CN109134350A - Donepezil-BHT heterozygote, preparation method and its for treating Alzheimer's disease - Google Patents

Abstract

The invention relates to the field of medicinal chemistry, in particular to a class of donepezil-BHT hybrids, which are used as multifunctional acetylcholinesterase (AChE) inhibitors and monoamine oxidase B (MAO-B) inhibitors, and have good in vitro anti- Oxidation, for hydrogen peroxide (H ₂ O ₂ )-induced PC12 neuronal oxidative damage and bacterial lipopolysaccharide (LPS)-stimulated BV-2 inflammatory damage, it has a good neuroprotective effect, and it has been proved in animal experiments. The effect of improving cognitive function can be considered as a candidate drug for the treatment of Alzheimer's disease. The most preferred compound structure is as follows:

Description

Donepezil-BHT hybrid, its preparation method and its use in the treatment of Alzheimer's disease mutism

technical field

The invention relates to the field of medicinal chemistry, in particular to a class of donepezil-BHT hybrids, which are used as multifunctional acetylcholinesterase (AChE) inhibitors and monoamine oxidase B (MAO-B) inhibitors, and have good in vitro resistance Oxidation, has a good neuroprotective effect on the oxidative damage of PC12 nerve cells induced by hydrogen peroxide (H ₂ O ₂ ) and the inflammatory damage of BV-2 stimulated by bacterial lipopolysaccharide (LPS). Good cognitive function improvement can be considered as a candidate drug for the treatment of Alzheimer's disease.

Background technique

Alzheimer's disease (AD), also known as senile dementia, is a common neurodegenerative disease in the elderly. The pathogenesis of AD is complex. So far, the pathological mechanism of Alzheimer's disease is not fully understood. Its causes are not only related to the reduction of choline levels in the brain, but also to oxidative stress, neuroinflammation, and amyloid. (Aβ) aggregation, metal ion metabolism disorders and calcium balance disorders and other factors are closely related. Among them, the cholinergic hypothesis, that is, changes in the cholinergic system are closely related to the degree of cognitive impairment in Alzheimer's disease. Based on this theory, a lot of research has been done on agonists of acetylcholine receptors and inhibitors of acetylcholinesterase (AChE). In addition, the main solution to the problem of oxidative stress in the brain is to develop antioxidants, and to reduce the damage to nerve cells caused by oxidative stress and exert neuroprotective effects by inhibiting monoamine oxidase.

With the continuous elucidation of the pathogenic mechanism of Alzheimer's disease, we found that the occurrence and development of Alzheimer's disease is the result of the multiple actions of complex regulatory networks and regulatory factors in the organism, involving the association between multiple genes. Drugs targeting a single target can often only regulate one of the physiological pathways, but cannot fundamentally inhibit the pathological process of Alzheimer's disease. Therefore, finding drugs targeting multiple targets or having multiple therapeutic effects has become a new trend in drug development for Alzheimer's disease.

As an acetylcholinesterase inhibitor approved by the FDA for the treatment of patients with moderate Alzheimer's disease, donepezil's obvious cholinesterase inhibitory activity and the advantages of no obvious toxic and side effects have attracted the attention of the majority of researchers. Develop application prospects. 2,6-Di-tert-butyl-4-methylphenol (BHT), as a food additive commonly used in Europe and the United States, has attracted widespread attention because of its non-toxicity and good antioxidant activity, and its derivatives have been found in recent years. Has good anti-inflammatory activities. Therefore, the present invention designs and synthesizes a series of donepezil-BHT hybrids as multifunctional anti-Alzheimer's disease preparations. These _compounds are multifunctional acetylcholinesterase inhibitors and monoamine oxidase B (MAO _- B) inhibitors, and have good antioxidative effects in vitro. Bacterial lipopolysaccharide (LPS) stimulates BV-2 inflammatory injury, has a good neuroprotective effect, and has a good effect on improving cognitive function in animal experiments. The multifunctional preparation not only meets the requirements of anti-Alzheimer's disease, but also has a good market prospect.

SUMMARY OF THE INVENTION

The invention discloses a kind of donepezil-BHT hybrid. Pharmacodynamic experiments prove that the compounds of the present invention can be used as multifunctional preparations for the treatment of Alzheimer's disease.

The compound structure of the present invention is as follows:

Wherein n=0, 1, 2; R is CH ₃ , OCH ₃ , NO ₂ , CN, halogen atom, etc. respectively.

The most preferred compound structure is as follows:

The pharmaceutically acceptable salts of the compounds of the present invention are the hydrochloride, hydrobromide, sulfate, phosphate, mesylate, benzenesulfonate, p-toluenesulfonate, naphthalenesulfonate of the compound of the general formula , citrate, tartrate, lactate, pyruvate, acetate, maleate, succinate, fumarate, salicylate, phenylacetate, mandelicate, base salts of alkaline earth metal cations or ammonium cations. Most preferred are alkali metal cation salts.

The preparation method of the example compound of the present invention is as follows:

1) The preparation method of example compounds 3a-i of the present invention is as follows:

2) The preparation method of the example compound 3j-1 of the present invention is as follows:

3) The preparation method of example compounds 7a-m of the present invention is as follows:

The definition of R is the same as before.

The compounds of the present invention can be prepared by the above or similar preparation methods above, and the corresponding raw materials can be selected according to the change of the chain length, the difference of the substituent or the difference of the position of the substituent.

Basic reaction process: the preparation method of the donepezil-BHT hybrid of the present invention includes acid-amine condensation reaction, substitution reaction, de-Boc protecting group process and post-processing unit processes.

The synthesis of compound 3a-i is preferred: add BHT-type acids (1a-c) (1.1eq) with different chain lengths into a single-necked flask, dissolve with an appropriate amount of anhydrous dichloromethane, and then add a condensing agent 1-hydroxybenzotrimine oxazole (HOBT) (1.1eq), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) (1.1eq), triethylamine (3eq) dropwise, ice After stirring the reaction under the bath for 1 hour, the dichloromethane solution of amine (1 eq) was added dropwise, and the reaction was performed overnight. Phase and mix samples to pass through a silica gel column, and a dichloromethane/methanol product system is used to obtain the target product.

The preferred synthesis of compound 3j-1: 2-bromo-3', 5'-di-tert-butyl-4'-hydroxyacetophenone (1d) (2eq) and different amines (2a-c) (1eq) in carbonic acid Under the catalyst of potassium (K ₂ CO ₃ ) and potassium iodide (KI), react with acetonitrile as solvent overnight, monitor the reaction, and purify the target product through silica gel column.

The synthesis of compound 7a-m is preferably: aminoethylpiperidine Boc reacts with different benzyl bromide in ethanol as solvent and triethylamine as acid binding agent to produce tert-butoxycarbonylaminoethylbenzylpiperidine, and further in triethylamine. The tert-butoxycarbonyl group (Boc protecting group) is removed under the action of fluoroacetic acid, and finally acid-amine condensation is carried out, the method is the same as above.

The following are some pharmacological experiments and data of the compounds of the present invention:

1. Cholinesterase activity:

experimental method:

The test method of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitory activity was Ellman's method. The compound was dissolved in dimethyl sulfoxide (DMSO), and then diluted with buffer to the desired concentration, and the content of DMSO in the prepared solution was controlled to be less than 1%. To the 96 blank, 160 μl of 1.5 mM 5,5’-dithiobis(2-nitrobenzoic acid) (DTNB), 50 μl of AChE (0.22 U/mL, prepared with buffer B) and 10 μl of different concentrations of inhibitors. After incubation at 37°C for 6 minutes, 30 μl of acetylcholine iodide (15 mM) was added rapidly. Absorbance changes at 0, 60, 120 and 180 seconds were measured at 405 nm. The assay method of butyrylcholinesterase is similar to that of acetylcholinesterase. The acetylcholinesterase used is replaced by butyrylcholinesterase (0.12U/mL) and the substrate acetylcholine iodide is replaced by thioiodide butyrylcholinesterase. Base (15mM). The inhibition rate was calculated as: [1-(absorbance change in experimental group/absorbance change in blank group)]*100%. Select five to seven concentrations of the compound to measure the inhibition rate of the enzyme (0.001-100 μM) and perform linear regression with the negative logarithm of the molar concentration of the compound and the enzyme inhibition rate, and obtain the molar concentration when 50% inhibition is obtained. _IC50 value. Each experiment was repeated three times, and the experimental results were expressed as mean ± SEM.

Table 1 Cholinesterase inhibitory activity of the compounds of the present invention

a) IC ₅₀ : the concentration of the compound at which the inhibition rate reaches 50%;

b) Selectivity coefficient = _IC50 of human butyrylcholinesterase / _IC50 of human acetylcholinesterase

It can be seen from Table 1 that the compounds 3a-i have certain activity on acetylcholine as a whole, but the activity on butyrylcholine is poor, and the inhibitory activity of compound 3i is the best (IC ₅₀ =0.53 μM). The structure-activity relationship showed that the activity of acetylcholinesterase was closely related to the change of chain length, and the longer the chain length, the better the activity. On the basis of 3i, it was further modified, and the following is the activity result of its further modification. The acetylcholine activity of compounds 7a-m is comparable to that of 3i, basically between 0-1 μM. In addition, compound 7d inhibited the activity of human acetylcholine significantly, reaching IC ₅₀ =0.075μM, with poor inhibitory effect on butyrylesterase and good selectivity. This result is basically equivalent to the effect of the positive drug of donepezil, which shows the rationality of further modification, and also shows that the compound disclosed in the present invention has a good inhibitory effect on acetylcholinesterase.

2. Kinetic study of acetylcholinester

experimental method:

The enzyme kinetics of compounds 3i and 7d were studied by Ellman method. Three different concentrations of the compound were separately studied for kinetics. To 96 empty plates, 160 μl of 1.5 mM DTNB, 50 μl of AChE (0.22 U/mL, prepared with buffer B) and 10 μl of different concentrations of compounds were added sequentially. After incubation at 37°C for 6 minutes, 30 μl of various concentrations of acetylcholine iodide were rapidly added. Absorbance changes at 0, 60, 120 and 180 seconds were measured at 405 nm. Draw the first double-reciprocal curve by plotting the inverse of the concentration as the X-axis and the rate of change of absorbance as the Y-axis. According to this method, different concentrations of compounds are added, and the second, third, and fourth double-reciprocal curves are drawn, and the action mode of the compound and the enzyme is judged by the intersection of the double-reciprocal curves.

The results are shown in Figure 1, which shows that: with the increase of the concentration of the compounds disclosed in the present invention, the slope and intercept of the double-reciprocal curve of Lineweaver-Burk also increase continuously. This model shows that the compound is a mixed-type inhibitor, indicating that the compound binds to different sites of the enzyme, that is, it can simultaneously bind to the peripheral anion site (PAS) and the catalytic site (CAS) of the enzyme. Dual-site inhibitors are more beneficial for the treatment of Alzheimer's disease. At the same time, the activity of compound 7d (Ki=1.4μM) was further verified than that of 3i (Ki=3.2μM) by its inhibition constant Ki.

3. Monoamine oxidase inhibitory activity

experimental method:

The monoamine oxidase inhibitory activity was tested according to the method reported in the literature. Compounds were dissolved in DMSO and sequentially diluted with buffer to the desired concentration (the DMSO content in the prepared solution was controlled to be less than 1%). Add 80 μl of enzyme (diluted with buffer) and 20 μl of compounds of different concentrations to the black 96-well microtiter plate. After incubation at 37°C for 15 minutes, add Amplex Red solution with a final concentration of 200 μM, 1 U/mL horseradish peroxide. Enzyme, 1 mM tyramine solution. Fluorescence absorption was measured at excitation light wavelength 545 nm and absorption light wavelength 590 nm. The inhibition rate was calculated as: [1-F _{experimental group} /F _{blank group} ]*100%. Select five to seven concentrations of the compound to measure the inhibition rate of the enzyme (0.001-100 μM) and perform linear regression with the negative logarithm of the molar concentration of the compound and the enzyme inhibition rate, and obtain the molar concentration when 50% inhibition is obtained. _IC50 value. Each experiment was repeated three times, and the experimental results were expressed as mean ± SEM.

Table 2 Monoamine oxidase inhibitory activity of the compounds of the present invention

It can be seen from Table 2 that the monoamine oxidase activity of compound 3a-1 is average. For anti-AD drugs, we focus on the activity of monoamine oxidase B, among which compounds 3h (IC ₅₀ =8.5 μM) and 3i (IC ₅₀ =11.7 μM) have the best activities. , and has better selectivity than monoamine oxidase A. The monoamine oxidase activities of compounds 7a-m were basically the same as those of 3i.

4. Antioxidant effect

The in vitro antioxidant activity of the compound is reflected by testing the free radical scavenging ability, and the present invention adopts the ABTS test, the DPPH test and the ORAC test. The three methods are as follows:

1. ABTS test method: water-soluble vitamin E analog (Trolox) standard: prepare a stock solution of 5mM Trolox standard with absolute ethanol, and then dilute it with absolute ethanol to several different concentrations to make it. They have 5% to 85% free radical scavenging capacity, respectively. According to the preliminary experimental results, the appropriate concentration of Trolox standard solution is: 0, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6 and 1.8mM. ABTS working solution: Mix 5 mL of 7 mM ABTS with 88 μl of 140 mM potassium persulfate solution and let stand overnight at room temperature in the dark to form ABTS·+ stock solution, which is stored at room temperature in the dark in the dark stable down. Before use, it was diluted with anhydrous methanol into a working solution, incubated at 30°C for 6 minutes in the dark, and the absorption value at 415nm wavelength was measured. The experiment was repeated three times.

2. DPPH test method: Dissolve an appropriate amount of DPPH in methanol to 400 μM, ultrasonicate for 5 minutes, and shake well. Compounds and controls resveratrol and curcumin were dissolved in methanol to different concentrations. During the test, 20 μl of compound was added to each well of the 96-well plate, and then 180 μl of DPPH solution was added, and it was placed in the dark at room temperature for 30 minutes to measure the absorption value at a wavelength of 517 nm. The experiment was repeated three times. The free radical scavenging rate was calculated as: 1-[(A _DPPH + A _compound )-A _DPPH /A _compound )]*100%. Five to seven concentrations of the compound were selected to determine the inhibition rate, and the molar concentration at which 50% inhibition was obtained was the IC50 value of the compound. Each experiment was repeated three times, and the experimental results were expressed as mean ± SEM.

3. ORAC test method: Before the test, prepare fluorescein (FL) and free radical releasing agent (APPH) to prepare a stock solution, and prepare a 110nM FL and 40mM AAPH solution with PBS. Compounds and Trolox were formulated at different concentrations. During the test, add 20 μl of compound to each well of a black 96-well plate, then add 120 μl of FL, shake at 37°C for 15 minutes in the dark, and then quickly add 60 μl of AAPH to detect and read the plate under excitation light of 485 nm and emission light of 538 nm. Read once a minute and scan for 120 minutes. The area under the fluorescence curve (AUC) was calculated, and the result was expressed as the equivalent value of trolox, ORAC-FL=(AUC _sample -AUC _blank )/(AUC _trolox -AUC _blank )*(C _trolox /C _sample )

Table 5 Free radical scavenging activity of the compounds of the present invention

^a IC ₅₀ : the concentration of the compound at which the inhibition rate reaches 50%;

^b The antioxidant activity of the compound is expressed as the multiple of trolox at the same concentration (mmol trolox/mmol of the compound)

The results of antioxidant effect are shown in Table 3. BHT and resveratrol were used as positive control drugs to compare the antioxidant effect of the compounds of the present invention. The antioxidant activity of the compounds in the ABTS test was expressed as the multiple of trolox at the same concentration (mmoltrolox/mmol of the measured compound). The assay results showed that most of the compounds exhibited moderately good antioxidant effects, and their antioxidant capacity was slightly weaker than that of trolox. The antioxidant capacity of the optimal compounds 3i and 7d was 0.91 and 0.82 times that of trolox, respectively. The antioxidant activity of the compounds in the DPPH test was expressed by IC ₅₀ , and the antioxidant activity of most compounds was better than that of the control drug resveratrol and parent BHT, among which the IC ₅₀ of the best compounds 3i and 7d were 71.7 μM and 55.9 μM, respectively. . The ORAC test results are expressed as trolox multiples at the same concentration (mmol trolox/mmol test compound). The results show that the antioxidant activity of the compounds is mostly equivalent to that of the parent BHT, and some compounds are slightly higher than the parent, but all are better than Trolox. These results show that this These compounds have good antioxidant activity, can fight against oxidative stress, and then be used in AD treatment.

5. Cytotoxicity of PC12 and its protective effect on H ₂ O ₂ -induced injury of PC12 cells

PC12 cells were seeded in 25 ml culture flasks containing a 1:1 mixture of Dulbecco's modified Eagle's medium (EMEM) and ham's F-12 medium supplemented with 10% fetal bovine serum, 100 U/mL penicillin and 100 μg/mL chain and cultured in a 5% carbon dioxide incubator at 37°C. Cells were then grown in 96-well plates at a density of 20,000 per well. When the cell growth reached the requirement, the cells were placed in serum-free medium with the addition of 200 μM H ₂ O ₂ and various concentrations of compounds and incubated for 24 hours. After the incubation, 20 μl of MTT was added and incubated at 37° C. for 4 hours. After completion, 200 μl of DMSO was added to dissolve the formazan crystals, and the absorbance was measured at 570 nm.

The best compound 3i and 7d were selected to evaluate their cytotoxicity to nerve cells by MTT method. The results are shown in Figure 2. After exposure to cells for 24 hours at a concentration of 30 μM, compounds 3i and 7d showed acceptable cytotoxicity, but had no effect on cell viability at 20 μM. To further study its effect _on H2O2 _- induced PC12 cell injury, the experimental results are shown in Figure _3. Compared with the H2O2 cell injury group, the _compounds at 5-20 μM can significantly improve the cell survival rate. Compounds all had obvious protective effect on H ₂ O ₂ -induced PC12 cell injury at the concentration of 5 μM. The degree of protective effect of these compounds on H ₂ O ₂ -induced PC12 cell injury was consistent with their in vitro antioxidant activity, indicating that the neuroprotective effect of our designed compounds may be related to their free radical scavenging effect.

6. Inhibition of LPS-induced release of inflammatory cytokine nitric oxide (NO) from BV-2 cells

The BV-2 cell culture method is similar to that of PC12 cells. BV-2 cells were seeded in Dulbecco's modified Eagle's medium supplemented with 10% Gemini fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin, and placed at 37°C, Cultured in a 5% carbon dioxide incubator. Then the cells were planted in a 96-well plate at a density of 30,000 per well for 18 hours, and then cultured with different concentrations of compounds. After 1 hour, 10 μL of LPS with a concentration of 1 ng/ml was added for 18 hours, and 50 μl of the supernatant was taken with NO The detection kit measures the inhibition rate of NO. The experimental results are shown in Figure 4. Compared with the parent drugs BHT and donepezil, it has better NO inhibitory activity.

7. Acute toxicity test and behavioral cognitive improvement test

According to the guidelines of acute toxicity, 20 KM mice were randomly divided into 4 groups, namely the normal group and the administration group (677, 1333, 2000 mg/kg). After 14 days of observation, no obvious damage was observed after being sacrificed, indicating that the safety of compound 7d is high.

After evaluating the safety, a pharmacodynamic evaluation was carried out. The memory loss model caused by scopolamine was used to study the effect of the compound on its behavioral improvement by passive dark avoidance experiment. After the behavioral experiment was completed, it was combined with Pathological sections were observed and the activities of alanine aminotransferase and aspartate aminotransferase (ALT/AST) were measured, and the effect on the liver was evaluated. The results are shown in Figure 5, indicating that the compound has a better effect on improving cognition, and the concentration depends on the tolerance. . And there is no hepatotoxicity under the administered dose, and the results can be obtained from FIG. 6 and FIG. 7 .

The Donepezil-BHT hybrid of the present invention is proved by molecular docking that it can bind to acetylcholinesterase and monoamine oxidase B, thereby showing better inhibition of acetylcholinesterase activity and inhibition of monoamine oxidase B activity; in addition, through the in vitro blood-brain barrier permeability test It was shown that the best compounds 3i and 7d could penetrate the blood-brain barrier. In conclusion, the donepezil-BHT hybrid showed good inhibition of acetylcholinesterase activity, inhibition of monoamine oxidase B activity and antioxidant effects in vitro experiments, and had the ability to inhibit hydrogen peroxide (H ₂ O ₂ )-induced oxidative damage in PC12 neurons and Bacterial lipopolysaccharide (LPS) stimulated BV-2 inflammatory injury has better neuroprotective effect and better cognitive function improvement in animal experiments, so they can be used as multifunctional preparations to control the development of AD.

Description of drawings

Figure 1 is a kinetic study of compounds 3i and 7d inhibiting acetylcholinesterase

Figure 2 is the cytotoxic effect of compound PC12

Figure 3 shows that compounds 3i and 7d are compounds PC12 neuroprotective

Figure 4 is the effect of compound 7d on inhibiting the release of inflammatory factor NO from LPS-induced BV-2 cells

Figure 5 is the behavioral improvement of cognition and body weight change of compound 7d

Figure 6 is the change of serum alanine aminotransferase and aspartate aminotransferase (ALT/AST) measured after compound 7d administration

Fig. 7 is the result of liver pathological section after administration of compound 7d

Figure 8 is a drawing of the patent abstract

Specific Synthetic Embodiments

Example 1

Preparation of 4-(N-1-benzylpiperidine)amino-2,4-di-tert-butyl-p-hydroxybenzamide (3a).

Dissolve 100 mg (0.39 mmol, 1 eq) of raw material 2,4-di-tert-butyl-p-hydroxybenzoic acid (1a) in 6 ml of anhydrous dichloromethane, add 49.74 mg (0.36 mg) of condensing agent 1-hydroxybenzotriazole (HOBT) mmol, 1.1eq) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) 69.62mg (0.36mmol, 1.1eq), add dropwise triethylamine 151μl (1.09 mmol, 3eq), after stirring the reaction under ice bath for 1 hour, add dropwise a solution of 4-amino-1-benzylpiperidine (2a) 69.1mg (0.36mmol, 1eq) in dichloromethane to react overnight, monitor the reaction until the amine is complete , washed three times with saturated sodium bicarbonate, washed three times with saturated sodium chloride until HOBT was washed, the organic phase was combined and the samples were passed through a silica gel column, and the dichloromethane/methanol product system was about 140 mg of the target product white solid 3a, and the yield was about 140 mg. 80%. mp 220-222°C. ¹ H NMR (500 MHz, DMSO) δ 8.02 (d, J=7.7 Hz, 1H), 7.78 (s, 1H), 7.57 (s, 1H), 7.37-7.31 (m, 4H), 3.78(d, J=7.2Hz, 1H), 3.49(s, 2H), 2.84(d, J=8.2Hz, 2H), 2.03(s, 2H), 1.77(d, J=10.9Hz, 1H), 1.62(d, J=10.3Hz, 2H), 1.41(s, 18H), 1.25(s, 1H); ¹³ C NMR (126MHz, DMSO) δ 167.13, 156.95, 138.65, 129.29, 128.65, 127.44, 126.57, 124.51, 62.47, 52.76, 34.94, 30.69, 30.52. ESI-MS m/z: 423.3[M+H] ⁺ ; HRMS(ESI) m/z 423.3004[M+H] ⁺ (calcd for 423.3006, C ₂₇ H ₃₉ N ₂ O ₂ ).

Example 2

Preparation of 4-(N-1-benzylpiperidine)aminomethyl-2,4-di-tert-butyl-p-hydroxybenzamide (3b)

The preparation process is the same as 1, except that the raw material 4-amino-1-benzylpiperidine (2a) is replaced by 4-aminomethyl-1-benzylpiperidine (2b), the yield is 82%; white solid, mp217-218℃ , ¹ H NMR (500MHz, DMSO) δ8.27(d, J=7.7Hz, 1H), 7.60(s, 2H), 7.31(m, 5H), 3.45(s, 2H), 3.13(t, J= 6.2Hz, 2H), 2.81 (d, J=11.0Hz, 2H), 1.90 (t, J=11.0Hz, 2H), 1.64 (d, J=12.2Hz, 2H), 1.54 (d, J=6.2Hz) , 1H), 1.41(s, 18H), 1.22-1.11(m, 2H); ¹³ C NMR (126MHz, DMSO) δ 167.51, 156.51, 139.24, 138.64, 129.20, 128.56, 127.24, 126.45, 124.43, 62.91, 53.46, 45.20, 36.33, 35.03, 30.71, 30.36. ESI-MS m/z: 437.3[M+H] ⁺ ; HRMS(ESI) m/z: 437.3164[M+H] ⁺ (calcd for 437.3163, C ₂₈ H ₄₁ N ₂ O ₂ ).

Example 3

Preparation of 4-(N-1-benzylpiperidine)aminoethyl-2,4-di-tert-butyl-p-hydroxybenzamide (3c)

The preparation process is the same as 1, except that the raw material 4-amino-1-benzylpiperidine (2a) is replaced by 4-aminoethyl-1-benzylpiperidine (2c), the yield is 75%; white solid, mp182-183°C , ¹ H NMR (500MHz, CDCl3)δ7.61(s, 2H), 7.42-7.33(m, 4H), 7.32(s, 1H), 5.99(s, 1H), 5.57(s, 1H), 3.60( s, 2H), 3.51 (dd, J=14.1, 6.4Hz, 2H), 2.98 (d, J=10.3Hz, 2H), 2.06 (s, 2H), 1.78 (d, J=10.3Hz, 2H), 1.61 (d, J=6.4 Hz, 2H), 1.50 (s, 18H), 1.43 (s, 2H), 1.32 (d, J=14.1 Hz, 1H); ¹³ C NMR (126 MHz, DMSO) δ 167.95, 156.51, 135.96, 129.56, 128.31, 126.04, 123.98, 53.30, 37.76, 36.50, 34.43, 33.62, 31.54, 30.20. ESI-MS m/z: 451.3[M+H] ⁺ ; HRMS(ESI) m/z: 451.3317 [M+H] ⁺ (calcd for 451.3319, C ₂₉ H ₄₃ N ₂ O ₂ ).

Example 4

Preparation of 4-(N-1-benzylpiperidine)amino-2,4-di-tert-butyl-p-hydroxyphenylpropanamide (3d)

The preparation process is the same as 1, except that the raw material 2,4-di-tert-butyl-p-hydroxybenzoic acid (1a) is replaced with 2,4-di-tert-butyl-p-hydroxybenzoic acid (1b), the yield is 76%; white solid, mp142 -144°C, ¹ H NMR (500MHz, DMSO) δ 7.71(d, J=7.6Hz, 1H), 7.36-7.22(m, 5H), 6.69(s, 1H), 3.54(m, 1H), 3.45 (s, 2H), 2.71 (dd, J=11.1, 7.7Hz, 4H), 2.30 (t, J=7.7Hz, 2H), 1.99 (t, J=10.4Hz, 2H), 1.68 (d, J= 10.4Hz, 2H), 1.42 (s, 1H), 1.37 (s, 18H), 1.33 (d, J=11.1Hz, 2H), 1.25 (m, 1H); ¹³ C NMR (126MHz, DMSO) δ 171.37 , 152.33, 139.20, 132.67, 129.19, 128.60, 127.30, 124.62, 62.62, 52.40, 46.37, 38.01, 34.89, 32.04, 31.67, 30.83.ESI-MS m/z: 451.3[M+H] ⁺ ; HRMS(ESI) m/z: 451.3320[M+H] ⁺ (calcd for 451.3319, C ₂₉ H ₄₃ N ₂ O ₂ ).

Example 5

Preparation of 4-(N-1-benzylpiperidine)aminomethyl-2,4-di-tert-butyl-p-hydroxyphenylpropanamide (3e)

The preparation process is the same as 1, except that the raw materials 2,4-di-tert-butyl-p-hydroxybenzoic acid (1a) and 4-amino-1-benzylpiperidine (2a) are replaced with 2,4-di-tert-butyl-p-hydroxybenzene respectively Propionic acid (1b) and 4-aminomethyl-1-benzylpiperidine (2b), 62% yield; brown oil, ¹ H NMR (500 MHz, DMSO) δ 7.36-7.21 (m, 5H), 6.91(s, 2H), 3.45(s, 2H), 2.94(t, J=6.2Hz, 2H), 2.78(d, J=9.2Hz, 2H), 2.70(s, 2H), 2.33(s, 2H ), 1.87(s, 2H), 1.56(s, 2H), 1.37(s, 18H), 1.25(s, 1H), 1.10(m, 2H); ¹³ C NMR (126MHz, DMSO) δ 172.06, 152.30 , 139.42, 132.65, 129.28, 128.57, 127.37, 124.60, 62.79, 53.22, 44.54, 37.90, 35.99, 34.89, 31.67, 30.77, 29.89.ESI-MS m/z: 465.3[M+H] ⁺ ; HRMS(ESI) m/z: 465.3474[M+H] ⁺ (calcd for 465.3476, C ₃₀ H ₄₅ N ₂ O ₂ ).

Example 6

Preparation of 4-(N-1-benzylpiperidine)aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylpropanamide (3f)

The preparation process is the same as 1, except that the raw materials 2,4-di-tert-butyl-p-hydroxybenzoic acid (1a) and 4-amino-1-benzylpiperidine (2a) are replaced with 2,4-di-tert-butyl-p-hydroxybenzene respectively Propionic acid (1c) and 4-aminomethyl-1-benzylpiperidine (2c), 64% yield; light brown oil, ¹ H NMR (500 MHz, DMSO) δ 7.75 (t, J=7.6 Hz) , 1H), 7.35-7.23(m, 5H), 6.69(s, 1H), 3.45(s, 4H), 3.07(d, J=6.4Hz, 2H), 2.78(d, J=11.1Hz, 2H) , 2.70(t, J=7.6Hz, 2H), 2.30(t, J=7.7Hz, 2H), 1.90(t, J=12.4Hz, 2H), 1.61(d, J=12.4Hz, 2H), 1.38 (s, 18H), 1.25 (d, J=6.4Hz, 1H), 1.12 (m, 2H); ¹³ C NMR (126 MHz, DMSO) δ 171.89, 152.35, 139.47, 132.71, 129.24, 128.55, 127.26, 124.62 , 62.62, 53.64, 38.01, 36.65, 36.45, 34.90, 33.32, 32.24, 31.67, 30.93. ESI-MS m/z: 479.3[M+H] ⁺ ; HRMS(ESI) m/z: 479.3635[M+H] ⁺ (calcd for 479.3632, C ₃₁ H ₄₇ N ₂ O ₂ ).

Example 7

Preparation of 4-(N-1-benzylpiperidine)amino-2,4-di-tert-butyl-p-hydroxyphenylacrylamide (3g)

The preparation process is the same as that in 1, except that the raw material 2,4-di-tert-butyl-p-hydroxybenzoic acid (1a) is replaced with 2,4-di-tert-butyl-p-hydroxybenzoic acid (1c), yield 70%; brown solid, mp214- 215°C, ¹ H NMR (500 MHz, DMSO) δ 7.91 (d, J=15.7 Hz, 1H), 7.37-7.31 (m, 7H), 6.48 (d, J=15.7 Hz, 1H), 3.67 (d, J=6.6Hz, 1H), 3.35(s, 2H), 2.78(d, J=11.1Hz, 2H), 2.06(s, 2H), 1.78(m, 2H), 1.41(s, 18H), 1.30( dd, J=11.1, 6.6 Hz, 2H); ¹³ C NMR (125 MHz, DMSO) δ 165.03, 155.74, 140.04, 139.70, 129.25, 128.53, 127.24, 126.81, 124.66, 119.56, 62.66, 52.21, 46.65, 34.96 32.15, 30.60. ESI-MS m/z: 449.3 [M+H] ⁺ ; HRMS(ESI) m/z: 449.3165 [M+H] ⁺ (calcd for 449.3163, C ₂₉ H ₄₁ N ₂ O ₂ ).

Example 8

Preparation of 4-(N-1-benzylpiperidine)aminomethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide (3h)

The preparation process is the same as 1, except that the raw materials 2,4-di-tert-butyl-p-hydroxybenzoic acid (1a) and 4-amino-1-benzylpiperidine (2a) are replaced with 2,4-di-tert-butyl-p-hydroxybenzene respectively Propionic acid (1c) and 4-aminomethyl-1-benzylpiperidine (2b), 65% yield; brown solid, ¹ H NMR (500 MHz, DMSO) δ 7.96 (d, J=15.7 Hz, 1H ), 7.35(d, J=6.8Hz, 1H), 7.32-7.28(m, 5H), 7.26(d, J=6.8Hz, 1H), 6.51(d, J=15.7Hz, 1H), 3.45(s , 2H), 3.08 (t, J=6.1Hz, 2H), 2.81 (d, J=11.2Hz, 2H), 1.98-1.83 (m, 2H), 1.64 (d, J=11.8Hz, 2H), 1.41 (s, 18H), 1.19 (d, J=6.1 Hz, 2H), 1.06 (s, 1H); ¹³ C NMR (126 MHz, DMSO) δ 165.90, 155.89, 139.97, 139.69, 129.24, 129.24, 128.46, 128.46 , 127.27, 126.82, 124.64, 119.53, 62.87, 53.30, 53.30, 44.73, 44.73, 36.28, 36.28, 34.91, 34.91, 30.58, 30.58, 30.17, 30.17.ESI-MS m/z: 463.3 [M ⁺ H]; HRMS (ESI) m/z: 463.3321 [M+H] ⁺ (calcd for 463.3319, C ₃₀ H ₄₃ N ₂ O ₂ ).

Example 9

Preparation of 4-(N-1-benzylpiperidine)aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide (3i)

The preparation process is the same as 1, except that the raw materials 2,4-di-tert-butyl-p-hydroxybenzoic acid (1a) and 4-amino-1-benzylpiperidine (2a) are replaced with 2,4-di-tert-butyl-p-hydroxybenzene respectively Propionic acid (1c) and 4-aminomethyl-1-benzylpiperidine (2c), 70% yield; brown solid, mp 126-128°C. ¹ H NMR (500MHz, DMSO) δ 7.93 (d, J=15.7Hz, 1H), 7.33-7.28 (m, 7H), 6.46 (d, J=15.7Hz, 1H), 3.35 (s, 4H), 3.20(dd, J=12.8, 6.6Hz, 2H), 2.81(d, J=6.6Hz, 2H), 1.93(s, 2H), 1.66(d, J=12.9Hz, 2H), 1.42(s, 18H ^The , 36.68, 36.38, 34.96, 33.23, 32.22, 30.68. ESI-MS m/z: 477.3[M+H] ⁺ ; HRMS(ESI) m/z: 477.3475[M+H] ⁺ (calcd for 477.3476, C ₃₁ H ₄₅ N ₂ O ₂ ).

Example 10

Preparation of 4-(N-1-benzylpiperidine)amino-2,4-di-tert-butyl-p-hydroxyphenethylamine-2-one (3j)

The raw material 2,4-di-tert-butyl-p-hydroxyacetophenone (1d) 100 mg (0.31 mmol, 1.1 eq) was dissolved in acetonitrile, potassium carbonate (K ₂ CO ₃ ) 115 mg (0.833 mmol, 3 eq) was added, and then 4-Amino-1-benzylpiperidine (2a) was added dropwise and stirred overnight, monitored until the reaction of the amine was completed, the solvent was spin-dried, extracted with dichloromethane and water, and the sample was mixed and passed through a silica gel column to obtain the target product as a brown oil. The yield was 32%, ¹ H NMR (500 MHz, DMSO) δ 7.34 (t, J=5.3 Hz, 5H), 7.14 (d, J=2.3 Hz, 1H), 6.79 (d, J=2.4 Hz, 1H), 5.78 (s, 1H), 3.56-3.49 (m, 2H), 2.86 (d, J=10.0Hz, 2H), 2.23 (d, J=6.3Hz, 2H), 1.78-1.63 (m, 4H), 1.26 ( m, 18H), 1.19-1.00 (m, 2H), 0.86 (dd, J=10.0, 6.1 Hz, 1H); ¹³ CNMR (126 MHz, DMSO) δ 188.05, 157.19, 152.34, 150.75, 138.75, 136.37, 129.12 , 128.61, 127.33, 124.51, 119.03, 62.71, 51.55, 35.83, 35.03, 33.84, 30.71, 29.55.ESI-MS m/z: 437.3.[M+H] ⁺ ; HRMS(ESI) m/z: 437.3165[M +H] ⁺ (calcd for 437.3163, C ₂₈ H ₄₁ N ₂ O ₂ ).

Example 11

Preparation of 4-(N-1-benzylpiperidine)aminomethyl-2,4-di-tert-butyl-p-hydroxyphenethylamine-2-one (3k)

The process was the same as in 10, except that the raw material 4-amino-1-benzylpiperidine (2a) was replaced with 4-aminomethyl-1-benzylpiperidine, yield 34%; brown oil. ¹ H NMR (500MHz, DMSO) δ 8.06-7.94 (m, 1H), 7.77 (s, 1H), 7.43-7.15 (m, 5H), 5.23-4.82 (m, 1H), 4.74 (s, 1H) , 3.47(s, 2H), 3.18(d, J=7.4Hz, 2H), 2.82(d, J=10.2Hz, 2H), 1.91(d, J=10.2Hz, 2H), 1.64-1.55(m, 2H), 1.49-1.27 (m, 18H), 1.26 (s, 1H), 1.19-0.72 (m, 2H); ¹³ C NMR (126MHz, DMSO) δ 193.94, 163.78, 138.80, 129.21, 128.57, 127.29, 125.55, 62.78, 53.20, 49.58, 34.99, 34.01, 31.43, 30.47, 29.72. ESI-MS m/z: 451.3[M+H] ⁺ ; HRMS(ESI) m/z: 451.3321[M+H] ⁺ (calcd for 451.3319, C ₂₉ H ₄₃ N ₂ O ₂ ).

Example 12

Preparation of 4-(N-1-benzylpiperidineaminoethyl)-2,4-di-tert-butyl-p-hydroxyphenethylamine-2-one (3l)

The preparation process was the same as that in 10, except that the raw material 4-amino-1-benzylpiperidine (2a) was replaced with 4-aminoethyl-1-benzylpiperidine, the yield was 30%; the brown oil was obtained. ¹ H NMR (500MHz, DMSO) δ 7.99(s, 1H), 7.53(s, 1H), 7.32(m, 6H), 3.49(s, 2H), 3.10(m, 2H), 2.83(s, 3H) ), 2.11(s, 1H), 1.90(d, J=9.8Hz, 2H), 1.56(m, 3H), 1.46-1.18(m, 18H), 1.16-0.92(m, 3H), 0.87(d, J=9.2Hz, 2H); ¹³ C NMR (126MHz, DMSO) δ 193.94, 163.78, 138.80, 129.21, 128.57, 127.29, 125.55, 62.89, 53.29, 36.68, 36.38, 34.96, 33.23, 32.22, 30.68.ESI- MS m/z: 465.3 [M+H] ⁺ ; HRMS (ESI) m/z: _465.3479 [M+H] ⁺ ( _calcd for _465.3476 , _C30H45N2O2 ).

Example 13

Preparation of 4-(N-1-(4-methoxy-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7a

Raw material 100mg aminoethylpiperidine Boc (0.5mmol, 1.1eq) and 104.24mg of 4-methoxybenzyl bromide (0.45mmol, 1eq) in ethanol as solvent, triethylamine (1.36mmol, 3eq) as acid binding agent The reaction produces tert-butoxycarbonylaminoethylbenzylpiperidine under the conditions of , and the tert-butoxycarbonyl group (Boc protecting group) is further removed under the action of trifluoroacetic acid, and finally the acid-amine condensation method is the same as above 1.

A white solid was obtained, the yield was 58%; mp197-198°C, ¹ H NMR (500MHz, DMSO) δ7.99(s, 1H), 7.53(s, 1H), 7.32(m, 6H), 3.89(s, 3H) ), 3.10(m, 2H), 2.83(s, 3H), 2.11(s, 1H), 1.90(d, J=9.2Hz, 2H), 1.56(m, 3H), 1.46-1.18(m, 18H) , 1.16-0.92 (m, 3H), 0.87 (d, J=9.2Hz, 2H); ¹³ CNMR (126MHz, DMSO) δ 165.65, 158.61, 155.95, 139.88, 139.64, 130.90, 130.41, 126.70, 124.67, 119.411 , 113.92, 62.39, 55.43, 53.60, 36.62, 36.42, 34.97, 33.30, 32.31, 30.64. ESI-MS m/z: 507.3[M+H] ⁺ ; HRMS(ESI) m/z: 507.3581[M+H] ⁺ (calcd for 507.3579, C ₃₂ H ₄₇ N ₂ O ₃ ).

Example 14

Preparation of 4-(N-1-(2-methyl-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7b

The preparation process was the same as that in 13, except that the raw material 4-methoxybenzyl bromide was replaced with 2-methyl benzyl bromide, the yield was 48%; the brown solid, mp182-184°C, ¹ H NMR (500MHz, DMSO)δ7.95(d, J=15.7Hz, 1H), 7.32(s, 3H), 7.19(m, 4H), 6.47(d, J=15.7Hz, 1H), 3.39(s, 2H), 3.19(s, 2H), 2.78( d, J=11.1Hz, 2H), 2.32(s, 3H), 1.91(t, J=7.1Hz, 2H), 1.65(d, J=11.1Hz, 2H), 1.40(s, 18H), 1.32( d, J=7.2Hz, 1H), 1.11 (s, 2H), 0.99 (s, 2H). ¹³ C NMR (151 MHz, DMSO) δ 165.66, 155.92, 139.87, 139.65, 137.40, 137.24, 130.46, 129.88, HRMS( ^ESI )m /z: 491.3632[M+H] ⁺ (calcd for 491.3629, C ₃₂ H ₄₇ N ₂ O ₂ ).

Example 15

Preparation of 4-(N-1-(4-methyl-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7c

The preparation process is the same as in 13, the raw material 4-methoxybenzyl bromide is replaced with 4-methyl benzyl bromide, the yield is 44%; the brown solid, mp190-191℃, 1H NMR (500MHz, DMSO)δ7.95(d, J =15.7Hz, 1H), 7.32(s, 3H), 7.19(m, 4H), 6.47(d, J=15.7Hz, 1H), 3.39(s, 2H), 3.19(s, 2H), 2.78(d , J=11.1Hz, 2H), 2.32(s, 3H), 1.93(t, J=7.1Hz, 2H), 1.65(d, J=11.1Hz, 2H), 1.40(s, 18H), 1.32(d , J=7.2Hz, 1H), 1.11(s, 2H), 0.99(s, 2H). ¹³ C NMR (151MHz, DMSO) δ165.66, 155.92, 139.87, 139.65, 137.40, 137.24, 130.46, 129.88, 127.21 , 126.73, 125.79, 124.67, 119.43, 61.02, 53.96, 46.16, 36.64, 36.34, 34.97, 32.40, 30.64, 28.75, 19.28.ESI-MS m/z: 491.3[M+H]+; HRMS(ESI)m/ z: 491.3632[M+H]+(calcd for 491.3629, C ₃₂ H ₄₇ N ₂ O ₂ ).

Example 16

Preparation of 4-(N-1-(2-Fluoro-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7d

The preparation process is the same as that in 13, except that the raw material 4-methoxybenzyl bromide is replaced with 2-fluorobenzyl bromide, the yield is 62%; the brown solid, mp 177-178°C. ¹ H NMR (500 MHz, DMSO) δ 7.97 (d, J=15.7 Hz, 1H), 7.38-7.30 (m, 6H), 7.14 (t, J=8.8 Hz, 2H), 6.46 (d, J=15.7 Hz, 1H), 3.52 (s, 2H), 3.22 (d, J=6.7Hz, 2H), 2.78 (d, J=11.3Hz, 2H), 2.11 (dd, J=10.8, 6.7Hz, 1H), 1.89 (t, J=10.8Hz, 2H), 1.65 (d, J=11.3Hz, 2H), 1.41 (s, 18H), 1.34-1.26 (m, 4H). ¹³ C NMR (126MHz, DMSO) δ 165. 69, 155.91, 139.91, 139.66, 132.01, 129.36, 126.74, 124.63, 119.43, 115.54, 55.34, 53.53, 36.61, 36.36, 34.97, 33.09, 32.23, 30.65.ESI-HMS] m/z: 495.3 ; HRMS (ESI) m/z: 495.3381 [M+H]+ (calcd for 491.3382, C ₃₁ H ₄₄ FN ₂ O ₂ ).

Example 17

Preparation of 4-(N-1-(3-Fluoro-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7e

The preparation process is the same as in 13, the raw material 4-methoxybenzyl bromide is replaced with 3-fluorobenzyl bromide, the yield is 51%; the brown solid, mp167-169℃, ¹ H NMR (500MHz, DMSO)δ7.93(d, J =15.7Hz, 1H), 7.38-7.30(m, 6H), 7.14(t, J=8.8Hz, 2H), 6.46(d, J=15.7Hz, 1H), 3.42(s, 2H), 3.20(d , J=6.7Hz, 2H), 2.77 (d, J=11.3Hz, 2H), 2.13 (dd, J=10.8, 6.7Hz, 1H), 1.89 (t, J=10.8Hz, 2H), 1.65 (d , J=11.2Hz, 2H), 1.41 (s, 18H), 1.34-1.26 (m, 4H). ¹³ C NMR (126MHz, DMSO) δ 165.69, 155.91, 139.91, 139.66, 132.01, 129.36, 126.74, 124.63 , 119.43, 115.54, 55.34, 53.53, 36.61, 36.36, 34.97, 33.09, 32.23, 30.65. ESI-MS m/z: 495.3[M+H]+; HRMS(ESI) m/z: 495.3381[M+H] +(calcd for 491.3382, C ₃₁ H ₄₄ FN ₂ O ₂ ).

Example 18

Preparation of 4-(N-1-(4-Fluoro-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7f

The preparation process is the same as that in 13, except that the raw material 4-methoxybenzyl bromide is replaced with 4-fluorobenzyl bromide, the yield is 54%; the brown solid, mp181-183°C. ¹ H NMR (500MHz.DMSO) δ 7.93 (d, J=15.7Hz, 1H), 7.38-7.30 (m, 6H), 7.14 (t, J=8.8Hz, 2H), 6.46 (d, J=15.7 Hz, 1H), 3.42 (s, 2H), 3.20 (d, J=6.7Hz, 2H), 2.77 (d, J=11.3Hz, 2H), 2.13 (dd, J=10.8, 6.7Hz, 1H), 1.89 (t, J=10.8Hz, 2H), 1.65 (d, J=11.0Hz, 2H), 1.41 (s, 18H), 1.34-1.26 (m, 4H). ¹³ C NMR (126MHz, DMSO) δ 165. 69, 155.91, 139.91, 139.66, 132.01, 129.36, 126.74, 124.63, 119.43, 115.54, 55.34, 53.53, 36.61, 36.36, 34.97, 33.09, 32.23, 30.65.ESI-HMS] m/z: 495.3 ; HRMS (ESI) m/z: 495.3381 [M+H]+ (calcd for 491.3382, C ₃₁ H ₄₄ FN ₂ O ₂ ).

Example 19

Preparation of 4-(N-1-(2,4-difluoro-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7g

The preparation process is the same as that in 13, except that the raw material 4-methoxybenzyl bromide is replaced with 2,4-difluorobenzyl bromide, the yield is 44%; the brown solid, mp217-218°C. ¹ H NMR (500 MHz, DMSO) δ 7.93 (t, J=10.1 Hz, 1H), 7.44 (dd, J=15.7, 8.5 Hz, 1H), 7.33 (m, 4H), 7.19 (td, J=10.1 , 2.5Hz, 1H), 7.06 (td, J=8.5, 2.3Hz, 1H), 6.46 (d, J=15.7Hz, 1H), 3.47 (s, 2H), 3.19 (d, J=6.1Hz, 2H) ), 2.79(d, J=11.2Hz, 2H), 1.94(t, J=10.7Hz, 2H), 1.65(d, J=10.7Hz, 2H), 1.47-1.37(m, 18H), 1.34-1.23 (m, 3H), 1.10 (dd, J=11.2, 6.4Hz, 2H). ¹³ C NMR (151MHz, DMSO) δ 165.66, 155.90, 139.89, 139.64, 133.06, 126.70, 124.66, 119.39, 111.66, 103.82, 54.83, 53.39, 46.13, 36.62, 36.33, 35.03, 33.04, 32.19, 30.50. ESI-MS m/z: 513.3[M+H]+; HRMS(ESI)m/z: 513.3287[M+H]+(calcd for 513.3290, C ₃₁ H ₄₃ F ₂ N ₂ O ₂ ).

Example 20

Preparation of 4-(N-1-(3,4-difluoro-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7h

The preparation process was the same as that in 13, except that the raw material 4-methoxybenzyl bromide was replaced with 3-fluorobenzyl bromide, the yield was 46%; the brown solid, mp 224-226°C, ¹ H NMR (500 MHz, DMSO) δ7.93 (t, J=10.1Hz, 1H), 7.44 (dd, J=15.7, 8.5Hz, 1H), 7.33 (m, 4H), 7.19 (td, J=10.1, 2.5Hz, 1H), 7.06 (td, J=8.5 , 2.5Hz, 1H), 6.46 (d, J=15.7Hz, 1H), 3.47 (s, 2H), 3.19 (d, J=6.1Hz, 2H), 2.79 (d, J=11.2Hz, 2H), 1.94(t, J=10.7Hz, 2H), 1.65(d, J=10.7Hz, 2H), 1.47-1.37(m, 18H), 1.34-1.23(m, 3H), 1.10(dd, J=11.8, 6.1Hz, , 2H). ¹³ C NMR (151MHz, DMSO) δ 165.66, 155.90, 139.89, 139.64, 133.06, 126.70, 124.66, 119.39, 111.66, 103.82, 54.83, 53.39, 46.13, 36.62, 3.04.33 , 32.19, 30.50. ESI-MS m/z: 513.3 [M+H]+; HRMS(ESI) m/z: 513.3287 [M+H]+ (calcd for 513.3290, C ₃₁ H ₄₃ F ₂ N ₂ O ₂ ).

Example 21

Preparation of 4-(N-1-(2-chloro-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7i

The preparation process was the same as in 13, except that the raw material 4-methoxybenzyl bromide was replaced with 2-chlorobenzyl bromide, the yield was 48%; the brown solid, mp 203-205°C, ¹ H NMR (500MHz, DMSO) δ7.93 (d, J=15.7Hz, 1H), 7.40-7.25(m, 7H), 6.45(d, J=15.7Hz, 1H), 3.45(s, 2H), 3.35(s, 5H), 2.78(d, J=11.3 Hz, 2H), 1.92 (t, J=10.6Hz, 2H), 1.66 (d, J=11.0Hz, 2H), 1.41 (s, 18H), 1.16 (dd, J=11.8, 10.6Hz, 2H). ¹³ C NMR (126MHz, DMSO) δ 165.69, 155.91, 139.91, 139.66, 132.01, 129.36, 126.74, 124.63, 119.43, 115.54), 52.34, 53.53, 36.61, 36.36, 34.97, 33.5.09, 32.3 MS-3 m/z: 511.3 [M+H]+; HRMS (ESI) m/z: 511.3086 [M+H] ₊ ( _calcd for _511.3088 , _C31H43ClN2O2 ).

Example 22

Preparation of 4-(N-1-(3-chloropiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7j

The preparation process was the same as that in 13, except that the raw material 4-methoxybenzyl bromide was replaced with 3-chlorobenzyl bromide, the yield was 64%; the brown solid, mp194-196°C, ¹ H NMR (500MHz, DMSO)δ7.93(d, J =15.7Hz, 1H), 7.40-7.25(m, 7H), 6.45(d, J=15.7Hz, 1H), 3.45(s, 2H), 3.35(s, 5H), 2.78(d, J=11.3Hz , 2H), 1.92 (t, J=10.6Hz, 2H), 1.66 (d, J=11.0Hz, 2H), 1.41 (s, 18H), 1.16 (dd, J=11.8, 10.6Hz, 2H). ¹³ C NMR (126MHz, DMSO) δ165.69, 155.91, 139.91, 139.66, 132.01, 129.36, 126.74, 124.63, 119.43, 115.54, 55.34, 53.53, 36.61, 36.36, 34.97, 33.09, 32.23 z: 511.3 [M+H]+; HRMS (ESI) m/z: 511.3086 [M+H] ₊ ( _calcd for _511.3088 , _C31H43ClN2O2 ).

Example 23

Preparation of 4-(N-1-(4-bromo-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7k

The preparation process is the same as in 13, the raw material 4-methoxybenzyl bromide is replaced with 4-bromobenzyl bromide, the yield is 64%; the brown solid, mp154-155℃, ¹ H NMR (500MHz, DMSO)δ7.94(d, J =15.7Hz, 1H), 7.49(s, 1H), 7.47-7.44(m, 1H), 7.35-7.29(m, 5H), 6.45(d, J=15.7Hz, 1H), 4.13(s, 2H) , 3.45(s, 2H), 2.77(d, J=11.3Hz, 2H), 1.92(t, J=10.6Hz, 2H), 1.66(d, J=10.8Hz, 2H), 1.41(m, 20H) ^The 33.23, 32.22, 30.68. ESI-MS m/z: 555.2 [M+H]+; HRMS (ESI) m/z: 555.2509 [M+H]+ (cal cd for 555.2508, C ₃₁ H ₄₃ BrN ₂ O ₂ ) .

Example 24

Preparation of 4-(N-1-(4-nitro-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 71

The preparation process was the same as that in 13, except that the raw material 4-methoxybenzyl bromide was replaced with 4-nitrobenzyl bromide, the yield was 64%; the brown solid, mp 127-128°C, ¹ H NMR (500MHz, DMSO) δ8.18-8.10 ( m, 2H), 7.94 (d, J=15.7Hz, 1H), 7.77 (d, J=7.6Hz, 1H), 7.64 (t, J=7.9Hz, 1H), 7.36-7.30 (m, 4H), 6.46(d, J=15.7Hz, 1H), 3.60(s, 2H), 3.21(dd, J=12.8, 6.7Hz, 2H), 2.80(d, J=12.8Hz, 2H), 1.97(t, J =11.3Hz, 2H), 1.67 (d, J=11.3Hz, 2H), 1.41 (m, 20H), 1.34 (s, 3H). ¹³ C NMR (151 MHz, DMSO) δ 165.69, 155.91, 148.26, 141.77 , 139.91, 139.64, 135.81, 130.12, 126.70, 124.66, 123.38, 122.33, 119.37, 61.66, 53.64, 36.62, 36.33, 34.95, 33.11, 32.25, 31.64, 30.63..3 [SIH-MS m/z ]+; HRMS(ESI) m/z: 522.3326 [M+H]+ (calcd for 522.3325, C ₃₁ H ₄₄ N ₃ O ₄ ).

Example 25

Preparation of 4-(N-1-(4-cyano-benzylpiperidine))aminoethyl-2,4-di-tert-butyl-p-hydroxyphenylacrylamide 7m

The preparation process is the same as that in 13, except that the raw material 4-methoxybenzyl bromide is replaced with 4-cyanobenzyl bromide, the yield is 64%; the brown solid, mp215-217°C. ¹ H NMR (500MHz, DMSO) δ 8.18-8.10 (m, 2H), 7.94 (t, J=7.9Hz, 1H), 7.77 (d, J=15.6Hz, 1H), 7.64 (t, J=7.9 Hz, 1H), 7.36-7.30(m, 4H), 6.46(d, J=15.7Hz, 1H), 3.60(s, 2H), 3.21(dd, J=12.8, 6.7Hz, 2H), 2.80(d , J=12.8Hz, 2H), 1.97(t, J=11.0Hz, 2H), 1.67(d, J=11.3Hz, 2H), 1.41(m, 20H), 1.34(s, 3H). ¹³ C NMR (151MHz，DMSO)δ165.69，155.91，148.26，141.77，139.91，139.64，135.81，130.12，126.70，124.66，123.38，122.33，119.37，61.66，53.64，36.62，36.33，34.95，33.11，32.25，31.64，30.63 .ESI-MS m/z: 502.3[M+H]+; HRMS(ESI) m/z: 502.3357[M+H]+ (calcd for 522.3355, C32H43N3O2).

Claims (7)

1. a kind of donepezil-BHT heterocomplex or its pharmaceutically acceptable salt that general formula is following:

Wherein n=0,1,2, R is respectively CH₃, OCH₃, NO₂, CN, halogen atom etc..

2. the compound of claim 1 or its pharmaceutically acceptable salt, wherein optimal compound is:

3. the compound of claim 1 or its pharmaceutically acceptable salt, wherein pharmaceutically acceptable salt is general formula compound Hydrochloride, hydrobromate, sulfate, phosphate, mesylate, benzene sulfonate, tosilate, naphthalene sulfonate, lemon Hydrochlorate, tartrate, lactate, acetonate, acetate, maleate, succinate, fumarate, salicylate, phenyl Acetate, mandelate, alkali metal cations salt, alkaline earth metal cation salt or ammonium cation salt.

4. a kind of pharmaceutical composition, the compound of the general formula containing claim 1 or its pharmaceutically acceptable salt and pharmaceutically Acceptable carrier.

5. preparing claim

A kind of preparation method of claim formula (1) compound, this method comprises:

1) acid with the benzyl piepridine amine of different chain length of the BHT class of different chain length in condensing agent I-hydroxybenzotriazole (HOBT) and Target product 3a-i is generated under 1- ethyl-(3- dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDCI)；

2) bromo- 3 ', 5 '-di-t-butyl-the 4 '-hydroxy acetophenone of 2- and the amine of different chain length are in K₂CO₃With acetonitrile as solvents under effect Generate target product 3j-1；

3) the different bromobenzyls replaced and t-butoxycarbonylaminoethyl piperidines are in ethanol as solvent, and triethylamine is under conditions of acid binding agent Then the t-butoxycarbonylaminoethyl benzyl piepridine of generation takes off tertbutyloxycarbonyl (Boc protection under the action of trifluoroacetic acid Base), then reacted with 2,5- di-t-butyl -4- hydroxy-cinnamic acid and generate target product 7a-m.

6. the compound of the general formula of claim 1 or its pharmaceutically acceptable salt are used to be used as acetylcholinesterase inhibitor, With preferable Antioxidation in vitro, apparent neuroprotection is shown on cell, anti-inflammatory effect and in animal row Prove to have the preferable purposes for improving related disease drug to learn.

7. the purposes of claim 6, wherein disease relevant to such activity is Alzheimer disease.