WO2023029814A1 - Matrine crystal, pharmaceutical composition, therapeutic agent for soil-borne diseases, pesticide and uses thereof - Google Patents

Abstract

Provided in the embodiments of the present invention are a matrine crystal, a pharmaceutical composition, a therapeutic agent for soil-borne diseases and the uses thereof, which can reduce the solubility of matrine and a drug using matrine as an active ingredient so as to play a slow release role. In an X-ray powder diffraction pattern using Cu Kα radiation for detection, the matrine crystal has diffraction peaks at diffraction angles 2θ of 17.951°±0.2°; 12.146°±0.2° and 14.858°±0.2°; 20.333°±0.2° and 26.414°±0.2°; and 10.754°±0.2°, 13.794°±0.2°, 18.369°±0.2°, 21.054°±0.2° and 21.348°±0.2°. The matrine crystal has a lower solubility and a slower diffusion in water because of the characteristic of the low solubility thereof. When used for soil-borne diseases, the persistence period of the matrine crystal can be prolonged, the action speed thereof can be effectively reduced, and the action time thereof can be prolonged, so that the pesticide effect is generated for a longer time and the pesticide requirement of long-acting release is met.

Description

Matrine crystal, pharmaceutical composition, therapeutic agent for soil-borne diseases, insecticide and use technical field

The invention relates to a new crystal of matrine.

Background technique

Matrine is a common species of botanical pesticides. At present, the research on matrine mainly focuses on the field of pest control, and no one studies the crystal form of its original drug. Matrine is used in the prevention and control of common soil-borne diseases such as blight, damping-off, fusarium wilt, root rot, blight, root-knot nematode, etc. Because it dissolves quickly, its duration of action is short.

The XRPD spectrum of the conventional matrine crystal form shows that the 2θ (±0.2°) of crystal form I (anhydrous matrine) has diffraction peaks at 11.678°, 14.090°, and 23.006°, and the relative abundance exceeds 50%; There are diffraction peaks at 7.024° and 11.453°, and the relative abundance is between 50% and 30%; there is a diffraction peak at 16.676°, and the relative abundance is between 20% and 10%.

Contents of the invention

Embodiments of the present invention provide a matrine crystal, a pharmaceutical composition, a therapeutic agent for soil-borne diseases, an insecticide, and its use, so as to reduce the solubility of matrine and drugs with matrine as an active ingredient, so as to relieve role of interpretation.

Embodiments of the present invention are realized through the following technical solutions:

In the first aspect, the embodiment of the present invention provides a matrine crystal, which shows a diffraction peak at the following diffraction angle 2θ in the X-ray powder diffraction pattern detected by Cu Kα radiation:

There is a diffraction peak at 17.951°±0.2°;

There are diffraction peaks at 12.146°±0.2° and 14.858°±0.2°;

There are diffraction peaks at 20.333°±0.2° and 26.414°±0.2°;

There are diffraction peaks at 10.754°±0.2° and 13.794°±0.2°, 18.369°±0.2°, 21.054°±0.2°, 21.348°±0.2°.

In the second aspect, the embodiment of the present invention provides a matrine crystal, the preparation method of the matrine crystal is a suspension crystallization method; the suspension crystallization method includes:

Maintain matrine and solvent in a state of suspension and stirring, stir, and filter to obtain matrine crystals;

The solvent is n-heptane.

Further, the stirring temperature of the suspension crystallization method is 50°C.

Further, the stirring time is 5 days.

Further, the ratio of matrine to solvent is 50.53mg:0.2mL.

In the third aspect, the embodiment of the present invention provides a pharmaceutical composition, which contains the matrine crystals as an active ingredient.

In a fourth aspect, embodiments of the present invention provide a therapeutic agent for soil-borne diseases, which contains the matrine crystals described in claim 1 as an active ingredient.

Further, the therapeutic agents for soil-borne diseases include those for blight, damping-off, fusarium wilt, root rot, blight or symptoms associated with root-knot nematodes.

In the fifth aspect, the embodiment of the present invention provides a use of the matrine crystal as an active ingredient of a slow-release drug.

In the sixth aspect, the embodiment of the present invention provides an insecticide, which contains the matrine crystal (crystal form III) as an active ingredient.

Compared with the prior art, the embodiment of the present invention has the following advantages and beneficial effects:

A matrine crystal, a pharmaceutical composition, a therapeutic agent for soil-borne diseases, an insecticide, and its use according to an embodiment of the present invention, due to the low solubility of the matrine crystal, has a lower solubility in water and slower diffusion , when it is used for soil-borne diseases, it can prolong its duration of action, effectively reduce the speed of action of matrine, and prolong the action time, thereby producing drug action in a longer period of time and meeting the demand for long-acting release drugs; at the same time, the The control effect of matrine crystals on aphids is better, and has a good insecticidal effect. At the same concentration, the control effect of matrine crystals (crystal form III) on aphids is significantly higher than that of the original crystal form (crystal form I ).

Description of drawings

In order to more clearly illustrate the technical solutions of the exemplary embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention. Therefore, it should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can also be obtained according to these drawings without creative work.

Figure 1 is the XRPD pattern of conventional matrine crystals (form I).

Fig. 2 is an XRPD pattern of matrine crystals (form III).

Fig. 3 is a TGA diagram of matrine crystals (form III).

Fig. 4 is the result data analysis data of Fig. 3 .

Fig. 5 is the HNMR chart of matrine crystal (crystal form III).

FIG. 6 is the detection condition data of FIG. 5 .

Fig. 7 is a DSC chart of matrine crystals (form III).

FIG. 8 is the detection parameter data of FIG. 7 .

Fig. 9 is a PLM diagram of matrine crystals (form III). Wherein, Fig. 9 (a) is a detection diagram of 90° polarized light; Fig. 9 (b) is a detection diagram of 0° polarized light;

Fig. 10 is the broad bean leaf after conventional matrine crystallization (crystal form I);

Fig. 11 broad bean leaves after matrine crystallization (crystal form II);

Fig. 12 shows broad bean leaves treated with different concentrations of matrine crystallization.

Detailed ways

In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the examples and accompanying drawings. As a limitation of the present invention.

In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. It will be apparent, however, to one of ordinary skill in the art that these specific details need not be employed to practice the present invention.

Example 1

Referring to Fig. 2, an embodiment of the present invention provides a matrine crystal, which is detected by Cu Kα radiation (hereinafter referred to as crystal III).

In the X-ray powder diffraction pattern, the diffraction peaks are displayed at the following diffraction angle 2θ:

There is a diffraction peak at 17.951°±0.2°;

There are diffraction peaks at 12.146°±0.2° and 14.858°±0.2°;

There are diffraction peaks at 20.333°±0.2° and 26.414°±0.2°;

There are diffraction peaks at 10.754°±0.2° and 13.794°±0.2°, 18.369°±0.2°, 21.054°±0.2°, 21.348°±0.2°.

As shown in Figure 3, the crystal form III was analyzed by TGA in NETZSCH TG 209F3TGA209F3A-0652-L, and the crucible used was made of Al ₂ O ₃ ; as shown in Figure 4, the decomposition temperature started at 40.9°C-50°C, and the mass decreased The temperature range of 7% is 50-150°C; the temperature range of 92.5% mass reduction is 150-344.8°C.

Referring to Figure 5, the structure of matrine in crystal form III was determined under HNMR. The detection conditions are shown in Figure 6.

Referring to Figure 7 and Figure 8, Figure 7 is the DSC analysis of Form III.

Referring to FIG. 9 , FIG. 9 is a PLM diagram of matrine crystals of crystal form III (form III).

In the second aspect, the embodiment of the present invention provides a matrine crystal, the preparation method of the matrine crystal is a suspension crystallization method; the suspension crystallization method includes:

Maintain matrine and solvent in a state of suspension and stirring, stir, and filter to obtain matrine crystals;

The solvent is n-heptane.

Further, the stirring temperature of the suspension crystallization method is 50°C.

Further, the stirring time is 5 days.

Further, the ratio of matrine to solvent is 50.53mg:0.2mL.

Matrine crystal form II was studied by suspension crystallization. Weigh about 50 mg of raw materials into a sample bottle, add a certain amount of solvent, keep the system in a state of suspension and stirring at a certain temperature, stir for five days and filter, and analyze the obtained solid. The results are shown in Table 1 below.

Table 1

In the third aspect, the embodiment of the present invention provides a pharmaceutical composition, which contains the matrine crystals as an active ingredient.

In the fourth aspect, an embodiment of the present invention provides a therapeutic agent for soil-borne diseases, which contains the matrine crystals described in claim 1 as an active ingredient.

Further, the therapeutic agent for said soil-borne disease includes a therapeutic agent for blight, damping-off, fusarium wilt, root rot, blight or symptoms associated with root-knot nematode.

In the fifth aspect, the embodiment of the present invention provides a use of the matrine crystal as an active ingredient of a slow-release drug.

The solubility data of each crystal form are compared in Table 2 below.

Table 2

crystal form	Solubility (mg/mL)
Form I	113-117
Form II	130-134
Form III	105-108

*Test conditions: temperature 25°C, pH value of water 5.8-7.4

It can be seen from the above table 2 that the solubility of crystal form III is much lower than that of matrine in crystal form I and II, thus, when slow release of matrine is required for treatment, crystal form III can slowly release the drug to play a role The delayed and sustained effect can be used in the preparation and use of slow-release medicines that require matrine as an active ingredient. For example, the crystal form III can be used as a slow-release drug in the treatment of soil-borne diseases such as blight, damping-off, fusarium wilt, root rot, blight or symptoms associated with root-knot nematodes.

Example 2

The embodiments of the present invention first analyze the influence of 40% matrine solutions with different concentrations on the indoor activity of aphids

1. Experimental design

Drug name and concentration

Table 3 Test design for test agents

Note: The test sample has poor spreadability, and the sample needs to be diluted with 0.05% Tween water.

2. Test method

Insect soaking method:

(1) Preparation of test insect sources: transfer 15 adult aphids (black and shiny, larger in size) to 6cm-10cm broad bean twigs (the lower end is fixed and vertically cultivated with soaked hydroponics planting cotton), and then use Cover the cultivation with a transparent plastic tank with a hole in the bottom, remove the adult aphids after 24 hours, and then carry out chemical treatment after 48 hours of cultivation (ambient temperature is about 25°C);

(2) Investigation of the base number of insect populations: before dipping, investigate the base number of insect populations of each treatment, remove smaller and dead insects, and ensure that the size of aphids in each treatment group is consistent and the number is more than 30.

(3) Soaking medicine: Prepare the medicine solution according to Table 1, and treat it according to different settings. Soak the broad bean twigs with aphids in the medicine for 5 seconds. The wells were covered with transparent plastic jars and placed in an environment of 25°C for cultivation.

3. Outcome Survey

3.1 Investigation time and method

48 hours after the treatment, lightly touch the worms with a small brush, and those that do not move are regarded as dead worms.

Survey Metrics:

(1) Investigate and record the number of dead insects and live insects in each treatment group.

(2) Take photos to record whether the test plants have phytotoxicity.

(3) Record the behavior of the test insect body, such as whether it falls from the plant if it is not dead, and cannot attach to the plant.

3.2 Calculation method

According to the survey data, calculate the control effect of each treatment, calculate according to the formula (1) and (2), the calculation results are kept to two decimal places.

Note: 1) The total number of insect populations after the treatment group refers to the base number of insect populations after the treatment (the sum of the number of live insects and the number of dead insects), not the base number of insect populations recorded before the medicine soaking (some aphids will fall into the treatment solution during the medicine soaking process) In this way, the pre-drug insect population base is larger than the actual treated insect population base).

2) When the population reduction rate of the blank group is more than or equal to 5%, the control effect of each chemical treatment group should be corrected by formula (2).

4. Test results and analysis

4.1 Test results

Table 4 Aphid control effect after each group of liquid treatment

4.2 Result analysis

Adopt Duncan's new compound range method to carry out statistical analysis to test data, as can be seen from Table 4, after test 48h, 40% matrine solution is at 25 times, 50 times, 100 times, 200 times, 400 times, 800 times dilution degree The following control effects on broad bean aphids were 90.86%, 65.46%, 49.25%, 35.99%, 25.68%, 22.13%, respectively, and the 40% matrine solution was diluted 25 times and 50 times to treat broad bean leaves after serious phytotoxicity. 100-fold dilution only has slight phytotoxicity; 0.05% Tween 80 has a control effect on aphids of 5.82%, and 70% imidacloprid (chemical control) diluted 10,000 times has a control effect on faba bean aphids of 98.43%.

5 Test conclusion

To sum up, after 48 hours of the test, the 40% matrine solution diluted 25 times has the highest activity against faba bean aphids, with a control effect of 90.86%. The next best activity is the dilution of 50 times and 100 times, with a control effect of 65.46% respectively. , 49.25%; 0.05% Tween 80 has low activity to aphids, and the control effect is 5.82%. The control effect of 70% imidacloprid (chemical control) diluted 10000 times to faba bean aphids is 98.43%.

Sample safety: 40% matrine solution diluted 25 times and 50 times to treat broad bean leaves has serious phytotoxicity to the leaves, and 100 times dilution has only slight phytotoxicity (as shown in Figure 12, from left to right is successively diluted 25 times, 50 times, 100 times).

As can be seen from the above results, the 40% matrine solution diluted 100 times has a good control effect on broad bean aphids and has low phytotoxicity to leaves. 40% matrine solution was used to dilute the solution 100 times for detection of indoor activity.

Example 3

An embodiment of the present invention provides an insecticide, which contains the matrine crystal (form III) as an active ingredient.

In order to prove the insecticidal performance of matrine crystals (crystal form III) of the present invention, adopt existing crystal form matrine and matrine crystals of the present invention (crystal form III) to carry out the indoor activity determination test of broad bean aphid, specifically as follows:

1. Experimental design

Drug name and concentration

Table 5 Test design for test agents

Note: The test sample has poor spreadability, and the sample needs to be diluted with 0.05% Tween water.

2. Test method

Insect soaking method:

(1) Preparation of test insect sources: transfer 15 adult aphids (black and shiny, larger in size) to 6cm-10cm broad bean twigs (the lower end is fixed and vertically cultivated with soaked hydroponics planting cotton), and then use Cover the cultivation with a transparent plastic tank with a hole in the bottom, remove the adult aphids after 24 hours, and then carry out chemical treatment after 48 hours of cultivation (ambient temperature is about 25°C);

(2) Investigation of the base number of insect populations: before dipping, investigate the base number of insect populations of each treatment, remove smaller and dead insects, and ensure that the size of aphids in each treatment group is consistent and the number is more than 30.

(3) Soaking medicine: Prepare the medicine solution according to Table 5, and treat it according to different settings. Soak the broad bean twigs with aphids in the medicine for 5 seconds. The wells were covered with transparent plastic jars and placed in an environment of 25°C for cultivation.

3. Outcome Survey

3.1 Investigation time and method

48 hours after the treatment, lightly touch the worms with a small brush, and those that do not move are regarded as dead worms.

Survey Metrics:

(1) Investigate and record the number of dead insects and live insects in each treatment group.

(2) Take photos to record whether the test plants have phytotoxicity.

(3) Record the behavior of the test insect body, such as whether it falls from the plant if it is not dead, and cannot attach to the plant.

3.2 Calculation method

According to the survey data, the control effect of each treatment is calculated according to the formulas (1) and (2), and the calculation results are kept to two decimal places.

Note: 1) The total number of insect populations after the treatment group refers to the base number of insect populations after the treatment (the sum of the number of live insects and the number of dead insects), not the base number of insect populations recorded before the medicine soaking (some aphids will fall into the treatment solution during the medicine soaking process) In this way, the pre-drug insect population base is larger than the actual treated insect population base).

2) When the population reduction rate of the blank group is more than or equal to 5%, the control effect of each chemical treatment group should be corrected by formula (2).

4. Test results and analysis

4.1 Test results

Table 6 Aphid control effect after each group of liquid treatment

Note: The control effect in the above table is the average value of each repetition, and the different lowercase letters in the data in the same column indicate the significance of the difference at the 0.05 level.

4.2 Result Analysis

Adopt Duncan's new compound range method to carry out statistical analysis to test data, as can be seen from Table 6, after test 48h, 40% matrine solution (crystal form I), 40% matrine solution (crystal form III) in 100 times The control effects on faba bean aphids were 57.27% and 78.36% respectively at dilutions. 40% matrine solution (crystal form I) and 40% matrine solution (crystal form III) were diluted 100 times to treat broad bean leaves. Slight phytotoxicity; the control effect of 0.05% Tween 80 on aphids is 6.25%, and the control effect of 70% imidacloprid (chemical control) diluted 10000 times on broad bean aphids is 94.28%.

5 Test conclusion

In summary, after 48 hours of testing, the control effects of 40% matrine solution (crystal form I) and 40% matrine solution (crystal form III) on broad bean aphids at 100 times dilution were 57.27%, 78.36%, respectively. %; 0.05% Tween 80 is less active to aphids, and the control effect is 6.25%. The control effect of 70% imidacloprid (chemical control) diluted 10000 times to broad bean aphids is 94.28%; The insecticidal effect of the invented matrine solution (crystal form III) is obviously higher than that of the original crystal form, and the aphid control effect is good.

Sample safety: 40% matrine solution (crystal form I), 40% matrine solution (crystal form III) dilutes 100 times and all has only slight phytotoxicity to blade after treating broad bean leaf (as shown in Figure 10 and Figure 11 ).

The specific embodiments described above have further described the purpose, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above descriptions are only specific embodiments of the present invention and are not intended to limit the scope of the present invention. Protection scope, within the spirit and principles of the present invention, any modification, equivalent replacement, improvement, etc., shall be included in the protection scope of the present invention.

Claims (10)

1. Matrine crystal, it is characterized in that, described matrine crystal shows diffraction peak at following diffraction angle 2θ place in the X-ray powder diffraction pattern that uses Cu Kα radiation detection:

   There is a diffraction peak at 17.951°±0.2°;

   There are diffraction peaks at 12.146°±0.2° and 14.858°±0.2°;

   There are diffraction peaks at 20.333°±0.2° and 26.414°±0.2°;

   There are diffraction peaks at 10.754°±0.2° and 13.794°±0.2°, 18.369°±0.2°, 21.054°±0.2°, 21.348°±0.2°.
2. The matrine crystal is characterized in that, the preparation method of the matrine crystal is a suspension crystallization method; the suspension crystallization method comprises:

   Maintain matrine and solvent in a state of suspension and stirring, stir, and filter to obtain matrine crystals;

   The solvent is n-heptane.
3. The crystallization of matrine according to claim 2, wherein the stirring temperature of the suspension crystallization method is 50°C.
4. matrine crystallization as claimed in claim 2, is characterized in that, the time of described stirring is 5 days.
5. The matrine crystallization as claimed in claim 2, is characterized in that, the ratio of described matrine and solvent is 50.53mg:0.2mL.
6. The pharmaceutical composition is characterized in that the pharmaceutical composition contains the matrine crystals described in claim 1 as an active ingredient.
7. A therapeutic agent for soil-borne diseases, characterized in that the therapeutic agent for soil-borne diseases contains matrine crystals according to claim 1 as an active ingredient.
8. The therapeutic agent for soil-borne diseases according to claim 3, characterized in that, the therapeutic agent for said soil-borne diseases comprises blight, damping-off, fusarium wilt, root rot, blight or root-knot nematode with symptoms therapeutic agent.
9. The use of the matrine crystals described in claim 1 as an active ingredient of a slow-release medicine.
10. An insecticide, characterized in that the insecticide contains matrine crystals as claimed in claim 1 as an active ingredient.

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