Polycyclic polyisopentenyl cyclopentanone compound as well as preparation method and application thereof

Abstract

The invention relates to the technical field of biological medicines, in particular to a polycyclic polyisopentenyl cyclopentanone compound as well as a preparation method and application thereof. The polycyclic polyisopentenyl cyclopentanone compound provided by the invention has a structural formula as shown in a formula 1; the preparation method comprises the following steps: by taking dry fruits of hypericum berry as raw materials, carrying out percolation extraction and concentration to obtain an extract, suspending the extract in water to obtain a suspension, and extracting and concentrating the suspension with petroleum ether to obtain a crude extract; the crude extract is taken and subjected to positive and negative phase silica gel column separation, MCI medium-pressure column chromatography, Sephadex LH-20 gel column chromatographic separation, high performance liquid chromatography preparation and CHIRALPAK IG chiral column resolution, and the polycyclic polyisopentenyl cyclopentanone compound is obtained. The protection activity of the compound on AC16 myocardial cell injury induced by cold ischemia is evaluated, and the compound can be used for preparing the polycyclic polyisopentenyl cyclopentanone compound. The compounds can be used as lead compounds for developing medicines for preventing and treating myocardial cold ischemia injury.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, and particularly relates to a polycyclic polyisoprenyl cyclopentanone compound and a preparation method and application thereof. BACKGROUND

[0002] Heart transplantation is still the definitive treatment for end-stage heart failure, but its clinical efficacy is significantly limited due to cold ischemic injury during donor heart preservation. Current myocardial protection strategies, including cryopreservation solutions and adenosine receptor agonists, calcium channel blockers and other drugs, have limited protective effects on ischemia-reperfusion injury (IRI), especially when the cold ischemic time exceeds 4-6 hours. This limitation significantly increases the incidence of primary graft dysfunction (PGD). According to the International Society for Heart and Lung Transplantation registry data, PGD causes more than 30% of early post-transplant mortality. With the continued exacerbation of the imbalance between organ supply and demand, extending the effective preservation time of the heart beyond the current 6-hour threshold has become an urgent clinical challenge. Therefore, there is an urgent need to develop new therapeutic agents that can provide strong protection against cold ischemic injury at the level of myocardial cells to improve long-term survival outcomes after heart transplantation.

[0003] Hypericum ascyron, a perennial semi-deciduous shrub of the genus Hypericum in the family Guttiferae. Multiple species of the genus Hypericum are medicinal plants. Traditional Chinese medicine believes that plants in this genus have the effects of clearing heat and resolving toxicity, astringing and hemostasis, and dampness. They are used to treat hemoptysis, blood, enterovirus, trauma, and rheumatic pain. In recent years, studies have found that plants in this genus have antidepressant, antitumor, antiviral, analgesic, antibacterial, and anti-inflammatory activities. Studies have shown that the chemical components of Hypericum ascyron mainly include cyclopolyprenyl ketones (phenols), including m-trihydroxybenzene and ketones, which are also considered to be the main material basis for the numerous pharmacological activities of Hypericum ascyron. So far, there has been no patent or literature reported on a new polycyclic polyisoprenyl cyclopentanone compound and its activity.

[0004] Therefore, whether a lead compound for the development of a drug for preventing and treating myocardial cold ischemic injury can be extracted from plants has become a technical problem to be solved by those skilled in the art. SUMMARY

[0005] The purpose of the present application is to provide a polycyclic polyisoprenyl cyclopentanone compound and a preparation method and application thereof, which provides a new therapeutic agent for providing strong protection against cold ischemic injury at the level of myocardial cells to improve long-term survival outcomes after heart transplantation.

[0006] In order to achieve the above-mentioned purpose of the application, the present application provides the following technical solutions.

[0007] The present application provides a polycyclic polyisoprenyl cyclopentanone compound, the structural formula of which is shown in formula 1.

[0008]

[0009] The application also provides a preparation method of the above polycyclic polyisoprenyl cyclopentanone compound, comprising the following steps:

[0010] (1) taking dried fruits of Heuchus monggolicus as raw material, obtaining an extract by percolation extraction and concentration, and obtaining a suspension by suspending the extract in water, and then obtaining a crude extract by petroleum ether extraction and concentration;

[0011] (2) performing gradient elution on the crude extract by a silica gel column chromatography to obtain A-H; the mobile phase is petroleum ether-ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 50:1-1:1;

[0012] (3) selecting E for gradient elution by an MCI medium-pressure column chromatography to obtain E1-E6; the mobile phase is methanol-water, and the volume ratio of the methanol to the water is 10:90-100:0;

[0013] (4) selecting E5 for isocratic elution by a Sephadex LH-20 gel column chromatography to obtain E5-1-E5-9; the mobile phase is methanol;

[0014] (5) selecting E5-7 for gradient elution by an ODS reverse-phase medium-pressure column chromatography to obtain E5-7A-E5-7G; the mobile phase is methanol-water, and the volume ratio of the methanol to the water is 70:30-90:10;

[0015] (6) selecting E5-7E for purification by a preparative high-performance liquid chromatography column to obtain a racemic mixture; the mobile phase is acetonitrile-water, and the volume ratio of the acetonitrile to the water is 70:30;

[0016] (7) performing chiral compound resolution on the racemic mixture by a CHIRALPAK IG liquid chromatography to obtain compounds (+)-1 and (–)-1; the mobile phase is acetonitrile-water, and the volume ratio of the acetonitrile to the water is 63:37.

[0017] Alternatively, the solvent for the percolation extraction is ethanol, the volume fraction of the ethanol is 95%, and the percolation extraction time is 120 h; and the percolation extraction flow rate of the ethanol is 0.5 L / h.

[0018] Alternatively, the volume ratio of the petroleum ether to the suspension is 1:1.

[0019] Alternatively, the E is an eluate obtained when the volume ratio of the petroleum ether to the ethyl acetate is 5:1; and the E5 is an eluate obtained when the volume ratio of the methanol to the water is 70:30.

[0020] Optionally, the flow rate of the mobile phase in step (4) is 20 mL / h, the elution time is 18 h, and the eluate is collected in segments every 2 h by isocratic elution of the mobile phase; the E5-7 is the eluate collected in the seventh elution time segment.

[0021] Optionally, the volume ratio of methanol and water in step (5) is increased in a gradient of 70:30 to 90:10 at a rate of 20 / 7 per hour, the total elution time is 7 h, the flow rate is 25 mL / min, and the E5-7E is the eluate collected in the fifth time segment.

[0022] The application also provides the use of the above polycyclic polyisoprenyl cyclopentanone compound in the preparation of a drug for preventing and treating cold ischemia-induced myocardial injury.

[0023] Optionally, the polycyclic polyisoprenyl cyclopentanone compound is used as a raw material to prepare a drug composition or any pharmaceutically acceptable salt for preventing and treating cold ischemia-induced myocardial injury.

[0024] Optionally, the drug composition or any pharmaceutically acceptable salt is in the form of a tablet, a capsule, a granule, an oral solution, a powder, a dripping pill or a pellet; and the drug composition further comprises a pharmaceutically acceptable excipient.

[0025] Compared with the prior art, the application has the following beneficial effects:

[0026] The application separates and purifies ethanol extracts of Heuchera micrantha to obtain two new compounds. The structure of the compounds is determined to be polycyclic polyisoprenyl cyclopentanone by using various spectral analysis methods and other methods. Through evaluation of the protective activity of the compound (±)-1 on cold ischemia-induced AC16 myocardial cell injury, it is found that the compound (±)-1 significantly reduces myocardial cell injury at 24 hours and 36 hours after cold ischemia. In the concentration-effect curve study, (±)-1 shows a dose-dependent protective effect, can reduce the apoptosis rate of cold-stressed myocardial cells, can significantly inhibit the excessive generation of ROS during ischemia, and can effectively inhibit the dephosphorylation of GSK3β Ser9 at 12 hours after ischemia, proving that these compounds can be used as lead compounds for the development of drugs for preventing and treating cold ischemia-induced myocardial injury.

[0027] The application first realizes the extraction of polycyclic polyisoprenyl cyclopentanone compounds with anti-cold ischemia-induced myocardial injury from Heuchera micrantha. The compounds have great development value as new natural heart protectants, and are the first heart protectants from traditional Chinese medicines. The design idea of the compounds also provides a new idea and approach for the development of new heart protectants. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1 is the X-ray single crystal diffraction structure of formula 1.

[0029] Figure 2 Equation 1 shows the experimental and calculated ECD spectra.

[0030] Figure 3 The diagram shows the protective effects of the compounds against cold ischemia injury in cardiomyocytes. In the diagram, A represents the protective effect of compound (±)-1 on AC16 cells 24 hours after cold ischemia injury; B represents the protective effect of compound (±)-1 on AC16 cells 36 hours after cold ischemia injury; C represents the dose-dependent protective effect of compound (+)-1; D represents the dose-dependent protective effect of compound (–)-1; E represents that compound (±)-1 significantly reduces the apoptosis rate of cold-stressed cardiomyocytes; F represents that compound (±)-1 significantly inhibits excessive ROS production during ischemic stimulation; and G represents that treatment with compound (±)-1 effectively inhibits dephosphorylation of GSK3β serine 9 after 12 hours of cold ischemia stimulation. Detailed Implementation

[0031] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0032] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0033] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0034] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be obvious to those skilled in the art. This application specification and embodiments are merely exemplary.

[0035] As used herein, the terms "comprising", "including", "containing", "having" and the like are to be construed to be open-ended terms (i.e., comprising but not limited to).

[0036] The raw materials used in the present application can be obtained commercially or prepared by prior art.

[0037] The present application provides a polycyclic polyisoprenyl cyclopentanone compound, whose structural formula is shown as formula 1:

[0038]

[0039] The present application also provides a preparation method of the polycyclic polyisoprenyl cyclopentanone compound, comprising the following steps:

[0040] (1) taking the dried fruits of Heuchus sanguinea as raw material, obtaining an extract after percolation extraction and concentration, suspending the extract in water to obtain a suspension, and obtaining a crude extract after petroleum ether extraction and concentration;

[0041] (2) gradient elution of the crude extract by a silica gel column chromatography to obtain A-H; the mobile phase is petroleum ether-ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 50:1-1:1;

[0042] (3) gradient elution of E by an MCI medium pressure column chromatography to obtain E1-E6; the mobile phase is methanol-water, and the volume ratio of the methanol to the water is 10:90-100:0;

[0043] (4) isocratic elution of E5 by a Sephadex LH-20 gel column chromatography to obtain E5-1-E5-9; the mobile phase is methanol;

[0044] (5) gradient elution of E5-7 by an ODS reverse phase medium pressure column chromatography to obtain E5-7A-E5-7G; the mobile phase is methanol-water, and the volume ratio of the methanol to the water is 70:30-90:10;

[0045] (6) purification of E5-7E by a preparative high performance liquid chromatography column to obtain a racemic mixture; the mobile phase is acetonitrile-water, and the volume ratio of the acetonitrile to the water is 70:30;

[0046] (7) chiral compound resolution of the racemic mixture by a CHIRALPAK IG liquid chromatography to obtain compound (+)-1 and (–)-1; the mobile phase is acetonitrile-water, and the volume ratio of the acetonitrile to the water is 63:37.

[0047] The present application first takes the dried fruits of Heuchus sanguinea as raw material, obtains an extract after percolation extraction and concentration, suspends the extract in water to obtain a suspension, and obtains a crude extract after petroleum ether extraction and concentration;

[0048] In the embodiment of the present application, the solvent for the percolation extraction is ethanol, the volume fraction of the ethanol is 95%, the time for the percolation extraction is 120 h; the flow rate of the percolation extraction of the ethanol is 0.5 L / h; the volume ratio of the petroleum ether to the suspension is 1:1.

[0049] In the step (2) of the present application, the crude extract is subjected to gradient elution through a silica gel column to obtain eight main fractions A to H; the mobile phase is petroleum ether-ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 50:1 to 1:1;

[0050] In the embodiment of the present application, the gradient volume ratio of the petroleum ether to the ethyl acetate is specifically 50:1, 20:1, 10:1, 8:1, 5:1, 3:1, 2:1 or 1:1.

[0051] In the step (3) of the present application, E is selected and subjected to gradient elution through an MCI medium-pressure column to obtain six sub-fractions E1 to E6; the mobile phase is methanol-water, and the volume ratio of the methanol to the water is 10:90 to 100:0;

[0052] In the embodiment of the present application, the gradient volume ratio of the methanol to the water is specifically 10:90, 20:80, 40:60, 60:40, 70:30 or 100:0; and the E is an eluate obtained when the volume ratio of the petroleum ether to the ethyl acetate in the step (2) is 5:1.

[0053] In the step (4) of the present application, E5 is selected and subjected to isocratic elution through a Sephadex LH-20 gel column to obtain nine main fractions E5-1 to E5-9;

[0054] In the embodiment of the present application, the mobile phase in the step (4) is methanol, the flow rate of the mobile phase is 20 mL / h, the elution time is 18 h, and the eluate is collected in sections every 2 h after isocratic elution of the mobile phase; and the E5 is an eluate obtained when the volume ratio of the methanol to the water in the step (3) is 70:30.

[0055] In the step (5) of the present application, E5-7 is selected and subjected to gradient elution through an ODS reverse-phase medium-pressure column to obtain E5-7A to E5-7G; the mobile phase is methanol-water;

[0056] In the embodiment of the present application, the volume ratio of the methanol to the water in the step (5) is gradiently increased at a rate of 20 / 7 proportions per hour from 70:30 to 90:10, the total elution time is 7 h, and the flow rate is 25 mL / min; and the E5-7 is an eluate collected in the seventh elution time section in the step (4).

[0057] The step (6) of the present application selects E5-7E to be purified by a preparative high performance liquid chromatography column to obtain a racemic mixture; the mobile phase is acetonitrile-water;

[0058] In the embodiment of the present application, the volume ratio of acetonitrile to water in the step (6) is 70:30; the flow rate of the mobile phase is 8 mL / min; and the E5-7E is an eluate obtained by elution in the fifth time period.

[0059] In the step (7) of the present application, the racemic mixture is subjected to chiral compound resolution by a CHIRALPAK IG liquid chromatography column to obtain compounds (+)-1 and (–)-1; the mobile phase is acetonitrile-water, the volume ratio of acetonitrile to water is 63:37; and the flow rate of the mobile phase is 2 mL / min.

[0060] The present application also provides application of the above polycyclic polyisoprenyl cyclopentanone compound in preparation of a drug for preventing and treating myocardial cold ischemic injury.

[0061] In the present application, the polycyclic polyisoprenyl cyclopentanone compound is used as a raw material to prepare a drug composition or any pharmaceutically acceptable salt for preventing and treating myocardial cold ischemic injury.

[0062] In the present application, the dosage form of the drug composition or any pharmaceutically acceptable salt is a tablet, a capsule, a granule, an oral liquid, a powder, a dripping pill or a pellet; and the drug composition further comprises a pharmaceutically acceptable excipient.

[0063] The technical solutions provided by the present application are described in detail below in combination with examples, but they should not be understood as limitations to the protection scope of the present application.

[0064] Example 1

[0065] (1) Take the fruit (10 kg) of dry berry Hypericum as a raw material, perform 95% ethanol infiltration extraction for 120 h at a flow rate of 0.5 L / h, concentrate to obtain an extract, and suspend the extract in water to obtain a suspension, and then extract the suspension with petroleum ether (PE) in an equal volume to the suspension, and concentrate to obtain a crude extract after three times of extraction.

[0066] (2) Perform silica gel column chromatography on the crude extract, use petroleum ether (PE)-ethyl acetate (EtOAc) as a mobile phase, and set the volume ratio of petroleum ether to ethyl acetate to be 50:1, 20:1, 10:1, 8:1, 5:1, 3:1, 2:1 and 1:1 respectively to obtain eight main flow fractions (A-H).

[0067] (3) The fraction E eluted with petroleum ether and ethyl acetate in a volume ratio of 5:1 was separated by MCI column chromatography with methanol-water as the mobile phase, and the volume ratio of methanol to water was 10:90, 20:80, 40:60, 60:40, 70:40 and 100:0, respectively, to obtain six sub-fractions (E1-E6).

[0068] (4) The fraction E5 eluted with methanol and water in a volume ratio of 70:40 was separated by Sephadex LH-20 column chromatography with methanol as the mobile phase, and the flow rate of the mobile phase was 20 mL / h, and the elution time was 18 h. The fraction E5 was separated into nine main fractions (E5-1-E5-9) by collecting every 2 h.

[0069] (5) The fraction E5-7 eluted in the seventh time period was separated by gradient elution with a medium-pressure reverse-phase column, and the mobile phase was methanol-water with a volume ratio of 70:30 to 90:10. The ratio was increased by 20 / 7 every hour, and the flow rate of the mobile phase was 25 mL / min. The elution time was 7 h, and seven sub-fractions (E5-7A-E5-7G) were obtained.

[0070] (6) The fraction E5-7E eluted in the fifth time period was purified by preparative high-performance liquid chromatography with acetonitrile-water as the mobile phase, and the volume ratio of acetonitrile to water was 70:30. The flow rate of the mobile phase was 8 mL / min, and a racemic mixture (6.1 mg, t R 22.4 min) was obtained.

[0071] (7) The racemic mixture was separated by CHIRALPAK IG liquid chromatography to obtain compounds (+)-1 and (-)-1 with acetonitrile-water as the mobile phase, and the volume ratio of acetonitrile to water was 63:37. The flow rate of the mobile phase was 2 mL / min.

[0072] Example 2

[0073] The compound (±)-1 prepared in Example 1 was structurally identified as follows:

[0074] The compound (±)-1 was tested for nuclear magnetic resonance, mass spectrometry, optical rotation, infrared spectroscopy, ultraviolet spectroscopy, circular dichroism, etc. The compound (±)-1 was tested for X-ray single crystal diffraction data (as shown in Figure 1 ), and the compound (±)-1 was calculated for ECD data (as shown in Figure 2 ), so as to determine the structure of the compound (±)-1 as follows:

[0075] Compound 1 ((±)-hypericandrone A): colorless crystals; mp: 181-183 °C; UV (methanol) λmax(log ε): 203 (3.86), 220 (3.58), 242 (3.62) nm; IR (potassium bromide) νmax: 3432, 2969, 2930, 2856, 1709, 1672, 1451, 1383, 1240, 1205, 1180, 1142, 1040, 942 cm –1 ; HRESIMS m / z 519.2342 [M + Na] + (C 26 H 36 O7Na + calculated for m / z 519.2359); 1 H and 13 C NMR data are reported in Table 1; Compound (+)-1 ((+)-hypericandrone A): white powder; +32.4 (methanol, c 0.1); ECD (MeOH) λmax(Δε): 202 (-31.7), 225 (+12.5), 262 (-11.9); Compound (-)-1 ((-)-hypericandrone A): white powder; -32.4 (methanol, c 0.1); ECD (MeOH) λmax(Δε): 202 (+33.7), 225 (-17.6), 262 (+11.0).

[0076] Table 1 H and C NMR data (δ in ppm, J in Hz) of compound 1 1 HNMR and 13 C NMR data (δ in ppm, J in Hz)

[0077]

[0078]

[0079] Example 3

[0080] (1) Determination of the protective effect of compound (±)-1 on AC16 cells cold ischemia injury

[0081] AC16 human cardiomyocytes preserved in 4°C HTK (histidine-tryptophan-ketoglutarate) cardioplegic solution were treated with 20 μΜ of compound (+)-1 and DMSO at the same ratio, respectively, and incubated for 24 hours or 36 hours. Cell viability was assessed using a commercial CCK-8 kit based on the manufacturer's operating procedures, and the absorbance value was measured at 450 nm. The protective effect of the compound on AC16 cells after cold ischemic injury was evaluated according to the survival rate (%) = OD compound / OD control x 100%.

[0082] The results are shown in Table 1. Figure 3 As shown in Table 1, 20 μΜ of compound (+)-1 improved the survival rate of cold-injured AC16 cells, indicating that it protected AC16 cells from cold ischemic injury after 24 hours to some extent. Compound (+)-1 also protected AC16 cells from cold ischemic injury after 36 hours at 20 μΜ.

[0083] (2) Protective effect of compound (+)-1 and (-)-1 on AC16 cells after 24 hours of cold ischemic injury

[0084] AC16 human cardiomyocytes preserved in 4°C HTK (histidine-tryptophan-ketoglutarate) cardioplegic solution were treated with 0 μΜ, 10 μΜ, 20 μΜ, and 50 μΜ of compound (+)-1, respectively, and incubated for 24 hours. Cell viability was assessed using a commercial CCK-8 kit based on the manufacturer's operating procedures, and the absorbance value was measured at 450 nm. The protective effect of the compound on AC16 cells after cold ischemic injury was evaluated according to the survival rate (%) = OD compound / OD control x 100%.

[0085] The results are shown in Table 2. Figure 3 As shown in Table 2, compound (+)-1 showed a dose-dependent protective effect on AC16 cells after 24 hours of cold ischemic injury in the concentration-effect curve study.

[0086] (3) Effect of compound (+)-1 and (-)-1 on the apoptosis rate of cold ischemic myocardial cells

[0087] AC16 cells were placed in HTK solution (4°C preservation) with (sample group) or without (control group) test compound and incubated with 20 μΜ PI solution for 15 minutes. The flow cytometer was used to analyze the apoptosis rate of cold ischemic injury AC16 cells treated with the compound.

[0088] The results are shown in Table 3. Figure 3 As shown in Table 3, compound (+)-1 and (-)-1 can reduce the apoptosis rate of cold ischemic myocardial cells.

[0089] (4) Effect of compound (+)-1 and (-)-1 on ROS production during cold ischemia

[0090] Intracellular reactive oxygen species (ROS) levels were determined using the fluorescent probe DCFH-DA (Sigma). Briefly, AC16 cells stored in 4°C HTK solution with and without test compounds were washed and incubated with DCFH-DA for 25 min, washed again, and then analyzed for the effect of the compounds on ROS production during cold ischemic injury using a flow cytometer.

[0091] Results are shown in Figure 3 As shown in Figure F, compounds (+)-1 and (-)-1 significantly inhibited the excessive production of ROS during cold ischemic injury.

[0092] (5) Effect of compounds (+)-1 and (-)-1 on the expression of specific proteins during cold ischemia

[0093] Protein extraction was performed using RIPA lysis buffer from Biyun Tian Biotechnology Co., Ltd., and proteinase and phosphatase inhibitor cocktail from MCE Co. The protein concentration in the lysate was quantified using the BCA kit from Biyun Tian. The steps of Western blot analysis were as follows: 4-12% gradient SDS-PAGE gels (Thermo) were selected according to the molecular weight of the target protein, and an equal amount of protein was taken from each group for electrophoretic separation. After electrophoresis, the protein was transferred to a PVDF membrane (polyvinylidene fluoride membrane) from Merck, and 5% skim milk powder was prepared in TBS-T solution containing 0.5% Triton X-100 for blocking. The membrane was incubated with the following primary antibodies overnight at 4°C: p-GSK3β, GSK3β, β-actin (Proteintech, 60008-1-Ig). Then, the secondary antibody labeled with horseradish peroxidase (HRP) was incubated at room temperature for 1 hour. The enhanced chemiluminescence (ECL) detection kit from Biyun Tian was used for development, and the band quantitative analysis was performed by ImageJ software (National Institutes of Health, USA).

[0094] Results are shown in Figure 3 As shown in Figure G, the treatment of compounds (+)-1 and (-)-1 effectively inhibited the dephosphorylation of GSK3β Ser9 site induced 12 hours after cold ischemic injury.

[0095] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A polycyclic polyisoprenylcyclopentanone compound, characterized by, The structural formula is shown as formula 1:

2. A method for producing the polycyclic polyisoprenylcyclopentanone compound according to claim 1, characterized by, It comprises the following steps: (1) taking the dried fruit of Heuchus japonicus as raw material, extracting by percolation, concentrating to obtain extract, mixing the extract in water to obtain suspension, extracting by petroleum ether, concentrating to obtain crude extract; (2) gradient elution of the crude extract by silica gel column chromatography to obtain A-H; the mobile phase is petroleum ether-ethyl acetate, and the volume ratio of the petroleum ether and ethyl acetate is 50:1-1:1; (3) selecting E for gradient elution by MCI medium pressure column chromatography to obtain E1-E6; the mobile phase is methanol-water, and the volume ratio of the methanol and water is 10:90-100:0; (4) selecting E5 for isocratic elution by Sephadex LH-20 gel column chromatography to obtain E5-1-E5-9; the mobile phase is methanol; (5) selecting E5-7 for gradient elution by ODS reverse phase medium pressure column chromatography to obtain E5-7A-E5-7G; the mobile phase is methanol-water, and the volume ratio of the methanol and water is 70:30-90:10; (6) selecting E5-7E for purification by preparative high performance liquid chromatography to obtain a racemic mixture; the mobile phase is acetonitrile-water, and the volume ratio of the acetonitrile and water is 70:30; (7) chiral compound resolution of the racemic mixture by CHIRALPAK IG liquid chromatography to obtain compounds (+)-1 and (-)-1, and the mobile phase is acetonitrile-water, and the volume ratio of the acetonitrile and water is 63:

37.

3. The method of claim 2, wherein, The solvent for the percolation extraction is ethanol, the volume fraction of the ethanol is 95%, and the percolation extraction time is 120 h; the percolation extraction flow rate of the ethanol is 0.5 L / h.

4. The preparation method according to claim 2, characterized in that, The volume ratio of the petroleum ether and the suspension is 1:

1.

5. The preparation method according to claim 2, characterized in that, The E is an eluate obtained when the volume ratio of the petroleum ether and ethyl acetate is 5:1; and the E5 is an eluate obtained when the volume ratio of the methanol and water is 70:

30.

6. The preparation method according to claim 2, characterized in that, In step (4), the flow rate of the mobile phase is 20 mL / h, the elution time is 18 h, and each 2 h is collected by section after isocratic elution of the mobile phase; and the E5-7 is an eluate collected in the seventh elution time period.

7. The preparation method according to claim 2, characterized in that, In step (5), the volume ratio of the methanol and water is gradiently increased at a rate of 20 / 7 per hour, the total elution time is 7 h, the flow rate is 25 mL / min, and the E5-7E is an eluate collected in the fifth time period.

8. The polycyclic polyisoprenyl cyclopentanone compound of claim 1 is used for preparing a drug for preventing and treating cold ischemia injury of myocardium.

9. Use according to claim 8, characterized in that, The polycyclic polyisoprenyl cyclopentanone compound is used as raw material to prepare a drug composition or any pharmaceutically acceptable salt for preventing and treating cold ischemia injury of myocardium.

10. Use according to claim 9, characterized in that, The dosage form of the drug composition or any pharmaceutically acceptable salt is tablet, capsule, granule, oral liquid, powder, dripping pill or pellet; and the drug composition further comprises a pharmaceutically acceptable excipient.

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