Open-loop resin glycoside derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicines, and discloses a ring-opened resin glycoside derivative as well as a preparation method and application thereof. According to the method disclosed by the invention, the ring-opening resin glycoside derivative with a novel chemical structure is obtained by taking a sedge tricolor seed as a separation target and comprehensively applying a plurality of separation technologies and means. In-vitro activity experiments prove that the ring-opened resin glycoside derivative provided by the invention has remarkable anti-neuroinflammation activity, can be used as an active ingredient for treating neuroinflammation related diseases, and has wide application.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and relates to a ring-opening resin glycoside derivative, a preparation method and application thereof. Background Art

[0002] The incidence of neurological diseases such as neurodegenerative diseases, traumatic brain injury and stroke has increased year by year, bringing a heavy burden to society and families. Studies have shown that neuroinflammation plays an important role in the pathogenesis of a variety of neurological diseases. Excessive activation of microglia is considered a typical sign of neuroinflammation. Excessive activation of microglia can produce and secrete a large number of neurotoxic factors, including chemokines and proinflammatory factors, leading to damage to the nervous system. Therefore, inhibiting neuroinflammation mediated by abnormal activation of microglia is an important strategy for developing drugs to treat neurological diseases.

[0003] Resin glycosides are characteristic components of Convolvulaceae plants and are divided into macrocyclic resin glycosides and open-ring resin glycoside compounds. Open-ring resin glycosides are composed of three parts: oligosaccharide chains, long-chain fatty acid aglycones, and modified fatty acyl groups, and the terminal carboxyl group of the aglycone usually exists in a free state or in the form of methyl ester. In recent years, many open-ring resin glycoside derivatives have been reported to have significant biological activities, such as anti-epileptic, sedative and vasodilatory activities, providing a variety of template molecules for drug development. Therefore, these novel structures are increasingly attracting the attention of drug researchers.

[0004] Therefore, it is necessary to study the active ingredients of some plants and develop active resin glycoside compounds with novel structures. Summary of the invention

[0005] In view of this, the object of the present invention is to provide a ring-opening resin glycoside derivative and a preparation method and application thereof.

[0006] To solve the above problems, the present invention provides the following technical solutions:

[0007] In the first aspect of the present invention, the present invention provides a ring-opening resin glycoside derivative, wherein the ring-opening resin glycoside derivative has a chemical structure shown in any of the following structural formulas:

[0008]

[0009] In the second aspect of the present invention, the present invention provides a method for preparing the above-mentioned open-ring resin glycoside derivatives, specifically: isolating and obtaining the resin glycoside derivatives from the seeds of Convolvulus tricolor L.

[0010] In some embodiments of the present invention, the preparation method is specifically to select the seeds of Convolvulus tricolor, perform reflux extraction with 95%, and then obtain the resin glycoside derivatives of Formula I to Formula III through various separation techniques and means, and have significant anti-neuroinflammatory activity, can be used as an active ingredient for treating nervous system diseases, and has a wide range of uses.

[0011] In some embodiments of the present invention, the above preparation method comprises the following steps:

[0012] The invention uses seeds of Convolvulus tricolor to carry out reflux extraction with ethanol aqueous solution with a volume fraction of 95%, and prepares the compound of the invention through various separation techniques and means. The seeds of Convolvulus tricolor are purchased in Beijing.

[0013] 1) Extraction

[0014] The seeds of Convolvulus tricolor are subjected to heat reflux extraction for several times with an ethanol aqueous solution, and the extracts are combined and concentrated under reduced pressure to obtain an extract.

[0015] 2) Silica gel chromatography separation

[0016] The extract was subjected to silica gel column chromatography, and gradient elution was performed using a dichloromethane / methanol mixed solvent (1:0, 10:1, 5:1, 2:1, 1:1, 0:1, v / v) to obtain components 1 to 8 in sequence;

[0017] 3) Reverse phase ODS column chromatography

[0018] Component 2 was purified by reverse phase ODS column chromatography using methanol-water solution (50:50, 70:30, 80:20, 85:15, 90:10, 95:5, 100:0, v / v) as eluent to obtain components 2.1 to 2.3.

[0019] 4) Amino silica gel column chromatography

[0020] Component 2.2 was purified by amino silica gel column chromatography using dichloromethane-methanol (5:1, 2:1, 1:1, 0:1, v / v) as eluent to obtain components 2.2.1 to 2.2.4.

[0021] 5) Reverse phase ODS column chromatography

[0022] Component 2.2.2 was purified by reverse phase ODS column chromatography using methanol-water solution (60:40, 70:30, 80:20, 85:15, 90:10, 95:5, 100:0, v / v) as eluent to obtain components 2.2.2.1 to 2.2.2.8.

[0023] 6) Reverse-phase HPLC purification

[0024] Component 2.2.2.5 was purified by reverse phase high performance liquid chromatography. The preparation conditions were: preparative chromatography column YMC RP-C18 (10×200 mm), methanol-water (79:21, V / V), flow rate 2.0 mL / min, detector Shimadzu RID 10A, and compounds of Formula I to Formula III were prepared.

[0025] In the third aspect of the present invention, the present invention provides a use of the above-mentioned resin glycoside derivatives or pharmaceutically acceptable salts thereof as active ingredients in the preparation of anti-neuroinflammatory drugs.

[0026] In the fourth aspect of the present invention, the present invention provides a pharmaceutical composition, which comprises the above-mentioned ring-opening resin glycoside derivative or a pharmaceutically acceptable salt thereof.

[0027] Compared with the prior art, the present invention selects the seeds of Convolvulus tricolor as the object, performs reflux extraction with ethanol, and then obtains the above-mentioned resin glycoside derivatives of formula I to formula III through various separation techniques and means. Compounds of formula I to formula III are all open-ring resin glycoside acid derivatives, among which formula I and formula II are rare resin glycoside derivatives containing eight sugar residues. Such novel structural resin glycoside compounds have significant anti-neuroinflammatory activity, can be used as an active ingredient for anti-neuroinflammatory, and have a wide range of uses. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a hydrogen spectrum data diagram of compound I obtained in Example 1 of the present invention;

[0029] Figure 2 This is a carbon spectrum data diagram of compound I obtained in Example 1 of the present invention;

[0030] Figure 3 This is a hydrogen spectrum data diagram of compound II obtained in Example 1 of the present invention;

[0031] Figure 4 This is the carbon spectrum data diagram of compound II obtained in Example 1 of the present invention;

[0032] Figure 5 This is a hydrogen spectrum data diagram of compound III obtained in Example 1 of the present invention;

[0033] Figure 6 This is the carbon spectrum data diagram of compound III obtained in Example 1 of the present invention. DETAILED DESCRIPTION

[0034] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only intended to illustrate the present invention and are not intended to limit the scope of the present invention. The operating methods in the following examples where no specific conditions are indicated are usually carried out under conventional conditions or under conditions recommended by the manufacturer.

[0035] Example 1: Preparation of compounds

[0036] The invention uses seeds of Convolvulus tricolor to carry out reflux extraction with ethanol aqueous solution with a volume fraction of 95%, and prepares the compound of the invention through various separation techniques and means. The seeds of Convolvulus tricolor are purchased in Beijing.

[0037] 1) Extraction

[0038] Tricolor bindweed seeds (5 kg) were crushed, and 5 times the volume of 95% ethanol aqueous solution was added, and hot reflux extraction was performed 3 times, each time for 3 hours. After the extracts were combined, they were concentrated under reduced pressure to obtain an extract (500 g).

[0039] 2) Silica gel chromatography separation

[0040] The extract was subjected to silica gel column chromatography and gradient elution was performed using a dichloromethane / methanol mixed solvent (1:0, 10:1, 5:1, 2:1, 1:1, 0:1, v / v) to obtain components 1 to 8, respectively.

[0041] 3) Reverse phase ODS column chromatography

[0042] Component 2 was purified by reverse phase ODS column chromatography using methanol-water solution (50:50, 70:30, 80:20, 85:15, 90:10, 95:5, 100:0, v / v) as eluent to obtain components 2.1 to 2.3.

[0043] 4) Amino silica gel column chromatography

[0044] Component 2.2 was purified by amino silica gel column chromatography using dichloromethane-methanol (5:1, 2:1, 1:1, 0:1, v / v) as eluent to obtain components 2.2.1 to 2.2.4.

[0045] 5) Reverse phase ODS column chromatography

[0046] Component 2.2.2 was purified by reverse phase ODS column chromatography using methanol-water solution (60:40, 70:30, 80:20, 85:15, 90:10, 95:5, 100:0, v / v) as eluent to obtain components 2.2.2.1 to 2.2.2.8.

[0047] 6) Reverse-phase HPLC purification

[0048] Component 2.2.2.5 was purified by reverse phase high performance liquid chromatography. The preparation conditions were preparative chromatography column YMC RP-C18 (10×200 mm), methanol-water (79:21, V / V), flow rate 2.0 mL / min, detector Shimadzu RID 10A, and compounds I to III were prepared.

[0049] The hydrogen spectrum data and carbon spectrum data of compounds I to III are as follows: Figure 1-6 As shown. Figure 1-Figure 6 The mass spectra and spectroscopy data of compounds I-III are shown below.

[0050] Compound I: controlin V; main absorption peak of infrared spectrum (KBr) ν max :3420, 2932, 1734, 1650, 1077cm -1 ; [α] 25 D -12.9(c 0.11,MeOH); UV(MeOH)λmax(logε)221(3.42)nm; HRESIMS m / z931.4070[M+2Na] 2+ (C 80 H 136 Na2O 45 , calculated value 931.4069); the H NMR and C NMR data are shown in Table 1.

[0051] Compound II: controlin VI; main absorption peak of infrared spectrum (KBr) ν max :3421,2932,1733,1650,1075 -1 ; [α] 25 D -30.2(c 0.09,MeOH); UV(MeOH)λmax(logε)220(3.56)nm; HRESIMS m / z981.4335[M+2Na] 2+ (C 85 H 144 Na2O 47 , calculated value 981.4331); the H NMR and C NMR data are shown in Table 1.

[0052] Compound III: controlin VIII; main absorption peak of infrared spectrum (KBr) ν max :3444,2930,1734,1650,1075 -1 ; [α] 25 D-21.8(c 0.12,MeOH); UV(MeOH)λmax(logε)218(3.32)nm; HRESIMS m / z850.3804[M+2Na] 2+ (C 74 H 126 Na2O 40 , calculated value 850.3805); the H NMR and C NMR data are shown in Table 1.

[0053] Table 1. H NMR and C NMR data of compounds of formula I to formula III

[0054]

[0055]

[0056]

[0057] a Overlapped signals are reported without designating multiplicity; b Abbreviations: Glc=glucopyranosyl, Rha=rhamnopyranosyl, Fuc=fucopyranosyl, Qui=quinovopyranosyl, Ag=12-hydroxyhexadecanoyl aglycone, Nla=3-hydroxy-2-methylbutanoyl, Tga=2-methyl-2-butenoyl.

[0058] The above results show that the structural formulas of the obtained compounds I-III are respectively chemical formulas I-III.

[0059] Example 2: Anti-neuroinflammatory activity of compounds I to III

[0060] 1) Experimental Materials

[0061] Instruments and reagents: CO2 incubator (Thermo Fisher Scientific, USA); DMEM / RPMI 1640 culture medium (Gibco, Thermo Fisher Scientific, USA); DMSO and trypsin (Sigma, USA); CCK-8 and nitric oxide (NO) kits (Biyuntian Biotechnology Research Institute); fetal bovine serum (Hangzhou Sijiqing Biomaterials Research Institute); other commonly used inorganic salts and other reagents were of analytical grade.

[0062] Test cell line: mouse microglial BV2 cell line.

[0063] Test samples: resin glycoside derivatives of formula I to formula III obtained from seeds of Convolvulus tricolor; L-arginine was selected as a positive control drug, and each compound was dissolved in DMSO and then diluted.

[0064] 2) Experimental methods

[0065] BV2 cells in the logarithmic growth phase were seeded in a 96-well culture plate at a cell density of 1.5 × 10 4 After 24 hours of culture, lipopolysaccharide (LPS) was added to a final concentration of 100 ng / mL and culture was continued for 3 hours. Subsequently, samples or positive drugs of different concentrations were added and culture was continued for 24 hours. According to the instructions of the kit, the level of NO in the culture medium of each group was determined by the Griess method, and the half inhibitory concentration (IC50) of each test sample to inhibit NO release was calculated. 50 ); CCK-8 method was used to determine the survival rate of cells in each group.

[0066] 3) Experimental results

[0067] According to the Griess method and CCK-8 test results, the effects of the compounds of Formula I to Formula III of the present invention on the NO release of BV-2 cells induced by LPS were calculated. The results are shown in Table 2.

[0068] Table 2. Results of the test samples inhibiting LPS-induced NO release in BV-2 cells

[0069]

[0070] a IC 50 Values ​​are expressed as mean ± SD; b The cell viability was tested at a concentration of 50 μM

[0071] The results showed that the IC values ​​of compounds I to III of the present invention on NO inhibition induced by LPS in BV-2 microglia were 50 The concentration of β-catenin was 6.73-19.10 μM, so it can be used as an active ingredient for treating neuroinflammation-related diseases.

[0072] Those skilled in the art should understand that the present invention is not limited to the above embodiments, and the above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, and these changes and improvements fall within the scope of the present invention to be protected. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. A ring-opening resin glycoside derivative, characterized in that: The ring-opening resin glycoside derivative has a chemical structure shown in any of the following structural formulas:

2. A method for preparing the ring-opening resin glycoside derivative according to claim 1, characterized in that: The preparation method comprises the following steps: separating and obtaining the resin glycoside derivatives from seeds of Convolvulus tricolor.

3. Use of the open-ring resin glycoside derivative according to claim 1 as an active ingredient in the preparation of anti-neuroinflammatory drugs.

4. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the ring-opening resin glycoside derivative according to claim 1 or a pharmaceutically acceptable salt thereof.

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