PPD derivative with cardiac protection activity as well as preparation method and application of PPD derivative

Abstract

The invention discloses application of PPD derivatives as shown in formulas (I), (II) and (III) and pharmaceutically acceptable salts thereof in preparation of anti-hypoxia reoxygenation injury drugs, anti-ischemic injury drugs or health care products. The derivatives of the ginsenoside PPD are found to have myocardial ischemia-reperfusion injury resisting activity for the first time and show good ischemic injury resisting effect and ischemia-reperfusion injury resisting effect, compared with a model group, the survival rate of cells is remarkably increased, the relative activity of the ginsenoside PPD is superior to that of PPD, cytotoxicity does not occur, and the ginsenoside PPD has good development and application value. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a class of PPD derivatives with novel structures and a preparation method thereof, and the application of the derivatives as heart protection drugs or health products. Background Art

[0002] Myocardial ischemia-reperfusion injury (MIRI) is one of the important causes of myocardial damage in coronary heart disease. It mainly refers to the fact that after a long period of ischemia, the myocardial tissue is restored to blood perfusion through surgery and other means, resulting in more significant functional disorders and damage than before blood perfusion, such as myocardial cell necrosis, decreased cardiac contractile function and other cardiac pathological symptoms. At present, although the drugs used in clinical practice have certain efficacy, they have certain limitations such as safety. Therefore, it is necessary to seek new structural types of drugs with anti-MIRI activity.

[0003] Natural products have advantages such as multiple skeletons and high biological activity, and are an important source for drug discovery. Therefore, finding lead compounds from natural products and optimizing their structures has become one of the important ways of modern drug discovery. Ginsenosides are the main active ingredients of ginseng and have a wide range of pharmacological activities. In particular, 20S-protopanaxadiol (PPD) has a good effect on resisting myocardial ischemia-reperfusion injury, but its toxicity is relatively large, which affects its clinical use. The present invention is expected to reduce its toxicity by structural modification of PPD, and obtain PPD derivatives with higher resistance to myocardial ischemia-reperfusion injury or myocardial ischemia and novel structures. Summary of the invention

[0004] In order to find drugs with low cytotoxicity and novel structure that have anti-myocardial ischemia-reperfusion injury or anti-myocardial ischemia activity, the present invention explores through a series of constructions based on the PPD parent structure, in order to develop novel compounds with low cytotoxicity and good anti-myocardial ischemia-reperfusion injury or anti-myocardial ischemia activity. The present invention provides a series of derivatives with structural characteristics of general formula (I), (II), and (III), and pharmaceutically acceptable salts thereof. Compound 10 with structural characteristics of general formula (I) and compounds 12 to 16 with structural characteristics of (II) and (III) are all new compounds discovered and synthesized for the first time.

[0005] The technical problem to be solved by the present invention is to find new structural types of low cytotoxic compounds with excellent anti-MIRI or anti-myocardial ischemia properties. Another object of the present invention is to provide compounds 12 to 16 having the structural characteristics of formula (II) and (III) and their preparation methods.

[0006] Another object of the present invention is to provide a series of derivatives having the structural characteristics of general formula (I), (II), (III), and pharmaceutically acceptable salts thereof, and their use in the preparation of drugs for preventing or treating IRI or myocardial ischemia, as well as their use in health products for enhancing resistance to myocardial ischemia.

[0007] In order to achieve the object of the invention, the present invention adopts the following technical scheme:

[0008] The use of PPD derivatives represented by formula (I), (II) and (III) and pharmaceutically acceptable salts thereof in the preparation of anti-hypoxia-reoxygenation injury drugs or health products; preferably, the anti-hypoxia-reoxygenation injury drugs are drugs for treating myocardial ischemia-reperfusion injury, but are not limited to the treatment and prevention of myocardial ischemia-reperfusion injury, and are used to improve the ability to resist myocardial ischemia.

[0009]

[0010] In formula (I), when R3 is hydroxyl and R2 is 12R-hydroxyl, R1 is amino or carbonyl; when R3 is hydroxyl and R2 is 12S-hydroxyl, carbonyl or oxime, R1 is hydroxyl; when R3 is hydroxyl and R2 is carbonyl, R1 is carbonyl; when R3 is tert-butyldimethylsilyloxy, R2 and R1 are both hydroxyl;

[0011] In formula (III), R4 is 2-methyl-vinyl or 2-hydroxy-isopropyl.

[0012] The present invention also provides the use of PPD derivatives represented by formula (I), (II) and (III) and pharmaceutically acceptable salts thereof in the preparation of anti-ischemic injury drugs or health products; the anti-ischemic injury drugs are preferably drugs for treating and preventing myocardial ischemic injury, but are not limited to the treatment and prevention of myocardial ischemic injury, and are preferably used to improve the ability to resist myocardial ischemia.

[0013] Furthermore, it is preferred that the compound represented by formula (I) is one of the following compounds 1 to 11; it is preferred that the compound represented by formula (II) is one of the following compounds 12 to 13; it is preferred that the compound represented by formula (III) is one of the following compounds 14 to 16.

[0014] In the present invention, compound 1 is [(20S, 24R)-epoxy dammarane-12β, 25-diol-3β-amine]; compound 2 is [(20S, 24R)-epoxy dammarane-12β, 25-diol-3α-amine]; compound 3 is [(20S, 24S)-epoxy dammarane-12β, 25-diol-3β-amine]; compound 4 is [(20S, 24R)-epoxy dammarane-12β, 25-diol-3-ketone]; compound 5 is [(20S, Compound 6 is [(20S,24S)-epoxy dammarane-12β,25-diol-3-one]; Compound 7 is [(20S,24S)-epoxy dammarane-25-ol-3,12-one]; Compound 8 is [(20S,24S)-epoxy dammarane-3β,25-diol-12-one]; Compound 9 is [(20S,24R)-epoxy dammarane-3β,25-diol-1 2-keto-oxime]; Compound 10 is [(20S,24S)-epoxy-dammarane-3β,12α,25-triol]; Compound 11 is 25-O-tert-butyldimethylsilyl-20S,24R-epoxy-dammarane-3β,12β-diol; Compound 12 is (20S,24R)-epoxy-3-oxo-4-oxa-A-homo-dammarane-12β,25-diol; Compound 13 is (20S,24S)-epoxy-3-oxo-4-oxa-A -homo-dammarane-12β,25-diol; compound 14 is (20S,24R)-epoxy-3,4-opening-3-carboxy-4(28)-ene-dammarane-12β,25-diol; compound 15 is (20S,24R)-epoxy-3,4-opening-3-carboxy-dammarane-4,12β,25-triol; compound 16 is (20S,24S)-epoxy-3,4-opening-3-carboxy-dammarane-4,12β,25-triol.

[0015]

[0016]

[0017] The pharmaceutically acceptable salts described in the present invention refer to conventional acid addition salts, which have the same pharmaceutical efficacy as the derivatives and are salts formed with suitable non-toxic organic or inorganic acids.

[0018] Furthermore, preferably, compound 1, 8, 11, 12, 14 or 16 is used for preparing drugs or health products against hypoxia-reoxygenation injury.

[0019] Preferably, compound 7, 10, 12 or 16 is used for preparing anti-ischemic injury drugs or health products.

[0020] The present invention also provides a pharmaceutical composition for resisting hypoxia-reoxygenation injury or ischemia injury, wherein the pharmaceutical composition comprises a PPD derivative represented by formula (I), (II), or (III) and a pharmaceutically acceptable salt thereof.

[0021] Preferably, the pharmaceutical composition comprises one or more of compounds 1 to 16 and pharmaceutically acceptable salts thereof.

[0022] Furthermore, the pharmaceutical composition for resisting hypoxia-reoxygenation injury preferably comprises one or more of Compounds 1, 8, 11, 12, 14 or 16 and pharmaceutically acceptable salts thereof.

[0023] The pharmaceutical composition for preventing ischemic injury preferably comprises one or more of compounds 7, 10, 12 or 16 and pharmaceutically acceptable salts thereof.

[0024] The pharmaceutical composition can be added with pharmaceutically acceptable carriers to prepare common pharmaceutical preparations, such as tablets, capsules, powders, syrups, liquids, suspensions, injections, and can also be added with common pharmaceutical excipients such as sweeteners, diluents, and fillers.

[0025] The pharmaceutical composition provided by the present invention can be administered clinically by oral administration, injection or the like.

[0026] The clinical dosage of the pharmaceutical composition of the present invention is 0.01 mg to 1000 mg / day of the active ingredient, and may deviate from this range depending on the severity of the disease or the dosage form.

[0027] The present invention also provides a compound having cardioprotective activity, which is one of compounds 12 to 16:

[0028]

[0029] Furthermore, the preparation method of compounds 10 and 12 to 16 of the present invention is as shown in the following method:

[0030] The method comprises the following steps:

[0031] (1) adding 24S-pyxinol, NaHCO3, Dess-Martin reagent, and tert-butanol in a molar ratio of 1:5-10:5-6:1:3-5 to carry out an oxidation reaction to obtain compound 7;

[0032] Further, the reaction solvent of step (1) is an aprotic solvent, preferably CH2Cl2;

[0033] The reaction temperature of the oxidation reaction is -20°C to 80°C; preferably, the reaction temperature is -10°C to 30°C, and more preferably, it is room temperature.

[0034] The reaction time of the oxidation reaction is 3 to 8 hours; preferably, the reaction time is 5 to 7 hours.

[0035] After the reaction is completed, the reaction solution is post-treated to obtain compound 7. The post-treatment method of the reaction solution is as follows: adding Na2SO3 aqueous solution to the reaction solution to quench the reaction, extracting with ethyl acetate, filtering and concentrating the organic phase after drying, and purifying the concentrated sample by silica gel column chromatography to obtain compound 7.

[0036] (2) Compound 7 is subjected to reduction reaction with NaBH4 to obtain compound 10;

[0037] The molar ratio of compound 7 to NaBH4 is 1:3-10;

[0038] Furthermore, the reaction solvent of step (2) is an aprotic solvent, preferably methanol (MeOH);

[0039] The temperature of the reduction reaction is room temperature, and the reaction time is 4 to 10 hours.

[0040] After the reaction is completed, the reaction solution is post-treated to obtain compound 10. The post-treatment method of the reaction solution is as follows: adding water to the reaction solution to quench the reaction, extracting with ethyl acetate, washing and drying the organic phase, filtering and concentrating it, and purifying the concentrated sample by silica gel column chromatography to obtain compound 10.

[0041] (3) adding 24R-Pyxinol or 24S-Pyxinol and pyridinium chlorochromate (PCC) in a molar ratio of 1:0.5 to 2 to carry out an oxidation reaction to obtain compound 4 or compound 6 respectively;

[0042] The reaction solvent of step (3) is an aprotic solvent, preferably dichloromethane (DCM);

[0043] The reaction temperature of the oxidation reaction is -20°C to 80°C; preferably, the reaction temperature is -10°C to 30°C, and more preferably, it is room temperature.

[0044] The reaction time of the oxidation reaction is 3 to 8 hours, preferably 3 to 5 hours. TLC is usually used to monitor the reaction until the reaction is complete.

[0045] After the reaction is completed, the reaction solution is post-treated to obtain compound 4 or compound 6. The post-treatment method of the reaction solution is: filtering the reaction solution, vacuum concentrating, and then using silica gel column chromatography (SGCC) to obtain compound 4 or compound 6.

[0046] (4) Compound 4 or Compound 6 is reacted with m-chloroperbenzoic acid to undergo a Baeyer-Villiger reaction to obtain Compound 12 or Compound 13, respectively;

[0047] The molar ratio of compound 4 or compound 6 to m-chloroperbenzoic acid (m-CPBA) is 1:3 to 15. Further, the reaction solvent of step (4) is an aprotic solvent, preferably dichloromethane (DCM);

[0048] The reaction temperature of the Baeyer-Villiger reaction is 10° C. to 40° C., preferably room temperature; and the reaction time is 5 to 12 hours.

[0049] After the reaction is completed, the reaction solution is post-treated to obtain compound 12 or compound 13. The post-treatment method of the reaction solution is as follows: adding water to the reaction solution to quench the reaction, extracting with ethyl acetate, washing and drying the organic phase, filtering and concentrating it, and purifying the concentrated sample by silica gel column chromatography to obtain compound 12 or compound 13, respectively.

[0050] (5) Compound 12 is subjected to an acidic ring-opening reaction with p-toluenesulfonamide (P-TSA) to obtain compound 14;

[0051] The molar ratio of the compound 12 to p-toluenesulfonamide (P-TSA) is 1:3-9.

[0052] Further, the reaction solvent of step (5) is an aprotic solvent, preferably dichloromethane (DCM);

[0053] The reaction temperature of the acidic ring-opening reaction is 10° C. to 40° C., preferably room temperature; and the reaction time is 5 to 18 hours.

[0054] After the reaction is completed, the reaction solution is post-treated to obtain compound 14. The post-treatment method of the reaction solution is as follows: adding water to the reaction solution to quench the reaction, extracting with ethyl acetate, washing and drying the organic phase, filtering and concentrating it, and purifying the concentrated sample by silica gel column chromatography to obtain compound 14.

[0055] (6) Compound 12 or Compound 13 undergoes an alkaline ring-opening reaction with potassium hydroxide (KOH) to obtain Compound 15 or Compound 16, respectively;

[0056] The molar ratio of the compound 12 or compound 13 to potassium hydroxide is 1:1.5-5.

[0057] Furthermore, the reaction solvent of step (6) is an aprotic solvent, preferably a tetrahydrofuran aqueous solution (THF / H2O); the volume ratio of tetrahydrofuran to water in the tetrahydrofuran aqueous solution is preferably 10 to 20:1.

[0058] The reaction temperature of the alkaline ring-opening reaction is 10° C. to 40° C., preferably room temperature; and the reaction time is 5 to 18 hours.

[0059] After the reaction is completed, the reaction solution is post-treated to obtain compound 15 or compound 16. The post-treatment method of the reaction solution is: the solvent of the reaction solution is evaporated under vacuum, and the concentrated sample is purified by silica gel column chromatography to obtain compound 15 or compound 16 respectively.

[0060] Compared with the prior art, the present invention has the following advantages:

[0061] The compounds represented by formula (I), (II) and (III) provided by the present invention and their pharmaceutically acceptable salts are all found for the first time to have activity against myocardial ischemia-reperfusion injury, wherein the derivatives having the structural characteristics of general formula (II) and (III) are all new compounds synthesized for the first time. The experimental results show that in the anti-ischemic injury model experiment, compared with the H / R group, the derivatives of ginsenoside PPD all showed good cell protection activity, significantly improved the survival rate of cells, and the relative activity was better than PPD, and at a concentration of 30 μM, PPD had obvious cytotoxicity to ischemic cells, while compounds 1 to 16 did not show cytotoxicity and showed good anti-ischemic injury effect. In the model experiment of anti-ischemia reperfusion injury, compared with the H / R group, the derivatives of ginsenoside PPD showed better cell protection activity, significantly improved the survival rate of cells, and the relative activity was better than PPD. At a concentration of 30 μM, PPD had obvious cytotoxicity to ischemia reperfusion cells, while compounds 1 to 16 showed no cytotoxicity and showed better anti-ischemia reperfusion injury effect. The compounds of the present invention have better anti-ischemic injury and anti-ischemia reperfusion injury effects, and have low cytotoxicity, and have good development and application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1 The graph shows the experimental results of the protective activity of PPD and compounds 1 to 8 on H9c2 cells in a cell model experiment against ischemic injury.

[0063] Figure 2 The graph shows the experimental results of the protective activity of compounds 9 to 16 on H9c2 cells in a cell model experiment against ischemic injury.

[0064] Figure 3 The graph shows the experimental results of the protective activity of PPD and compounds 1 to 8 on H9c2 cells in a cell model experiment against ischemia-reperfusion injury.

[0065] Figure 4 The graph shows the experimental results of the protective activity of compounds 9 to 16 on H9c2 cells in a cell model experiment against ischemia-reperfusion injury.

[0066] Figure 5The graph shows the cytotoxicity test results of compound PPD and compounds 1 to 16 on normal H9c2 cells. DETAILED DESCRIPTION

[0067] The technical solution of the present invention is further described below in conjunction with specific embodiments, but the protection scope of the present invention is not limited to these embodiments.

[0068] Example 1: [(20S,24R)-epoxy dammarane-12β,25-diol-3β-amine];

[0069] Using protopanaxadiol (20S-PPD) as the raw material, m-CPBA was first used for oxidative cyclization reaction, and 24R-Pyxinol was obtained at room temperature. 24R-Pyxinol (200 mg, 0.42 mmol) was dissolved in anhydrous dichloromethane (16 mL), and pyridinium chlorochromate (174 mg, 0.80 mmol) was added. After stirring at room temperature for 3 h, it was filtered and concentrated in vacuo, and then silica gel column chromatography (SGCC) was used to obtain compound 4. Compound 4 (4.000 g, 8.426 mmol) was dissolved in 80 mL of pyridine, and hydroxylamine hydrochloride (0.878 g, 12.634 mmol) was added. The mixture was stirred at 80 ° C for 3 h to obtain the compound [(20S, 24R)-epoxydammarane-12β,25-diol-3-ketoxime]. After mixing [(20S,24R)-epoxy dammarane-12β,25-diol-3-ketoxime] (72 mg, 0.147 mmol) and AcONH4 (99 mg, 1.284 mmol) in MeOH (6.45 mL), NaCNBH3 (81 mg, 1.284 mmol) and 15% TiCls solution (450 μL) were added under argon at 0°C. Stir at room temperature for 6 hours, dilute with ethyl acetate, rinse with water, and vacuum dry and concentrate. Silica gel column chromatography (SGCC) was used to obtain compound 1 [(20S,24R)-epoxy dammarane-12β,25-diol-3β-amine]

[0070] 1H NMR(400 MHz, CDCl3)d 3.84(dd,J=8.4,6.8 Hz,1H),3.51(td,J=10.4,4.3Hz,1H),2.85(dd,J=10.6,5.8 Hz,1H),2.18(td,J=10.0,3.4 Hz,1H),2.09-1.96(m,2H),1.92-1.26(m,16H),1.27(s,3H),1.26(s,3H),1.11-1.05(m,1H),1.13(s,3 H),1.09(s,3H),0.98(s,3H),0.98-0.92(m,1H),0.95(s,3H),0.89(s,3H),0.87(s,3H),0.78(d,J=9.2 Hz,1H)

[0071] Example 2: [(20S,24R)-epoxy dammarane-12β,25-diol-3α-amine];

[0072] After mixing [(20S,24R)-epoxy dammarane-12β,25-diol-3-ketoxime] (24 mg, 0.049 mmol) and AcONH4 (33 mg, 0.428 mmol) in MeOH (2.15 mL), NaCNBH3 (27 mg, 0.428 mmol) and 15% TiCls solution (150 μL) were added under argon at 0°C. Stir at room temperature for 6 hours, dilute with ethyl acetate, rinse with water, and vacuum dry and concentrate. Silica gel column chromatography (SGCC) was used to obtain compound 2 [(20S,24R)-epoxy dammarane-12β,25-diol-3α-amine].

[0073] 1 H NMR (400 MHz, CDCl3) δ3.83 (dd, J = 8.0, 6.8 Hz, 1H), 3.53 (td, J = 9.9, 4.0 Hz, 1H), 3.08 (m, 1H), 2.17 (td, J = 9.8, 2.6 Hz,1H),2.04–1.81(m,7H),1.59(m,6H),1.45-1.36(m,3H),1.32–1.23(m,3H),1.28(s,3H),1.26(s,3H),1.26 (s,3H),1.14-1.10(m,1H),1.10(s,3H),1.10(s,3H),0.96-0.90(m,1H),0.96(s,3H),0.92(s,3H),0.87(s,3H)

[0074] Example 3: [(20S,24S)-epoxy dammarane-12β,25-diol-3β-amine];

[0075] NaCNBH3 (86 mg, 1.37 mmol) and AcONH4 (105 mg, 1.36 mmol) were added to a solution of (20S, 24S)-epoxy-12β, 25-dihydroxydammarane-3-one oxime (266.5 mg, 0.55 mmol) in i-PrOH (20 mL), and 15% TiCl3 aq (0.52 mL) was added at 0°C. After stirring at room temperature for 15 h, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The residue was recrystallized from hot methanol to obtain compound 3 [(20S, 24S)-epoxydammarane-12β, 25-diol-3β-amine].

[0076] 1 H NMR(400 MHz, CDCl3)d 5.78(s,1H),3.89(dd,J=10.7,5.3 Hz,1H),3.53(td,J=10.4,4.7 Hz,1H),2.98(ddd,J=15.6,5.7,3.9 Hz,1H),2.34-2.22(m,2H),2.06(td,J=11.1,4.7 Hz,1H),2.01-1.82(m,5H),1.77(dd,J=11.4,6.9 Hz,1H),1.70(t,J=10.1 Hz,1H),1.56-1.49(m,5H),1.34-1.09(m,6H),1.28(s,3H),1.24(s,3H),1. 15(s,3H),1.11(s,3H),1.07(s,3H),1.04(s,3H),0.98(s,3H),0.90(s,3H).

[0077] Example 4: [(20S,24R)-epoxydammarane-12β,25-diol-3-one];

[0078] Pyxinol (150 mg, 0.315 mmol) was dissolved in anhydrous dichloromethane (8 mL), and pyridinium chlorochromate (43.5 mg, 0.20 mmol) was added. After stirring at room temperature for 3 h, the mixture was filtered and concentrated in vacuo. Silica gel column chromatography (SGCC) was used to obtain compound 4 [(20S, 24R)-epoxydammarane-12β, 25-diol-3-one].

[0079] 1H NMR(CDCl3,500 MHz)d 3.85(dd,J=8.6 Hz,6.0 Hz,1H),3.53(td,J=10.5Hz,4.5 Hz,1H),2.48-2.54(m,1H),2.42(ddd,J=11.5 Hz,7.5 Hz,4.0 Hz,1H),2.21(td,J=11.5 Hz,3.5 Hz,1H),1.28(s,6H),1.10(s,3H),1.07(s,3H),1.04(s,3H),1.02(s,3H),0.96(s,3H),0.91(s,3H).

[0080] Example 5: [(20S,24R)-epoxydammarane-3β,25-diol-12-one];

[0081] Under argon at 0℃, sodium borohydride (12 mg, 0.32 mmol) was added to the compound [(20S,24R)-epoxydammarane-25-ol-3,12-one] (62 mg, 0.13 mmol) dissolved in isopropanol.

[0082] After stirring for 1 h, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. Silica gel column chromatography (SGCC) was then used to obtain compound 5 [(20S, 24R)-epoxydammarane-3β, 25-diol-12-one].

[0083] 1 H NMR (400 MHz, CDCl3) δ3.69 (dd, J = 8.6, 6.1 Hz, 1H), 3.19 (dd, J = 11.2, 4.9 Hz, 1H), 2.90 (d, J = 9.5 Hz, 1H), 2.56 (td, J = 10.5, 4.5 Hz,1H),2.25–2.15(m,2H),1.89–1.49(m,15H),1.46–0.73(m,3H),1.21(s,3H) ,1.20(s,3H),1.11(s,6H),0.99(s,3H),0.93(s,3H),0.80(s,3H),0.76(s,3H).

[0084] Example 6: [(20S,24S)-epoxydammarane-12β,25-diol-3-one];

[0085] 24S-Pyxinol (130 mg, 0.273 mmol) was dissolved in anhydrous dichloromethane (10.4 mL), and pyridinium chlorochromate (113.1 mg, 0.52 mmol) was added. After stirring at room temperature for 8 h, the mixture was filtered and concentrated in vacuo. Silica gel column chromatography (SGCC) was used to obtain compound 6 [(20S, 24S)-epoxydammarane-12β,25-diol-3-one].

[0086] 1 H NMR(CDCl3,500 MHz)d 3.89(dd,J=10.5 Hz,5.0 Hz,1H),3.54(td,J=10.0Hz,4.5 Hz,1H),2.49-2.54(m,1H),2.45(ddd,J=11.0 Hz,8.0 Hz,3.5 Hz,1H),2.27(td,J=10.5 Hz,4.5 Hz,1H),1.28(s,3H),1.23(s,3H),1.11(s,3H),1.08(s,3H),1.053(s,3H),1.046(s,3H),0.96(s,3H),0.93(s,3H)

[0087] Example 7: [(20S,24S)-epoxydammarane-25-ol-3,12-one];

[0088] A mixture of 24S-Pyxinol (22.0 mg, 0.0461 mmol) and NaHCO3 (23.2 mg, 0.277 mmol) was added to dehydrated CH2Cl2 (1.0 mL), and Dess-Martin reagent (98 mg, 0.23 mmol) and tert-butyl alcohol (17.6 μL, 0.1844 mmol) were added at 0°C. After stirring at room temperature for 6 hours, an aqueous Na2SO3 solution was added, extracted with ethyl acetate, dried over Na2SO4, and purified by column chromatography to obtain compound 7 [(20S, 24S)-epoxydammarane-25-ol-3,12-one].

[0089] 1 H NMR(CDCl3,300 MHz)d 3.69-3.73(m,1H),3.01(d,J=9.8 Hz,1H),2.49-2.58(m,1H),2.45-2.47(m,1H),2.24-2.29(m,1H),2.20-2.30(m,2H),1.4 0(s,3H),1.25(s,6H),1.20(s,3H),1.10(s,3H),1.04(s,3H),0.88(s,3H),0.87(s,3H).

[0090] Example 8: [(20S,24S)-epoxydammarane-3β,25-diol-12-one];

[0091] Compound 7 (62 mg, 0.13 mmol) was dissolved in isopropanol, and NaBH4 (12 mg, 0.32 mmol) was added at 0°C.

[0092] After stirring for 1 h, aqueous NH4Cl solution was added to the reaction mixture, extracted with ethyl acetate, dried over Na2SO4, and purified by column chromatography to obtain compound 8 [(20S,24S)-epoxydammarane-3β,25-diol-12-one].

[0093] 1 H NMR (CDCl3, 500 MHz) d 3.71 (dd, J = 10.5 Hz, 6.0 Hz, 1H), 3.20 (dd, J = 11.0 Hz, 4.5 Hz, 1H), 2.96 (d, J = 9.5 Hz, 1H), 2.53 (td, J = 10.5 Hz, 4.5 Hz,1H),2.20-2.22(m,2H),1.20(s,3H),1.19(s,3H),1.09(s,3H),1.04(s,3H),0.99(s,3H),0.94(s,3H),0.80(s,3H),0.76(s,3H).

[0094] Example 9: [(20S,24R)-epoxydammarane-3β,25-diol-12-ketoxime];

[0095] At room temperature, hydroxylamine hydrochloride (33 mg, 0.475 mmol) was added to compound 5 (28 mg, 0.59 mmol) dissolved in pyridine. After stirring at 80 °C for 5 h, the mixture was purified by column chromatography to obtain compound 9 [(20S,24R)-epoxydammarane-3β,25-diol-12-ketoxime].

[0096] 1H NMR (400 MHz, CDCl3) δ3.71 (dd, J = 8.8, 6.1 Hz, 1H), 3.33 (d, J = 10.4 Hz, 1H), 3.20 (dd, J = 11.5, 4.6 Hz, 1H), 2.60 (td, J = 10.2, 4.2 Hz,1H),2.52(d,J=9.7 Hz,1H),2.06-2.16(m,1H),1.92-0.73(m,18H),1.25(s,3H),1.21(s,3H),1. 20(s,3H),1.09(s,3H),0.98(s,3H),0.92(s,3H),0.80(s,3H),0.79(s,3H).

[0097] Example 10: [(20S,24S)-epoxydammarane-3β,12α,25-triol];

[0098] Compound 7 (321 mg, 0.676 mmol) was dissolved in methanol (7.5 mL), and NaBH4 (255.5 mg, 6.755 mmol) was added at 0°C. The mixture was stirred at room temperature for 4 h. Deionized water was added to quench the mixture, and the mixture was extracted with ethyl acetate, washed with saturated NH4Cl solution, dried with Na2SO4, filtered, concentrated, and subjected to silica gel column chromatography to obtain compound 10 [(20S,24S)-epoxydammarane-3β,12α,25-triol].

[0099] 1 H NMR (400 MHz, CDCl3) δ4.16 (s, 1H), 3.64 (dd, J = 9.8, 5.1 Hz, 1H), 3.22 (dd, J = 11.0, 5.1 Hz, 1H), 2.37 (td, J = 10.4, 7.1 Hz,1H),2.01–0.97(m,21H),1.19(s,3H),1.19(s,3H),1.12(s,3H),1.11(s,3H),0.99(s,3H),0.95(s,3H),0.87(s,3H),0.79(s,3H).

[0100] Example 11: 25-O-tert-butyldimethylsilyl-20S,24R-epoxydammarane-3β,12β-diol

[0101] Pyxinol (64 mg, 0.114 mmol) was dissolved in anhydrous DMF, 2,6-dimethylpyridine (40 μL) and TBSOTf (108 μL) were added at 0°C under nitrogen, stirred at room temperature for 6 h, quenched with deionized water, extracted with ethyl acetate, dried over Na2SO4, filtered, and concentrated in vacuo. Then, the remaining solution was dissolved in THF (1.2 mL) and added to MeOH (2.4 mL) containing 10% KOH at 0°C. After 24 h, it was neutralized with Amberlite IR120B, filtered, concentrated in vacuo, and then silica gel column chromatography (SGCC) was used to obtain compound 1125-O-tert-butyldimethylsilyl-20S,24R-epoxydammarane-3β,12β-diol.

[0102] 1 H NMR (400 MHz, CDCl3) δ3.67 (dd, J = 8.2, 6.4 Hz, 1H), 3.66 (td, J = 10.4, 4.0 Hz, 1H), 3.20 (dd, J = 11.4, 4.6 Hz, 1H), 2.16 (ddd, J = 12.8, 8.5, 4.7 Hz,1H),2.05(td,J=9.9,3.4 Hz,1H),1.87–0.69(m,20H),1.21(s,3H),1.18(s,3H),1.09(s,3H),0.97(s,3H) ,0.97(s,3H),0.91(s,9H),0.89(s,3H),0.86(s,3H),0.77(s,3H),0.07(s,6H).

[0103] Example 12: (20S, 24R)-epoxy-3-oxo-4-oxa-A-homo-dammarane-12β, 25-diol;

[0104] Compound 4 (100 mg, 0.211 mmol) was dissolved in anhydrous DCM (2 mL), and m-CPBA (388 mg, 2.249 mmol) was slowly added thereto. The mixture was stirred at room temperature overnight. An appropriate amount of deionized water was then added for quenching, the organic layer was extracted with ethyl acetate, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in sequence, and then dried over anhydrous Na2SO4. Anhydrous Na2SO4 was removed by filtration, and the filtrate was concentrated. The concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain compound 12 (20S, 24R)-epoxy-3-oxo-4-oxa-A-homo-dammarane-12β, 25-diol.

[0105] 1H NMR (400 MHz, CDCl3) δ3.86(dd,J=8.6,6.7 Hz,1H),3.50(td,J=10.5,4.4Hz,1H),2.64(ddd,J=14.3,12.3,4.7 Hz,1H),2.52(ddd,J=14.4,6.2,3.7 Hz,1H),2.21(ddd,J=11.0,9.3,3.9 Hz,1H),2.12–0.88(m,19H),1.48(s,3H),1.41(s,3H),1.28(s,3H),1.27(s,3H),1.10(s,3H),1.08(s,3H),1.04(s,3H),0.90(s,3H).

[0106] Example 13: (20S, 24S)-epoxy-3-oxo-4-oxa-A-homo-dammarane-12β, 25-diol;

[0107] Compound 6 (100 mg, 0.211 mmol) was dissolved in anhydrous DCM (2 mL), and m-CPBA (388 mg, 2.249 mmol) was slowly added thereto. The mixture was stirred at room temperature overnight. An appropriate amount of deionized water was then added for quenching, the organic layer was extracted with ethyl acetate, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in sequence, and then dried over anhydrous Na2SO4. Anhydrous Na2SO4 was removed by filtration, and the filtrate was concentrated. The concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain compound 13 (20S, 24S)-epoxy-3-oxo-4-oxa-A-homo-dammarane-12β, 25-diol.

[0108] 1 HNMR (400 MHz, CD3OD) δ3.82(dd,J=10.5,5.4 Hz,1H),3.50(td,J=10.5,4.5Hz,1H),2.78(ddd,J=14.3,12.5,5.1 Hz,1H),2.47(ddd,J=14.4,5.6,3.6 Hz,1H),2.25(ddd,J=11.1,9.5,4.2 Hz,1H),2.05–0.77(m,19H),1.50(s,3H),1.41(s,3H),1.26(s,3H),1.17(s,3H),1.11(s,3H),1.11(s,3H),1.10(s,3H),0.94(s,3H).

[0109] Example 14: [(20S, 24R)-epoxy-3,4-opening-3-carboxy-4(28)-ene-dammarane-12β,25-diol];

[0110] Compound 12 (100 mg, 0.204 mmol) was dissolved in anhydrous DCM (2 mL), and P-TSA (281 mg, 1.641 mmol) was slowly added thereto. The mixture was stirred at room temperature overnight. An appropriate amount of deionized water was then added for quenching, the organic layer was extracted with ethyl acetate, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in sequence, and then dried over anhydrous Na2SO4. Anhydrous Na2SO4 was removed by filtration, and the filtrate was concentrated. The concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain compound 14 [(20S, 24R)-epoxy-3,4-opened-3-carboxyl-4(28)-ene-dammarane-12β,25-diol].

[0111] 1 H NMR (400 MHz, CDCl3) δ4.86 (t, J = 1.8 Hz, 1H), 4.67 (d, J = 1.3 Hz, 1H), 3.85 (dd, J = 8.6, 6.6 Hz, 1H), 3.57 (td, J = 10.6, 4.4 Hz,1H),2.41–2.27(m,1H),2.27–2.16(m,2H),2.09–1.13(m,19H),1.73(s,3H) ,1.28(s,3H),1.27(s,3H),1.11(s,3H),1.02(s,3H),0.92(s,3H),0.86(s,3H).

[0112] Example 15: (20S, 24R)-epoxy-3,4-opening-3-carboxy-dammarane-4,12β,25-triol;

[0113] Compound 12 (100 mg, 0.204 mmol) was dissolved in a stirred solution of THF / H2O (2 mL, 20:1, v / v), and KOH (57.2 mg, 1.020 mmol) was slowly added thereto. The mixture was stirred at room temperature overnight, and the solvent was evaporated under vacuum and purified by silica gel column chromatography to obtain compound 15 [(20S, 24R)-epoxy-3,4-opened-3-carboxy-dammarane-4,12β,25-triol].

[0114] 1H NMR (400 MHz, CDCl3) δ3.85 (dd, J=8.7, 6.6 Hz, 1H), 3.54 (td, J=10.6, 4.4Hz, 1H), 2.47 (qd, J=14.8, 4.1 Hz,2H),2.28–2.17(m,2H),2.12–0.83(m,18H),1.29(s,3H),1.28(s,3H),1. 27(s,3H),1.26(s,3H),1.10(s,3H),1.02(s,3H),1.00(s,3H),0.89(s,3H).

[0115] Example 16: (20S, 24S)-epoxy-3,4-opening-3-carboxy-dammarane-4,12β,25-triol;

[0116] Compound 13 (100 mg, 0.204 mmol) was dissolved in a stirred solution of THF / H2O (2 mL, 20:1, v / v), and KOH (57.2 mg, 1.020 mmol) was slowly added thereto. The mixture was stirred at room temperature overnight, and the solvent was evaporated under vacuum and purified by silica gel column chromatography to obtain compound 16 [(20S,24S)-epoxy-3,4-opened-3-carboxyl-dammarane-4,12β,25-triol].

[0117] 1 H NMR (400 MHz, CDCl3) δ3.89 (dd, J=10.7,5.2 Hz,1H),3.58(td,J=10.4,4.5Hz,1H),2.55–2.36(m,2H),2.31–2.17(m,2H),2.08–0.84(m,18H),1.3 0(s,3H),1.27(s,3H),1.25(s,3H),1.22(s,3H),1.11(s,3H),1.04(s,3H),1.03(s,3H),0.91(s,3H).

[0118] Example 17: Protective activity of PPD derivatives on H9c2 cells in a cell model of ischemic injury Experimental method:

[0119] (1) Cell plating: H9c2 cells in good condition during the logarithmic growth phase were digested and counted, and 8 × 10 3 / well were inoculated in a 96-well plate and cultured in a 37°C, 5% CO2 cell culture incubator;

[0120] (2) Cell administration: After the cells were plated and attached for 24 hours, 3 μM, 10 μM, and 30 μM PPD derivatives were added for pretreatment for 1 hour, and 3 parallel wells were set up for each group. After pretreatment, the cells were placed steadily in an anaerobic box for hypoxia treatment. After 6 hours of hypoxia, the cells were taken out and returned to the incubator for reoxygenation at 37°C for 12 hours;

[0121] (3) MTT assay: After reoxygenation, add 20 μL of 5 mg / mL MTT solution and continue to culture in a 37°C incubator for 2 to 4 h. Discard the supernatant. Do not destroy the crystals formed at the bottom of the well during the process. Add 150 μL DMSO to each well and place it on a shaker in the dark for 10 min to fully dissolve the formazan crystals. Finally, use an enzyme reader to detect the absorbance at a wavelength of 570 nm. The H9c2 cell group that was not treated with H / R was used as the control group, and the group that was not treated with drugs was used as the model group. The cell survival rate was calculated. Cell survival rate = (experimental group / normal group) × 100%.

[0122] Experimental results: In the anti-ischemic injury model experiment, compared with the hypoxia-reoxygenation model group (H / R group), the cell protection activity of ginsenoside PPD derivatives was better, which could significantly improve the survival rate of cells. The relative activity was better than PPD, among which compounds 7, 10, 12, and 16 showed the best anti-ischemic injury effect. At a concentration of 30 μM, PPD had obvious cytotoxicity to ischemic cells, while compounds 1 to 16 did not show cytotoxicity (see experimental results). Figure 5 ) and showed good anti-ischemic damage effect. The experimental results are shown in the attached Figure 1 and Figure 2 .

[0123] Example 18: Protective activity of PPD derivatives on H9c2 cells in a cell model experiment against ischemia-reperfusion injury

[0124] Experimental methods:

[0125] (1) Cell plating: H9c2 cells in good condition during the logarithmic growth phase were digested and counted, and 8 × 10 3 / well were inoculated in a 96-well plate and cultured in a 37°C, 5% CO2 cell culture incubator;

[0126] (2) Cell administration: After the cells were plated and attached to the wall for 24 hours, they were placed in an anaerobic box for hypoxia. After 6 hours of hypoxia, the cells were taken out and pretreated with 3 μM, 10 μM, and 30 μM PPD derivatives for 1 hour. Three parallel wells were set for each group. After the addition of drugs, the cells were returned to the incubator for reoxygenation at 37°C for 12 hours.

[0127] (3) MTT assay: After reoxygenation, add 20 μL of 5 mg / mL MTT solution and continue to culture in a 37°C incubator for 2 to 4 h. Discard the supernatant. Do not destroy the crystals formed at the bottom of the well during the process. Add 150 μL DMSO to each well and place it on a shaker in the dark for 10 min to fully dissolve the formazan crystals. Finally, use an enzyme reader to detect the absorbance at a wavelength of 570 nm. The H9c2 cell group that was not treated with H / R was used as the control group, and the group that was not treated with drugs was used as the model group. The cell survival rate was calculated. Cell survival rate = (experimental group / normal group) × 100%.

[0128] Experimental results: In the model experiment of anti-ischemia-reperfusion injury, compared with the hypoxia-reoxygenation model group (H / R group), the derivatives of ginsenoside PPD showed better cell protection activity, significantly improved the survival rate of cells, and the relative activity was better than PPD. Among them, compounds 1, 8, 11, 12, 14, and 16 showed the best anti-ischemia-reperfusion injury effect. And at a concentration of 30μM, PPD had obvious cytotoxicity to ischemia-reperfusion cells, while compounds 1 to 16 did not show cytotoxicity and showed better anti-ischemia-reperfusion injury effect. The experimental results are attached. Figure 3 and Figure 4 .

[0129] Example 19: Cytotoxicity assay of PPD derivatives in normal H9c2 cells

[0130] (1) Cell plating: H9c2 cells in good condition during the logarithmic growth phase were digested and counted, and 8 × 10 3 / well were inoculated in a 96-well plate and cultured in a 37°C, 5% CO2 cell culture incubator;

[0131] (2) Cell administration: After the cells were plated and adhered for 24 h, 30 μM PPD derivatives were added and cultured for 1 h. Three parallel wells were set up for each group.

[0132] (3) MTT assay: After the culture is completed, add 20 μL of 5 mg / mL MTT solution and continue to culture in a 37°C incubator for 2 to 4 h. Discard the supernatant. Do not destroy the crystals formed at the bottom of the well during the process. Add 150 μL DMSO to each well and place it on a shaker in the dark for 10 min to fully dissolve the formazan crystals. Finally, use an enzyme marker to detect the absorbance at a wavelength of 570 nm. The H9c2 cell group treated with H / R was used as the control group, and the group without drug treatment was used as the model group. The cell survival rate was calculated. Cell survival rate = (experimental group / normal group) × 100%.

[0133] Experimental results: The MTT method was used to detect the effects of ginsenoside PPD and its derivatives on the survival rate of normal H9c2 cells. The experimental results showed that ginsenoside PPD had a significant effect on the survival rate of normal H9c2 cells, indicating that there was cytotoxicity, while PPD derivatives had no significant effect on the survival rate of normal H9c2 cells (P<0.05), indicating that no cytotoxicity was detected. Therefore, the cytotoxicity of these derivatives was significantly lower than that of PPD. The experimental results are shown in the attached Figure 5 .

[0134] The above description is only a partial embodiment of the present application and an illustration of the technical principles used.

[0135] Those skilled in the art should understand that the scope of the invention involved in this application is not limited to the technical solutions formed by the combination of the above technical features, but should also cover other technical solutions formed by any combination of the above technical features or their equivalent features without departing from the inventive concept. For example, the above features are replaced with but not limited to the technical features with similar functions disclosed in this application.

Claims

1. Use of the PPD derivatives represented by formula (I), (II) or (III) and pharmaceutically acceptable salts thereof in the preparation of anti-hypoxia-reoxygenation injury drugs, anti-ischemic injury drugs or health products; In formula (I), when R3 is hydroxyl and R2 is 12R-hydroxyl, R1 is amino or carbonyl; when R3 is hydroxyl and R2 is 12S-hydroxyl, carbonyl or oxime, R1 is hydroxyl; when R3 is hydroxyl and R2 is carbonyl, R1 is carbonyl; when R3 is tert-butyldimethylsilyloxy, R2 and R1 are both hydroxyl; In formula (III), R4 is 2-methyl-vinyl or 2-hydroxy-isopropyl.

2. The use according to claim 1, characterized in that The compound represented by formula (I) is one of the following compounds 1 to 11; the compound represented by formula (II) is one of the following compounds 12 to 13; the compound represented by formula (III) is one of the following compounds 14 to 16:

3. The use according to claim 2, characterized in that The use of the compound 1, 8, 11, 12, 14 or 16 in the preparation of an anti-hypoxia-reoxygenation injury drug or health product.

4. The use according to claim 2, characterized in that Use of compound 7, 10, 12 or 16 in the preparation of anti-ischemic injury drugs or health products.

5. The use according to claim 1 or 2, characterized in that The anti-hypoxia-reoxygenation injury drug is a drug for treating myocardial ischemia-reperfusion injury; the anti-ischemic injury drug is a drug for treating and preventing myocardial ischemic injury.

6. A pharmaceutical composition for resisting hypoxia-reoxygenation injury or ischemia injury, the pharmaceutical composition comprising a PPD derivative represented by formula (I), (II), (III) and a pharmaceutically acceptable salt thereof.

7. The pharmaceutical composition according to claim 6, characterized in that The pharmaceutical composition comprises one or more of compounds 1 to 16 and pharmaceutically acceptable salts thereof.

8. The pharmaceutical composition according to claim 7, characterized in that The pharmaceutical composition for resisting hypoxia-reoxygenation injury comprises one or more of compounds 1, 8, 11, 12, 14 or 16 and pharmaceutically acceptable salts thereof; The pharmaceutical composition for resisting ischemic injury comprises one or more of compounds 7, 10, 12 or 16 and pharmaceutically acceptable salts thereof.

9. A compound having cardioprotective activity, which is one of compounds 12 to 16:

10. The method for preparing compounds 12 to 16 according to claim 9, characterized in that The method is: (1) adding 24R-Pyxinol or 24S-Pyxinol and pyridinium chlorochromate (PCC) in a molar ratio of 1:0.5 to 2 to carry out an oxidation reaction to obtain compound 4 or compound 6 respectively; (2) Compound 4 or Compound 6 is reacted with m-chloroperbenzoic acid to undergo a Baeyer-Villiger reaction to obtain Compound 12 or Compound 13, respectively; (3) Compound 12 is subjected to an acidic ring-opening reaction with p-toluenesulfonamide to obtain compound 14; The molar ratio of the compound 12 to p-toluenesulfonamide is 1:3-9; (6) Compound 12 or Compound 13 undergoes an alkaline ring-opening reaction with potassium hydroxide to obtain Compound 15 or Compound 16, respectively; The molar ratio of the compound 12 or compound 13 to potassium hydroxide is 1:1.5-5.

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