CN111303222A - Salidroside derivative and preparation method and application thereof - Google Patents

Abstract

The invention relates to a salidroside derivative and a preparation method and application thereof. Specifically provided is a compound shown in formula I, or a salt thereof, or a stereoisomer thereof, or a crystal form thereof. The salidroside derivative shown in the formula I improves the problem of poor lipid solubility of salidroside, and can overcome the defects of low bioavailability and rapid metabolism of salidroside in clinical application; at the same time, the red sceneryThe astragaloside derivative can effectively resist CoCl₂Induced apoptosis of PC12 on CoCl₂The induced PC12 cell hypoxia injury model has excellent neuroprotective effect, can be used for preparing medicines for preventing and/or treating various nervous system diseases (including Parkinson disease, Alzheimer disease, cerebral apoplexy and the like) related to cell hypoxia injury, and has wide application prospect.

Description

A kind of salidroside derivative and its preparation method and use

technical field

The invention belongs to the technical field of medicine, and in particular relates to a salidroside derivative and a preparation method and use thereof.

Background technique

Salidroside is an extract of Chinese traditional Chinese medicine Rhodiola, and it is also the main active ingredient of Rhodiola and a marker of its pharmacodynamic material basis. Numerous studies have shown that salidroside has anti-inflammatory, antioxidant and anti-apoptotic effects, and has broad application prospects in the treatment of atherosclerosis and cardiovascular and cerebrovascular diseases. The document "Neuroprotective effect of salidroside on hypoxic injury of PC12 cells induced by CoCl ₂ " discloses that salidroside has neuroprotective effect on the model of PC12 cell injury induced by CoCl ₂ , and inhibits the complement component C3, promotes the expression of Egrs, and then It is related to inhibiting the expression of Bax protein and the activity of caspase-3 and promoting the expression of Bcl-xl. Studies have shown that salidroside has neuroprotective effects and a treatment time window of up to 48 hours in the treatment of neurological diseases (such as ischemic stroke); it has a strong effect on the apoptosis of vascular endothelial cells under oxidative stress. good inhibition.

Although salidroside has many of the above-mentioned pharmacological activities, it is limited in clinical application. This is because salidroside is a polyhydroxy compound with very high molecular polarity, excellent water solubility, but poor fat solubility. The characteristics of salidroside molecules with strong hydrophilicity and weak lipophilicity are not conducive to the passage of drugs through lipid membranes, and are not easily absorbed by the gastrointestinal tract and enter tissue cells, and are also difficult to pass through the blood-brain barrier; After the drug enters the body, it is easily metabolized in large quantities, which reduces the amount of the drug, reduces the efficacy of the drug, and weakens the pharmacological activity. Studies have shown that the bioavailability of salidroside is only 33.7% (see Wu Hao et al. Establishment of UPLC-MS/MS detection method for salidroside in rat plasma and its application in pharmacokinetic research. Chinese patent medicine , 2014, 36(6): 1176-1181.), the half-life in blood is about 40min (see Guo Na. Pharmacokinetic study of salidroside and its metabolite tyrosol in rats. Northeast Forestry University Doctoral dissertation, 2012). After one administration, almost all salidroside in blood and brain tissue was eliminated within 3 hours.

Therefore, in order to overcome the shortcomings of low bioavailability and fast metabolism of salidroside in clinical applications, proper modification of salidroside molecules to improve the lipid solubility of the molecules has become a research hotspot in the development of salidroside drugs. However, the effect of molecular modification on the pharmacological activity of the obtained derivatives is unpredictable, and how to obtain salidroside derivatives with improved lipid solubility and pharmacological activity at the same time has become an urgent problem to be solved in the development of salidroside drugs.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a salidroside derivative and its preparation method and use.

The present invention provides a compound shown in formula I, or its salt, or its stereoisomer, or its crystal form:

where R is

X is CO or SO ₂ ;

R ₁ is selected from the following groups substituted by 0-2 R ₂ : phenyl, naphthyl, C1-4 alkyl, n-hexyl, C2-4 alkenyl, cyclohexyl; R ₂ is independently selected from C1- 4 alkyl, C1-4 alkoxy, halogenated C1-4 alkyl, halogen;

And when X is CO, R ₁ is not phenyl.

Further, X is CO or SO ₂ ; R ₁ is selected from phenyl substituted by 1-2 R ₂ ; R ₂ is selected from C1-4 alkoxy.

Further, R is one of the following structures:

Further, the compound is selected from:

The present invention also provides a method for preparing the above compound, or its salt, or its stereoisomer, or its crystalline form. The method is to react Compound A with X-R to obtain;

The above X is halogen, preferably chlorine; R is as described above.

Further, the method includes the following steps:

(1) compound A, strong base and weak acid salt, phase transfer catalyst are dissolved in water to obtain an aqueous solution;

(2) X-R is dissolved in the organic solvent, then joins in the aqueous solution of step (1) gained, reacts;

The temperature of the reaction is below 10°C, preferably -5°C to 5°C;

The molar ratio of compound A to strong base and weak acid salt is 1:0.8～1:1.2, preferably 1:1; the molar ratio of compound A to X-R is 1:0.7～1:1.3, preferably 1:1: 1; the molar ratio of the compound A to the phase transfer catalyst is 1:0.01˜1:0.1, preferably 1:0.0155.

Further, the strong base weak acid salt is selected from K ₂ CO ₃ , Na ₂ CO ₃ , KHCO ₃ , NaHCO ₃ , preferably Na ₂ CO ₃ ; and/or, the phase transfer catalyst is a quaternary ammonium salt phase transfer Catalyst, quaternary ammonium salt phase transfer catalyst is preferably benzyl triethyl ammonium chloride, tetrabutyl ammonium bromide, tetrabutyl ammonium chloride, tetrabutyl ammonium hydrogen sulfate, trioctyl methyl ammonium chloride, ten Dialkyltrimethylammonium chloride, tetradecyltrimethylammonium chloride, more preferably tetrabutylammonium bromide; and/or, the organic solvent is selected from ethyl acetate, dichloromethane, diethyl ether , isopropyl ether, tetrahydrofuran, preferably ethyl acetate; and/or, the structure of the compound A is

Further, the method further includes a purification step, and the purification method is as follows: adding the above-mentioned organic solvent to the obtained system after the reaction, extracting, and retaining the organic phase;

Or, the purification method further comprises after drying the above-mentioned organic phase, performing gradient elution by rapid preparative liquid chromatography, the eluent is a mixed solvent of chloroform and methanol, and the volume percentage of methanol in the mixed solvent is 0-20%, The elution gradient was as follows: the volume percentage of methanol in the eluent increased linearly from 0% to 20% over 3 column volumes.

The present invention also provides the use of the above compound, or a salt thereof, or a stereoisomer thereof, or a crystalline form thereof in the preparation of a medicament for preventing and/or treating hypoxic injury of nerve cells.

The present invention also provides the use of the above compound, or a salt thereof, or a stereoisomer thereof, or a crystalline form thereof, in the preparation of a medicament for preventing and/or treating nervous system diseases;

The neurological disease is preferably a neurological disease related to hypoxic injury of nerve cells, more preferably Parkinson's disease, Alzheimer's disease, and stroke.

The salidroside derivatives provided by the invention improve the problem of poor lipid solubility of salidroside, and can overcome the shortcomings of low bioavailability and fast metabolism of salidroside in clinical applications; Astragaloside derivatives can effectively resist CoCl ₂ -induced PC12 cell apoptosis, and have excellent neuroprotective effects on the CoCl ₂ -induced PC12 cell hypoxic injury model. It is a drug for various neurological diseases (including Parkinson's disease, Alzheimer's disease, stroke, etc.), and has broad application prospects.

In the substituents of the present invention, Me means methyl group, Et means ethyl group, nPr means n-propyl group, and n-Hexyl means n-hexyl group.

In the present invention, the structures of salidroside Sal, salidroside isomer α-Sal, and aglycone tyrosol Tyrosol are as follows:

In the present invention, the minimum and maximum values of carbon atoms in the hydrocarbon group are indicated by prefixes, for example, C1-4 alkyl refers to a straight-chain or branched alkyl group containing 1-4 carbon atoms.

Obviously, according to the above-mentioned content of the present invention, according to the common technical knowledge and conventional means in the field, without departing from the above-mentioned basic technical idea of the present invention, other various forms of modification, replacement or change can also be made.

The above content of the present invention will be further described in detail below through the specific implementation in the form of examples. However, this should not be construed as limiting the scope of the above-mentioned subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention belong to the scope of the present invention.

Description of drawings

Figure 1 shows the inhibitory effect of various drugs on CoCl ₂ -induced PC12 cell apoptosis; C: CoCl ₂ -induced hypoxic injury model control group; Sal: CoCl ₂ + salidroside treatment group; α-Sal: CoCl ₂ + salidroside isomer α-Sal treatment group; tyrosol: CoCl ₂ + aglycone tyrosol Tyrosol treatment group; compd1a～2g: CoCl ₂ + each salidroside derivative treatment group (N=3; with the model *0.01<p<0.05, **p<0.01 compared to control group; #0.01<p<0.05, ##p<0.01 compared to Sal).

Detailed ways

The raw materials and equipment used in the present invention are all known products, obtained by purchasing commercially available products.

Example 1. Preparation of salidroside derivative 1a of the present invention

3g (0.01mol) of salidroside sal, 1.06g (0.01mol) of sodium carbonate, 0.05g (0.155mmol) of tetrabutylammonium bromide, and 10ml of water were added to a 50ml three-necked flask. Ice bath, stir until the solid is completely dissolved, and the temperature of the system drops to -5°C to 5°C. 0.01 mol of benzoyl chloride was dissolved in 10 ml of ethyl acetate and added to the system. The reaction was monitored by TLC (CHCl ₃ :CH ₃ OH=4:1), and the reaction was stopped when the starting point disappeared. Ethyl acetate was added to the system after the reaction, and the liquid was separated and extracted three times with 10 ml of ethyl acetate each time. The organic phases were combined, dried with anhydrous sodium sulfate, and the solvent was evaporated to dryness under reduced pressure to obtain a white solid. Using silica gel as the stationary phase and (CHCl ₃ -CH ₃ OH) mixed solvent as the eluent, the rapid preparative liquid chromatography (model: Biotageisolera one) was used for gradient elution. The elution gradient was as follows: within 3 column volumes, The methanol concentration increased linearly from 0% to 20%. After the product peaks, keep the methanol concentration unchanged, and collect the eluate at this time. The salidroside derivative 1a can be obtained: p-benzoyloxyphenethyl-β-D-glucopyranoside 1.6g, the yield is 40%, it is a white solid, the purity is 97% detected by HPLC, MS The molecular weight was determined, and the structure was confirmed by combining NMR carbon spectrum and hydrogen spectrum. ¹ H NMR (400MHz, MeOD) δ 8.18 (d, J=7.2Hz, 2H), 7.70 (t, J=7.5Hz, 1H), 7.57 (t, J=7.7Hz, 2H), 7.38 (d, J=8.5Hz, 2H), 7.15(d, J=8.5Hz, 2H), 4.34(d, J=7.8Hz, 1H), 4.15(dd, J=16.9, 7.2Hz, 1H), 3.89(d, J=12.2Hz, 1H), 3.82 (dd, J=16.8, 7.2Hz, 1H), 3.69 (dd, J=11.9, 5.2Hz, 1H), 3.38 (dd, J=12.2, 5.8Hz, 1H), 3.25–3.17 (m, 1H), 3.00 (t, J=7.1 Hz, 2H); ¹³ C NMR (101 MHz, MeOD) δ 165.39, 149.43, 136.69, 133.50, 129.75, 129.63, 129.44, 128.45, 121.21, 103.01, 76.72 , 76.57, 73.72, 70.27, 70.12, 61.40, 35.19; ESI MS (m/z): (M+Na ⁺ )427.53, (M+K ⁺ )442.95.

Example 2. Preparation of salidroside derivative 1b of the present invention

With reference to the method of Example 1, benzoyl chloride was replaced with p-toluoyl chloride to obtain the salidroside derivative 1b of the present invention: p-(p-methylbenzoyloxy)phenethyl-β-D - Glucopyranoside 2.4g, light yellow solid, yield 55%, purity was 96% by peak area detected by HPLC, molecular weight was determined by MS, and structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ HNMR(400MHz,MeOD)δ8.05(d,J=8.2Hz,2H),7.37(s,2H),7.35(t,J=2.0Hz,2H),7.21-7.03(m,2H),4.41 –4.28(m,1H),4.21-4.06(m,1H),3.90(dd,J=12.0,1.4Hz,1H),3.81(dt,J=9.7,7.2Hz,1H),3.75-3.66(m ,1H),3.39(dd,J＝12.8,5.1Hz,1H),3.33(ddd,J＝7.5,5.7,2.7Hz,2H),3.26–3.20(m,1H),3.07–2.88(m,2H ^The 70.14, 61.40, 35.19, 20.36. ESI MS (m/z): (M+Na ⁺ )441.52, (M+K ⁺ )456.99

Example 3. Preparation of salidroside derivative 1c of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with p-ethylbenzoyl chloride to obtain the salidroside derivative 1c of the present invention: p-(p-ethylbenzoyloxy)phenethyl-β-D - Glucopyranoside 0.9 g, white solid, yield 42%, purity was detected by HPLC, the peak area was 95%, the molecular weight was determined by MS, and the structure was confirmed by combining NMR carbon spectrum and hydrogen spectrum. ¹ HNMR(500MHz,MeOD)δ8.12-8.05(m,2H),7.39(d,J=8.4Hz,2H),7.36(d,J=8.5Hz,2H),7.16-7.09(m,2H) ,4.35(d,J＝7.8Hz,1H),4.19–4.10(m,1H),3.89(dt,J＝9.5,4.9Hz,1H),3.81(dt,J＝9.7,7.2Hz,1H), 3.75–3.66 (m, 1H), 3.43–3.35 (m, 1H), 3.33 (ddd, J=7.9, 6.4, 4.5Hz, 2H), 3.23 (dd, J=9.1, 7.8Hz, 1H), 3.03– 2.95(m, 2H), 2.79-2.71(m, 2H), 1.29(t, J=7.6Hz, 3H). ¹³ C NMR(126MHz, MeOD)δ165.47,150.83,149.48,136.59,129.86,129.73,127.93, 126.88,121.24,103.01,76.73,76.58,73.73,70.27,70.14,61.40,35.20,28.55,14.34.ESI MS(m/z):(M+Na ⁺ )455.15,(M+K ⁺ )471.07.

Example 4. Preparation of salidroside derivative 1d of the present invention

With reference to the method of Example 1, benzoyl chloride is replaced with p-propylbenzoyl chloride to obtain the salidroside derivative 1d of the present invention: p-(p-propylbenzoyloxy)phenethyl-β-D - Glucopyranoside 1.1 g, white solid, yield 49%, purity was detected by HPLC, the peak area was 96%, the molecular weight was determined by MS, and the structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ HNMR(500MHz,MeOD)δ8.14-8.02(m,2H),7.40-7.37(m,2H),7.37-7.35(m,2H),7.16-7.06(m,2H),4.35(d,J =7.8Hz,1H),4.14(dt,J=9.7,7.2Hz,1H),3.90(dd,J=11.9,1.8Hz,1H),3.81(dt,J=9.7,7.2Hz,1H),3.75 –3.67(m,1H),3.39(t,J=8.9Hz,1H),3.33(ddd,J=7.9,4.4,2.5Hz,2H),3.23(dd,J=9.1,7.9Hz,1H), 3.04–2.96 (m, 2H), 2.74–2.67 (m, 2H), 1.71 (dq, J=14.9, 7.4Hz, 2H), 0.98 (t, J=7.4Hz, 3H). ¹³ C NMR (126MHz, MeOD) δ165.48, 149.23, 136.60, 129.75, 129.72, 128.55, 126.92, 121.24, 103.01, 76.73, 76.58, 73.73, 70.27, 70.13, 61.40, 37.62, 35.20, 23.m99, 12.68 MS(ESI) +Na ⁺ ) 469.24, (M+K ⁺ ) 485.05.

Example 5. Preparation of salidroside derivative 1e of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with p-methoxybenzoyl chloride to obtain the salidroside derivative 1e of the present invention: p-(p-methoxybenzoyloxy)phenethyl-β -D-glucopyranoside 0.4g, white solid, yield 18%, purity was detected by HPLC, the peak area was 97%, the molecular weight was determined by MS, and the structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ H NMR(500MHz,MeOD)δ8.18-8.08(m,2H),7.35(d,J=8.5Hz,2H),7.14-7.09(m,2H),7.09-7.03(m,2H),4.34 (d, J=7.8Hz, 1H), 4.14 (dt, J=9.7, 7.3Hz, 1H), 3.93–3.87 (m, 4H), 3.84–3.78 (m, 1H), 3.70 (dd, J=11.9 , 5.3Hz, 1H), 3.39 (t, J=8.9Hz, 1H), 3.35–3.30 (m, 2H), 3.23 (dd, J=9.1, 7.9Hz, 1H), 3.02–2.96 (m, 2H) . ¹³ C NMR(126MHz,MeOD)δ165.24,164.30,149.51,136.50,131.83,129.70,121.47,121.28,113.70,103.01,76.73,71.57,73.73,70.26,70.14,35.40,54 z): (M+Na ⁺ )457.40, (M+K ⁺ )472.88.

Example 6. Preparation of salidroside derivative 1f of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with p-trifluoromethylbenzoyl chloride to obtain salidroside derivative 1f of the present invention: p-(p-trifluoromethylbenzoyloxy)phenethyl -β-D-glucopyranoside 2.22g, white solid, yield 94%, purity was detected by HPLC, peak area was 96%, molecular weight was determined by MS, and structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ H NMR(500MHz,MeOD)δ8.35(d,J=8.2Hz,2H),7.88(d,J=8.3Hz,2H),7.38(t,J=5.6Hz,2H),7.22-7.11( m, 2H), 4.33 (dd, J=18.5, 7.8Hz, 1H), 4.21–4.04 (m, 1H), 3.94–3.86 (m, 1H), 3.82 (dt, J=9.7, 7.1Hz, 1H) , 3.74–3.67 (m, 1H), 3.43–3.36 (m, 1H), 3.33–3.30 (m, 2H), 3.23 (dd, J=9.1, 7.9Hz, 1H), 3.04–2.97 (m, 2H) . ¹³ C NMR(126MHz,MeOD)δ164.05,149.24,137.00,134.64,134.38,133.10,130.32,129.82,125.41,121.07,103.01,76.73,76.58,73.72,70.27,5.40.09,6 z): (M+Na ⁺ ) 495.21, (M+K ⁺ ) 511.15.

Example 7. Preparation of 1 g of salidroside derivatives of the present invention

With reference to the method of Example 1, benzoyl chloride was replaced with o-toluoyl chloride to obtain 1 g of the salidroside derivative of the present invention: p-(o-toluoyloxy)phenethyl-β-D - Glucopyranoside 1.2g, white solid, yield 31%, purity was detected by HPLC, and the peak area was 92%, the molecular weight was determined by MS, and the structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ HNMR(400MHz,MeOD)δ8.13-8.09(m,1H),7.52(td,J=7.5,1.4Hz,1H),7.37(ddd,J=3.6,2.0,1.4Hz,2H),7.35( t, J=2.9Hz, 1H), 7.28–7.23 (m, 1H), 7.15–7.12 (m, 2H), 4.35 (d, J=7.8Hz, 1H), 4.15 (dt, J=9.7, 7.2Hz) ,1H),3.94-3.88(m,1H),3.81(dt,J=9.7,7.1Hz,1H),3.73-3.67(m,1H),3.44-3.37(m,1H),3.23(dd,J =9.1,7.8Hz,1H),3.00(t,J=7.2Hz,2H),2.64(s,3H). ^13C NMR(101MHz,MeOD)δ166.12,149.39,140.60,136.59,132.52,131.59,131.06, 130.64,129.74,125.75,121.32,103.00,76.72,76.58,73.73,70.26,70.14,61.39,35.20,20.56.ESI MS(m/z):(M+Na ⁺ )441.56,(M+K ⁺ )457.00.

Example 8. Preparation of salidroside derivatives of the present invention for 1 h

With reference to the method of Example 1, benzoyl chloride was replaced with m-toluoyl chloride to obtain the salidroside derivative 1h of the present invention: p-(m-tolyloxy)phenethyl-β-D -Glucopyranoside 1.9g, white solid, yield 49%, purity was 96% by peak area detected by HPLC, molecular weight was determined by MS, and structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ H NMR(400MHz,MeOD)δ8.05-7.99(m,1H),7.99-7.95(m,1H),7.52(dd,J=5.0,3.1Hz,1H),7.48-7.40(m,1H) ,7.40–7.34(m,2H),7.20–7.06(m,2H),4.33(dd,J=14.0,7.8Hz,1H),4.23–4.05(m,1H),3.90(dd,J=11.9, 1.7Hz, 1H), 3.81 (dt, J=9.7, 7.1Hz, 1H), 3.73–3.65 (m, 1H), 3.42–3.35 (m, 1H), 3.35–3.28 (m, 3H), 3.22 (dd , J=9.0, 7.8Hz, 1H), 3.05–2.96 (m, 2H), 2.42 (d, J=21.5Hz, 3H). ¹³ C NMR (101MHz, MeOD) δ165.52, 149.47, 138.53, 136.65, 134.17, 130.04,129.73,129.39,128.33,126.81,121.21,103.01,76.72,76.59,73.72,70.25,70.12,61.38,35.19,19.91.ESI MS(m/z):(M+Na ⁺ )441.46,(M+K ⁺ )457.01.

Example 9. Preparation of salidroside derivative 1i of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with m-fluorobenzoyl chloride to obtain salidroside derivative 1i of the present invention: p-(m-fluorobenzoyloxy)phenethyl-β-D-pyridine Glucopyranoside 1.21g, white solid, yield 57%, purity detected by HPLC, peak area was 96%, molecular weight was determined by MS, and structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ H NMR(500MHz,MeOD)δ8.03-7.97(m,1H),7.89-7.83(m,1H),7.59(td,J=8.0,5.6Hz,1H),7.49-7.41(m,1H) ,7.40–7.35(m,2H),7.19–7.13(m,2H),4.35(d,J=7.8Hz,1H),4.15(dt,J=9.7,7.2Hz,1H),3.90(dd,J =11.9,1.7Hz,1H),3.82(dt,J=9.7,7.1Hz,1H),3.74–3.66(m,1H),3.39(t,J=9.0Hz,1H),3.32(tt,J= 2.6, 1.6Hz, 2H), 3.23 (dd, J=9.1, 7.8Hz, 1H), 3.04–2.95 (m, 2H). ¹³ C NMR (126MHz, MeOD) δ 163.66, 161.71, 149.28, 136.89, 131.75, 130.45 ,129.78,125.59,121.10,120.27,116.05,103.01,76.73,76.58,73.72,70.27,70.10,61.40,35.19.ESI MS(m/z):(M+Na ⁺ )445.58,(M+K ⁺ )460.98

Example 10. Preparation of salidroside derivative 1j of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with cinnamoyl chloride to obtain salidroside derivative 1j of the present invention: p-cinnamoyloxyphenethyl-β-D-glucopyranoside 0.80g, white solid , the yield was 37%, the purity was detected by HPLC, the peak area was 96%, the molecular weight was determined by MS, and the structure was confirmed by combining NMR carbon spectrum and hydrogen spectrum. ¹ H NMR (500MHz, MeOD) δ 7.88 (d, J=16.0Hz, 1H), 7.69 (dt, J=7.8, 3.3Hz, 2H), 7.46 (dd, J=6.6, 3.6Hz, 3H), 7.37–7.32 (m, 2H), 7.12–7.07 (m, 2H), 6.75 (d, J=16.0Hz, 1H), 4.34 (d, J=7.8Hz, 1H), 4.13 (dt, J=9.7, 7.2Hz, 1H), 3.93–3.87 (m, 1H), 3.80 (dt, J=9.7, 7.1Hz, 1H), 3.73–3.65 (m, 1H), 3.38 (t, J=9.0Hz, 1H), 3.34–3.30 (m, 2H), 3.22 (dd, J=9.1, 7.9Hz, 1H), 2.98 (dd, J=14.4, 7.3Hz, 2H). ¹³ C NMR (126MHz, MeOD) δ 165.76, 149.32, 146.51 ,136.53,134.20,130.49,129.65,128.72,128.10,121.13,116.82,103.01,76.73,76.58,73.72,70.27,70.12,61.40,35.18.ESI MS(m/z):(M+Na ⁺ )453 M+K ⁺ )469.01.

Example 11. Preparation of salidroside derivative 1k of the present invention

With reference to the method of Example 1, benzoyl chloride was replaced with m-chloromethylbenzoyl chloride to obtain the salidroside derivative 1k of the present invention: p-(m-chloromethylbenzoyloxy)phenethyl-β -D-glucopyranoside 1.43g, white solid, yield 63%, purity was detected by HPLC, peak area was 97%, molecular weight was determined by MS, and structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ H NMR (500MHz, MeOD) δ 8.22 (d, J=1.4Hz, 1H), 8.17-8.10 (m, 1H), 7.75 (d, J=7.7Hz, 1H), 7.57 (t, J=7.7 Hz, 1H), 7.37 (d, J=8.5Hz, 2H), 7.19–7.11 (m, 2H), 4.75 (s, 2H), 4.35 (d, J=7.8Hz, 1H), 4.15 (dt, J ＝9.7,7.2Hz,1H),3.91(dd,J＝10.4,8.5Hz,1H),3.82(dt,J＝9.7,7.1Hz,1H),3.75-3.65(m,1H),3.44-3.36( m, 1H), 3.34–3.30 (m, 2H), 3.30–3.19 (m, 1H), 3.05–2.96 (m, 2H). ¹³ C NMR (126MHz, MeOD) δ164.90, 149.37, 138.91, 136.77, 133.63, 129.96,129.77,129.74,129.47,128.92,121.19,103.01,76.73,76.58,73.72,70.26,70.12,61.40,44.72,35.20.ESI MS(m/z):(M+Na ⁺ )475.26,(M+K ⁺ )490.98

Example 12. Preparation of salidroside derivative 11 of the present invention

With reference to the method of Example 1, benzoyl chloride was replaced with p-chloromethylbenzoyl chloride to obtain salidroside derivative 11 of the present invention: p-(p-chloromethylbenzoyloxy)phenethyl-β -D-glucopyranoside 2.7g, white solid, yield 60%, purity was detected by HPLC, the peak area was 97%, the molecular weight was determined by MS, and the structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ H NMR(400MHz,MeOD)δ8.25-8.11(m,2H),7.72-7.54(m,2H),7.44-7.32(m,2H),7.20-7.08(m,2H),4.74(s, 2H), 4.35(d, J=7.8Hz, 1H), 4.15(dt, J=9.7, 7.2Hz, 1H), 3.93–3.87(m, 1H), 3.81(dt, J=9.7, 7.1Hz, 1H) ), 3.74–3.67 (m, 1H), 3.43–3.36 (m, 1H), 3.34–3.29 (m, 2H), 3.26–3.20 (m, 1H), 3.03–2.96 (m, 2H). ¹³ C NMR (101MHz, MeOD)δ164.91,149.38,143.80,136.74,130.03,129.77,129.24,128.63,121.19,103.01,76.73,76.58,73.72,70.26,70.12,61.40,44.57,35.19 ESI MS(m/z.)( M+Na ⁺ )475.13,(M+K ⁺ )490.97

Example 13. Preparation of salidroside derivative 1m of the present invention

With reference to the method of Example 1, benzoyl chloride was replaced with m-chlorobenzoyl chloride to obtain the salidroside derivative 1m of the present invention: p-(m-chlorobenzoyloxy)phenethyl-β-D-pyridine Glucopyranoside 2.6g, white solid, yield 59%, purity was detected by HPLC, the peak area was 98%, the molecular weight was determined by MS, and the structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ H NMR (400MHz, MeOD) δ 8.15 (t, J=1.8Hz, 1H), 8.13-8.09 (m, 1H), 7.72 (ddd, J=8.1, 2.2, 1.1Hz, 1H), 7.57 (dd , J=12.0, 4.1Hz, 1H), 7.43–7.30 (m, 2H), 7.21–7.09 (m, 2H), 4.34 (d, J=7.8Hz, 1H), 4.15 (dt, J=9.7, 7.2 Hz, 1H), 3.90 (dd, J=11.9, 1.6Hz, 1H), 3.82 (dt, J=9.7, 7.1Hz, 1H), 3.72–3.66 (m, 1H), 3.42–3.35 (m, 1H) ,3.34–3.31(m,2H),3.25–3.18(m,1H),2.99(q,J=7.2Hz,2H). ^13C NMR(101MHz,MeOD)δ164.02,149.27,136.92,134.45,133.35,131.44 ,130.12,129.78,129.39,127.96,121.09,103.01,76.72,76.59,73.72,70.26,70.09,61.38,35.19.ESI MS(m/z):(M+Na ⁺ )461.28,(M+K ⁺ )476.94 .

Example 14. Preparation of salidroside derivative 1n of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with 3,4-dichlorobenzoyl chloride to obtain the salidroside derivative 1n of the present invention: p-(3,4-dichlorobenzoyloxy)benzene Ethyl-β-D-glucopyranoside 1.23g, white solid, yield 52%, the purity was detected by HPLC, the peak area was 97%, the molecular weight was determined by MS, and the structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ H NMR (500MHz, MeOD) δ 8.27 (d, J=2.0Hz, 1H), 8.13-8.02 (m, 1H), 7.74 (dd, J=8.4, 4.3Hz, 1H), 7.38 (d, J = 8.6Hz, 2H), 7.21–7.12 (m, 2H), 4.34 (d, J=7.8Hz, 1H), 4.15 (dt, J=9.7, 7.2Hz, 1H), 3.92–3.87 (m, 1H) ,3.82(dt,J=9.7,7.1Hz,1H),3.73–3.67(m,1H),3.38(t,J=9.0Hz,1H),3.32(ddd,J=9.9,4.8,2.8Hz,2H ^The 76.73, 76.58, 73.72, 70.26, 70.08, 61.40, 35.19. ESI MS (m/z): (M+Na ⁺ ) 497.03, (M+K ⁺ ) 512.87.

Example 15. Preparation of salidroside derivative 1o of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with 2,3-dichlorobenzoyl chloride to obtain the salidroside derivative 1o of the present invention: p-(2,3-dichlorobenzoyloxy)benzene Ethyl-β-D-glucopyranoside 1.49g, white solid, yield 63%, the purity was detected by HPLC, the peak area was 98%, the molecular weight was determined by MS, and the structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ H NMR (500MHz, MeOD) δ 7.91 (dd, J=7.8, 1.5Hz, 1H), 7.78 (dd, J=8.1, 1.5Hz, 1H), 7.50-7.44 (m, 1H), 7.39 (d , J=8.5Hz, 2H), 7.21–7.13 (m, 2H), 4.35 (d, J=7.8Hz, 1H), 4.15 (dt, J=9.7, 7.2Hz, 1H), 3.92–3.87 (m, 1H), 3.82 (dt, J=9.7, 7.1Hz, 1H), 3.69 (dt, J=11.1, 6.6Hz, 1H), 3.41–3.36 (m, 1H), 3.32 (ddd, J=7.8, 4.1, 1.4Hz, 2H), 3.23 (dd, J=9.1, 7.9Hz, 1H), 3.05–2.93 (m, 2H). ¹³ C NMR (126MHz, MeOD) δ 163.87, 149.10, 137.16, 134.15, 133.46, 132.44, 130.99 ,129.85,129.22,127.74,120.98,103.02,76.73,76.59,73.72,70.26,70.07,61.39,35.19.ESI MS(m/z):(M+Na ⁺ )497.10,(M+K ⁺ )512.88.

Example 16. Preparation of salidroside derivative 1p of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with 2,6-dichlorobenzoyl chloride to obtain the salidroside derivative 1p of the present invention: p-(2,6-dichlorobenzoyloxy)benzene Ethyl-β-D-glucopyranoside 0.51g, white solid, yield 22%, purity was detected by HPLC, peak area was 96%, molecular weight was determined by MS, and structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ H NMR (500MHz, MeOD) δ 7.56-7.48 (m, 3H), 7.42 (dd, J=7.8, 5.5Hz, 2H), 7.21-7.17 (m, 2H), 4.34 (d, J=7.8Hz) ,1H),4.15(dt,J=9.7,7.2Hz,1H),3.92-3.87(m,1H),3.82(dt,J=9.8,7.1Hz,1H),3.72-3.67(m,1H), 3.41–3.36 (m, 1H), 3.32 (ddd, J=7.7, 4.4, 2.1Hz, 2H), 3.22 (dd, J=9.1, 7.9Hz, 1H), 3.01 (q, J=7.2Hz, 2H) . ¹³ C NMR(126MHz,MeOD)δ163.21,148.88,137.50,132.83,131.77,131.43,129.97,127.99,120.84,103.01,76.73,76.59,73.72,70.25,70.03,61.38,35 MSSI(m/z.8) : (M+Na ⁺ )497.13, (M+K ⁺ )512.89.

Example 17. Preparation of salidroside derivative 2a of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with acetyl chloride to obtain salidroside derivative 2a of the present invention: p-acetoxyphenethyl-β-D-glucopyranoside 3.1 g, white solid, The yield is 90%, the purity is detected by HPLC, and the peak area is 98%. The molecular weight is determined by MS, and the structure is confirmed by combining NMR carbon spectrum and hydrogen spectrum. ¹ H NMR(500MHz,MeOD)δ7.38-7.26(m,2H),7.04-6.97(m,2H),4.33(d,J=7.8Hz,1H),4.11(dt,J=9.7,7.2Hz ,1H),3.88(dd,J=11.8,1.6Hz,1H),3.78(dt,J=9.7,7.1Hz,1H),3.71–3.66(m,1H),3.38(dd,J=11.3,6.5 Hz, 1H), 3.34–3.29(m, 2H), 3.21(dd, J=9.1, 7.9Hz, 1H), 2.95(dd, J=13.9, 6.8Hz, 2H), 2.27(s, 3H). ¹³ C NMR(126MHz,MeOD)δ169.99,149.29,136.48,129.60,121.11,102.98,76.71,76.56,73.70,70.25,70.11,61.38,35.15,19.56.ESI MS(m/z):(M+Na ⁺ )365.58 , (M+K ⁺ )380.88.

Example 18. Preparation of salidroside derivative 2b of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with n-heptanoyl chloride to obtain salidroside derivative 2c of the present invention: p-n-heptanoyloxyphenethyl-β-D-glucopyranoside 0.5g, White solid, the yield is 12%, the purity is detected by HPLC, the peak area is 90%, the molecular weight is determined by MS, and the structure is confirmed by combining NMR carbon spectrum and hydrogen spectrum. ¹ H NMR (500MHz, MeOD) δ 7.36-7.26 (m, 2H), 7.02-6.94 (m, 2H), 4.33 (dd, J=7.8, 2.4Hz, 1H), 4.12 (dt, J=9.7, 7.2Hz, 1H), 3.91–3.86 (m, 1H), 3.82–3.75 (m, 1H), 3.69 (dd, J=11.9, 5.4Hz, 1H), 3.41–3.35 (m, 1H), 3.33–3.29 (m, 2H), 3.21 (dd, J=9.1, 7.8Hz, 1H), 3.01–2.93 (m, 2H), 2.57 (t, J=7.4Hz, 2H), 1.79–1.69 (m, 2H), 1.41–1.34 (m, 6H), 0.94 (dd, J=9.6, 4.5Hz, 3H). ¹³ C NMR (126MHz, MeOD) δ 172.73, 149.29, 136.44, 129.62, 121.09, 102.98, 76.71, 76.56, 73.70, 70.25 ,70.11,61.38,35.15,33.66,31.24,28.44,24.57,22.17,12.99.ESI MS(m/z):(M+Na ⁺ )435.10,

(M+K ⁺ )451.06.

Example 19. Preparation of salidroside derivative 2c of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with acryloyl chloride to obtain the salidroside derivative 2d of the present invention: p-acryloyloxyphenethyl-β-D-glucopyranoside 1.40g, white solid , the yield was 79%, the purity was detected by HPLC, the peak area was 96%, the molecular weight was determined by MS, and the structure was confirmed by combining NMR carbon spectrum and hydrogen spectrum. ¹ H NMR(500MHz,MeOD)δ7.36-7.31(m,2H),7.07-7.03(m,2H),6.57(dd,J=17.3,1.3Hz,1H),6.41-6.32(m,1H) ,6.09–6.05(m,1H),4.33(d,J=7.8Hz,1H),4.12(dt,J=9.7,7.2Hz,1H),3.91–3.86(m,1H),3.80(dt,J ＝9.7,7.1Hz,1H),3.69(dd,J＝11.9,5.3Hz,1H),3.39(dd,J＝11.1,6.8Hz,1H),3.34–3.29(m,2H),3.22(ddd, J=10.8,7.9,3.0Hz,1H),3.00–2.94(m,2H) ^.13C NMR(126MHz,MeOD)δ164.86,149.08,136.65,131.91,129.69,127.68,121.02,102.98,76.71,76.56,73.71 , 70.25, 70.11, 61.38, 35.16. ESI MS (m/z): (M+Na ⁺ )377.00, (M+K ⁺ )392.92.

Example 20. Preparation of salidroside derivative 2d of the present invention

Referring to the method of Example 1, replace benzoyl chloride with crotonyl chloride to obtain salidroside derivative 2e of the present invention: p-crotonyloxyphenethyl-β-D-glucopyranoside 0.40g, white solid , the yield was 22%, the purity was detected by HPLC, the peak area was 97%, the molecular weight was determined by MS, and the structure was confirmed by combining NMR carbon spectrum and hydrogen spectrum. ¹ H NMR(500MHz,MeOD)δ7.34-7.30(m,2H),7.24-7.15(m,1H),7.04-7.00(m,2H),6.15-5.99(m,1H),4.40-4.28( m, 1H), 4.20–4.06 (m, 1H), 3.89 (dd, J=11.9, 1.8Hz, 1H), 3.79 (dt, J=9.7, 7.1Hz, 1H), 3.72–3.65 (m, 1H) ,3.39(dd,J＝17.1,8.2Hz,1H),3.35–3.28(m,2H),3.21(dd,J＝9.1,7.8Hz,1H),2.97(q,J＝7.2Hz,2H), 2.01–1.88(m, 3H). ¹³ C NMR(126MHz, MeOD)δ165.21,149.20,147.19,136.44,129.63,121.51,121.12,102.98,76.71,76.56,73.71,70.25,70.12,61.38,35. MS (m/z): (M+Na ⁺ ) 391.46, (M+K ⁺ ) 406.95.

Example 21. Preparation of salidroside derivative 2e of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with cyclohexanyl chloride to obtain the salidroside derivative 2f of the present invention: p-cyclohexanyloxyphenethyl-β-D-glucopyranoside 0.8 g, white solid, the yield is 39%, the purity is detected by HPLC, the peak area is 94%, the molecular weight is determined by MS, and the structure is confirmed by combining NMR carbon spectrum and hydrogen spectrum. ¹ H NMR(500MHz,MeOD)δ7.41-7.22(m,2H),7.02-6.92(m,2H),4.32(t,J=7.3Hz,1H),4.11(dt,J=9.7,7.2Hz ,1H),3.88(dd,J＝11.9,1.9Hz,1H),3.78(dt,J＝9.7,7.2Hz,1H),3.71–3.65(m,1H),3.38(t,J＝9.0Hz, 1H), 3.34–3.29 (m, 2H), 3.24–3.18 (m, 1H), 2.99–2.93 (m, 2H), 2.59 (tt, J=11.0, 3.7Hz, 1H), 1.83–1.25 (m, 11H). ¹³ C NMR(126MHz, MeOD)δ174.89,149.38,136.39,129.62,121.06,102.98,76.71,76.56,73.71,70.25,70.12,61.38,42.89,35.15,28.73,25.50,25.m z): (M+Na ⁺ ) 433.43, (M+K ⁺ ) 449.02.

Example 22. Preparation of salidroside derivative 2f of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with 2-naphthoyl chloride to obtain 2g of salidroside derivatives of the present invention: p-(2-naphthoyloxy)phenethyl-β-D-pyridine Glucopyranoside 1.45g, white solid, yield 64%, the purity was detected by HPLC, the peak area was 97%, the molecular weight was determined by MS, and the structure was confirmed by NMR carbon spectrum and hydrogen spectrum. ¹ H NMR (500MHz, MeOD) δ 8.80 (s, 1H), 8.16 (dd, J=8.6, 1.7Hz, 1H), 8.07 (d, J=8.1Hz, 1H), 8.02 (d, J=8.6 Hz, 1H), 7.99 (d, J=8.3Hz, 1H), 7.70–7.66 (m, 1H), 7.64–7.60 (m, 1H), 7.40 (d, J=8.5Hz, 2H), 7.23–7.18 (m, 2H), 4.35 (d, J=7.8Hz, 1H), 4.16 (dt, J=9.7, 7.2Hz, 1H), 3.93–3.88 (m, 1H), 3.83 (dt, J=9.7, 7.2 Hz, 1H), 3.69 (dt, J=10.3, 6.5Hz, 1H), 3.42–3.36 (m, 1H), 3.35–3.32 (m, 2H), 3.23 (dd, J=9.1, 7.9Hz, 1H) ,3.05–2.98(m,2H) ^.13C NMR(126MHz,MeOD)δ165.52,149.55,136.73,135.94,132.59,131.39,129.76,129.14,128.50,128.19,127.51,126.70,126.30,126.70,126.30,124.124 76.74, 76.60, 73.74, 70.28, 70.13, 61.40, 35.21. ESI MS (m/z): (M+Na ⁺ ) 477.38, (M+K ⁺ ) 493.05.

Example 23. Preparation of salidroside derivative 2g of the present invention

Referring to the method of Example 1, benzoyl chloride was replaced with benzenesulfonyl chloride to obtain the salidroside derivative 2h of the present invention: p-benzenesulfonyloxyphenethyl-β-D-glucopyranoside 1.3g, White solid, the yield is 59%, the purity is detected by HPLC, the peak area is 97%, the molecular weight is determined by MS, and the structure is confirmed by combining NMR carbon spectrum and hydrogen spectrum. ¹ H NMR(500MHz,MeOD)δ7.83(dt,J=8.6,1.5Hz,2H),7.77-7.73(m,1H),7.68-7.53(m,2H),7.31-7.18(m,2H) ,6.96-6.82(m,2H),4.30(d,J=7.8Hz,1H),4.08(dt,J=9.8,7.1Hz,1H),3.91-3.85(m,1H),3.74(dt,J ＝9.8,7.0Hz,1H),3.67(dt,J＝10.5,5.3Hz,1H),3.37(t,J＝8.9Hz,1H),3.33–3.27(m,2H),3.19(dd,J＝ 9.1, 7.9Hz, 1H), 2.91 (q, J=7.3Hz, 2H). ¹³ C NMR (126MHz, MeOD) δ148.07, 138.40, 135.30, 134.23, 129.98, 129.11, 128.20, 121.73, 102.97, 76.70, 76.56, 73.67, 70.24, 69.78, 61.37, 35.03. ESI MS (m/z): (M+Na ⁺ )463.51, (M+K ⁺ )478.98

The beneficial effects of the salidroside derivatives of the present invention are demonstrated below through experimental examples.

Experimental Example 1. Inhibitory effect of salidroside derivatives on CoCl ₂ -induced apoptosis of PC12 cells

(1) The purpose of the experiment

Using CoCl ₂ -induced chemical hypoxic injury model of PC12 cells, in vitro pharmacodynamic experiments were conducted to evaluate the neuroprotective effect of salidroside derivatives on CoCl ₂ -induced hypoxic injury of PC12 cells.

(2) cell culture

The PC12 cell line was obtained from the National Infectious Disease Diagnosis and Vaccine Engineering Technology Research and Development Center of Xiamen University. PC12 cells were inoculated into 25cm ² culture flasks, and DMEM high-glucose medium containing 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin was added, and the cells were incubated at 37°C in a 5% CO ₂ constant temperature incubator. Cultivation and passage.

(3) Test method

Drugs to be tested: 1a-2g of each salidroside derivative prepared in the embodiment of the present invention; commercial products: salidroside Sal, salidroside isomer α-Sal, and aglycone tyrosol Tyrosol.

Take PC12 cells in logarithmic growth phase, add 100 μL per well at 2×10 ⁵ cells/mL, inoculate in a 96-well culture plate, and culture at 37°C in a 5% CO ₂ incubator. The cells were divided into the following groups: blank control group, CoCl ₂ model control group, and CoCl ₂ + test drug group. After the cells had grown to 80%, DMEM containing 400 μM CoCl ₂ and 1% fetal bovine serum was added to each group, cultured at 37°C and 5% CO ₂ for 8 h, and then the culture medium was aspirated and washed with 100 μL PBS for two times. After administration, the concentration of the drug to be tested is 1 μM, which is dissolved in DMEM high-glucose medium containing 1% fetal bovine serum. After the cells were cultured for 24 h, 10 μL of 5 mg/mL MTT solution was added to each well of a 96-well plate, incubated at 37 °C in a 5% CO ₂ incubator for 4 h, and then the culture medium was aspirated, and 200 μL of DMSO was added to each well. The absorbance was measured by a functional microplate reader, and the results were statistically analyzed.

表示，各组间比较采用单因素方差分析(one-way ANOVA)。p<0.05表示差异有统计学意义。SPSS 21.0 was used for statistical analysis, and the data were expressed as mean ± standard error

The comparison between groups was performed by one-way analysis of variance (one-way ANOVA). p<0.05 indicates a statistically significant difference.

(4) Experimental results

Compared with the blank control group, the cell survival rate of the CoCl ₂ model control group was significantly reduced, indicating that the modeling was successful; compared with the CoCl ₂ model control group, the cell survival rate of most of the salidroside derivatives of the present invention was significantly increased. However, the treatment of salidroside isomer α-Sal and aglycone tyrosol had no effect on cell viability. It shows that the salidroside derivatives of the present invention have a significant inhibitory effect on the apoptosis of PC12 cells induced by CoCl ₂ . In addition, among the salidroside derivatives of the present invention, the cell survival rate after treatment with compound 1e was the highest.

Therefore, the salidroside derivatives of the present invention, especially compound 1e, can effectively resist the apoptosis of PC12 cells induced by CoCl ₂ , and have excellent neuroprotective effects on the hypoxic injury model of PC12 cells induced by CoCl ₂ , and can be used to prepare therapeutic Drugs for various neurological disorders associated with cellular hypoxic injury.

In conclusion, the present invention provides a class of salidroside derivatives, which improves the problem of poor lipid solubility of salidroside and can overcome the low bioavailability of salidroside in clinical applications. , the disadvantage of fast metabolism; at the same time, the salidroside derivatives can effectively resist the apoptosis of PC12 cells induced by CoCl ₂ , and have excellent neuroprotective effects on the hypoxic injury model of PC12 cells induced by CoCl ₂ , and can be used to prepare prophylactic And/or drugs for treating various neurological diseases (including Parkinson's disease, Alzheimer's disease, stroke, etc.) related to cellular hypoxic injury, have broad application prospects.

Claims (10)

1. A compound represented by formula I, or a salt thereof, or a stereoisomer thereof, or a crystal form thereof:

wherein R is

X is CO or SO₂；

R₁Selected from the group consisting of 0 to 2R₂Substituted of the following groups: phenyl, naphthyl, C1-4 alkyl, n-hexyl, C2-4 alkenyl and cyclohexyl; r₂Each independently selected from C1-4 alkaneA C1-4 alkoxy group, a halogenated C1-4 alkyl group, and a halogen;

and when X is CO, R₁Is not phenyl.

2. The compound, or a salt thereof, or a stereoisomer thereof, or a crystalline form thereof, according to claim 1, characterized in that: x is CO or SO₂；R₁Selected from 1 to 2R₂Substituted phenyl; r₂Selected from C1-4 alkoxy.

3. The compound, or a salt thereof, or a stereoisomer thereof, or a crystalline form thereof, according to claim 1, characterized in that: r is one of the following structures:

4. the compound according to claim 3, or a salt thereof, or a stereoisomer thereof, or a crystalline form thereof, characterized in that: the compound is selected from:

5. a process for preparing a compound according to any one of claims 1 to 4, or a salt thereof, or a stereoisomer thereof, or a crystalline form thereof, characterized in that: the method comprises the steps of reacting a compound A with X-R to obtain the compound A;

x is halogen, preferably chlorine; r is as defined in any one of claims 1 to 4.

6. The method of claim 5, wherein: the method comprises the following steps:

(1) dissolving a compound A, strong base and weak acid salt and a phase transfer catalyst into water to obtain an aqueous solution;

(2) dissolving X-R into an organic solvent, adding into the aqueous solution obtained in the step (1), and reacting;

the reaction temperature is below 10 ℃, preferably-5 ℃;

the molar ratio of the compound A to the strong base weak acid salt is 1: 0.8-1: 1.2, preferably 1: 1; the molar ratio of the compound A to the X-R is 1: 0.7-1: 1.3, preferably 1: 1; the molar ratio of the compound A to the phase transfer catalyst is 1: 0.01-1: 0.1, and preferably 1: 0.0155.

7. The method of claim 6, wherein: the strong base weak acid salt is selected from K₂CO₃、Na₂CO₃、KHCO₃、NaHCO₃Preferably Na₂CO₃(ii) a And/or the phase transfer catalyst is a quaternary ammonium salt phase transfer catalyst, the quaternary ammonium salt phase transfer catalyst is preferably benzyltriethylammonium chloride, tetrabutylammonium bromide, tetrabutylammonium chloride, tetrabutylammonium hydrogen sulfate, trioctylmethylammonium chloride, dodecyltrimethylammonium chloride and tetradecyltrimethylammonium chloride, and more preferably tetrabutylammonium bromide; and/or the organic solvent is selected from ethyl acetate, dichloromethane, diethyl ether, isopropyl ether and tetrahydrofuran, and is preferably ethyl acetate; and/or the structure of the compound A is

8. The method according to claim 6 or 7, characterized in that: the method also comprises a purification step, and the purification method comprises the following steps: adding the organic solvent of claim 6 or 7 into the system obtained after the reaction, and extracting to keep an organic phase;

or, the purification method further comprises drying the organic phase, and performing gradient elution by using rapid preparative liquid chromatography, wherein the eluent is a mixed solvent of chloroform and methanol, the volume percentage of the methanol in the mixed solvent is 0-20%, and the elution gradient is as follows: the percentage of methanol volume in the eluent increased linearly from 0% to 20% over 3 column volumes.

9. Use of the compound of any one of claims 1 to 4, or a salt thereof, or a stereoisomer thereof, or a crystalline form thereof, for the preparation of a medicament for preventing and/or treating hypoxic injury of nerve cells.

10. Use of a compound according to any one of claims 1 to 4, or a salt thereof, or a stereoisomer thereof, or a crystalline form thereof, for the manufacture of a medicament for the prevention and/or treatment of a neurological disease;

the nervous system disease is preferably a nervous system disease associated with hypoxic injury of nerve cells, more preferably Parkinson's disease, Alzheimer's disease, stroke.

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