Preparation and antimalarial applications of a 7,7'-epoxy lignan

Abstract

The present application relates to a preparation of a 7,7'-epoxy lignan compound and its application in preparing anti-malaria products, and belongs to the field of traditional Chinese medicine and natural medicine pharmacy. The anti-malaria medicine with the compound as an active ingredient. The present application expands the medicinal value of 7,7'-epoxy lignan compounds in Rhodiola henryi.

Description

Technical Field

[0001] The present invention belongs to the field of traditional Chinese medicine and natural medicine pharmaceuticals, and specifically relates to the preparation of a 7,7'-epoxylignan compound, a drug combination with the compound as an active ingredient, and the application of the compound in an antimalarial agent. Background Art

[0002] Rhodiola rosea ( Rhodiola fastigiate (Hook. f. et Thoms.) SH Fu), also known as broad-leaved Rhodiola rosea, vertical branched Sedum, Dali Sedum, belongs to the genus Rhodiola ( Rhodiola ) plant, a perennial herb, the whole herb is used for medicinal purposes; it is produced in Tibet, Yunnan, Sichuan and other places in my country, grows in high mountain gravel at an altitude of 2500-5400 meters, and is also distributed in Kashmir, Nepal, Sikkim and Bhutan. The "Sichuan Tibetan Medicinal Materials Standard (2020 Edition)" records that Rhodiola rosea tastes astringent, sweet, bitter, and cool in nature; it has the function of clearing heat and benefiting the lungs, and is used to treat lung heat caused by colds, tracheitis, and bad breath. It is recorded in the "Yunnan Traditional Chinese Medicine Resources Catalog" that it has the effect of treating trauma. Literature reports that Rhodiola rosea can also treat blood stasis, swelling and trauma (Application of microscopy in authentication of traditional Tibetan medicinal plants of five Rhodiola (Crassulaceae) alpine species by comparative anatomyand micromorphology. Microscopy Research and Technique , 2008, 71(6): 448-458). In recent years, there has been little research on the chemical components of Rhodiola rosea, and studies on its pharmacological activities are also rare. Modern pharmaceutical research shows that Rhodiola rosea contains a variety of chemical components, including organic phenolic acids, glycosides, flavonoids, volatile oils, steroids, and trace elements. Some of these chemical components have the effects of delaying aging, scavenging free radicals, preventing and treating senile diseases, and providing antioxidant, anti-fatigue, antibacterial, antiviral, and immunomodulatory effects (Research Overview of Rhodiola rosea. Tibet Sci-Tech , 2009, 26(4): 73-78). However, there is no report on the antimalarial activity of Rhodiola rosea to date. Summary of the Invention

[0003] The present invention aims to provide a 7,7'-epoxylignan compound and a preparation method thereof, a pharmaceutical composition containing the 7,7'-epoxylignan compound as an active ingredient, and use of the 7,7'-epoxylignan compound in the preparation of an antimalarial agent.

[0004] The above-mentioned object of the present invention is achieved through the following technical solutions:

[0005] 7,7'-epoxy lignan compounds shown in the following structure,

[0006]

[0007] The preparation method of the compound of the present application is to extract the roots or rhizomes or whole plants of the genus Rhodiola with organic solvents chloroform or ethyl acetate or acetone or methanol or ethanol or water directly by cold soaking or hot refluxing, or first extracting by cold soaking or refluxing with the above-mentioned organic solvents or water and then extracting with ethyl acetate to obtain total extract, and the total extract is repeatedly column chromatographed to obtain the compound of the present application. Rhodiola The preparation method of the compound of the present application is more specifically to extract the roots or rhizomes or whole plants of the genus Rhodiola with:

[0008] The preparation method of the compound of the present application is more specifically to extract the roots or rhizomes or whole plants of the genus Rhodiola with:

[0009] A: total extract obtained by cold soaking or hot refluxing extraction of the roots or rhizomes or whole plants of the genus Rhodiola with acetone or methanol or ethanol or water, and ethyl acetate extract obtained by extraction with ethyl acetate, which can be repeatedly column chromatographed to obtain the compound of the present application.

[0010] B: total extract obtained by directly cold soaking or hot refluxing extraction of the roots or rhizomes or whole plants of the genus Rhodiola with organic solvents (such as methanol, ethanol, acetone, ethyl acetate, chloroform), which can be repeatedly column chromatographed to obtain the compound of the present application.

[0011] More specifically, the preparation method of the compound of the present application is to shade dry the roots or rhizomes or whole plants of the genus Rhodiola, crush them to 20-30 mesh, extract them with 95% ethanol at room temperature for 3 times, each time for 24 h, combine the extract, concentrate the extract under reduced pressure to obtain extract, and then extract with ethyl acetate as the extraction solvent by solid phase extraction or liquid-liquid extraction method to obtain ethyl acetate extract, dissolve the ethyl acetate extraction part with an appropriate amount of chloroform / acetone, then mix the sample with 80-100 mesh silica gel, and then perform column chromatography with 200-300 mesh silica gel, and gradient elution is performed with 1:0-0:1 chloroform / acetone or 1:0-0:1 chloroform / methanol to obtain 8 main parts, and the 9:1 chloroform / acetone and 8:2 chloroform / acetone parts are repeatedly column chromatographed with silica gel, MCI column chromatographed and Sephadex LH-20 column chromatographed to obtain the compound of the present application.

[0012] The antimalarial agent contains the compound of the present application and conventional adjuvants.

[0013] The pharmaceutical composition contains a therapeutically effective amount of the compound of the present application and a pharmaceutically acceptable carrier.

[0014] The compound of the present application is used for preparing an antimalarial agent.

[0015] The pharmaceutical composition for anti-malaria contains the compound of the present application and a pharmaceutically acceptable carrier. ​

[0016] The pharmaceutically acceptable carrier in the pharmaceutical composition of the present application means a pharmaceutical carrier which is conventionally used in the pharmaceutical field. The compound of the present application can be administered to a patient in need of such treatment in the form of a composition by oral, nasal inhalation, rectal or parenteral administration. For oral administration, it can be formulated into a conventional solid preparation such as a tablet, powder, granule, capsule, etc., a liquid preparation such as an oil suspension, syrup, elixir, etc., and for parenteral administration, it can be formulated into a solution for injection, etc. The preferred form is a tablet, capsule and injection.

[0017] The various dosage forms of the pharmaceutical composition of the present application can be prepared according to the conventional production method in the pharmaceutical field. For example, the active ingredient is mixed with one or more carriers, and then it is formulated into a desired dosage form.

[0018] The pharmaceutical composition of the present application preferably contains 0.1-99.5% by weight of the active ingredient, and most preferably contains 0.5-95% by weight of the active ingredient.

[0019] The amount of the compound of the present application to be administered can vary depending on the route of administration, the age, body weight, type and severity of the disease to be treated, etc., and the daily dose can be 0.01-10 mg / kg of body weight, preferably 0.1-5 mg / kg of body weight. It can be administered once or more.

[0020] The compound of the present application shows good anti-malaria activity.

[0021] The resulting compound of the present application is screened for anti-malaria activity, and the compound shows good anti-malaria activity. In the application of anti-malaria activity, the compound of the present application is administered to a substrate or a group in an amount ranging from 1 to 1000 µM, preferably from 10 to 200 µM, optionally in combination with a carrier and / or medium. DETAILED DESCRIPTION

[0022] The essential content of the present application is further illustrated by the following examples of the present application, and a person skilled in the art can more fully understand the present application, but the present application is not limited in any way by the present application. Example 1:

[0023] Extraction, separation and purification of the compound of the present application:

[0024] The whole plant of Rhodiola henryi Hemsl. (9.36 kg) was dried, crushed to 20-30 mesh, extracted with 95% ethanol for 3 times at room temperature, 50 L for each time, 24 h, the extract was combined, concentrated under reduced pressure to get extract, then extracted with ethyl acetate to get ethyl acetate extract (326 g), the ethyl acetate extract was dissolved with chloroform / acetone, then mixed with silica gel of 80-100 mesh, then column chromatography was carried out with silica gel of 200-300 mesh, 8 main parts were obtained by gradient elution with chloroform / acetone (1:0-0:1), the 9:1 chloroform / acetone part and the 8:2 chloroform / acetone part were subjected to silica gel column chromatography, MCI column chromatography and Sephadex LH-20 column chromatography to obtain the compound of the present application. Example 2:

[0025] Physical and spectral data of the compound of the present application:

[0026] Light yellow solid. [α] 20 D +117.8 ( c = 0.1, MeOH); UV (MeOH) λ max (log ε): 201 (4.23), 228 (4.09), 277 (3.25) nm; CD (MeOH) λ max (Δε): 201 (+8.2), 232 (+9.2), 279 (-0.1) nm. HR-ESI-MS: m / z 297.1500 [M-H] - (calcd for C 19 H 21 O3297.1496). 1 H-NMR (400 MHz, CDCl3) δ : 7.24 (2H, d, J = 8.3 Hz, H-2, 6), 7.22(2H, d, J = 8.3 Hz, H-2', 6'), 6.87 (2H, d, J = 8.3Hz, H-3, 5), 6.76 (2H, d, J = 8.5 Hz, H-3', 5'), 5.46 (1H, d, J = 4.2 Hz, H-7), 4.64 (1H, d, J= 9.0Hz, H-7'), 2.43 (2H, m, H-8, 8'), 0.97 (3H, d, J = 6.5 Hz, 8'-Me), 0.60 (3H,d, J = 7.0 Hz, 8-Me), 3.80 (3H, s, 4-OMe); 13 C-NMR (100 MHz, CDCl3) δ : 132.7(s, C-1), 134.9 (s, C-1'), 127.2 (d, C-2, 6), 127.7 (d, C-2', 6'), 113.4 (d,C-3, 5), 115.3 (d, C-3', 5'), 158.5 (s, C-4), 155.2 (s, C-4'), 84.8 (d, C-7),85.6 (d, C-7'), 43.4 (d, C-8), 47.6 (d, C-8'), 9.6 (q, 8-Me), 11.8 (q, 8'-Me), 55.4 (q, 4-OMe)。 Example 3:

[0027] Detection of anti-malaria activity of the compounds of the present application:

[0028] The anti-malaria activity of the compounds of the present application was evaluated in vitro against P. falciparum (3D7) using SYBR Green I method. The parasites were cultured in RPMI 1640 medium with 2% hematocrit of human red blood cells at 37°C and 5% CO2. The parasites were synchronized to ring stage with 5% sorbitol and the final test system contained 1% infected red blood cells with 2% hematocrit. After 72 h of co-culturing the ring stage parasites with the test compounds in 96-well plates, the fluorescent dye SYBR Green I was added. Finally, the fluorescence value (excitation: 485 nm, emission: 535 nm) was measured by a microplate reader to detect the growth of the parasites. The inhibition rate was calculated according to the following formula: Plasmodium falciparum Parasite inhibition rate (%) = (fluorescence value of negative control group - fluorescence value of test sample group) / (fluorescence value of negative control group - fluorescence value of blank group) x 100%.

[0029] The activity data are shown in Table 1.

[0030] The activity data are shown in Table 1.

[0031]

[0032] Example 4:

[0033] Tablets: 10 mg of the compound obtained in Examples 1 and 2, 180 mg of lactose, 55 mg of starch, and 5 mg of magnesium stearate;

[0034] Preparation method: The compound of the present invention, lactose and starch are mixed and uniformly moistened with propylene glycol. The moistened mixture is sieved and dried, sieved again, magnesium stearate is added, and the mixture is compressed into tablets. Each tablet weighs 250 mg and contains 10 mg of the compound of the present invention.

[0035] Example 5:

[0036] Ampoule: 2 mg of the compound obtained in Examples 1 and 2;

[0037] Preparation method: The compounds obtained in Examples 1 and 2 were dissolved in 3 mL of propylene glycol. The resulting solution was filtered and filled into an ampoule under sterile conditions.

[0038] Example 6:

[0039] Capsules: 10 mg of the compound obtained in Examples 1 and 2, 187 mg of lactose, and 3 mg of magnesium stearate;

[0040] Preparation method: The compound of the present invention is mixed with an adjuvant, sieved, and evenly mixed. The resulting mixture is filled into hard gelatin capsules. Each capsule weighs 200 mg and contains 10 mg of active ingredient.

Claims

1. The compound described by the following structural formula, 2. The method for preparing the compound according to claim 1, characterized in that The root, rhizome or whole plant of the genus Rhodiola is dried in the shade and crushed into 20-30 mesh size. The extract is extracted three times with 95% ethanol at room temperature for 24 hours each time. The extracts are combined, and the extract is concentrated under reduced pressure to obtain an extract. The extract is then extracted using ethyl acetate as an extraction solvent by solid phase extraction or liquid-liquid extraction to obtain an ethyl acetate extract. The ethyl acetate extracted portion is dissolved with an appropriate amount of chloroform / acetone and then mixed with 80-100 mesh silica gel. The sample is then subjected to column chromatography using 200-300 mesh silica gel for coarse fractionation. Gradient elution is performed with 1:0-0:1 chloroform / acetone or 1:0-0:1 chloroform / methanol to obtain eight major fractions. The 9:1 chloroform / acetone and 8:2 chloroform / acetone fractions are repeatedly subjected to silica gel column chromatography, MCI column chromatography and Sephadex LH-20 column chromatography to obtain the compound of the present invention.

3. A pharmaceutical composition comprising a therapeutically effective amount of the compound according to claim 1 and a pharmaceutically acceptable carrier.

4. An antimalarial agent comprising the compound according to claim 1 and conventional adjuvants.

5. Use of the compound according to claim 1 in the preparation of antimalarial agents.

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