Compound with neuroprotective activity, preparation method and application

Abstract

The invention relates to the technical field of medical chemistry, and discloses a compound PX5-9 with neuroprotective activity as well as a preparation method and application thereof. The compound PX5-9 is obtained by introducing an N-Oxide group on the basis of the structure of a compound 5, and has remarkable neuroprotective activity, improved solubility and equivalent biological activity. The compound PX5-9 is used for preparing drugs for treating central nervous system degenerative diseases or traumatic brain injury, such as Alzheimer's disease or Parkinson's disease, solves the problems of poor solubility and affected druggability of the compound 5, and has potential clinical application value.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical chemistry, and specifically relates to a compound with neuroprotective activity, a preparation method and an application thereof. Background Art

[0002] Neurodegenerative diseases (NDDs) are the most important components of disabling diseases in the world. Globally, the number of patients with NDDs such as Alzheimer's disease and Parkinson's disease has exceeded 100 million, and it shows an accelerating growth trend with the aging of the population. At present, most clinical treatment drugs are limited to symptom relief, lacking radical therapies that can reverse the disease process. Although many compounds with high-efficiency neuroprotective effects have been verified in vitro, the vast majority of compounds have poor in vivo effects due to their poor physicochemical properties. More than 40% of the candidate drugs have a bioavailability of less than 10% due to insufficient solubility. The problem of blood-brain barrier (BBB) penetration further exacerbates the difficulty of developing central nervous system drugs - the drug needs to be highly lipophilic to penetrate the BBB and also need to have sufficient water solubility to maintain plasma stability.

[0003] Chinese Patent CN 112209848 A discloses a cyclopentane cinnamide derivative (Compound 5) with neuroprotective activity, and its structural formula is as follows:

[0004]

[0005] Although Compound 5 has shown good neuroprotective effects, its solubility is very poor, seriously affecting its drugability. Therefore, it is necessary to improve the solubility, stability and biological activity of Compound 5 to enhance its clinical application value. Summary of the Invention

[0006] Aiming at the deficiencies of the prior art, the present invention provides a compound with neuroprotective activity, a preparation method and an application thereof. On the basis of the structure of Compound 5, the present invention introduces an N-Oxide group to obtain a new cyclopentane cinnamide N-Oxide compound PX5-9. This compound PX5-9 has significant neuroprotective activity, improved solubility and equivalent biological activity, and has potential clinical application value.

[0007] To achieve the above object, the present invention adopts the following technical solutions:

[0008] A compound PX5-9 with neuroprotective activity, and its general structural formula is as follows:

[0009]

[0010] An embodiment of the present invention also discloses a pharmaceutical composition, comprising compound PX5-9 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0011] An embodiment of the present invention also discloses a preparation method of compound PX5-9, which is as follows: Dissolve compound 5 in a solvent to form a compound 5 solution, dissolve an oxidant in a solvent to form an oxidant solution, and under stirring conditions from 0 °C to room temperature, slowly drop the oxidant solution into the compound 5 solution. Monitor the reaction process by thin-layer chromatography (TLC). After the raw material spots disappear, remove the solvent by reduced pressure distillation; The crude product is purified to obtain the target compound PX5-9 in the form of a white powder; The molar ratio of compound 5 to the oxidant is 1:(1 to 1.5).

[0012] In some embodiments, the oxidant is m-chloroperoxybenzoic acid, hydrogen peroxide or p-chloroperoxybenzoic acid.

[0013] In some embodiments, the solvent is dichloromethane, ethyl acetate, acetonitrile or methanol.

[0014] In some embodiments, the purification method is column chromatography, recrystallization or HPLC purification.

[0015] An embodiment of the present invention also discloses the use of compound PX5-9 or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating central nervous system degenerative diseases or traumatic brain injury.

[0016] Preferably, the central nervous system degenerative diseases include Alzheimer's disease or Parkinson's disease.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0018] Compound PX5-9 obtained by N-Oxide modification of compound 5 disclosed in CN 112209848 A of the present invention has significant neuroprotective activity, improved solubility and equivalent biological activity, solves the problems of poor solubility and affected drugability of compound 5, and has potential clinical application value. Description of the Drawings

[0019] Figure 1 is the 1H nuclear magnetic resonance spectrum of compound PX5-9 prepared in Example 1;

[0020] Figure 2 is the 13C nuclear magnetic resonance spectrum of compound PX5-9 prepared in Example 1;

[0021] Figure 3 is the high-resolution mass spectrum of compound PX5-9 prepared in Example 1;

[0022] Figure 4It is the liquid chromatogram of the compound PX5-9 prepared in Example 1;

[0023] Figure 5 It is a schematic diagram comparing the neuroprotective effects of compound 5 and PX5-9 in a glutamate-induced HT22 cell injury model. Detailed implementation manners

[0024] The present invention will be further described below in conjunction with the detailed implementation manners, but the embodiments do not limit the present invention in any form. The reagents and instruments used in the embodiments are all conventional products that can be obtained through commercial purchase without special instructions. The experimental methods used in the embodiments are all conventional experimental methods or technical means in the art without special instructions.

[0025] An embodiment of the present invention provides a compound PX5-9 having neuroprotective activity, and its structural general formula is as follows:

[0026]

[0027] The embodiment of the present invention also discloses a pharmaceutical composition, which includes the compound PX5-9 or its pharmaceutically acceptable salt and a pharmaceutically acceptable carrier.

[0028] The embodiment of the present invention also discloses a preparation method of the compound PX5-9, specifically as follows: Dissolve the compound 5 in a solvent to form a compound 5 solution, dissolve the oxidant in a solvent to form an oxidant solution, and under the stirring condition of 0 °C to room temperature, slowly drop the oxidant solution into the compound 5 solution, monitor the reaction process by thin layer chromatography (TLC), and after the raw material spots disappear, remove the solvent by reduced pressure distillation; The crude product is purified to obtain the target compound PX5-9 in the form of a white powder; the molar ratio of the compound 5 to the oxidant is 1:(1-1.5).

[0029] In some embodiments, the oxidant is m-chloroperoxybenzoic acid, hydrogen peroxide or p-chloroperoxybenzoic acid.

[0030] In some embodiments, the solvent is dichloromethane, ethyl acetate, acetonitrile or methanol.

[0031] In some embodiments, the purification method uses column chromatography, recrystallization or HPLC purification.

[0032] Example 1

[0033] The preparation method of compound PX5-9 is as follows: Dissolve compound 5 (3.00 g, 7.88 mmol, 1.0 equiv) in dichloromethane (20 mL); under stirring at room temperature, slowly add a dichloromethane solution (10 mL) of m-chloroperbenzoic acid (m-CPBA, 1.37 g, 7.96 mmol, 1.01 equiv) dropwise to the reaction system; monitor the reaction progress by thin-layer chromatography (TLC), and after the disappearance of the starting material spot, remove the solvent by distillation under reduced pressure; the crude product is rapidly purified by column chromatography on basic alumina, using dichloromethane as the eluent, and finally the target compound PX5-9 in the form of a white powder is obtained with a yield of 83%. 1 H NMR (500 MHz, MeOD) δ 7.93 (d, J = 9.1 Hz, 2H), 7.86–7.79 (m, 2H), 7.62 (d, J = 15.6 Hz, 1H), 7.21–7.12 (m, 2H), 6.98–6.92 (m, 1H), 6.64 (dd, J = 15.6, 1.3 Hz, 1H), 4.86 (d, J = 2.9 Hz, 5H), 3.86 (d, J = 1.9 Hz, 3H), 3.58 (s, 6H), 1.98–1.85 (m, 2H), 1.88–1.75 (m, 5H), 1.69–1.59 (m, 2H). 13 C NMR (126 MHz, MeOD) δ 167.19, 151.63, 151.18, 150.15, 143.54, 141.19, 128.99, 123.47, 121.64, 121.17, 119.47, 115.82, 112.36, 81.81, 63.13, 56.65, 33.73, 24.93. HRMS (ESI) calcd for C 23 H 29 N 2 O 3 + , [M - O] + : 380.2094, found: 380.2087。

[0034] Figure 1 is the 1H NMR spectrum of the compound PX5-9 prepared in Example 1.

[0035] Figure 2 is the 13C NMR spectrum of the compound PX5-9 prepared in Example 1.

[0036] Figure 3 is the high-resolution mass spectrum of the compound PX5-9 prepared in Example 1.

[0037] Figure 4 is the liquid chromatography spectrum of the compound PX5-9 prepared in Example 1.

[0038] Example 2

[0039] Determination of Solubility and LogP of Compound 5 and PX5-9 in Example 1

[0040] The solubility of Compound 5 and PX5-9 in water was determined by adding 10 mg of Compound 5 / PX5-9 to 3 ml of distilled water in a 5 ml centrifuge tube respectively. The test tube was sealed and stirred at 25 °C for 24 hours. After centrifuging at 10000 rpm for 10 minutes, the supernatant was filtered off and analyzed by HPLC / UV. By measuring the standard samples of the compound with known concentration gradients, a calibration curve was plotted, and thus the solubility of Compound 5 and PX5-9 was calculated.

[0041] The oil-water partition coefficient (LogP) was determined using the shake-flask method. Briefly, 2 mg of each compound was added to a mixture of 1 ml of n-octanol and 1 ml of water. The mixture was shaken at 25 °C for 24 hours, and then the undissolved solids were separated by centrifugation. The concentrations of the two phases were measured by HPLC / UV, and LogP was calculated as the ratio of the octanol concentration to the water concentration. The calculation formula is as follows:

[0042]

[0043] The solubility and LogP of Compound 5 and PX5-9 were detected using HPLC / UV technology. The experimental results showed that the solubility of PX5-9 (37.10 ± 0.33 μg / mL) was increased by more than 2400 times compared with Compound 5 (<0.015 μg / mL), and the LogP value decreased from >4.52 to 2.55 ± 0.03, indicating a significant increase in its hydrophilicity. This improvement can be attributed to the polar oxygen atoms introduced by N-Oxideation, indicating a better lipophilicity balance, which is beneficial to drug absorption and distribution. The detection data are shown in the following table:

[0044]

[0045] HT22 cells were cultured in DMEMs containing 10% fetal bovine serum and incubated under the conditions of 37 °C and 5% CO 2 HT22 cells were cultured in a 96-well plate at a concentration of 1×10 5 / mL. When the cell confluence reaches 40 - 50%, the cells are incubated with the medium containing compound 5 / PX5 - 9 (10 μM) for 30 minutes, and then treated with glutamate (6 mM) for 24 hours. 10 μL of 5 mg / mL MTT solution is added and incubated at 37 °C for 2 hours. The MTT reagent is removed and replaced with DMSO (100 μL). After shaking at room temperature for 10 minutes, the absorbance is measured by a UV spectrophotometer at 570 nm. The results are expressed as a percentage of untreated cells. There are at least 3 replicate experiments for each concentration. All data are expressed as the mean ± SEM of three independent experiments. In the glutamate - induced HT22 cell injury experiment, at a concentration of 10 μM, the neuroprotective effect of PX5 - 9 is comparable to that of compound 5 at the same concentration of 10 μM, as Figure 5 shown.

[0046] Figure 5 is a schematic diagram comparing the neuroprotective effects of compound 5 and PX5 - 9 in the glutamate - induced HT22 cell injury model.

[0047] It can be seen that compared with compound 5, PX5 - 9 has higher solubility and a more suitable LogP value, and has greater development potential.

[0048] The above embodiments have introduced the technical solutions provided by the present invention in detail. Specific examples are used in this article to elaborate on the principles and implementation manners of the embodiments of the present invention. The descriptions of the above embodiments are only applicable to helping understand the principles of the embodiments of the present invention; at the same time, for those of ordinary skill in the art, according to the embodiments of the present invention, there will be changes in the specific implementation manners and application scopes. In summary, the content of this specification should not be construed as a limitation to the present invention.

Claims

1. A compound having neuroprotective activity, characterized in that Its general structure is as follows:

2. A pharmaceutical composition, characterized in that It comprises the compound according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

3. The method for preparing a compound having neuroprotective activity according to claim 1, characterized in that: Specifically as follows: Compound 5 is dissolved in a solvent to form a compound 5 solution, an oxidant is dissolved in a solvent to form an oxidant solution, the oxidant solution is slowly dropped into the compound 5 solution under stirring conditions from 0°C to room temperature, the reaction progress is monitored by thin layer chromatography (TLC), and the solvent is removed by distillation under reduced pressure after the raw material point disappears; the crude product is purified to obtain the target compound PX5-9 as a white powder; the molar ratio of the compound 5 to the oxidant is 1:(1 to 1.5).

4. The method for preparing a compound having neuroprotective activity according to claim 3, characterized in that: The oxidant is m-chloroperbenzoic acid, hydrogen peroxide or p-chloroperbenzoic acid.

5. The method for preparing a compound having neuroprotective activity according to claim 3, characterized in that: The solvent is dichloromethane, ethyl acetate, acetonitrile or methanol.

6. The method for preparing a compound having neuroprotective activity according to claim 3, characterized in that: The purification method is column chromatography, recrystallization or HPLC purification.

7. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating degenerative diseases of the central nervous system or traumatic brain injury.

8. The use according to claim 7, wherein the central nervous system degenerative disease comprises Alzheimer's disease or Parkinson's disease.

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