A resveratrol imine derivative, and a preparation method and application thereof

Abstract

The application belongs to the technical field of anti-inflammatory and antioxidant drug development, and discloses a resveratrol imine derivative, a preparation method and application thereof. The resveratrol imine derivative is a compound with the structure shown in formula 2 or a pharmaceutically acceptable salt thereof, and a solvate of the compound with the structure shown in formula 2 or the pharmaceutically acceptable salt thereof. The resveratrol derivative is a novel compound, and the compounds are designed and successfully synthesized for the first time, and the structures thereof are characterized. The preparation method of the resveratrol derivative is simple, convenient to operate, and capable of rapidly synthesizing the compounds.

Description

Technical Field

[0001] This invention belongs to the field of anti-inflammatory and antioxidant drug development technology, and specifically relates to a resveratrol imine derivative, its preparation method and application. Background Technology

[0002] Resveratrol, chemically named 3,4',5-trihydroxy-1,2-diphenylethylene, has the molecular formula C2. 14 H 12 O3, with a molecular weight of 228.23 and CAS number 501-36-0, has the structure shown in Formula 1. Resveratrol was first discovered and extracted by Japanese scientists in 1939 from the roots of the poisonous plant *Resveratrol alatus*. Resveratrol is widely found in various plants, such as grapes, *Polygonum cuspidatum*, peanuts, and mulberries, and is a natural antitoxin secreted by plants in response to adverse conditions or pathogen invasion. Resveratrol exists in plants in both free and glycoside-bound forms, with the trans isomer exhibiting stronger biological activity.

[0003]

[0004] Resveratrol, a non-flavonoid polyphenol containing a stilbene structure, possesses various biological activities, including anti-aging, antioxidant, anti-inflammatory, anti-cancer, and cardiovascular protection, and its development and utilization are becoming increasingly sophisticated. Studies have shown that 600 mg / kg resveratrol can maintain gut health in laying hens under tBHP-induced oxidative stress by downregulating the mRNA expression levels of NF-κB and TNF-α, thereby mitigating the decline in laying performance and ovarian function caused by oxidative stress. Meng et al., through transcriptome and 16S rRNA sequencing analysis, found that adding resveratrol to the diet of sows may alleviate weaning-induced diarrhea and intestinal inflammation in weaned piglets by improving intestinal gene expression and regulating gut microbiota. These studies indicate that high-dose resveratrol can exert its anti-stress enteritis effect in livestock and poultry by regulating oxidative stress, inflammation-related signaling pathways, and gut microbiota.

[0005] However, resveratrol has an unstable structure, is easily oxidized, and its double bonds are highly photosensitized, readily transforming into a less active cis structure under ultraviolet light irradiation. It also has low polarity, poor water solubility, and low oral bioavailability (approximately 12%), requiring relatively large doses to achieve effective concentrations in plasma or tissues. Therefore, rapid absorption, low bioavailability, and poor water solubility are key factors limiting the clinical use of resveratrol. Given that resveratrol possesses certain anti-stress enteritis activity in livestock and poultry, the high in vivo dosage is only due to its low bioavailability and poor water solubility. If its structure can be modified by introducing characteristic groups into its core, it is hoped that resveratrol derivatives with better biological activity and higher bioavailability can be obtained.

[0006] Molecular hybridization refers to the chemical synthesis of two active molecular fragments to form a new active compound. Molecular hybridization can enhance the pharmacological activity of active molecules, improve drug stability, and reduce toxic side effects; it also provides more possibilities for drug design. Summary of the Invention

[0007] In order to overcome the shortcomings and deficiencies of the existing technology, the primary objective of this invention is to provide a resveratrol imine derivative.

[0008] Another object of the present invention is to provide a method for preparing the above-mentioned resveratrol imine derivative; the method improves structural stability and bioavailability by branching an aldehyde group on the benzene ring of resveratrol and connecting it with glycine or benzylamine derivatives.

[0009] Another object of the present invention is to provide an application of the above-mentioned resveratrol imine derivative. Because this resveratrol derivative has good in vitro antioxidant activity, it is suitable as a novel drug for oxidative stress diseases.

[0010] The objective of this invention is achieved through the following technical solution:

[0011] A resveratrol imine derivative, said derivative being a compound with the structure shown in Formula 2 or a pharmaceutically acceptable salt thereof, and a solvent compound of said compound with the structure shown in Formula 2 or a pharmaceutically acceptable salt thereof:

[0012]

[0013] Wherein, R is one of the following groups:

[0014]

[0015] The specific functional groups of the above compounds are summarized in Table 1:

[0016] Table 1. Compound Numbers and Structures

[0017]

[0018]

[0019] The pharmaceutically acceptable salt is a salt formed by the compound of formula 2 with lithium ions, sodium ions, potassium ions, calcium ions, magnesium ions and ammonium ions.

[0020] The preparation method of the above-mentioned resveratrol imine derivative includes the following steps:

[0021] (1) Resveratrol was reacted with N,N-dimethylformamide and phosphorus oxychloride to obtain intermediate I with the structure shown in Formula 3;

[0022]

[0023] (2) One of glycine, 2-F benzylamine, 4-F benzylamine and 4-OH benzylamine, together with intermediate I, were used as raw materials. Through condensation reaction and purification, a resveratrol imine derivative with the structure shown in Formula 2 was obtained.

[0024] The molar ratio of resveratrol, N,N-dimethylformamide and phosphorus oxychloride in step (1) is 1:1.1:1.1; the reaction is carried out at room temperature for 16 hours.

[0025] The specific steps (1) are as follows: using acetonitrile as solvent, N,N-dimethylformamide and phosphorus oxychloride are added under stirring in an ice-salt bath. After stirring for 5 minutes, resveratrol is added and the reaction is carried out overnight. The red solid is obtained by filtration and washed with cold acetonitrile. The obtained solid is transferred to a new reaction flask, water is added, and the reaction is carried out at 55°C for 3-5 hours. The solid is then filtered, washed, and dried to obtain intermediate I.

[0026] In step (2), one of glycine, 2-F benzylamine, 4-F benzylamine and 4-OH benzylamine is present in a molar ratio of 1:1 with intermediate I; the condensation reaction is carried out at room temperature for 16 hours under nitrogen protection.

[0027] The above-mentioned resveratrol imine derivatives are used in the preparation of drugs for preventing and treating oxidative stress.

[0028] The synthetic route for the resveratrol imine derivative is shown in the following formula, where all R groups are groups listed in Table 1:

[0029]

[0030] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0031] (1) The resveratrol imine derivatives provided by this invention are novel compounds that have not been reported before. This invention designs and successfully synthesizes these compounds for the first time, and characterizes their structures.

[0032] (2) The preparation method of the resveratrol imine derivatives of the present invention is simple and convenient to operate, and can quickly synthesize these compounds. Attached Figure Description

[0033] Figure 1 This is the NMR spectrum of compound 1.

[0034] Figure 2 This is the NMR spectrum of compound 2.

[0035] Figure 3 This is the NMR spectrum of compound 3.

[0036] Figure 4 This is the NMR spectrum of compound 4.

[0037] Figure 5 This test uses the DPPH method to determine the scavenging ability of resveratrol imine derivatives against DPPH free radicals.

[0038] Figure 6 This is the NMR spectrum of intermediate I. Detailed Implementation

[0039] The present invention will be further described in detail below with reference to the embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto.

[0040] Example 1 Preparation of Intermediate I

[0041] Add 30 mL of acetonitrile to a single-diameter round-bottom flask, incubate in an ice-salt bath for 5 min, then add N,N-dimethylformamide (1.76 g, 24.1 mmol) and phosphorus oxychloride (3.69 g, 24.1 mmol). Stir for 5 min, then add resveratrol (5 g, 21.9 mmol). Stir overnight, then filter to collect the solid and wash with cold acetonitrile. Transfer the obtained solid to a new single-diameter round-bottom flask, add 30 mL of water, and react at 55 °C for 3–5 h. Filter to collect the solid and wash with cold water, then dry to obtain intermediate I with the structure shown in Formula 3, in 80% yield. The NMR spectrum of intermediate I is shown below. Figure 6 As shown, the proton NMR integral and chemical shift of intermediate I are similar to the predictions.

[0042]

[0043] Example 2 Synthesis of 2-(((E)-2,4-dihydroxy-6-((E)-4-hydroxystyrene)benzylidene)amino)acetic acid (compound 1)

[0044] Glycine (0.16 g, 2.14 mmol) was added to a single-diameter round-bottom flask, followed by 5 mL of anhydrous methanol and potassium hydroxide powder (0.12 g, 2.14 mmol). The mixture was stirred at room temperature until the solid dissolved. Intermediate I (0.5 g, 1.95 mmol) obtained in Example 1 was then added, and stirring continued for 16 h. The reaction was monitored by TLC. After the reaction was complete, the reaction solution was filtered to obtain a solid, which was then purified by slurry to obtain the resveratrol imine derivative. The NMR spectrum of the product is shown below. Figure 1 As shown, 1¹H NMR (600MHz, DMSO-d⁶) δ 14.62 (s, 1H), 10.37 (d, J = 285.8 Hz, 2H), 8.55 (s, 1H), 7.45 (d, J = 8.7 Hz, 2H), 7.29 (d, J = 15.9 Hz, 1H), 6.91 (d, J = 15.8 Hz, 1H), 6.79 (d, J = 8.7 Hz, 2H), 6.29 (d, J = 2.2 Hz, 1H), 5.90 (d, J = 2.2 Hz, 1H), 3.99 (s, 2H). The ¹H NMR integral and chemical shift of compound 1 were similar to the predictions, leading to the conclusion that the product structure is 2-(((E)-2,4-dihydroxy-6-((E)-4-hydroxystyrene)benzylidene)amino)acetic acid.

[0045] Example 3 Synthesis of compounds 2-4

[0046] Intermediate I (0.5 g, 1.95 mmol) obtained in Example 1 was added to a single-diameter round-bottom flask along with 2-F benzylamine, 4-F benzylamine, or 4-OH benzylamine (2.14 mmol), dissolved in 5 mL of anhydrous methanol. The mixture was stirred at room temperature for 16 h, and the reaction was monitored by TLC. After the reaction was complete, the methanol was removed under vacuum, and the mixture was purified by slurry to obtain resveratrol imine derivatives, namely compounds 2–4. The NMR spectra of compounds 2–4 are shown below. Figures 2 to 4 As shown.

[0047] The yields of the above compounds are summarized in Table 2.

[0048] Table 2 Yields of compounds 1–4

[0049]

[0050] Effect Example

[0051] DPPH free radical scavenging experiment

[0052] 190 μL of a pre-prepared 100 μmol / L DPPH solution was added to each well of a 96-well plate, followed by 10 μL of compounds 1-4 (resveratrol imine derivatives prepared in Examples 2-3) at a final concentration of 5–50 μmol / L. A control group was also included, maintaining a total volume of 200 μL. The reaction was allowed to proceed in the dark at room temperature for 30 min, and the absorbance was measured at 517 nm. The DPPH radical scavenging rate was calculated as follows: DPPH scavenging rate = [1 - (Am - An) / As] × 100%. Am represents the OD value of the sample group, An represents the OD value of the methanol solution, and As represents the OD value of the negative control group. A curve was plotted with the concentration of the resveratrol imine derivative as the x-axis and the scavenging rate corresponding to each resveratrol imine derivative concentration as the y-axis. The results are shown below.Figure 5 Compounds 1 through 4 all exhibited stronger DPPH free radical scavenging ability than water-soluble vitamin C.

[0053] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A resveratrol imine derivative, characterized in that: The derivative is a compound with the structure shown in Formula 2 or a pharmaceutically acceptable salt thereof: Wherein, R is one of the following groups: or .

2. The resveratrol imine derivative according to claim 1, characterized in that: The pharmaceutically acceptable salt is a salt formed by the compound of formula 2 with lithium ions, sodium ions, potassium ions, calcium ions, magnesium ions and ammonium ions.

3. The method for preparing the resveratrol imine derivative according to claim 1 or 2, characterized in that... It includes the following steps: (1) Resveratrol was reacted with N,N-dimethylformamide and phosphorus oxychloride to obtain intermediate I with the structure shown in Formula 3; (2) One of glycine and 4-OH benzylamine, together with intermediate I, is used as a raw material. Through condensation reaction and purification, a resveratrol imine derivative with the structure shown in Formula 2 is obtained.

4. The preparation method according to claim 3, characterized in that: The molar ratio of resveratrol, N,N-dimethylformamide and phosphorus oxychloride in step (1) is 1:1.1:1.1; the reaction is carried out at room temperature for 16 hours.

5. The preparation method according to claim 3, characterized in that: The specific steps (1) are as follows: using acetonitrile as solvent, N,N-dimethylformamide and phosphorus oxychloride are added under stirring in an ice-salt bath. After stirring for 5 minutes, resveratrol is added and the reaction is carried out overnight. The red solid is obtained by filtration and washed with cold acetonitrile. The obtained solid is transferred to a new reaction flask, water is added, and the reaction is carried out at 55°C for 3-5 hours. The solid is then filtered, washed, and dried to obtain intermediate I.

6. The preparation method according to claim 3, characterized in that: The molar ratio of glycine and 4-OH benzylamine mentioned in step (2) to intermediate I is 1:1; the condensation reaction is carried out at room temperature for 16 hours under nitrogen protection.

7. The application of the resveratrol imine derivative according to claim 1 in the preparation of drugs for preventing and treating oxidative stress.

Images