CN117586192A - Compound and preparation method and application thereof - Google Patents

Abstract

The invention provides a compound, a preparation method and application thereof, and relates to the field of natural pharmaceutical chemistry. The invention utilizes the particularity of the framework of the atractylenolide I to carry out chemical structure modification to the atractylenolide I, so as to obtain the atractylenolide I derivative, and the chemical structural formula of the atractylenolide I is shown as (I). The compound provided by the invention can improve Abeta _25‑35 The cell survival rate of the induced SH-SY5Y cell injury has the function of protecting nerve cell injury caused by Abeta, and has the prospect of being developed into an anti-Alzheimer disease drug.

Description

Compound and preparation method and application thereof

Technical Field

The invention relates to the field of natural pharmaceutical chemistry, in particular to a compound and a preparation method and application thereof.

Background

Alzheimer's Disease (AD) is a neurodegenerative Disease that severely threatens the physical health of elderly people and is clinically characterized by global dementia such as dysmnesia, aphasia and behavioral changes. The disease rate of AD is high, the course of disease is long, and the hazard is huge. The existing clinical commonly used AD therapeutic drugs mainly comprise acetylcholinesterase inhibitors (donepezil, rivastigmine, huperzine A, galantamine and the like) and NMDA receptor antagonists memantine, but the drugs can only improve related symptoms, and can not delay and prevent the development of diseases or achieve the effect of curing AD. The pathogenesis of AD is complex, the influence factors are numerous, and the currently accepted pathological characteristics are neurotoxicity caused by senile plaques formed by the excessive deposition of beta-Amyloid (Abeta) in brain, so that cholinergic neurons of the central nervous system die, lose and the like. Thus, inhibition of aβ deposition or alleviation of aβ damage to neuronal cells is one of the main directions for anti-AD drug development.

The pathology mechanism of the Alzheimer disease is complex, western medicine research and development are not good for a single target or a single action path, but the traditional Chinese medicine compound has the advantages of multiple targets, multiple channels, diversity and the like, and the clinical curative effect evaluation of the traditional Chinese medicine compound for treating the AD and the pharmacological mechanism research of the traditional Chinese medicine have positive results, so that the traditional Chinese medicine has wide prospect in the aspect of treating the Alzheimer disease. The bighead atractylodes rhizome is a common warm tonic, has the effects of preventing abortion, suppressing sweating, drying dampness, promoting diuresis, strengthening spleen, tonifying qi and the like, has various components, exerts main medicinal components such as volatile components, polysaccharides, lactones, glycosides and the like, and is concentrated in the research of chemical components of the bighead atractylodes rhizome at present. The atractylenolide I is taken as one of important components of the atractylenolide I, and pharmacological researches on the atractylenolide I are carried out in the field of treatment of different diseases, and the researches show that the atractylenolide I has a certain protection effect on blood brain barrier and a certain neuroprotection effect by inhibiting the apoptosis of cerebrovascular endothelial cells induced by oxidative stress injury. However, the atractylenolide I has weak AD resistance, is unstable in structure under alkaline conditions and is easy to decompose, and the application of the atractylenolide I in the development of AD resistance medicines is affected. Based on the background, the invention aims to utilize the particularity of the framework of the atractylenolide I to carry out chemical structure transformation modification on the framework of the atractylenolide I so as to expect to obtain a compound with better stability and higher anti-AD activity.

Disclosure of Invention

(one) solving the technical problems

Aiming at the defects of the prior art, the invention provides a compound for treating or preventing Alzheimer's disease and a preparation method thereof.

(II) technical scheme

In order to achieve the above purpose, the invention is realized by the following technical scheme:

in one aspect, the present invention provides a compound which is a derivative of atractylenolide I, having the chemical structural formula (I)

Further, a compound represented by formula (I) or a pharmaceutically acceptable salt or a solvent compound thereof.

In another aspect, the invention provides the use of a compound of formula (I) for the manufacture of a medicament for the treatment or prophylaxis of Alzheimer's disease.

In yet another aspect, the invention provides a pharmaceutical composition for the treatment of Alzheimer's disease comprising an effective amount of formula (I) and a pharmaceutically acceptable carrier.

Further, the pharmaceutical composition is a capsule, a tablet, an injection, a granule, a pill or a powder.

In yet another aspect, the present invention also provides a method for preparing a compound, comprising the steps of:

(1) Dissolving atractylenolide I and selenium dioxide in anhydrous dioxane solvent, heating to 80-90 ℃ under the protection of nitrogen, stirring for reaction for 2-3 hours, cooling to room temperature after the reaction is finished, distilling under reduced pressure to remove dioxane, then adding ethyl acetate, washing an organic phase with saturated sodium thiosulfate solution for 2-3 times, washing with deionized water for 3-5 times, separating the organic phase, concentrating and drying the organic phase to obtain an intermediate 1; intermediate 1 has a chemical structural formula of formula (II)

(2) Dissolving the intermediate 1 in dichloromethane, adding N, N' -Carbonyl Diimidazole (CDI), stirring at room temperature for 12-16h, adding ammonia water with mass fraction of 25-30%, and continuously stirring overnight; after the reaction is finished, adding dichloromethane into the reaction liquid, washing 3-5 times by deionized water, separating an organic phase, concentrating and drying the organic phase to obtain an intermediate 2; intermediate 2 has a chemical structural formula of formula (III)

(3) Dissolving the intermediate 2 in ethanol, adding hydrazine hydrate with the mass fraction of 80-85%, heating to 40-50 ℃ under the protection of nitrogen, reacting for 36-48h, cooling to room temperature after the reaction is finished, distilling under reduced pressure to remove ethanol, adding ethyl acetate, washing with deionized water for 3-5 times, separating an organic phase, concentrating and drying the organic phase, and obtaining the compound shown in the formula (I).

(III) beneficial effects

Alzheimer's disease is a progressive neurodegenerative disease with hidden disease. Clinically, irreversible deterioration of cognitive and memory functions is manifested by damage to neurons caused by the excessive deposition of aβ in the brain. The invention uses the particularity of the framework of the atractylenolide I to carry out chemical structure on the atractylenolide IAnd (3) modifying to obtain the derivatives of the atractylenolide I, wherein the chemical structural formula of the derivatives is shown as (I). The compound provided by the invention can improve Abeta ₂₅ The cell survival rate of SH-SY5Y cell injury induced by-35 has the function of protecting nerve cell injury caused by Abeta, and has the prospect of being developed into an anti-Alzheimer disease drug.

Drawings

FIG. 1 is a flow chart showing the steps for preparing the compound represented by formula (I).

Detailed Description

For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions in the embodiments of the present invention will be clearly and completely described in the following in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

Example 1

A compound is a derivative of atractylenolide I, and has a chemical structural formula (I)

The compound is prepared according to the steps shown in fig. 1, specifically:

(1) Bighead atractylodes lactone I (115 mg,0.5 mmol) and selenium dioxide (83 mg,0.75 mmol) were added to a dry round bottom flask, then 3mL of anhydrous dioxane was added for dissolution, and the mixture was heated to 80℃under nitrogen protection and stirred for 2h. After the reaction solution was cooled to room temperature, dioxane was distilled off under reduced pressure, and then 5mL of ethyl acetate was added, and the organic phase was washed twice with a saturated sodium thiosulfate solution and three times with water. The organic phase was concentrated by drying and then purified by column chromatography (petroleum ether/ethyl acetate=4:1-2:1) to give intermediate 1 (86 mg, 70% yield) as a pale yellow oily liquid;

intermediate 1 spectrum data: ¹ HNMR(500MHz,Chloroform-d)δ5.63(s,1H),5.15(s,1H),4.79(s,1H),4.37(t,J＝2.9Hz,1H),2.91-2.95(m,1H),2.64-2.68(m,1H),2.47-2.54(m,1H),2.02-2.08(m,1H),1.91(m,3H),1.86(m,2H),1.49(m,2H),0.93(s,3H).

(2) Intermediate 1 (80 mg,0.32 mmol) was dissolved in 2mL of Dichloromethane (DCM), then CDI (79 mg,0.49 mmol) was added, stirred at room temperature for 12h, then 25% aqueous ammonia 0.1mL was added and stirring continued overnight. After the reaction, 3mL of DCM was added to the reaction solution, which was washed three times with water, and then the organic phase was concentrated by drying and purified by column chromatography (petroleum ether/ethyl acetate=2:1-1:1) to give intermediate 2 (37 mg, yield 40%) as a colorless wax;

intermediate 2 spectrogram data: ¹ HNMR(500MHz,Chloroform-d)δ5.63(s,1H),5.30(s,1H),5.29(m,1H),4.91(s,1H),4.58(br,2H),2.72(m,2H),2.51(m,1H),1.95-2.01(m,1H),1.92(s,3H),1.89(m,2H),1.55(m,1H),0.95(s,3H)；ESI-MS(m/z):290.33[M+1] ⁺ .

(3) Intermediate 2 (35 mg,0.12 mmol) was dissolved in 2mL of ethanol, 0.01mL of 80% hydrazine hydrate was added, and then heated to 40℃under nitrogen protection for reaction for 36h. The reaction solution was cooled to room temperature, ethanol was then removed by distillation under the reduced pressure, 3mL of ethyl acetate was then added, and the mixture was washed three times with water. The organic phase was concentrated by dryness and purified by column chromatography (petroleum ether/ethyl acetate=1:5), the compound of formula (i) (14.5 mg, 40% yield) was a white solid;

spectrogram data of a compound shown in formula (I): ¹ HNMR(500MHz,Chloroform-d)δ11.54(br,1H),5.15(s,1H),4.83(s,1H),4.42(t,J＝2.9Hz,1H),2.73(m,1H),2.61(m,2H),2.45(m,2H),2.17(s,3H),1.90(m,2H),1.40-1.48(m,2H),0.71(s,3H)；ESI-MS(m/z):304.21[M+1] ⁺ .

experimental example 1

Protection of beta-amyloid induced SH-SY5Y cell injury by Compounds of formula (I)

1 Experimental materials

Aβ _25-35 Purchased from beijing boaosen biotechnology limited; dimethyl sulfoxide (DMSO) was purchased from Shanghai microphone Biochemical technologies Co., ltd; thiazole blue (MTT) was purchased from calico biotechnology limited; fetal bovine serum is purchased from Zhejiang Tianzhou biotechnologyStock limited; DMEM/F-12 medium is purchased from Gibco corporation.

Aβ _25-35 Preparation of the oligomer: 1mg of Abeta _25-35 1mL of sterile ddH ₂ In O, 1mg/mL of Abeta is arranged _25-35 The solution was incubated in a 37℃water bath for 7 days.

2 Experimental methods

2.1 cell culture

Taking out human neuroblastoma cells SH-SY5Y frozen in liquid nitrogen, rapidly placing in 37 ℃ water bath, shaking until the cells melt, centrifuging at 1000rpm for 5min, discarding supernatant, adding a small amount of culture medium which is DMEM/F-12 culture medium containing 10% fetal bovine serum and 1% diab, transferring the cells into a culture dish, and concentrating at 37 ℃ and 5% CO ₂ Is cultured in a cell culture incubator.

2.2 preparation of cell model and drug pair Abeta _25-35 Evaluation of protective action of induced neuronal injury

SH-SY5Y cells in logarithmic growth phase are prepared to have a concentration of 1.0X10 ⁵ Cell suspensions at each mL were seeded in 96-well plates at 100 μl per well and after 24h incubation, treated in groups of 5 multiplex wells:

control group: changing the culture medium to a fresh culture medium for continuous culture for 3 hours;

Aβ _25-35 group (model group): changing the culture medium to a fresh culture medium for continuous culture for 3 hours;

donepezil hydrochloride group (positive drug group): the culture was continued for 3 hours with a fresh medium containing 20. Mu.M donepezil hydrochloride;

atractylodes macrocephala lactone group I: changing to fresh culture medium containing 20 μm atractylenolide I, and culturing for 3 hr;

a group of compounds of formula (i): the culture was continued for 3 hours by changing to fresh medium containing 10, 20, 40. Mu.M of the compound of formula (I).

After culturing for 3 hours according to the above group, Aβ _25-35 Group, donepezil hydrochloride group, atractylenolide group I group, compound group of formula (I) added with condensed state Abeta with final concentration of 15 mu M _25-35 Adding the control group with the solvent with equal volume, culturing for 24 hr, adding 20 μl of each groupThe culture was continued at 37℃for 4 hours.

After the culture is continued for 4 hours, absorbing and discarding supernatant in the holes, adding 150 mu L of DMSO into each hole, and vibrating for 10 minutes to fully dissolve crystals; the wavelength of 490nm is selected, the light absorption value (OD value) of each hole is measured on an ELISA detector, the average of the OD values of 6 compound holes is taken, the cell survival rate is calculated, and the control group is recorded as 100%.

3 data processing and analysis

The cell viability of each group was calculated using SPSS19.0 software, the comparison between groups was tested using one-way anova, and the comparison between groups was tested using q. The difference of P <0.05 is statistically significant.

Results 4 results

Table 1 shows the values of Aβ in each group _25-35 Cell viability under induced SH-SY5Y cell damage. As can be seen from Table 1, abeta compared with the control group _25-35 The survival rate of the group cells is obviously reduced, which indicates that the cell modeling is successful; with Abeta _25-35 Compared with the group, the cell viability of the donepezil hydrochloride group and that of the atractylenolide I group are increased, the cell viability of the compound shown in the formula (I) is respectively increased by about 8.3 percent and 4.5 percent at 20 mu M, and the cell viability of the compound shown in the formula (I) is respectively increased by about 12.4 percent, 18.1 percent and 22.4 percent at 10 mu M, which are obviously superior to those of the donepezil hydrochloride group and the atractylenolide I group. To exclude false positives caused by the promotion of cell proliferation by the compounds of formula (I), we tested the absence of Abeta _25-35 The results of the intervention of the compound of the formula (I) in the cell viability are shown in Table 2, the compound of the formula (I) has no significant effect on cell viability at 10, 20 and 40. Mu.M (P>0.05 Compound 3) showed no cell proliferation.

TABLE 1 Aβ _25-35 Cell survival following different drug treatments under induced SH-SY5Y cell injury

Note that: comparison with the control group ^# P<0.05， ^## P<0.01， ^### P<0.001; comparison with model group P<0.05，**P<0.01，***P<0.001; the following is the same.

TABLE 2 Effect of Compounds of formula (I) on SH-SY5Y cell viability

The results show that the compound shown as the formula (I) provided by the invention can improve Abeta _25-35 The induced SH-SY5Y cell damage cell survival rate has the function of protecting nerve cell damage caused by Abeta, and has the prospect of being developed into an AD-resistant medicament.

The above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims (6)

1. A compound is characterized in that the compound is a derivative of atractylenolide I, and the chemical structural formula of the compound is (I)

2. A compound according to claim 1, wherein the compound of formula (i) is a pharmaceutically acceptable salt or solvate thereof.

3. The use of a compound according to claim 1, wherein the compound of formula (i) is for the preparation of a medicament for the treatment or prophylaxis of alzheimer's disease.

4. A pharmaceutical composition for the treatment or prophylaxis of alzheimer's disease, characterized in that the pharmaceutical composition comprises an effective amount of formula (i) and a pharmaceutically acceptable carrier.

5. The pharmaceutical composition for treating or preventing alzheimer's disease according to claim 4, wherein said pharmaceutical composition is in the form of a capsule, tablet, injection, granule, pill or powder.

6. A process for the preparation of a compound according to claim 1, wherein the compound of formula (i) is prepared according to the following steps:

(1) Dissolving atractylenolide I and selenium dioxide in anhydrous dioxane solvent, heating to 80-90 ℃ under the protection of nitrogen, stirring for reaction for 2-3 hours, cooling to room temperature after the reaction is finished, distilling under reduced pressure to remove dioxane, then adding ethyl acetate, washing an organic phase with saturated sodium thiosulfate solution for 2-3 times, washing with deionized water for 3-5 times, separating the organic phase, concentrating and drying the organic phase to obtain an intermediate 1;

(2) Dissolving the intermediate 1 in dichloromethane, adding N, N' -Carbonyl Diimidazole (CDI), stirring at room temperature for 12-16h, adding ammonia water with mass fraction of 25-30%, and continuously stirring overnight; after the reaction is finished, adding dichloromethane into the reaction liquid, washing 3-5 times by deionized water, separating an organic phase, concentrating and drying the organic phase to obtain an intermediate 2;

(3) Dissolving the intermediate 2 in ethanol, adding hydrazine hydrate with the mass fraction of 80-85%, heating to 40-50 ℃ under the protection of nitrogen, reacting for 36-48h, cooling to room temperature after the reaction is finished, distilling under reduced pressure to remove ethanol, adding ethyl acetate, washing with deionized water for 3-5 times, separating an organic phase, concentrating and drying the organic phase, and obtaining the compound shown in the formula (I).