Monoterpene coumarin compounds in Ailanthus altissima leaves and their preparation method and insecticide application

Abstract

The invention relates to monoterpene coumarin compounds in Ailanthus altissima leaves and a preparation method and application thereof, belonging to the field of medical technology, and specifically to 10 monoterpene coumarin compounds extracted and separated from Ailanthus altissima leaves of Ailanthus altissima, a plant of the genus Ailanthus in the family Simaroubaceae. Biological activity experimental results show that these compounds have good insecticidal activity. In addition, the preparation method of the invention is simple and easy to implement, has good reproducibility, and has high product purity.

Description

Technical Field

[0001] The invention belongs to the technical field of medicine, and in particular relates to new monoterpene coumarin compounds prepared from leaves of Ailanthus altissima and applications of the compounds in preparing insecticides. Background Art

[0002] Ailanthus altissima (Mill.) Swingle is a plant of the genus Ailanthus (Ailanthus Desf.) of the family Simaroubaceae. It is widely distributed in Shaanxi, Gansu, Sichuan, Yunnan and other places in my country. Its root bark has the effects of clearing away heat and dampness, astringing and strengthening the intestines. It is mainly used to treat chronic dysentery, intestinal bleeding, leucorrhea, nocturnal emission and spermatorrhea. Modern chemical and pharmacological studies have shown that Ailanthus contains bitter substances, steroids, coumarins, alkaloids, lignin and other components, and has multiple activities such as anti-tumor, anti-viral, anti-inflammatory and antioxidant.

[0003] After searching, there are currently few reports or related patents on the monoterpene coumarins in Ailanthus altissima leaves. Summary of the invention

[0004] The purpose of the present invention is to provide a monoterpene coumarin compound prepared from Ailanthus altissima leaves, a preparation method thereof and an application thereof in insecticide.

[0005] The structures of the 10 monoterpene coumarin compounds isolated from Ailanthus altissima leaves of the plant Ailanthus genus of the Simaroubaceae family are shown below:

[0006]

[0007] The preparation technology scheme of the present invention comprises the following steps:

[0008] The dried Ailanthus altissima leaves were extracted with ethanol, and the combined extracts were concentrated to obtain an extract, which was extracted with dichloromethane and n-butanol, and the obtained components were subjected to silica gel column chromatography, and isocratic gradient elution was performed with a dichloromethane-methanol system of 100:1-1:1, and a total of 6 components Fr.AF were collected;

[0009] Using HP20, ODS column chromatography, the component Fr.D was gradient eluted with an ethanol-water system of 20:80-90:10 to obtain five components Fr.D1-D5;

[0010] Fr.D2 was separated by silica gel column chromatography with a petroleum ether-ethyl acetate system of 100:1-10:1 and by preparative reverse phase high performance liquid chromatography using an acetonitrile-water mobile phase to obtain compound 1-10.

[0011] Preferably, in the method for preparing the monoterpene coumarin compounds in Ailanthus altissima leaves, the Ailanthus altissima used is the dried leaves of Ailanthus altissima of the genus Simarium in the family Simarthaceae.

[0012] Preferably, the method for preparing the monoterpene coumarin compounds in Ailanthus altissima leaves comprises taking dried Ailanthus altissima leaves and extracting them with 70-80% industrial ethanol for 3 times under reflux, each time for 2-3 hours.

[0013] Preferably, the method for preparing monoterpene coumarin compounds in Ailanthus altissima leaves separates Fr.D2 using an acetonitrile-water mobile phase of 70:30-30:70.

[0014] The obtained compounds were systematically identified and the results are as follows:

[0015] The structures of compounds 1-10 were identified by high-resolution mass spectrometry, one-dimensional NMR, two-dimensional NMR, computed NMR, computed ECD and X-ray single crystal diffraction. The corresponding structural characterization data are shown in Table 1-3 and the spectra are shown in Figure 1-2 shown.

[0016] Ailancoumarin A (1): yellow oil; By HRESIMS m / z 391.1666 [M+H] + (cacld for C 21 H 27 O 7 391.1673) The molecular formula is determined to be C 21 H 26 O 7 ; By analyzing the 1 HNMR, 13 CNMR, HSQC, HMBC and calculated optical rotation confirmed the structure of ailancoumarin A, which is a new compound.

[0017] Ailancoumarin B (2): yellow oil; By HRESIMS m / z 391.1645 [M+H] + (cacld for C 20 H 25 O 6 391.1646) The molecular formula is determined to be C 20 H 24 O 6 ; By analyzing the 1 HNMR, 13CNMR, HSQC, HMBC and calculated optical rotation confirmed the structure of ailancoumarin B, which is a new compound.

[0018] Ailancoumarin C (3): yellow oil; By HRESIMS m / z 391.1644 [M+H] + (cacld for C 20 H 25 O 6 391.1646) The molecular formula is determined to be C 20 H 24 O 6 ; By analyzing the 1 HNMR, 13 CNMR, HSQC, HMBC and calculated optical rotation confirmed the structure of ailancoumarin C, which is a new compound.

[0019] Ailancoumarin D (4): yellow oil; By HRESIMS m / z 429.1520 (cacld for C 20 H 26 NaO 6 429.1520) The molecular formula is determined to be C 20 H 26 O 6 ; By analyzing the 1 HNMR, 13 CNMR, HSQC, HMBC and calculated optical rotation confirmed the structure of ailancoumarin D, which is a new compound.

[0020] Ailancoumarin E (5): yellow oil; By HRESIMS m / z 363.1806 [M+Na] + (calcdfor C 21 H 26 NaO 8 :363.1802) The molecular formula is determined to be C 21 H 26 O 8 ; By analyzing the 1 HNMR, 13 CNMR, HSQC, HMBC, propylene method, and calculated optical rotation determined the structure of ailancoumarin E as a new compound.

[0021] Ailancoumarin F(6): yellow oil; By HRESIMS (m / z): 369.1544 [M+H] + (calcdforC 18 H 25 O 8 ,369.1544) to determine the molecular formula is C 18 H 24 O 8 ; By analyzing the 1 HNMR, 13 CNMR, HSQC, HMBC, propylene method, and calculated optical rotation determined the structure of ailancoumarin F, a new compound.

[0022] Ailancoumarin G (7): yellow oil; By HRESIMS (m / z): 577.2249 [M+H] + (calcd for C 27 H 38 NaO 12 ,577.2255) to determine the molecular formula is C 27 H 38 O 12 ; By analyzing the 1 HNMR, 13 CNMR, HSQC, HMBC and calculated optical rotation confirmed the structure of ailancoumarin G as a new compound.

[0023] Ailancoumarin H (8): yellow oil; By HRESIMS (m / z): 465.2085 [M+H] + (calcd for C 22 H 34 NaO 9 465.2095) The molecular formula is determined to be C 22 H 34 O 9 ; By analyzing the 1 HNMR, 13 CNMR, HSQC, HMBC, propylene method, and calculated optical rotation determined the structure of ailancoumarin H as a new compound.

[0024] Ailancoumarin I (9): yellow oil; By HRESIMS (m / z) 421.1469 [M+H] + (calcd for C 19 H 26 NaO 9 ,421.1469) to determine the molecular formula is C 19 H 26 O 9 ; By analyzing the 1 HNMR, 13 CNMR, HSQC, HMBC and calculated optical rotation confirmed the structure of ailancoumarin I, which is a new compound.

[0025] Ailancoumarin J (10): yellow oil; By HRESIMS (m / z) 611.2666 [M+Na] + (calcd for C 28 H 44 NaO 13 ,611.2674) to determine the molecular formula is C 28 H 44 O 13 ; By analyzing ailancoumarinJ 1 HNMR, 13 CNMR, HSQC, HMBC and calculated optical rotation confirmed the structure of ailancoumarin J, which is a new compound.

[0026] A pharmaceutical composition comprises the monoterpene coumarin compound prepared from Ailanthus altissima or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0027] The present invention also provides the use of the monoterpene coumarin compound prepared from Ailanthus altissima or its pharmaceutically acceptable salt or a pharmaceutical composition containing the compound in the preparation of insecticides.

[0028] The insecticidal activities of the 10 new compounds described in the present invention were investigated, and compound 5 showed good insecticidal activity against Plutella xylostella. Therefore, compound 5 described in the present invention has the prospect of being further developed as an insecticide.

[0029] Compared with the prior art, the present invention has the following beneficial effects: The advantages of the present invention are that the compounds are all new compounds with novel structures and are all optically pure compounds with definite stereo configurations. At the same time, they have good insecticidal activity and are worth further development.

[0030]

[0031]

[0032] BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 Compounds 1-10 1 H- 1 H COSY and HMBC are related;

[0034] Figure 2 Compounds 1-6, 8a were related to NOESY. DETAILED DESCRIPTION

[0035] The following examples are intended to help those skilled in the art to better understand the present invention, but are not intended to limit the present invention in any way.

[0036] Example 1

[0037] Preparation of Compound 1-10.

[0038] The dried Ailanthus altissima leaves were extracted with ethanol, and the combined extracts were concentrated to obtain an extract, which was extracted with dichloromethane and n-butanol, and the obtained components were subjected to silica gel column chromatography, and isocratic gradient elution was performed with a dichloromethane-methanol system of 100:1-1:1, and a total of 6 components Fr.AF were collected;

[0039] Using HP20, ODS column chromatography, the component Fr.D was gradient eluted with an ethanol-water system of 20:80-90:10 to obtain five components Fr.D1-D5;

[0040] Fr.D2 was separated by silica gel column chromatography with a petroleum ether-ethyl acetate system of 100:1-10:1 and by preparative reverse phase high performance liquid chromatography using an acetonitrile-water mobile phase to obtain compound 1-10.

[0041] Example 2

[0042] Investigation of antiparasitic activities of compounds 1-10

[0043] Prepare 200μg / mL of the drug solution, use a hole puncher to punch leaf discs with a diameter of 7mm from the feeding leaves corresponding to the test insects, soak the leaf discs in different concentrations of the drug solution for 10-20s, drain the excess solution and spread them horizontally in a 9cm culture dish with moisturizing filter paper, and dry them naturally until there is no water on the surface. A certain number of test insects are placed in each hole, and each drug solution concentration is repeated 3 times. Seal with plastic wrap and pierce holes for ventilation. Use the drug solvent as the negative control and the commercial insecticide as the positive control. Put the culture dish into the incubator: temperature 26±0.5℃, relative humidity 50%, light cycle L:D is 16h:24h, observe and record the number of deaths of the test insects after 48h, and consider it dead if there is no reaction when the test insect surface is touched lightly with a brush. Record the number of deaths, the number of survivals and the date of death of the test insects, and calculate the mortality rate and corrected mortality rate:

[0044] Mortality rate = number of dead insects / total number of insects treated × 100%;

[0045] Adjusted mortality rate = (treatment mortality rate - control mortality rate) / (1 - control mortality rate) x 100%.

[0046] Table 4 Insecticidal activity of compounds 1-10 against Plutella xylostella

[0047]

[0048]

[0049] The experimental results are shown in Table 4. The experimental results show that compound 5 exhibits good insecticidal activity against Plutella xylostella at a concentration of 200 μg / mL, so compound 4 of the present invention has the prospect of further development as an insecticide

[0050] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A monoterpene coumarin compound in Ailanthus altissima leaves, characterized in that: The compound is any one of the structures shown below:

2. The monoterpene coumarin compound in Ailanthus altissima leaves according to claim 1, characterized in that: The Ailanthus altissima is a plant of the genus Ailanthus in the family Simaroubaceae.

3. The method for preparing the monoterpene coumarin compounds in Ailanthus altissima leaves according to claim 1 or claim 2, characterized in that: The steps include: The dried Ailanthus altissima leaves were extracted with ethanol, and the combined extracts were concentrated to obtain an extract, which was extracted with dichloromethane and n-butanol, and the obtained components were subjected to silica gel column chromatography, and isocratic gradient elution was performed with a dichloromethane-methanol system of 100:1-1:1, and a total of 6 components Fr.AF were collected; Using HP20, ODS column chromatography, the component Fr.D was gradient eluted with an ethanol-water system of 20:80-90:10 to obtain 6 components Fr.D1-D5; Fr.D2 was separated by silica gel column chromatography with a petroleum ether-ethyl acetate system of 100:1-10:1 and by preparative reverse phase high performance liquid chromatography using an acetonitrile-water mobile phase to obtain compound 1-10.

4. The method for preparing monoterpene coumarin compounds from Ailanthus altissima leaves according to claim 3, characterized in that: The Ailanthus altissima used was a plant of the genus Ailanthus in the family Simosamineaceae.

5. The method for preparing monoterpene coumarin compounds in Ailanthus altissima leaves according to claim 3, characterized in that: Take the dried Ailanthus altissima leaves and extract them with 70-80% industrial ethanol for 3 times under reflux, each time for 2-3 hours.

6. The method for preparing monoterpene coumarin compounds from Ailanthus altissima leaves according to claim 3, characterized in that: Fr.D2 was separated with acetonitrile-water mobile phase of 70:30-30:

70.

7. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the monoterpene coumarin compound in the Ailanthus altissima leaf according to claim 1 or claim 2 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

8. Use of the monoterpene coumarin compounds or pharmaceutically acceptable salts thereof in Ailanthus altissima leaves according to claim 1 or claim 2 in the preparation of insecticides.

9. Use of the pharmaceutical composition according to claim 7 in the preparation of insecticides.

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