(E)-6-(4-carbonylphenyl) hex-2-ene-1-ketone compound as well as preparation method and application thereof

Abstract

The invention discloses an (E)-6-(4-carbonyl phenyl) hex-2-ene-1-ketone compound as well as a preparation method and application thereof, the compound can be used as a palmitoyl transferase inhibitor, and the palmitoylation process of substrate protein is further inhibited by inhibiting DHHC protein, so that the inhibition effect on tumor cells is achieved. Embodiments show that the compounds have different degrees of anti-proliferative activity to different cancer cells and different degrees of enzyme inhibitory activity to DHHC proteins, and part of the compounds have better inhibitory activity to cancer cells and DHHC proteins than positive control drugs, and can be further developed into a new generation of antitumor drugs.

Description

Technical Field

[0001] The present invention relates to the field of anti-tumor medicines, and particularly relates to a compound of (E)-6-(4-carbonylphenyl)hex-2-en-1-one, a preparation method thereof and an application thereof. Background Art

[0002] Protein palmitoylation is a common post-translational modification of proteins. It affects protein localization, function, signal transduction, etc. by connecting palmitoyl groups to cysteine residues of proteins through thioester bonds. Enzymes catalyzing palmitoylation all contain a highly conserved DHHC (Asp-His-His-Cys) motif, so this enzyme family is also called DHHC. There is overexpression of DHHC in most tumor cells, and DHHC inhibitors have an inhibitory effect on tumor growth. At present, only a few DHHC inhibitors such as 2-BP, CMA, cerulenin, tunicamycin, etc. have been published (Formula II). There are not only few in number, but also problems such as low activity and poor specificity.

[0003]

[0004] α,β-unsaturated carbonyl is one of the most common warheads of covalent drugs. It acts by targeting cysteine residues in proteins. When the drug interacts with the protein, the sulfhydryl group in cysteine first attacks the β-carbon of the α,β-unsaturated carbonyl, and then Michael addition occurs to form a covalent bond. α,β-unsaturated carbonyl not only has high reactivity with cysteine, but also can be modified by introducing substituents at both ends to adjust its reactivity. However, there are no DHHC inhibitors with related structures used in clinical anti-tumor. Summary of the Invention

[0005] An object of the present invention is to provide a compound of (E)-6-(4-carbonylphenyl)hex-2-en-1-one, and this kind of compound has an inhibitory effect on tumor growth through the selection and modification of α,β-unsaturated carbonyl and substituents at both ends.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] The compound shown in Formula I:

[0008]

[0009] Wherein, R1 is an alkoxy with 1-4 carbons, or phenyl, or phenyl substituted by an alkyl with 1-2 carbons; R2 is an amino substituted by a straight-chain alkyl with 1-8 carbons, or an amino substituted by a straight-chain alkyl with 1-8 carbons containing a hydroxyl group, or a straight-chain alkoxy with 1-8 carbons, or a straight-chain alkoxy with 1-8 carbons containing a hydroxyl group.

[0010] Preferably, R1 is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or sec-butoxy; or phenyl, or tolyl, or ethylphenyl, or dimethylphenyl; R2 is an amino group substituted by a straight-chain alkyl group with 2-7 carbons, or an amino group substituted by a straight-chain alkyl group with 2-7 carbons and having a hydroxyl group at the end, or a straight-chain alkoxy group with 2-7 carbons, or a straight-chain alkoxy group with 2-7 carbons and having a hydroxyl group at the end.

[0011] Further, R1 is ethoxy or phenyl, and R2 is n-propylamino, n-butylamino, n-pentylamino, n-hexylamino, 3-hydroxypropylamino, 4-hydroxybutylamino, 5-hydroxypentylamino, 6-hydroxyhexylamino, n-propoxy, n-butoxy, n-pentyloxy, n-hexyloxy, 3-hydroxypropoxy, 4-hydroxybutoxy, 5-hydroxypentyloxy or 6-hydroxyhexyloxy.

[0012] Another object of the present invention is to provide a preparation method of the above compound: When R1 is ethoxy, the synthesis route is as follows:

[0013]

[0014] Step i: 4-Phenyl-1-butanol reacts with pyridinium dichromate (PCC) in dichloromethane to form Compound 1;

[0015] Step ii: Compound 1 reacts with ethoxycarbonylmethylene triphenylphosphine in dichloromethane to form Compound 2;

[0016] Step iii: Compound 2 reacts with oxalyl chloride in dichloromethane in an ice-water bath in the presence of anhydrous aluminum chloride to form Compound 3;

[0017] Step iv: Compound 3 undergoes a condensation reaction with R2-H in the presence of O-benzotriazole-N,N,N,N-tetramethyluronium tetrafluoroborate (TBTU) and triethylamine to form Compounds I-1–I-16;

[0018] When R1 is phenyl, the synthesis route is as follows:

[0019]

[0020] Step v: 4-Phenyl-1-butanol reacts with acetyl chloride in dichloromethane to form Compound 4;

[0021] Step vi: Compound 4 reacts with oxalyl chloride in dichloromethane in an ice-water bath in the presence of anhydrous aluminum chloride to form Compound 5;

[0022] Step vii: Compound 5 reacts in a mixed solvent of methanol and water in the presence of potassium carbonate to form Compound 6;

[0023] Step viii: Compound 6 reacts with pyridinium dichromate in dichloromethane to form Compound 7;

[0024] Step ix: Compound 7 reacts with dimethyl (2-oxo-2-phenylethyl)phosphonate in dichloromethane in the presence of potassium carbonate to form Compound 8;

[0025] Step x: Compound 8 undergoes a condensation reaction with R2-H in the presence of O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium tetrafluoroborate and N,N-diisopropylethylamine to form Compounds II-1–II-16.

[0026] Preferably, in Step i, the molar ratio of 4-phenyl-1-butanol to pyridinium chlorochromate is 1:1.2; in Step ii, the molar ratio of Compound 1 to ethoxycarbonylmethylenetriphenylphosphine is 1:1.2; in Step iii, the molar ratio of Compound 2 to oxalyl chloride is 1:0.8; in Step iv, the molar ratio of Compound 3 to R2-H is 1:1.1.

[0027] Preferably, in Step v, the molar ratio of 4-phenyl-1-butanol to acetyl chloride is 1:1.2; in Step vi, the molar ratio of Compound 4 to oxalyl chloride is 1:0.85; in Step vii, the molar ratio of Compound 5 to potassium carbonate is 1:1.2, and the ratio of methanol to water in the mixed solvent is 3:1; in Step viii, the molar ratio of Compound 6 to pyridinium chlorochromate is 1:1.5; in Step ix, the molar ratio of Compound 7 to dimethyl (2-oxo-2-phenylethyl)phosphonate is 1:2; in Step x, the molar ratio of Compound 8 to R2-H is 1:1.2.

[0028] Another object of the present invention is to provide the use of the compound shown in Formula I in the preparation of anti-tumor drugs.

[0029] Preferably, the tumor is human breast cancer cell MCF-7, human colon cancer cell HCT116, mouse colon cancer cell MC38 or mouse liver cancer cell Hepa1-6.

[0030] The present invention also provides a palmitoyltransferase inhibitor, which comprises the compound shown in Formula I or a pharmaceutically acceptable salt thereof as an active ingredient, and one or more pharmaceutically acceptable carriers and / or excipients.

[0031] The present invention uses palmitoyltransferase (DHHC) as the target protein and conducts a large number of molecular simulation studies. According to the structure of the covalent ligand small molecule 2-BP in the DHHC structure and the distribution of amino acid residues in the active pocket, using the methods of computer-aided drug design and compound structure modification, (E)-6-(4-oxophenyl)hex-2-en-1-one compounds are designed and synthesized. This compound can be used as a palmitoyltransferase inhibitor. By inhibiting the DHHC protein, and then inhibiting the palmitoylation process of the substrate protein, it exerts an inhibitory effect on tumor cells. Examples show that the (E)-6-(4-oxophenyl)hex-2-en-1-one compounds described in the present invention have different degrees of anti-proliferative activity against different cancer cells. Some compounds have better inhibitory activity against cancer cells than the positive control drug cisplatin, and are expected to be further developed into a new generation of anti-tumor drugs. Detailed implementation mode

[0032] In the following examples, various processes and methods not described in detail are conventional methods well known in the art. Reagents used without indicating the source and specifications are all commercially available analytical pure or chromatographically pure reagents.

[0033] Example 1 Preparation of Series I Compounds Shown by Formula I

[0034] (i) Dissolve 10 mM of 4-phenylbutanol in 50 mL of dichloromethane, and successively add 2.0 g molecular sieve and 12 mM pyridinium chlorochromate and react for 1 h. After monitoring the reaction to completion by TLC, the reaction solution is extracted and then distilled under reduced pressure. The obtained product is purified by a thin-layer silica gel chromatographic column to obtain Compound 1 (4-phenylbutanal); the yield is 70%.

[0035]

[0036] (ii) Dissolve 10 mM of Compound 1 in 60 mL of dichloromethane, add 12 mM ethoxycarbonylmethylene triphenylphosphine and react for 12 h. After monitoring the reaction to completion by TLC, the reaction solution is extracted and then distilled under reduced pressure. The obtained product is purified by a thin-layer silica gel chromatographic column to obtain Compound 2 ((E)-ethyl 6-phenylhex-2-enoate); the yield is 63%.

[0037]

[0038] (iii) Under nitrogen protection, 10 mM of Compound 2 was dissolved in 60 mL of dry dichloromethane, cooled to 0 °C, then 8 mM of oxalyl chloride was added dropwise, and then 15 mM of anhydrous aluminum chloride was added in batches. After reacting for 1 h and monitoring the reaction equilibrium by TLC, 50 mL of 2 M ice-cold hydrochloric acid was added to the reaction solution for quenching. The obtained product was purified by thin-layer silica gel chromatography column to obtain Compound 3 ((E)-4-[6-ethoxy-6-oxohex-4-enyl]benzoic acid); the yield was 40%.

[0039]

[0040] (iv) 10 mM of Compound 3 was dissolved in 50 mL of dry DMF. 15 mM of triethylamine and 15 mM of O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU) were added successively and stirred for 0.5 h. Then 11 mM of amine or alcohol compound R2-H was added and reacted for 2 - 24 h. After monitoring the end of the reaction by TLC, the reaction solution was extracted and then distilled under reduced pressure. The obtained product was purified by thin-layer silica gel chromatography column to obtain the target compound, and the yield was 22 - 96%.

[0041]

[0042] In the above reaction, when R2 in the amine or alcohol compound R2-H described in step (iv) is any one of n-propylamino, n-butylamino, n-pentylamino, n-hexylamino, 3-hydroxypropylamino, 4-hydroxybutylamino, 5-hydroxypentylamino, 6-hydroxyhexylamino, n-propoxy, n-butoxy, n-pentyloxy, n-hexyloxy, 3-hydroxypropoxy, 4-hydroxybutoxy, 5-hydroxypentyloxy or 6-hydroxyhexyloxy, the specific structures of the obtained target products are shown in Table 1.

[0043] Table 1 Specific structures of the series of compounds shown by Formula I-1 - I-16

[0044]

[0045]

[0046]

[0047] The characterization data of the series of Compound I shown by Formula I prepared by the above method are as follows:

[0048] Compound I-1: Colorless liquid, yield 96%; 11H NMR (600 MHz, Chloroform-d) δ 7.68 (d, J = 8.2 Hz, 2H, Ar-H), 7.22 (d, J = 8.0 Hz, 2H, Ar-H), 6.95 (dt, J = 15.6, 6.9 Hz, 1H, =CH), 6.12 (s, 1H, -NH), 5.82 (d, J = 15.6 Hz, 1H, =CH), 4.18 (q, J = 7.1 Hz, 2H, -CH2), 3.44–3.39 (m, 2H, -CH2), 2.68 (t, J = 7.7 Hz, 2H, -CH2), 2.26–2.19 (m, 2H, -CH2), 1.80 (p, J = 7.5 Hz, 2H, -CH2), 1.67–1.62 (m, 2H, -CH2), 1.29 (t, J = 7.1 Hz, 3H, -CH3), 0.99 (t, J = 7.4 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 167.40, 166.62, 148.38, 145.37, 132.66, 128.59, 126.99, 121.89, 60.24, 41.72, 34.67, 31.48, 29.37, 22.98, 14.28, 11.45. HRMS (ESI): m / z [M+Na] + calculated for C 18 H 25 NO3: 326.1727; found: 326.1739.

[0049] Compound I-2: Colorless liquid, yield 93%; 1 1H NMR (600 MHz, Chloroform-d) δ 7.68 (d, J = 7.9 Hz, 2H, Ar-H), 7.22 (d, J = 8.0 Hz, 2H, Ar-H), 6.95 (dt, J = 15.6, 6.9 Hz, 1H, =CH), 6.07 (s, 1H, -NH), 5.82 (d, J = 15.6 Hz, 1H, =CH), 4.18 (q, J = 7.1 Hz, 2H, -CH2), 3.48–3.42 (m, 2H, -CH2), 2.68 (t, J = 7.7 Hz, 2H, -CH2), 2.25–2.17 (m, 2H, -CH2), 1.80 (p, J = 7.5 Hz, 2H, -CH2), 1.62–1.57 (m, 2H, -CH2), 1.45–1.38 (m, 2H, -CH2), 1.29 (t, J = 7.1 Hz, 3H, -CH3), 0.96 (t, J = 7.4 Hz, 3H, -CH3). 1313C NMR (150 MHz, Chloroform-d) δ 167.37, 166.62, 148.38, 145.36, 132.66, 128.59, 126.99, 121.89, 60.24, 39.76, 35.03, 31.80, 31.48, 29.37, 20.17, 14.28, 13.80. HRMS (ESI): m / z [M+Na] + calculated for C 19 H 27 NO3: 340.1883; found: 340.1895.

[0050] Compound I-3: Colorless liquid, yield 89%; 1 1H NMR (600 MHz, Chloroform-d) δ 7.70 (d, J = 8.2 Hz, 2H, Ar-H), 7.25 (d, J = 8.1 Hz, 2H, Ar-H), 6.98 (dt, J = 15.6, 7.0 Hz, 1H, =CH), 6.07 (s, 1H, -NH), 5.85 (d, J = 15.6 Hz, 1H, =CH), 4.21 (q, J = 7.1 Hz, 2H, -CH2), 3.45–3.48 (m, 2H, -CH2), 2.70 (t, J = 7.7 Hz, 2H, -CH2), 2.27–2.22 (m, 2H, -CH2), 1.83 (p, J = 7.6 Hz, 2H, -CH2), 1.66–1.60 (m, 2H, -CH2), 1.41–1.36 (m, 4H, -CH2), 1.31 (t, J = 7.1 Hz, 3H, -CH3), 0.96–0.92 (t, J = 7.1 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 167.37, 166.62, 148.38, 145.34, 132.65, 128.57, 127.01, 121.88, 60.23, 40.04, 35.02, 31.47, 29.41, 29.36, 29.16, 22.40, 14.27, 14.00. HRMS (ESI): m / z [M+Na] + calculated for C 20 H 29 NO3: 354.2040; found: 354.2051.

[0051] Compound I-4: Colorless liquid, yield 92%; 11H NMR (600 MHz, Chloroform-d) δ 7.68 (d, J = 8.2 Hz, 2H, Ar-H), 7.23 (d, J = 8.2 Hz, 2H, Ar-H), 6.96 (dt, J = 15.7, 7.0 Hz, 1H, =CH), 6.05 (s, 1H, -NH), 5.82 (dt, J = 15.6, 1.6 Hz, 1H, =CH), 4.19 (q, J = 7.1 Hz, 2H, -CH2), 3.47–3.42 (m, 2H, -CH2), 2.68 (t, J = 7.7 Hz, 2H, -CH2), 2.25–2.20 (m, 2H, -CH2), 1.80 (p, J = 7.6 Hz, 2H, -CH2), 1.63–1.58 (m, 2H, -CH2), 1.41–1.36 (m, 2H, -CH2), 1.34–1.30 (m, 4H, -CH2), 1.29 (t, J = 7.1 Hz, 3H, -CH3), 0.90 (t, J = 7.0 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 167.34, 166.62, 148.37, 145.37, 132.67, 128.60, 126.98, 121.89, 60.24, 40.07, 35.03, 31.53, 31.48, 29.70, 29.37, 26.68, 22.58, 14.28, 14.03. HRMS (ESI): m / z [M+Na] + calculated for C 21 H 31 NO3: 368.2196;found: 368.2208.

[0052] Compound I-5: Colorless liquid, yield 88%; 11H NMR(600 MHz, DMSO-d6) δ 8.35 (t, J = 5.6 Hz, 1H, -NH), 7.75 (d, J = 8.2 Hz, 2H, Ar-H), 7.27 (d, J = 8.1 Hz, 2H, Ar-H), 6.90 (dt, J = 15.6, 6.9 Hz, 1H, =CH), 5.85 (d, J = 15.6 Hz, 1H, =CH), 4.46 (t, J = 5.2 Hz, 1H, -OH), 4.10 (q, J = 7.1 Hz, 2H, -CH2), 3.47–3.43 (m, 2H, -CH2), 3.31–3.27 (m, 2H, -CH2), 2.63 (t, J = 7.7 Hz, 2H, -CH2), 2.24–2.16 (m, 2H, -CH2), 1.73 (p, J = 7.6 Hz, 2H, -CH2), 1.68–1.63 (m, 2H, -CH2), 1.19 (t, J = 7.1 Hz, 3H, -CH3). 13 13C NMR(150 MHz, DMSO-d6) δ 171.30, 170.87, 154.27, 150.12, 137.51, 133.38, 132.41, 126.50, 64.92, 63.84, 41.73, 39.57, 37.67, 36.22, 34.08, 19.37. HRMS(ESI): m / z [M+Na] + calculated for C 18 H 25 NO4: 342.1675; found: 342.1666.

[0053] Compound I-6: Colorless liquid, yield 90%; 11H NMR (600 MHz, DMSO-d6) δ 8.35 (t, J = 5.7 Hz, 1H, -NH), 7.75 (d, J = 8.1 Hz, 2H, Ar-H), 7.27 (d, J = 8.1 Hz, 2H, Ar-H), 6.90 (dt, J = 15.6, 6.9 Hz, 1H, =CH), 5.85 (d, J = 15.6 Hz, 1H, =CH), 4.39 (t, J = 5.1 Hz, 1H, -OH), 4.10 (q, 2H, -CH2), 3.42–3.39 (m, 2H, -CH2), 3.26–3.21 (m, 2H, -CH2), 2.62 (t, J = 7.7 Hz, 2H, -CH2), 2.24–2.16 (m, 2H, -CH2), 1.73 (p, J = 7.5 Hz, 2H, -CH2), 1.56–1.50 (m, 2H, -CH2), 1.47–1.41 (m, 2H, -CH2), 1.19 (t, J = 7.1 Hz, 3H, -CH3). 13 13C NMR (150 MHz, DMSO-d6) δ 171.15, 170.87, 154.27, 150.07, 137.60, 133.36, 132.42, 126.50, 65.72, 64.92, 44.27, 39.57, 36.23, 35.24, 34.09, 31.10, 19.37. HRMS (ESI): m / z [M+Na] + calculated for C 19 H 27 NO4: 356.1832; found: 356.1822.

[0054] Compound I-7: Colorless liquid, yield 67%; 11H NMR (600 MHz, DMSO-d6) δ 8.34 (t, J = 5.7 Hz, 1H, -NH), 7.75 (d, J = 8.0 Hz, 2H, Ar-H), 7.27 (d, J = 8.0 Hz, 2H, Ar-H), 6.90 (dt, J = 15.6, 6.9 Hz, 1H, =CH), 5.86 (d, J = 15.7 Hz, 1H, =CH), 4.35 (t, J = 5.1 Hz, 1H, -OH), 4.10 (q, J = 7.1 Hz, 2H, -CH2), 3.40–3.37 (m, 2H, -CH2), 3.25–3.20 (m, 2H, -CH2), 2.63 (t, J = 7.7 Hz, 2H, -CH2), 2.24–2.17 (m, 2H, -CH2), 1.74 (p, J = 7.5 Hz, 2H, -CH2), 1.53–1.48 (m, 2H, -CH2), 1.46–1.41 (m, 2H, -CH2), 1.34–1.28 (m, 2H, -CH2), 1.20 (t, J = 7.1 Hz, 3H, -CH3). 13 13C NMR (150 MHz, DMSO-d6) δ 165.92, 165.66, 149.05, 144.86, 132.39, 128.15, 127.22, 121.29, 60.66, 59.71, 54.91, 34.37, 32.28, 31.02, 29.09, 28.88, 23.06, 14.16. HRMS (ESI): m / z [M+Na] + calculated for C 20 H 29 NO4: 370.1988; found: 370.1978.

[0055] Compound I-8: Colorless liquid, yield 95%; 11H NMR (600 MHz, DMSO-d6) δ 8.34 (t, J = 5.7 Hz, 1H, -NH), 7.75 (d, J = 8.0 Hz, 2H, Ar-H), 7.27 (d, J = 8.0 Hz, 2H, Ar-H), 6.90 (dt, J = 15.5, 6.9 Hz, 1H, =CH), 5.86 (d, J = 15.7 Hz, 1H, =CH), 4.33 (t, J = 5.2 Hz, 1H, -OH), 4.10 (q, J = 7.1 Hz, 2H, -CH2), 3.39–3.36 (m, 2H, -CH2), 3.25–3.20 (m, 2H, -CH2), 2.63 (t, J = 7.7 Hz, 2H, -CH2), 2.25–2.16 (m, 2H, -CH2), 1.74 (p, J = 7.5 Hz, 2H, -CH2), 1.53–1.47 (m, 2H, -CH2), 1.44–1.38 (m, 2H, -CH2), 1.33–1.27 (m, 4H, -CH2), 1.20 (t, J = 7.1 Hz, 3H, -CH3). 13 13C NMR (150 MHz, DMSO-d6) δ 171.14, 170.87, 154.26, 150.06, 137.60, 133.35, 132.42, 126.49, 65.88, 64.92, 45.27, 39.57, 37.71, 36.22, 34.44, 34.08, 31.63, 30.50, 19.37. HRMS (ESI): m / z [M+Na] + calculated for C 21 H 31 NO4: 384.2145; found: 384.2134.

[0056] Compound I-9: Colorless liquid, yield 22%; 1 1H NMR (600 MHz, Chloroform-d) δ 7.97 (d, J = 8.1 Hz, 2H, Ar-H), 7.23 (d, J = 7.9 Hz, 2H, Ar-H), 6.96 (dt, J = 15.6, 6.9 Hz, 1H, =CH), 5.83 (d, J = 15.6 Hz, 1H, =CH), 4.27 (t, J = 6.7 Hz, 2H, -CH2), 4.19 (q, J = 7.1 Hz, 2H, -CH2), 2.70 (t, J = 7.7 Hz, 2H, -CH2), 2.27–2.20 (m, 2H, -CH2), 1.85–1.76 (m, 4H, -CH2), 1.29 (t, J = 7.1 Hz, 3H, -CH3), 1.03 (t, J = 7.4 Hz, 3H, -CH3).13 13C NMR (150 MHz, Chloroform-d) δ 166.67, 166.61, 148.33, 147.08, 129.74, 128.42, 128.38, 121.92, 66.42, 60.24, 35.22, 31.50, 29.31, 22.14, 14.28, 10.54. HRMS (ESI): m / z [M+Na] + calculated for C 18 H 24 O4: 327.1567; found: 327.1567.

[0057] Compound I-10: Colorless liquid, yield 47%; 1 1H NMR (600 MHz, Chloroform-d) δ 7.96 (d, J = 7.8 Hz, 2H, Ar-H), 7.23 (d, J = 7.9 Hz, 2H, Ar-H), 6.96 (dt, J = 15.9, 6.9 Hz, 1H, =CH), 5.83 (d, J = 15.6 Hz, 1H, =CH), 4.31 (t, J = 6.7 Hz, 2H, -CH2), 4.19 (q, J = 7.1 Hz, 2H, -CH2), 2.69 (t, J = 7.7 Hz, 2H, -CH2), 2.28–2.18 (m, 2H, -CH2), 1.81 (p, J = 7.6 Hz, 2H, -CH2), 1.78–1.72 (m, 2H, -CH2), 1.51–1.45 (m, 2H, -CH2), 1.29 (t, J = 7.1 Hz, 3H, -CH3), 0.98 (t, J = 7.5 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 166.68, 166.61, 148.33, 147.07, 129.74, 128.42, 128.39, 121.92, 64.71, 60.24, 35.22, 31.50, 30.81, 29.30, 19.29, 14.28, 13.78. HRMS (ESI): m / z [M+Na] + calculated for C 19 H 26 O4: 341.1723; found: 341.1714.

[0058] Compound I-11: Colorless liquid, yield 45%; 11H NMR (600 MHz, Chloroform-d) δ 7.96 (d, J = 8.2 Hz, 2H, Ar-H), 7.23 (d, J = 8.0 Hz, 2H, Ar-H), 6.96 (dt, J = 15.6, 6.9 Hz, 1H, =CH), 5.83 (d, J = 15.7 Hz, 1H, =CH), 4.30 (t, J = 6.7 Hz, 2H, -CH2), 4.19 (q, J = 7.1 Hz, 2H, -CH2), 2.70 (t, J = 7.7 Hz, 2H, -CH2), 2.27–2.19 (m, 2H, -CH2), 1.81 (p, J = 7.6 Hz, 2H, -CH2), 1.79–1.74 (m, 2H, -CH2), 1.45–1.36 (m, 4H, -CH2), 1.29 (t, J = 7.1 Hz, 3H, -CH3), 0.93 (t, J = 7.0 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 166.68, 166.61, 148.33, 147.07, 129.74, 128.42, 128.40, 121.92, 65.01, 60.24, 35.22, 31.50, 29.30, 28.46, 28.22, 22.38, 14.28, 14.00. HRMS (ESI): m / z [M+Na] + calculated for C 20 H 28 O4: 355.1880; found: 355.1870.

[0059] Compound I-12: Colorless liquid, yield 37%; 11H NMR (600 MHz, Chloroform-d) δ 7.96 (d, J = 8.1 Hz, 2H, Ar-H), 7.23 (d, J = 8.0 Hz, 2H, Ar-H), 6.96 (dt, J = 15.6, 6.9 Hz, 1H, =CH), 5.83 (d, J = 15.6 Hz, 1H, =CH), 4.30 (t, J = 6.7 Hz, 2H, -CH2), 4.19 (q, J = 7.1 Hz, 2H, -CH2), 2.70 (t, J = 7.7 Hz, 2H, -CH2), 2.27–2.18 (m, 2H, -CH2), 1.81 (p, J = 7.6 Hz, 2H, -CH2), 1.78–1.72 (m, 2H, -CH2), 1.47–1.41 (m, 2H, -CH2), 1.38–1.31 (m, 4H, -CH2), 1.29 (t, J = 7.1 Hz, 3H, -CH3), 0.91 (t, J = 7.1 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 166.68, 166.61, 148.34, 147.07, 129.74, 128.42, 128.40, 121.92, 65.02, 60.24, 35.22, 31.50, 31.49, 29.31, 28.72, 25.73, 22.57, 14.28, 14.02. HRMS (ESI): m / z [M+Na] + calculated for C 21 H 30 O4: 369.2036; found: 369.2035.

[0060] Compound I-13: Colorless liquid, yield 42%; 11H NMR (600 MHz, Chloroform-d) δ 7.96 (d, J = 7.8 Hz, 2H, Ar-H), 7.24 (d, J = 7.8 Hz, 2H, Ar-H), 7.00–6.92 (m, 1H, =CH), 5.83 (d, J = 15.7 Hz, 1H, =CH), 4.48 (t, J = 6.1 Hz, 2H, -CH2), 4.19 (q, J = 7.1 Hz, 2H, -CH2), 3.79–3.75 (m, 2H, -CH2), 2.70 (t, J = 7.7 Hz, 2H, -CH2), 2.27–2.19 (m, 2H, -CH2), 2.03–1.98 (m, 2H, -CH2), 1.93 (t, J = 5.5 Hz, 1H, -OH), 1.82 (p, J = 7.6 Hz, 2H, -CH2), 1.29 (t, J = 7.1 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 167.00, 166.61, 148.28, 147.42, 129.83, 128.50, 127.92, 121.94, 61.63, 60.25, 59.22, 35.23, 31.97, 31.49, 29.28, 14.28. HRMS (ESI): m / z [M+Na] + calculated for C 18 H 24 O5: 343.1516; found: 343.1514.

[0061] Compound I-14: Colorless liquid, yield 32%; 1 1H NMR (600 MHz, Chloroform-d) δ 7.96 (d, J = 7.7 Hz, 2H, Ar-H), 7.24 (d, J = 7.8 Hz, 2H, Ar-H), 7.00–6.92 (m, 1H, =CH), 5.83 (d, J = 15.6 Hz, 1H, =CH), 4.35 (t, J = 6.5 Hz, 2H, -CH2), 4.19 (q, J = 7.1 Hz, 2H, -CH2), 3.73 (t, J = 6.5 Hz, 2H, -CH2), 2.70 (t, J = 7.7 Hz, 2H, -CH2), 2.29–2.16 (m, 2H, -CH2), 1.90–1.84 (m, 2H, -CH2), 1.81 (p, J = 7.5 Hz, 2H, -CH2), 1.77–1.70 (m, 2H, -CH2), 1.40 (s, 1H, -OH), 1.29 (t, J = 7.1 Hz, 3H, -CH3). 1313C NMR (150 MHz, Chloroform-d) δ 166.62, 166.61, 148.31, 147.21, 129.75, 128.46, 128.19, 121.92, 64.62, 62.46, 60.24, 35.22, 31.49, 29.29, 29.27, 25.26, 14.27. HRMS (ESI): m / z [M+Na] + calculated for C 19 H 26 O5: 357.1672; found: 357.1673.

[0062] Compound I-15: colorless liquid, yield 48%; 1 1H NMR (600 MHz, Chloroform-d) δ 7.96 (d, J = 8.1 Hz, 2H, Ar-H), 7.24 (d, J = 7.9 Hz, 2H, Ar-H), 6.96 (dt, J = 15.5, 6.9 Hz, 1H, =CH), 5.83 (d, J = 15.6 Hz, 1H, =CH), 4.32 (t, J = 6.6 Hz, 2H, -CH2), 4.19 (q, J = 7.1 Hz, 2H, -CH2), 3.71–3.65 (m, 2H, -CH2), 2.70 (t, J = 7.7 Hz, 2H, -CH2), 2.27–2.19 (m, 2H, -CH2), 1.85–1.77 (m, 4H, -CH2), 1.65 (p, J = 6.8 Hz, 2H, -CH2), 1.57–1.50 (m, 2H, -CH2), 1.32 (s, 1H, -OH), 1.29 (t, J = 7.1 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 166.65, 166.62, 148.33, 147.16, 129.75, 128.44, 128.26, 121.92, 64.75, 62.76, 60.24, 35.22, 32.35, 31.49, 29.29, 28.59, 22.37, 14.27. HRMS (ESI): m / z [M+Na] + calculated for C 20 H 28 O5: 371.1829; found: 371.1830. Compound I-16: colorless liquid, yield 38%; 11H NMR (600 MHz, Chloroform-d) δ 7.96 (d, J = 8.2 Hz, 2H, Ar-H), 7.24 (d, J = 8.0 Hz, 2H, Ar-H), 6.96 (dt, J = 15.5, 7.0 Hz, 1H, =CH), 5.83 (d, J = 15.6 Hz, 1H, =CH), 4.31 (t, J = 6.6 Hz, 2H, -CH2), 4.19 (q, J = 7.1 Hz, 2H, -CH2), 3.66 (t, J = 6.6 Hz, 2H, -CH2), 2.70 (t, J = 7.7 Hz, 2H, -CH2), 2.27–2.20 (m, 2H, -CH2), 1.85–1.75 (m, 4H, -CH2), 1.63–1.61 (m, 2H, -CH2), 1.61 (s, 1H, -OH), 1.52–1.41 (m, 4H, -CH2), 1.29 (t, J = 7.1 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 166.68, 166.62, 148.34, 147.14, 129.74, 128.44, 128.31, 121.92, 64.81, 62.85, 60.25, 35.22, 32.64, 31.49, 29.29, 28.74, 25.86, 25.43, 14.27. HRMS (ESI): m / z [M+Na] + calculated for C 21 H 30 O5: 385.1985; found: 385.1985.

[0063] Example 2 Preparation of Series II Compounds of Formula I

[0064] (v) Dissolve 10 mM of 4-phenylbutanol in 60 mL of dichloromethane, add 12 mM of acetyl chloride, react at room temperature for 5 h. After monitoring the reaction to completion by TLC, extract the reaction solution and distill it under reduced pressure to obtain Compound 4 (4-phenylbutyl acetate); yield 90%.

[0065]

[0066] (vi) Under nitrogen protection, dissolve 10 mM of Compound 4 in 60 mL of dry dichloromethane, cool to 0 °C, then dropwise add 8.5 mM of oxalyl chloride, and then add 18 mM of anhydrous aluminum chloride in batches. React for 1.5 h. After monitoring the reaction equilibrium by TLC, quench the reaction solution by adding 50 mL of 2 M ice-cold hydrochloric acid. The obtained product is purified by silica gel column chromatography to obtain Compound 5 (4-[4-(acetyloxy)butyl]benzoic acid); yield 40%.

[0067]

[0068] (vii) Dissolve 10 mM of Compound 5 in 60 mL of a mixed solvent of CH3OH:H2O = 3:1, add 12 mM of potassium carbonate and react for 24 h. After monitoring the completion of the reaction by TLC, extract the reaction solution and then distill it under reduced pressure to obtain Compound 6 (4-[4-(hydroxy)butyl]benzoic acid); the yield is 82%.

[0069]

[0070] (viii) Under nitrogen protection, dissolve 10 mM of Compound 6 in 80 mL of dichloromethane, and successively add 3.0 g molecular sieve and 15 mM of pyridinium chlorochromate and react for 3 h. After monitoring the completion of the reaction by TLC, extract the reaction solution and then distill it under reduced pressure. The obtained product is purified by silica gel column chromatography to obtain Compound 7 (4-[3-(formyl)propyl]benzoic acid); the yield is 17%.

[0071]

[0072] (ix) Dissolve 10 mM of dimethyl (2-oxo-2-phenylethyl)phosphonate in 50 mL of dry dichloromethane, add 12.5 mM of anhydrous potassium carbonate and stir for 1 h, then add 5 mM of Compound 7 and react for 24 h. After monitoring the end of the reaction by TLC, extract the reaction solution and then distill it under reduced pressure. The obtained product is purified by thin-layer silica gel column chromatography to obtain Compound 8 ((E)-4-[-6-oxo-6-phenylhex-4-enyl]benzoic acid); the yield is 72%.

[0073]

[0074] (x) Dissolve 10 mM of Compound 8 in 50 mL of dry DMF, successively add 15 mM of N,N-diisopropylethylamine (DIPEA) and 15 mM of O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU) and stir for 0.5 h, add 12 mM of amine or alcohol compound R2-H and react for 2 - 24 h. After monitoring the end of the reaction by TLC, extract the reaction solution and then distill it under reduced pressure. The obtained product is purified by silica gel column chromatography to obtain the target compound; the yield is 17 - 82%.

[0075]

[0076] In the above reaction, when R2 in the amine or alcohol compound R2-H described in step (x) is n-propylamino, n-butylamino, n-pentylamino, n-hexylamino, 3-hydroxypropylamino, 4-hydroxybutylamino, 5-hydroxypentylamino, 6-hydroxyhexylamino, n-propoxy, n-butoxy, n-pentyloxy, n-hexyloxy, 3-hydroxypropoxy, 4-hydroxybutoxy, 5-hydroxypentyloxy or 6-hydroxyhexyloxy respectively, the specific structures of the obtained target products are shown in Table 2.

[0077] Table 2 Specific structures of the target compounds of Formula II-1-II-16

[0078]

[0079]

[0080]

[0081] The characterization data of the series II compounds of Formula I prepared by the above method are as follows:

[0082] Compound II-1: Colorless liquid, yield 34%; 1 H NMR(600MHz,Chloroform-d)δ7.92(d,J=7.3Hz,2H,Ar-H),7.70(d,J=7.8Hz,2H,Ar-H),7.56(t,J=7.4Hz,1H,Ar-H),7.47(t,J=7.6Hz,2H,Ar-H),7.24(d,J=7.9Hz,2H,Ar-H),7.03–7.08(m,1H,=CH),6.88(d,J=15.4Hz,1H,=CH),6.13(s,1H,-NH),3.40–3.44(m,2H,-CH2),2.72(t,J=7.6Hz,2H,-CH2),2.33–2.37(m,2H,-CH2),1.88(p,J=7.5Hz,2H,-CH2),1.61–1.67(m,2H,-CH2),0.99(t,J=7.4Hz,3H,-CH3). 13 C NMR(150MHz,Chloroform-d)δ190.75,167.44,148.97,145.36,137.88,132.72,132.69,128.64,128.56,128.53,127.02,126.34,41.73,35.16,32.14,29.54,22.97,11.46.HRMS(ESI):m / z[M+Na] + calculated for C 22 H25 NO2: 358.1778; found: 326.1772.

[0083] Compound II-2: colorless liquid, yield 46%; 1 H NMR (600 MHz, Chloroform-d) δ 7.92 (d, J = 8.2 Hz, 2H, Ar-H), 7.69 (d, J = 7.7 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.6 Hz, 2H, Ar-H), 7.24 (d, J = 7.8 Hz, 2H, Ar-H), 7.03–7.08 (m, 1H, =CH), 6.88 (d, J = 15.4 Hz, 1H, =CH), 6.11 (s, 1H, -NH), 3.44–3.47 (m, 2H, -CH2), 2.72 (t, J = 7.7 Hz, 2H, -CH2), 2.33–2.36 (m, 2H, -CH2), 1.87 (p, J = 7.6 Hz, 2H, -CH2), 1.57–1.62 (m, 2H, -CH2), 1.40–1.43 (m, 2H, -CH2), 0.96 (t, J = 7.4 Hz, 3H, -CH3). 13 C NMR (150 MHz, Chloroform-d) δ 190.74, 167.40, 148.97, 145.35, 137.88, 132.72, 132.70, 128.63, 128.56, 128.53, 127.02, 126.34, 39.78, 35.16, 32.14, 31.79, 29.55, 20.17, 13.80. HRMS (ESI): m / z [M+Na] + calculated for C 23 H 27 NO2: 372.1934; found: 372.1928.

[0084] Compound II-3: colorless liquid, yield 51%; 11H NMR (600 MHz, Chloroform-d) δ 7.92 (d, J = 7.3 Hz, 2H, Ar-H), 7.69 (d, J = 7.8 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.6 Hz, 2H, Ar-H), 7.24 (d, J = 7.9 Hz, 2H, Ar-H), 7.06 (dt, J = 15.2, 6.9 Hz, 1H, =CH), 6.88 (d, J = 15.4 Hz, 1H, =CH), 6.12 (s, 1H, -NH), 3.43–3.46 (m, 2H, -CH2), 2.72 (t, J = 7.6 Hz, 2H, -CH2), 2.33–2.37 (m, 2H, -CH2), 1.87 (p, J = 7.6 Hz, 2H, -CH2), 1.60–1.63 (m, 2H, -CH2), 1.35–1.38 (m, 4H, -CH2), 0.91 (t, J = 7.1 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 190.75, 167.41, 148.98, 145.36, 137.87, 132.73, 132.69, 128.63, 128.56, 128.53, 127.03, 126.34, 40.06, 35.16, 32.15, 29.54, 29.41, 29.16, 22.41, 14.01. HRMS (ESI): m / z [M+Na] + calculated for C 24 H 29 NO2: 386.2091; found: 386.2084.

[0085] Compound II-4: Colorless liquid, yield 18%; 11H NMR (600 MHz, Chloroform-d) δ 7.92 (d, J = 7.0 Hz, 2H, Ar-H), 7.69 (d, J = 8.0 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.6 Hz, 2H, Ar-H), 7.24 (d, J = 7.9 Hz, 2H, Ar-H), 7.06 (dt, J = 15.5, 7.0 Hz, 1H, =CH), 6.88 (d, J = 15.4 Hz, 1H, =CH), 6.09 (s, 1H, -NH), 3.43–3.46 (m, 2H, -CH2), 2.72 (t, J = 7.7 Hz, 2H, -CH2), 2.38–2.31 (m, 2H, -CH2), 1.88 (p, J = 7.5 Hz, 2H, -CH2), 1.58–1.63 (m, 2H, -CH2), 1.41–1.37 (m, 2H, -CH2), 1.30–1.34 (m, 4H, -CH2), 0.89 (t, J = 7.1 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 190.74, 167.38, 148.96, 145.35, 137.88, 132.72, 128.64, 128.56, 128.53, 128.07, 127.02, 126.34, 40.08, 35.16, 32.15, 31.52, 29.69, 29.55, 26.69, 22.58, 14.03. HRMS (ESI): m / z [M+Na] + calculated for C 25 H 31 NO2: 400.2247; found: 400.2240.

[0086] Compound II-5: Colorless liquid, yield 67%; 11H NMR (600 MHz, Chloroform-d) δ 7.91 (d, J = 7.0 Hz, 2H, Ar-H), 7.71 (d, J = 7.9 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.6 Hz, 2H, Ar-H), 7.25 (d, J = 7.9 Hz, 2H, Ar-H), 7.05 (dt, J = 15.6, 6.9 Hz, 1H, =CH), 6.88 (d, J = 15.4 Hz, 1H, =CH), 6.69 (s, 1H, -NH), 3.71 (t, J = 5.5 Hz, 2H, -CH2), 3.66–3.60 (m, 2H, -CH2), 3.25 (s, 1H, -OH), 2.72 (t, J = 7.6 Hz, 2H, -CH2), 2.37–2.31 (m, 2H, -CH2), 1.87 (p, J = 7.5 Hz, 2H, -CH2), 1.82–1.76 (m, 2H, -CH2). 13 13C NMR (150 MHz, Chloroform-d) δ 190.78, 168.45, 148.96, 145.72, 137.85, 132.75, 131.97, 128.70, 128.57, 128.53, 127.13, 126.35, 59.74, 37.06, 35.17, 32.25, 32.14, 29.52. HRMS (ESI): m / z [M+Na] + calculated for C 22 H 25 NO3: 374.1727; found: 374.1721.

[0087] Compound II-6: Colorless liquid, yield 51%; 11H NMR (600 MHz, Chloroform-d) δ 7.92 (d, J = 6.9 Hz, 2H, Ar-H), 7.70 (d, J = 8.1 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.8 Hz, 2H, Ar-H), 7.24 (d, J = 8.4 Hz, 2H, Ar-H), 7.05 (dt, J = 15.5, 7.0 Hz, 1H, =CH), 6.88 (d, J = 15.4 Hz, 1H, =CH), 6.48 (s, 1H, -NH), 3.73 (t, J = 6.0 Hz, 2H, -CH2), 3.53–3.47 (m, 2H, -CH2), 2.72 (t, J = 7.6 Hz, 2H, -CH2), 2.38–2.31 (m, 2H, -CH2), 1.87 (p, J = 7.5 Hz, 2H, -CH2), 1.76–1.71 (m, 2H, -CH2), 1.69–1.66 (m, 2H, -CH2). 13 13C NMR (150 MHz, Chloroform-d) δ 190.78, 167.54, 149.01, 145.42, 137.87, 132.73, 132.53, 128.65, 128.56, 128.53, 127.06, 126.34, 62.52, 39.75, 35.16, 32.15, 29.80, 29.53, 26.36. HRMS (ESI): m / z + calculated for C 23 H 27 NO3: 388.1883; found: 388.1876.

[0088] Compound II-7: Colorless liquid, yield 82%; 11H NMR (600 MHz, Chloroform-d) δ 7.92 (d, J = 7.1 Hz, 2H, Ar-H), 7.69 (d, J = 7.9 Hz, 2H, Ar-H), 7.56 (t, J = 7.3 Hz, 1H, Ar-H), 7.47 (t, J = 7.6 Hz, 2H, Ar-H), 7.24 (d, J = 7.9 Hz, 2H, Ar-H), 7.05 (dt, J = 15.4, 6.9 Hz, 1H, =CH), 6.88 (d, J = 15.4 Hz, 1H, =CH), 6.22 (s, 1H., -NH), 3.66 (t, J = 6.4 Hz, 2H, -CH2), 3.50–3.43 (m, 2H, -CH2), 2.72 (t, J = 7.6 Hz, 2H, -CH2), 2.37–2.32 (m, 2H, -CH2), 1.87 (p, J = 7.5 Hz, 2H, -CH2), 1.67–1.60 (m, 4H, -CH2), 1.51–1.44 (m, 2H, -CH2). 13 13C NMR (150 MHz, Chloroform-d) δ 190.77, 167.52, 148.99, 145.43, 137.87, 132.73, 132.55, 128.65, 128.56, 128.53, 127.05, 126.34, 62.66, 39.87, 35.16, 32.20, 32.15, 29.54, 29.47, 23.14. HRMS (ESI): m / z [M+Na] + calculated for C 24 H 29 NO3: 402.2040; found: 388.1876.

[0089] Compound II-8: Colorless liquid, yield 74%; 11H NMR (600 MHz, Chloroform-d) δ 7.92 (d, J = 7.7 Hz, 2H, Ar-H), 7.69 (d, J = 7.9 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.6 Hz, 2H, Ar-H), 7.24 (d, J = 7.9 Hz, 2H, Ar-H), 7.06 (dt, J = 14.4, 6.9 Hz, 1H, =CH), 6.88 (d, J = 15.4 Hz, 1H, =CH), 6.16 (s, 1H, -NH), 3.64 (t, J = 6.5 Hz, 2H, -CH2), 3.48–3.43 (m, 2H, -CH2), 2.72 (t, J = 7.6 Hz, 2H, -CH2), 2.38–2.32 (m, 2H, -CH2), 1.87 (p, J = 7.5 Hz, 2H, -CH2), 1.65–1.61 (m, 2H, -CH2), 1.60–1.56 (m, 2H, -CH2), 1.46–1.38 (m, 4H, -CH2). 13 13C NMR (150 MHz, Chloroform-d) δ 190.76, 167.49, 148.98, 145.42, 137.87, 132.73, 132.59, 128.65, 128.56, 128.53, 127.04, 126.34, 62.69, 39.81, 35.16, 32.55, 32.15, 29.71, 29.54, 26.56, 25.30. HRMS (ESI): m / z [M+Na] + calculated for C 25 H 31 NO3: 416.2196; found: 416.2190.

[0090] Compound II-9: Colorless liquid, yield 28%; 11H NMR (600 MHz, Chloroform-d) δ 7.98 (d, J = 8.4 Hz, 2H, Ar-H), 7.92 (d, J = 7.7 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.6 Hz, 2H, Ar-H), 7.26 (d, J = 7.8 Hz, 2H, Ar-H), 7.10–7.04 (m, 1H, =CH), 6.89 (d, J = 15.3 Hz, 1H, =CH), 4.27 (t, J = 6.5 Hz, 2H, -CH2), 2.74 (t, J = 7.7 Hz, 2H, -CH2), 2.39–2.32 (m, 2H, -CH2), 1.89 (p, J = 7.6 Hz, 2H, -CH2), 1.82–1.76 (m, 2H, -CH2), 1.03 (t, J = 7.4 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 190.73, 166.68, 148.91, 147.06, 137.89, 132.71, 129.77, 128.56, 128.53, 128.46, 128.42, 126.37, 66.44, 35.35, 32.16, 29.48, 22.15, 10.54. HRMS (ESI): m / z [M+Na] + calculated for C 22 H 24 O3: 359.1618; found: 359.1618.

[0091] Compound II-10: Colorless liquid, yield 48%; 11H NMR (600 MHz, Chloroform-d) δ 7.97 (d, J = 8.0 Hz, 2H, Ar-H), 7.92 (d, J = 7.4 Hz, 2H, Ar-H), 7.56 (t, J = 7.3 Hz, 1H, Ar-H), 7.47 (t, J = 7.6 Hz, 2H, Ar-H), 7.25 (d, J = 7.8 Hz, 2H, Ar-H), 7.07 (dt, J = 15.1, 6.9 Hz, 1H, =CH), 6.89 (d, J = 15.4 Hz, 1H, =CH), 4.32 (t, J = 6.6 Hz, 2H, -CH2), 2.74 (t, J = 7.7 Hz, 2H, -CH2), 2.40–2.31 (m, 2H, -CH2), 1.89 (p, J = 7.5 Hz, 2H, -CH2), 1.79–1.71 (m, 2H, -CH2), 1.52–1.45 (m, 2H, -CH2), 0.98 (t, J = 7.4 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 190.73, 166.68, 148.91, 147.06, 137.89, 132.71, 129.77, 128.56, 128.53, 128.45, 128.43, 126.37, 64.73, 35.35, 32.16, 30.81, 29.48, 19.29, 13.78. HRMS (ESI): m / z [M+Na] + calculated for C 23 H 26 O3: 373.1774; found: 373.1774.

[0092] Compound II-11: Colorless liquid, yield 31%; 11H NMR (600 MHz, Chloroform-d) δ 7.97 (d, J = 8.2 Hz, 2H, Ar-H), 7.92 (d, J = 6.9 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.7 Hz, 2H, Ar-H), 7.26 (d, J = 6.4 Hz, 2H, Ar-H), 7.07 (dt, J = 15.4, 6.9 Hz, 1H, =CH), 6.89 (d, J = 15.4 Hz, 1H, =CH), 4.30 (t, J = 6.7 Hz, 2H, -CH2), 2.74 (t, J = 7.7 Hz, 2H, -CH2), 2.38–2.33 (m, 2H, -CH2), 1.89 (p, J = 7.5 Hz, 2H, -CH2), 1.79–1.74 (m, 2H, -CH2), 1.46–1.36 (m, 4H, -CH2), 0.93 (t, J = 7.0 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 190.73, 166.68, 148.90, 147.05, 137.89, 132.71, 129.77, 128.55, 128.53, 128.46, 128.44, 126.37, 65.03, 35.36, 32.16, 29.48, 28.46, 28.22, 22.38, 14.01. HRMS (ESI): m / z [M+Na] + calculated for C 24 H 28 O3: 387.1931; found: 387.1931.

[0093] Compound II-12: Colorless liquid, yield 17%; 11H NMR (600 MHz, Chloroform-d) δ 7.97 (d, J = 8.1 Hz, 2H, Ar-H), 7.92 (d, J = 7.0 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.7 Hz, 2H, Ar-H), 7.26 (d, J = 6.8 Hz, 2H, Ar-H), 7.07 (dt, J = 15.5, 6.9 Hz, 1H, =CH), 6.89 (d, J = 15.4 Hz, 1H, =CH), 4.30 (t, J = 6.7 Hz, 2H, -CH2), 2.74 (t, J = 7.7 Hz, 2H, -CH2), 2.40–2.31 (m, 2H, -CH2), 1.89 (p, J = 7.6 Hz, 2H, -CH2), 1.78–1.73 (m, 2H, -CH2), 1.47–1.42 (m, 2H, -CH2), 1.37–1.32 (m, 4H, -CH2), 0.91 (t, J = 7.0 Hz, 3H, -CH3). 13 13C NMR (150 MHz, Chloroform-d) δ 190.72, 166.68, 148.90, 147.05, 137.89, 132.71, 129.80, 129.77, 128.56, 128.53, 128.46, 126.37, 65.04, 35.35, 32.16, 31.49, 29.48, 28.72, 25.73, 22.57, 14.02. HRMS (ESI): m / z [M+Na] + calculated for C 25 H 30 O3: 401.2087; found: 401.2087.

[0094] Compound II-13: Colorless liquid, yield 17%; 11H NMR (600 MHz, Chloroform-d) δ 7.97 (d, J = 8.2 Hz, 2H, Ar-H), 7.92 (d, J = 8.2 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.7 Hz, 2H, Ar-H), 7.27–7.25 (m, 2H, Ar-H), 7.07 (dt, J = 15.4, 6.9 Hz, 1H, =CH), 6.89 (d, J = 15.4 Hz, 1H, =CH), 3.71 (t, J = 5.5 Hz, 2H, -CH2), 3.66–3.60 (m, 2H, -CH2), 3.25 (s, 1H, -OH), 2.72 (t, J = 7.6 Hz, 2H, -CH2), 2.37–2.31 (m, 2H, -CH2), 1.87 (p, J = 7.5 Hz, 2H, -CH2), 1.82–1.76 (m, 2H, -CH2). 13 13C NMR (150 MHz, Chloroform-d) δ 190.73, 168.56, 148.88, 147.70, 137.85, 132.75, 131.97, 128.70, 128.57, 128.53, 127.13, 126.35, 59.74, 37.06, 35.17, 32.25, 32.14, 29.52. HRMS (ESI): m / z [M+Na] + calculated for C 22 H 24 O4: 375.1567; found: 375.1566.

[0095] Compound II-14: Colorless liquid, yield 21%; 1 1H NMR (600 MHz, Chloroform-d) δ 7.97 (d, J = 8.2 Hz, 2H, Ar-H), 7.92 (d, J = 8.2 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.7 Hz, 2H, Ar-H), 7.27–7.25 (m, 2H, Ar-H), 7.07 (dt, J = 15.4, 6.9 Hz, 1H, =CH), 6.89 (d, J = 15.4 Hz, 1H, =CH), 4.36 (t, J = 6.5 Hz, 2H, -CH2), 3.77–3.70 (m, 2H, -CH2), 2.74 (t, J = 7.6 Hz, 2H, -CH2), 2.39–2.32 (m, 2H, -CH2), 1.93–1.83 (m, 4H, -CH2), 1.77–1.71 (m, 2H, -CH2).13 13C NMR (150 MHz, Chloroform-d) δ 190.73, 166.62, 148.89, 147.20, 137.88, 132.72, 129.78, 128.56, 128.53, 128.50, 128.24, 126.38, 64.63, 62.48, 35.36, 32.16, 29.47, 29.28, 25.26. HRMS (ESI): m / z [M+Na] + calculated for C 23 H 26 O4: 389.1723; found: 389.1724.

[0096] Compound II-15: colorless liquid, yield 29%; 1 1H NMR (600 MHz, Chloroform-d) δ 7.97 (d, J = 8.0 Hz, 2H, Ar-H), 7.92 (d, J = 7.8 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.6 Hz, 2H, Ar-H), 7.27–7.25 (m, 2H, Ar-H), 7.07 (dt, J = 14.4, 6.9 Hz, 1H, =CH), 6.89 (d, J = 15.4 Hz, 1H, =CH), 3.66 (t, J = 6.4 Hz, 2H, -CH2), 3.50–3.43 (m, 2H, -CH2), 2.72 (t, J = 7.6 Hz, 2H, -CH2), 2.37–2.32 (m, 2H, -CH2), 1.87 (p, J = 7.5 Hz, 2H, -CH2), 1.67–1.60 (m, 4H, -CH2), 1.51–1.44 (m, 2H, -CH2). 13 13C NMR (150 MHz, Chloroform-d) δ 190.74, 167.64, 148.89, 147.13, 137.87, 132.73, 129.77, 128.55, 128.56, 128.53, 128.29, 126.37, 62.66, 39.87, 35.16, 32.20, 32.15, 29.54, 29.47, 23.14. HRMS (ESI): m / z [M+Na] + calculated for C 24 H 28 O4: 403.1880; found: 403.1879.

[0097] Compound II-16: colorless liquid, yield 23%;1 1H NMR (600 MHz, Chloroform-d) δ 7.97 (d, J = 8.0 Hz, 2H, Ar-H), 7.92 (d, J = 7.8 Hz, 2H, Ar-H), 7.56 (t, J = 7.4 Hz, 1H, Ar-H), 7.47 (t, J = 7.6 Hz, 2H, Ar-H), 7.27–7.25 (m, 2H, Ar-H), 7.07 (dt, J = 14.4, 6.9 Hz, 1H, =CH), 6.89 (d, J = 15.4 Hz, 1H, =CH), 4.31 (t, J = 6.6 Hz, 2H, -CH2), 3.66 (t, J = 6.6 Hz, 2H, -CH2), 2.74 (t, J = 7.7 Hz, 2H, -CH2), 2.39–2.32 (m, 2H, -CH2), 1.89 (p, J = 7.6 Hz, 2H, -CH2), 1.81–1.76 (m, 2H, -CH2), 1.63–1.59 (m, 2H, -CH2), 1.51–1.42 (m, 4H, -CH2). 13 13C NMR (150 MHz, Chloroform-d) δ 190.72, 166.65, 148.88, 147.11, 137.88, 132.70, 129.76, 128.55, 128.52, 128.47, 128.35, 126.37, 64.81, 62.85, 35.35, 32.63, 32.15, 29.47, 28.74, 25.86, 25.42. HRMS (ESI): m / z [M+Na] + calculated for C 25 H 30 O4: 417.2036; found: 417.2035.

[0098] Example 3 Determination of the anti-proliferative activity of the compounds of the present invention against cancer cells

[0099] 1. Experimental materials

[0100] The target compound and positive controls 2-BP and cisplatin, trypsin digestion solution, PBS buffer, fetal bovine serum, human breast cancer cell line MCF-7, human colon cancer cell line HCT116, mouse colon cancer cell line MC38, mouse hepatoma cell line Hepa1-6, microplate reader and 96-well plates, high-glucose DMEM medium and RPMI 1640 medium, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reagent, etc.

[0101] 2. Experimental method

[0102] The antitumor activities of the compounds against MCF-7, HCT116, MC38, and Hepa1-6 cells were detected by the MTT method. First, the cells were diluted to 4×10 4 cells / mL, and then seeded into 96-well plates, with 100 μL of the diluted cells added to each well. After incubation for 24 h, drugs at different concentrations were added to the wells. Three parallel groups were set for each concentration. After incubation at 37 °C for 48 h, MTT reagent (10 μL, 5 mg / mL) was added to each well of the culture plate (containing 100 μL of medium), and after further incubation for 3 h, the supernatant was aspirated with a syringe, 100 μL of DMSO was added to each well, and finally the absorbance was read at 570 nm using a Bio-Tek microplate reader.

[0103] The inhibition rate of the compound against the cells was calculated as follows:

[0104] Inhibition rate = (1 - A / A0) × 100%;

[0105] where A is the absorbance value measured after cell administration, and A0 is the absorbance value measured in the negative control.

[0106] Analysis was performed using the software GraphPad Prism 8.0.1. The logarithm of the sample concentration was linearly regressed against the cell inhibition rate, and the half-maximal inhibitory concentration IC 50 value (mean ± standard deviation) of the compound against each cell was calculated. The lower the half-maximal inhibitory concentration IC 50 value, the stronger the inhibitory ability of the compound against cancer cells. The detection results are shown in Tables 3 and 4.

[0107] Table 3 Comparison of IC 50 values of Series I compounds and control drugs against different cancer cells

[0108]

[0109]

[0110] Table 4 Comparison of IC 50 values of Series II compounds and control drugs against different cancer cells

[0111]

[0112] The experimental results showed that the series of compounds synthesized in this invention had good anti-proliferative activities against MCF-7 and HCT116 cells, and some compounds exhibited better anti-proliferative activities than the positive control.

[0113] Example 4 Determination of the inhibitory ability of the compounds of the present invention against DHHC activity

[0114] 1. Experimental materials

[0115] Target compounds, positive control 2-BP, dodecyl-β-D-maltoside (DDM) solution, palmitoyl-CoA solution, FAM-Kras4a solution, MES buffer, TCEP solution, DHHC protein, etc.

[0116] 2. Experimental methods

[0117] The inhibitory activity of the compound against DHHC enzyme was determined by the change in fluorescence anisotropy caused by the level of palmitoylation of the fluorescent substrate. First, 600 μL of DDM solution (2.5 mM), 120 μL of MES buffer (500 mM), 20 μL of TCEP solution (30 mM), and 6 μL of DHHC protein (3.5 mM) were mixed to prepare buffer A, and 100 μL of DDM solution (2.5 mM), 16.5 μL of palmitoyl-CoA solution (1.5 mM), and 25 μL of FAM-Kras4a solution (0.5 mM) were mixed to prepare buffer B. Then, 13.5 μL of buffer A and 6.5 μL of buffer B were added to each well of a 384-well plate, different concentrations of the drug were added to each well, and then incubated in the dark for 30 min. After the incubation, the detection was immediately carried out using a microplate reader.

[0118] The inhibition rate of the compound on cells was calculated as follows:

[0119] Inhibition rate = (1 - B0 / B) × 100%;

[0120] where B0 is the fluorescence anisotropy value measured for the negative control, and B is the fluorescence anisotropy value measured after drug administration.

[0121] Analysis was performed using the software GraphPad Prism 8.0.1. The logarithm of the sample concentration was linearly regressed with the protein inhibition rate, and the half-maximal inhibitory concentration IC50 value (mean ± standard deviation) of some compounds against DHHC protein was calculated. The lower the half-maximal inhibitory concentration IC 50 value, the stronger the inhibitory ability of the compound against DHHC protein. The detection results are shown in Table 4.

[0122] The experimental results show that the series of compounds synthesized in the present invention have good enzyme inhibitory activity against DHHC protein, and some compounds exhibit better enzyme inhibitory activity than the positive control.

[0123] Table 4 Comparison of the inhibition rate of the compounds of the present invention and the control drug against DHHC protein at 50 μM and the IC 50 value

[0124]

[0125] In summary, as demonstrated by the above embodiments, the (E)-6-(4-oxophenyl)hex-2-en-1-one compounds of the present invention have different degrees of anti-proliferative activity against different cancer cells and different degrees of enzyme inhibitory activity against DHHC proteins, and can be further developed into a new generation of anti-tumor drugs.

Claims

1. The compound shown in Formula I: Among them, R1 is an alkoxy group having 1 to 4 carbon atoms, or phenyl, or phenyl substituted with an alkyl group having 1 to 2 carbon atoms; R2 is an amino group substituted with a straight-chain alkyl group having 1 to 8 carbon atoms, or an amino group substituted with a straight-chain alkyl group having 1 to 8 carbon atoms and containing a hydroxyl group, or a straight-chain alkoxy group having 1 to 8 carbon atoms, or a straight-chain alkoxy group having 1 to 8 carbon atoms and containing a hydroxyl group.

2. The compound according to claim 1, wherein: R1 is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or sec-butoxy; or phenyl, or methylphenyl, or ethylphenyl, or dimethylphenyl; R2 is an amino group substituted with a straight-chain alkyl group having 2 to 7 carbon atoms, or an amino group substituted with a straight-chain alkyl group having 2 to 7 carbon atoms and having a hydroxyl group at the end, or a straight-chain alkoxy group having 2 to 7 carbon atoms, or a straight-chain alkoxy group having 2 to 7 carbon atoms and having a hydroxyl group at the end.

3. The compound shown in Formula I: Among them, R1 is ethoxy or phenyl, and R2 is n-propylamino, n-butylamino, n-pentylamino, n-hexylamino, 3-hydroxypropylamino, 4-hydroxybutylamino, 5-hydroxypentylamino, 6-hydroxyhexylamino, n-propoxy, n-butoxy, n-pentoxy, n-hexoxy, 3-hydroxypropoxy, 4-hydroxybutoxy, 5-hydroxypentoxy or 6-hydroxyhexoxy.

4. The preparation method of the compound according to claim 3, characterized in that, When R1 is ethoxy, the synthesis route is as follows: Step i: 4-Phenyl-1-butanol reacts with pyridinium dichromate in dichloromethane to form Compound 1; Step ii: Compound 1 reacts with ethoxycarbonylmethylene triphenylphosphine in dichloromethane to form Compound 2; Step iii: Compound 2 reacts with oxalyl chloride in dichloromethane in an ice-water bath in the presence of anhydrous aluminum chloride to form Compound 3; Step iv: Compound 3 undergoes a condensation reaction with R2-H in the presence of O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate and triethylamine to form Compounds I-1–I-16; When R1 is phenyl, the synthesis route is as follows: Step v: 4-Phenyl-1-butanol reacts with acetyl chloride in dichloromethane to form Compound 4; Step vi: Compound 4 reacts with oxalyl chloride in dichloromethane in an ice-water bath in the presence of anhydrous aluminum chloride to form Compound 5; Step vii: Compound 5 reacts in a mixed solvent of methanol and water in the presence of potassium carbonate to form Compound 6; Step viii: Compound 6 reacts with pyridinium dichromate in dichloromethane to form Compound 7; Step ix: Compound 7 reacts with dimethyl (2-oxo-2-phenylethyl)phosphonate in dichloromethane in the presence of potassium carbonate to form Compound 8; Step x: Compound 8 undergoes a condensation reaction with R2-H in the presence of O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate and N,N-diisopropylethylamine to form Compounds II-1–II-16.

5. The method according to claim 4, characterized in that, In step i, the molar ratio of 4-phenyl-1-butanol to pyridinium chlorochromate is 1:1.2; in step ii, the molar ratio of compound 1 to ethoxycarbonylmethylenetriphenylphosphine is 1:1.2; in step iii, the molar ratio of compound 2 to oxalyl chloride is 1:0.8; in step iv, the molar ratio of compound 3 to R2-H is 1:1.

1.

6. The method according to claim 4, wherein In step v, the molar ratio of 4-phenyl-1-butanol to acetyl chloride is 1:1.2; in step vi, the molar ratio of compound 4 to oxalyl chloride is 1:0.85; in step vii, the molar ratio of compound 5 to potassium carbonate is 1:1.2, and the ratio of methanol to water in the mixed solvent is 3:1; in step viii, the molar ratio of compound 6 to pyridinium chlorochromate is 1:1.5; in step ix, the molar ratio of compound 7 to dimethyl (2-oxo-2-phenylethyl)phosphonate is 1:2; in step x, the molar ratio of compound 8 to R2-H is 1:1.

2.

7. Use of the compound according to claim 1, 2 or 3 in the preparation of an anti-tumor drug.

8. The application according to claim 7, wherein The tumor is human breast cancer cell MCF-7, human colon cancer cell HCT116, mouse colon cancer cell MC38 or mouse liver cancer cell Hepa1-6.

9. A palmitoyltransferase inhibitor, characterized in that, Comprising the compound according to claim 1, 2 or 3 or a pharmaceutically acceptable salt thereof as an active ingredient, and one or more pharmaceutically acceptable carriers and / or excipients.

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