CN114853839B - A kind of panaxadiol compound and its preparation method and medical application - Google Patents

Abstract

The invention discloses a panaxadiol compound, a preparation method thereof and a medical application thereof. It is a novel compound having anti-Alzheimer's disease activity. The structure is modified and optimized on the basis of the natural product panaxadiol to improve its physical and chemical properties, enhance its pharmacological activity, and enhance its druggability. The preparation method of the panaxadiol compound provided by the invention has the advantages of mild reaction conditions, simple operation in the reaction process, and cheap and easy-to-obtain reagents. The prepared novel panaxadiol derivatives have a similar effect to panaxadiol. MTT experimental studies have shown that compound Q4 has a stronger neuroprotective effect on Aβ _25-35 induction than panaxadiol and the positive control donepezil, and does not show cytotoxicity to PC12 at 7.5 to 120 μM, and can be applied to the preparation of Alzheimer's disease drugs.

Description

A kind of panaxadiol compound and its preparation method and medical application

technical field

The invention discloses a ginsengdiol compound, which is a new compound; the invention also provides a preparation method and medical application of the compound, belonging to the technical field of biomedicine.

Background technique

Alzheimer's disease was first discovered and reported in 1906 by a German physician, Eloise Alzheimer. AD is the most common neurodegenerative disease, accounting for about 70% of the disease cases and is one of the main causes of human death. The latest official statistics released by the World Health Organization in 2020 show that more than 55 million people around the world are currently diagnosed with dementia, and the overall number of patients is growing at a rate of nearly 10 million cases per year. On average, one person suffers from dementia every three seconds. It is estimated that by 2050, the number of people living with dementia in the world will reach 139 million.

Although there are clinically available anti-Alzheimer's drugs, their effects are only to alleviate the symptoms of early patients and improve memory ability, but they cannot completely cure or prevent the serious condition.

Contents of the invention

The invention discloses a panaxadiol compound, which is a novel compound. The compound exhibits an anti-Alzheimer's disease effect in vitro, and can be applied to the preparation of anti-Alzheimer's disease medicine and a pharmaceutical composition containing it.

The invention discloses a preparation method of panaxadiol compounds, which has the advantages of mild reaction conditions, simple operation in the reaction process, and cheap and easy-to-obtain reagents.

A kind of Panaxadiol compound of the present invention, its chemical structural formula is:

The name of above-mentioned panaxadiol compound is:

(3 S ,5 R ,8 R ,9 R ,10 R ,12 R ,13 R ,14 R ,17 S )-12-hydroxy-4,4,8,10,14-pentamethyl-17-(( R )-2,6,6-trimethyltetrahydro-2 H -pyran-2-yl)hexadecahydro-1 H- cyclopenta[a] phenanthren-3-yl 4-methylpiperazine carbamate; Q4 for short; molecular formula: C ₃₆ H ₆₂ N ₂ O ₄ ; molecular weight: 586.47742.

The invention discloses a ginsengdiol compound and a preparation method thereof. The specific preparation steps are as follows:

1) Slowly add p-nitrophenyl chloroformate (5 mmol~7 mmol) in dichloromethane solution (6 mmol/10 mL) into the dichloromethane solution of ginseng diol (1 mmol~3 mmol) (1 mmol/1 mL) under ice bath, then add N,N-lutidine (DMAP) (1 mmol~3 mmol), triethylamine (2 mmol~4 mmol), and react overnight at room temperature;

2) After the reaction was completed, intermediate A was purified by silica gel chromatography with a yield of 93%;

3) Take intermediate A (1 mmol~2 mmol) and methylpiperazine (4 mmol~6 mmol), put them in 3 mL of ethanol, and react at room temperature for 8 hours;

4) After the reaction was completed, ethyl acetate was added for extraction, washed with saturated brine, filtered with suction, dried, and subjected to column chromatography to obtain compound Q4.

The nuclear magnetic resonance data of compound Q4 of the present invention are as follows:

M.p. 194-196 °C; yield 52%.¹H NMR (300 MHz, CDCl₃)δ 6.28 (s, 1H), 4.37 (dd,J ₁ = 9Hz,J ₂ =3 Hz, 1H), 3.51 (s, 5H), 2.39 (s, 4H), 2.32 (s, 3H), 2.14-1.83 (m, 3H), 1.83-1.66 (m, 4H), 1.65-1.53 (m, 4H), 1.53-1.39 (m, 5H), 1. 38-1.24 (m, 7H), 1.20 (d,J = 12 Hz, 6H), 1.09 (d,J = 15 Hz, 2H), 1.04-0.96 (m, 4H), 0.90 (s, 4H), 0.88 (s, 5H), 0.85 (s, 4H);¹³C NMR (126 MHz, CDCl₃)δ 155.48, 81.87, 76.66, 73.09, 69.88, 55.96, 54.85, 54.73, 51.22, 49.83, 49.20, 46.21, 39.82, 38.57, 38.21, 37.05, 36.46, 35 .76, 34.84, 33.03, 31.14, 30.59, 29.70, 28.10, 27.16, 25.17, 24.21, 19.44, 18.23, 17.08, 16.79, 16.29, 16.15, 15.65.

The positive effects of the present invention are:

Provided is a new compound-panaxadiol compound and its preparation method. The compound has a better neuroprotective effect, and it exhibits a stronger neuroprotective effect than the positive control drug donepezil in the Aβ _25-35- induced PC12 cell injury model; the cytotoxicity evaluation study shows that the compound Q4 is less toxic to PC12 cells, and can be applied to the preparation of Alzheimer's disease drugs in the later stage. The preparation method of the compound of the present invention has the advantages of mild reaction conditions, simple operation of the reaction process, and cheap and easy-to-obtain reagents.

Description of drawings

Figure 1 is the effect of compound Q4 of the present invention on the survival rate of PC12 cells at a concentration of 7.5 μM to 120 μM;

Fig. 2 shows the neuroprotective effect of compound Q4 of the present invention, panaxadiol (PD) (30 μM) and Donepezil (15 μM) in inhibiting PC12 cell damage induced by Aβ _25-35 .

Detailed ways

The present invention will be further described below in conjunction with examples, but the present invention is not limited to the above embodiments. Different embodiments can be realized with some modifications within the range shown in the claims, and such modifications should belong to the scope of the present invention.

Example 1:

1) Slowly add 6 mmol of p-nitrophenyl chloroformate in dichloromethane (6 mmol/10 mL) into 1 mmol of panaxadiol in dichloromethane (1 mmol/1 mL) in an ice bath, then add 2 mmol of N,N-lutidine (DMAP) and 3 mmol of triethylamine, and react overnight at room temperature;

2) After the reaction was completed, intermediate A was purified by silica gel chromatography with a yield of 93%;

3) Take 1 mmol of intermediate A and 5 mmol of methylpiperazine in 3 mL of ethanol, and react at room temperature for 8 hours;

4) After the reaction was completed, ethyl acetate was added for extraction, washed with saturated brine, filtered with suction, dried, and subjected to column chromatography to obtain compound Q4.

The resulting compound is named:

(3 S ,5 R ,8 R ,9 R ,10 R ,12 R ,13 R ,14 R ,17 S )-12-hydroxy-4,4,8,10,14-pentamethyl-17-(( R )-2,6,6-trimethyltetrahydro-2 H -pyran-2-yl)hexadecahydro-1 H- cyclopenta[a] phenanthren-3-yl 4-methylpiperazine carbamate; Q4 for short;

The chemical structural formula of gained compound is:

The molecular formula of the obtained compound is: C ₃₆ H ₆₂ N ₂ O ₄ ; the molecular weight: 586.47742.

The nuclear magnetic resonance data of gained compound is as follows:

M.p. 194-196 °C; yield 52%.¹H NMR (300 MHz, CDCl₃)δ 6.28 (s, 1H), 4.37 (dd,J ₁ = 9Hz,J ₂ =3 Hz, 1H), 3.51 (s, 5H), 2.39 (s, 4H), 2.32 (s, 3H), 2.14-1.83 (m, 3H), 1.83-1.66 (m, 4H), 1.65-1.53 (m, 4H), 1.53-1.39 (m, 5H), 1. 38-1.24 (m, 7H), 1.20 (d,J = 12 Hz, 6H), 1.09 (d,J = 15 Hz, 2H), 1.04-0.96 (m, 4H), 0.90 (s, 4H), 0.88 (s, 5H), 0.85 (s, 4H);¹³C NMR (126 MHz, CDCl₃)δ 155.48, 81.87, 76.66, 73.09, 69.88, 55.96, 54.85, 54.73, 51.22, 49.83, 49.20, 46.21, 39.82, 38.57, 38.21, 37.05, 36.46, 35 .76, 34.84, 33.03, 31.14, 30.59, 29.70, 28.10, 27.16, 25.17, 24.21, 19.44, 18.23, 17.08, 16.79, 16.29, 16.15, 15.65.

Example 2:

Pharmaceutical composition: 1000 tablets each containing 100 mg of active ingredient Formula: 100 g of compound Q4 of the present invention, 2 g of hydroxypropyl methylcellulose, 10 g of wheat starch, 100 g of sucrose, 6 g of magnesium stearate; the dose used should be adapted to the nature and severity of the disease, the route of administration, and the age and weight of the patient. The daily dose varies between 0.1 mg-1.0 g, and can be administered once or several times.

Further prove the medical purposes of compound of the present invention by following test:

Test example 1

MTT assay for cytotoxicity

Spread PC12 cells on a 96-well plate at a density of 1×10 ⁴ cells/well and incubate for 12 hours. After absorbing 10% of the complete medium, add 180 μL of 1% RPMI 1640 medium to the treatment group, add 20 μL of different concentrations of drugs to the treatment group, add 20 μL of 1% medium to the blank control group, and culture them in the incubator for 24 hours. On the second day, add 20 μL of 5 mg/mL MTT solution directly to the 96-well plate, place it in the dark for 4 hours, remove the medium, and then add 150 μL of dimethyl sulfoxide solution with a pipette gun. Wrap it in tin foil and shake it on a shaker for about 10 min. Set the wavelength of the microplate reader to 490 nm and then measure the absorbance of each well, and use Excel to calculate the cell viability of each well.

Test example 2

MTT method to detect the degree of cell damage

experimental method:

Spread PC12 cells on a 96-well plate at a density of 1×10 ⁴ cells/well and incubate for 12 hours. After 10% complete medium was sucked away, 178 μL of 1% medium was added to the treatment group. After starvation, 20 μL of different concentrations of drugs were added to the treatment group. After 1 hour of administration, 2 μL of 10 μM Aβ _25-35 was added to the control group and the treatment group, and 20 μL of 1% medium was added to the blank control group, and placed in the incubator Incubate for 24 hours. On the second day, add 20 μL of 5 mg/mL MTT solution directly to the 96-well plate, and place it in the dark for 4 hours, remove the medium, and then add 150 μL of dimethyl sulfoxide solution with a pipette gun. Wrap it in tin foil and shake it on a shaker for about 10 min. Set the wavelength of the microplate reader to 490 nm and then measure the absorbance of each well, and use Excel to calculate the cell viability of each well (see Table 1).

Table 1. Cell Viability of Q4 (15 μM)

Compd Cell protection( %) Compd Cell protection ( %) control 100±4.92 _Aβ25-35 65.41±3.11 ^### Donepezil 72.62±2.76 ^** PD 62.88±4.4 Q4 80.6±10.85 ^***

Conclusion: As shown in Table 1, Q4 of the present invention has a better neuroprotective effect (80.6±10.85%) at 15 μM.

Claims (4)

1. A ginseng diol derivative, which is characterized by having the following structural formula:

；

the molecular formula is: c (C) ₃₆ H ₆₂ N ₂ O ₄ The method comprises the steps of carrying out a first treatment on the surface of the Molecular weight: 586.47742.

2. The method for producing a ginseng diol derivative according to claim 1, comprising the steps of:

1) Slowly adding a 6 mmol/10 mL dichloromethane solution containing 5 mmol-7 mmol of p-nitrophenyl chloroformate into a 1 mmol/1 mL dichloromethane solution containing 1 mmol-3 mmol of ginseng glycol under an ice bath, then adding 1 mmol-3 mmol of N, N-lutidine (DMAP) and 2 mmol-4 mmol of triethylamine, and reacting at room temperature overnight;

2) After the reaction is completed, purifying by silica gel chromatography to obtain an intermediate A with the yield of 93%;

3) Taking 1-2 mmol of intermediate A, 4-6 mmol of methylpiperazine, placing in 3 mL ethanol, and reacting for 8 hours at room temperature;

4) And after the reaction is finished, adding ethyl acetate for extraction, washing with saturated saline water, carrying out suction filtration, drying and carrying out column chromatography to obtain the ginseng diol derivative.

3. Use of a ginseng diol derivative according to claim 1 for the preparation of a medicament for alzheimer's disease.

4. A pharmaceutical preparation characterized by comprising the ginseng diol derivative according to claim 1 as an active ingredient, together with one or more pharmaceutically acceptable auxiliary agents.