped and dosed, the tumor bearing mice were grouped and administered by gavage, and the grouping was as follows:

[0293] Test group: compounds LEE15 and LEE16, administered at a dose of 4 mg/kg/d, with a dose volume of 200 ?L/20 g per animal per dose;

[0294] Positive control group: positive drug panobinostat, administered at a dose of 4 mg/kg/d, intraperitoneal injection.

[0295] Blank control group: the same volume of PBS was given.

[0296] The mice were administered once a day, a tumor volume was measured every 3-4 days, an average value of each group was taken, and a tumor growth curve was drawn (see FIG. 6). On the day 23 of administration for the MV4-11 tumor bearing model, the nude mice was executed, the tumor and viscera were dissected, and at the end of the experiment, the tumor mass was weighed and a tumor inhibitory rate was calculated according to the formula. The maximum diameter (a) and minimum diameter (b) of the tumor were measured, the tumor volume (V): V=ab.sup.2/2 was calculated, and the relative tumor proliferation rate T/C (%) was calculated.

[00004]$\mathrm{Tumor}\mathrm{inhibitory}\mathrm{rate}\left(\%\right)=\frac{\begin{array}{c}\mathrm{average}\mathrm{tumor}\mathrm{weight}\mathrm{in}\mathrm{control}\mathrm{group}-\\ \mathrm{average}\mathrm{tumor}\mathrm{weight}\mathrm{in}\mathrm{treatment}\mathrm{group}\end{array}}{\mathrm{average}\mathrm{tumor}\mathrm{weight}\mathrm{of}\mathrm{control}\mathrm{group}}?100\%$

[0297] Relative tumor volume (RTV)=Vt/Vo

[0298] The evaluation index for anti-tumor activity is the relative tumor proliferation rate T/C (%)

[00005]$T/C\left(\%\right)=\frac{\mathrm{Treatment}\mathrm{group}\left(T\right)\mathrm{RTV}}{\mathrm{Negative}\mathrm{control}\mathrm{group}\left(C\right)\mathrm{RTV}}?100\%$

TABLE-US-00006 TABLE 8 In vivo study data results of compounds LEE15 and LEE16 on MV4-11 tumor bearing model Relative tumor Tumor inhibitory proliferation Compounds rate (TGI %) rate (T/C %) LEE15 (4 mg/kg) 81.3 22.4 LEE16 (4 mg/kg) 78.4 25.4 Panobinostat 18.3 72.9 Note: Panobinostat is a positive control drug, which is a marketed broad-spectrum HDAC inhibitor.

[0299] The experimental results show that at a dosage of 4 mg/kg, LEE15 and LEE16 had significant in vivo anti-acute myeloid leukemia effects, with tumor inhibitory rates of 81.3% and 78.4%, respectively. The in vivo anti-tumor activity is significantly higher than that of the positive control drug, panobinostat.

Example 49: Pharmacokinetic Properties of Target Compounds LEE15, LEE16 and LEE17

[0300] LEE15, LEE16 and LEE17 were dissolved in 40% PEG300 and 60% H.sub.2O. Three mice in each group were administered as a single dose by oral gavage (po) of 20 mg/kg and intravenous injection (iv) of 5 mg/kg, respectively. At the end of administration, blood samples were taken at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours, respectively. After the samples were prepared, parameters such as t1/2, C0, AUC, Vss, CLp, MRT, C.sub.max, t.sub.max and F % were measured, respectively.

TABLE-US-00007 Po administration po dose t? T.sub.max C.sub.max AUC.sub.last F (mg/kg) (hr) (hr) (ng/mL) (hr*ng/ml) (%) Comparative 20 7.45 0.500 45.1 242 19.8 example 2 LP97/13a LEE16 20 6.03 0.500 34400 129292 111 LEE17 20 4.46 0.250 9463 27585 64.6 LEE15 20 2.58 0.250 38800 128134 112 Pano(Biomedical 50 2.9 116 126 4.62 chromatography: BMC 2007, 21, 184-189)

[0301] 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 compounds in comparative example 2. The aforementioned compounds LEE16, LEE17 and LEE15 have unexpected technical effects in metabolism.

[0302] The above descriptions are only preferred embodiments of the present application, but not intended to limit the present application. For those skilled in the art, various modifications and changes can be made to the presennt application. Any modifications, equivalent substitutions, improvements and the like made within the spirit and principle of the present application shall be included within the scope of protection of the present application.