Application of dihydroorotate dehydrogenase inhibitor in preparation of insect growth regulator

Abstract

The invention discloses an application of a dihydroorotate dehydrogenase inhibitor in preparation of an insect growth regulator, the compound regulates mitochondrial pyruvic acid oxidation by inhibiting the activity of dihydroorotate dehydrogenase and inhibiting the de novo synthesis of pyrimidine, and finally, the activity of acetyl coenzyme A is influenced by influencing the biological synthesis of acetyl coenzyme A, so that the activity of the dihydroorotate dehydrogenase inhibitor is improved. The fluorine-containing structure compound with the dihydroorotate dehydrogenase inhibiting effect is selected to inhibit the biological synthesis of insect juvenile hormone and lead to abnormal growth and development of pests, while the fluorine-containing structure compound with the dihydroorotate dehydrogenase inhibiting effect is selected to further improve the activity, so that the compound can prevent and control various agricultural pests, especially has obvious effect on cotton bollworm, plutella xylostella, spodoptera litura, beet armyworm and prodenia litura, and has a wide application prospect. And a new choice is provided for the development of insecticides.

Description

Technical Field

[0001] The invention belongs to the technical field of pesticides, and specifically relates to the application of a dihydroorotate dehydrogenase inhibitor in the preparation of an insect growth regulator. Background Art

[0002] Insect Growth Regulators (IGRs) are a class of compounds that can disrupt the growth and development of insects. IGRs interfere with the formation of insect cuticles, including chitin formation, molting and metamorphosis, preventing insects from growing and developing normally, leading to incomplete metamorphosis or death of insect larvae. IGRs are highly selective, environmentally friendly, and safe for non-target organisms, thus meeting the requirements of integrated pest management (IPM) and sustainable agricultural development. Therefore, they have become one of the key areas of research and development and are known as the "pesticides of the 21st century." IGRs are usually divided into four categories: chitin synthesis inhibitors, juvenile hormone analogs, ecdysone analogs, and other IGRs.

[0003] Juvenile hormone (JH) is an important insect hormone. It regulates the growth and development of immature insects and the reproduction of adults. Using JH as a precursor, scientists have developed a large series of insect growth regulators (IGRs) - juvenile hormone analogs. The main varieties of existing juvenile hormone analogs are: methoprene, triprene, hydroprene, kinoprene, fenoxycard, pyriproxyfen and NC-170, etc. These commercialized pesticides have played an important role in integrated pest management. However, these pesticides are all juvenile hormone analogs in chemical structure, and there is a possibility of cross-resistance. Studying the JH biosynthesis pathway and developing JH biosynthesis inhibitors is a reliable way to solve this problem.

[0004] In medicine, the dihydroorotate dehydrogenase inhibitor Brequinar is a broad-spectrum antiviral agent that also has anti-SARS2 activity. The dihydroorotate dehydrogenase inhibitor Leflunomide has anti-rheumatic effects. The dihydroorotate dehydrogenase inhibitor BAY-2402234 is used in related research on myeloid malignancies. The dihydroorotate dehydrogenase inhibitor Farudodstat inhibits protein synthesis by activating the AP-1 transcription factor. Farudodstat can inhibit apoptosis and greatly prolong the survival of acute myeloid leukemia (AML) xenograft mice. The dihydroorotate dehydrogenase inhibitor hDHODH-IN-13 is used in the study of IBD.

[0005] In the field of pesticide research and development, dihydroorotate dehydrogenase has been used as a target for herbicide development. FMC discovered and developed Tetflupyrolimet for the prevention and control of weeds in rice fields, but its application in the field of insecticides has not been reported. Summary of the invention

[0006] The purpose of this section is to summarize some aspects of embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the specification abstract and the invention title of this application to avoid blurring the purpose of this section, the specification abstract and the invention title, and such simplifications or omissions cannot be used to limit the scope of the present invention.

[0007] In view of the above problems and / or the problems existing in the prior art, the present invention is proposed.

[0008] Therefore, the purpose of the present invention is to overcome the deficiencies in the prior art and provide the use of a dihydroorotate dehydrogenase inhibitor in the preparation of an insect growth regulator.

[0009] In order to solve the above technical problems, the present invention provides the following technical solution: the dihydroorotate dehydrogenase inhibitor satisfies the following conditions at the same time:

[0010] (i) having a dihydroorotate dehydrogenase inhibitory effect;

[0011] (ii) is a fluorine-containing heterocyclic compound.

[0012] As a preferred embodiment of the use of the dihydroorotate dehydrogenase inhibitor of the present invention in the preparation of an insect growth regulator, the dihydroorotate dehydrogenase inhibitor comprises one of Brequinar, Leflunomide, BAY-2402234, Farudodstat, and hDHODH-IN-13, the structural formulas of which are shown in formula (1) to formula (5) in sequence;

[0013]

[0014]

[0015] As a preferred embodiment of the use of the dihydroorotate dehydrogenase inhibitor of the present invention in the preparation of an insect growth regulator, the dihydroorotate dehydrogenase inhibitor affects the growth and development of insects by inhibiting the biosynthesis of insect juvenile hormone.

[0016] As a preferred embodiment of the use of the dihydroorotate dehydrogenase inhibitor of the present invention in the preparation of an insect growth regulator, the dihydroorotate dehydrogenase inhibitor inhibits the biosynthesis of insect juvenile hormone by affecting the biosynthesis of acetyl-CoA.

[0017] As a preferred embodiment of the use of the dihydroorotate dehydrogenase inhibitor of the present invention in the preparation of an insect growth regulator, the dihydroorotate dehydrogenase inhibitor affects the biosynthesis of acetyl-CoA by regulating mitochondrial pyruvate oxidation.

[0018] As a preferred embodiment of the use of the dihydroorotate dehydrogenase inhibitor of the present invention in the preparation of an insect growth regulator, the dihydroorotate dehydrogenase inhibitor regulates mitochondrial pyruvate oxidation by inhibiting the de novo synthesis of pyrimidine.

[0019] As a preferred embodiment of the use of the dihydroorotate dehydrogenase inhibitor of the present invention in the preparation of an insect growth regulator, the insects include agricultural pests.

[0020] As a preferred embodiment of the use of the dihydroorotate dehydrogenase inhibitor of the present invention in the preparation of an insect growth regulator, the agricultural pests include cotton bollworm, diamondback moth, fall armyworm, beet armyworm, and Spodoptera litura.

[0021] Another object of the present invention is to provide an insect growth regulator.

[0022] As a preferred embodiment of the insect growth regulator of the present invention, it comprises the dihydroorotate dehydrogenase inhibitor and an auxiliary material acceptable in pesticides.

[0023] Another object of the present invention is to provide an application of an insect growth regulator in agricultural pest control.

[0024] Beneficial effects of the present invention:

[0025] The present invention provides a new application of a dihydroorotate dehydrogenase inhibitor in the preparation of an insect growth regulator, and by selecting a compound containing a fluorine structure, the activity is further improved. The inhibitor can prevent and control a variety of agricultural pests, especially cotton bollworm, diamondback moth, fall armyworm, beet armyworm and cutworm, providing a new choice for the development of insecticides. DETAILED DESCRIPTION

[0026] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the embodiments of the specification.

[0027] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0028] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0029] The dihydroorotate dehydrogenase inhibitors used in the present invention are all purchased from MedChemExpress, including Breq uinar (CAS No.: 96187-53-0), Leflunomide (CAS No.: 75706-12-6), BAY-2402234 (CAS No.: 2225819-06-5), Farudodstat (CAS No.: 1035688-66-4), hDHODH-IN-13 (CAS No.: 3032611-13-2), Tenovin-6 (CAS No.: 1011557-82-6), and AG-636 (CAS No.: 1623416-31-8);

[0030] The structural formulas are shown in formula (1) to formula (7) in sequence:

[0031]

[0032]

[0033] Formula (7);

[0034] The insects used in the present invention were purchased from Henan Keyun Biological Pesticide Co., Ltd. and then bred and reproduced in the laboratory;

[0035] The breeding conditions were as follows: feed was purchased from Henan Keyun Biological Pesticide Co., Ltd., the water source was tap water, the breeding temperature was 28±1°C, the relative humidity was 50-70%, and the photoperiod was L14:D10.

[0036] The present invention adopts the GC-MS / MS method (Journal of Chromatography A, 2018, 1538: 67-74) to determine the inhibitory effect of the compounds of each embodiment and comparative example on the biosynthesis of dihydroorotate dehydrogenase (JH) of cotton bollworm, diamondback moth, fall armyworm, beet armyworm and Spodoptera litura. The specific operation is as follows:

[0037] After the test insects were anesthetized with ice, they were placed on a wax tray, and the lateral pharyngeal bodies (CA) were removed by microdissection, and the surrounding connective tissue was removed in M199 for bioassay;

[0038] CA was placed in 100 μL of M199 containing 20 nM internal standard citronellol and different concentrations of inhibitors (compounds of the examples and comparative examples), cultured in the dark for 3 hours at room temperature with slight shaking; then extracted with 200 μL of n-hexane, and 150 μL of the upper organic phase was taken to determine the dihydroorotate dehydrogenase content by GC-MS / MS.

[0039] The concentrations of the inhibitors were prepared at 0.001 μM, 0.01 μM, 0.1 μM, 1 μM, 10 μM, and 100 μM, respectively. The dihydrolactate dehydrogenase content corresponding to the different concentrations of the inhibitors was determined according to the above method, and the inhibition curve was drawn using GraphPad 5.0 to obtain the inhibitory median concentration.

[0040] The method for determining the stomach poison activity of the compounds of the embodiments and comparative examples of the present invention against cotton bollworm, diamondback moth, fall armyworm, beet armyworm and Spodoptera litura is as follows:

[0041] The newly hatched lepidopteran pest larvae were placed on feeds with different concentrations of inhibitors (compounds of the embodiments and comparative examples), and the feeds were changed once a day. The control group added a corresponding weight of solvent (pure water), and the other conditions were consistent with the experimental group. Each experimental group contained 10 repeated experiments, and 10 larvae were placed in each experiment. From the time of application, the number of dead insect bodies was recorded every day until the larvae stopped eating and pupated, and the mortality rate was finally recorded.

[0042] Example 1 (Brequinar)

[0043] This example tests the inhibitory activity of Brequinar on the biosynthesis of dihydroorotate dehydrogenase of different pests and the stomach poison activity on different pests. The results are shown in Table 1.

[0044] Table 1

[0045]

[0046] (Note: IC 50 The results represent the inhibitory activity on the biosynthesis of dihydroorotate dehydrogenase in pests, and the mortality results represent the stomach poison activity of compounds at different concentrations on pests, the same as in the following table)

[0047] Example 2 (Leflunomide)

[0048] In this example, the inhibitory activity of Leflunomide on the biosynthesis of dihydroorotate dehydrogenase of different pests and the stomach poison activity of Leflunomide on different pests were tested. The results are shown in Table 2.

[0049] Table 2

[0050]

[0051] Example 3 (BAY-2402234)

[0052] In this example, the inhibitory activity of BAY-2402234 on the biosynthesis of dihydroorotate dehydrogenase of different pests and the stomach poison activity of BAY-2402234 on different pests were tested. The results are shown in Table 3.

[0053] Table 3

[0054]

[0055] Example 4 (Farudodstat)

[0056] This example tests the inhibitory activity of Farudodstat on the biosynthesis of dihydroorotate dehydrogenase of different pests and the stomach poison activity of Farudodstat on different pests. The results are shown in Table 4.

[0057] Table 4

[0058]

[0059]

[0060] Example 5 (hDHODH-IN-13)

[0061] In this example, the inhibitory activity of hDHODH-IN-13 on the biosynthesis of dihydroorotate dehydrogenase of different pests and the stomach poison activity on different pests were tested. The results are shown in Table 5.

[0062] Table 5

[0063]

[0064] Comparative Example 1 (Tenovin-6 without fluorine structure)

[0065] In this example, the inhibitory activity of Tenovin-6 on the biosynthesis of dihydroorotate dehydrogenase of different pests and the stomach poison activity of Tenovin-6 on different pests were tested. The results are shown in Table 6.

[0066] Table 6

[0067]

[0068] Comparative Example 2 (AG-636 without fluorine structure)

[0069] This example tests the inhibitory activity of AG-636 on the biosynthesis of dihydroorotate dehydrogenase of different pests and the stomach poison activity of AG-636 on different pests. The results are shown in Table 7.

[0070] Table 7

[0071]

[0072]

[0073] It can be seen from the results in Tables 1 to 7 that the compounds of Examples 1 to 5 of the present application can effectively inhibit the biosynthesis of dihydroorotate dehydrogenase in cotton bollworm, diamondback moth, fall armyworm, beet armyworm and Spodoptera litura, inhibit the de novo synthesis of pyrimidine by inhibiting the activity of dihydroorotate dehydrogenase, and then regulate mitochondrial pyruvate oxidation, and finally inhibit the biosynthesis of insect juvenile hormone by affecting the biosynthesis of acetyl-CoA, resulting in abnormal growth and development of pests.

[0074] Although Tenovin-6 and AG-636 can effectively inhibit the biosynthesis of dihydroorotate dehydrogenase in cotton bollworm, diamondback moth, fall armyworm, beet armyworm and Spodoptera litura, their IC 50 The values ​​are higher than those of the compounds provided in this example, which means that the activities of Tenovin-6 and AG-636 are lower than those of the compounds of the present invention.

[0075] And from the determination experiment of stomach poison activity, it can be seen that the compounds of the present application embodiment have practical application value for pest control, and provide new options for the development of insecticides. Under the same experimental conditions, the toxicity of the commercialized juvenile hormone analog fenoxycarb and the non-fluorinated heterocyclic compounds to the above-mentioned 6 kinds of pests was determined. Under the above two concentrations, the insecticidal effects of fluorinated heterocyclic compounds and fenoxycarb are equivalent, and the structure of fluorinated heterocyclic compounds is simpler than that of fenoxycarb, making its synthesis process simpler, lower cost, and more competitive in the market. Under the same conditions, the insecticidal activity of fluorinated heterocyclic compounds is much higher than that of non-fluorinated heterocyclic compounds (Tenovin-6 and AG-636).

[0076] In summary, the present invention provides a new application of a dihydroorotate dehydrogenase inhibitor in the preparation of an insect growth regulator, and by selecting a fluorine-containing compound, the activity is further improved. The inhibitor can prevent and control a variety of agricultural pests, especially cotton bollworm, diamondback moth, fall armyworm, beet armyworm and cutworm, providing a new choice for the development of insecticides.

[0077] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. Use of a dihydroorotate dehydrogenase inhibitor in the preparation of an insect growth regulator, characterized in that: The dihydroorotate dehydrogenase inhibitor satisfies the following conditions at the same time: (i) having a dihydroorotate dehydrogenase inhibitory effect; (ii) is a fluorine-containing heterocyclic compound.

2. Use of the dihydroorotate dehydrogenase inhibitor according to claim 1 in the preparation of an insect growth regulator, characterized in that: The dihydroorotate dehydrogenase inhibitor includes one of Brequinar, Leflunomide, BAY-2402234, Farudodstat, and hDHODH-IN-13, the structural formulas of which are shown in Formula (1) to Formula (5) in sequence; 3. Use of the dihydroorotate dehydrogenase inhibitor according to claim 1 in the preparation of an insect growth regulator, characterized in that: The dihydroorotate dehydrogenase inhibitor affects the growth and development of insects by inhibiting the biosynthesis of insect juvenile hormone.

4. Use of the dihydroorotate dehydrogenase inhibitor according to claim 1 in the preparation of an insect growth regulator, characterized in that: The dihydroorotate dehydrogenase inhibitor inhibits the biosynthesis of insect juvenile hormone by affecting the biosynthesis of acetyl coenzyme A.

5. Use of the dihydroorotate dehydrogenase inhibitor according to claim 1 in the preparation of an insect growth regulator, characterized in that: The dihydroorotate dehydrogenase inhibitor affects acetyl-CoA biosynthesis by regulating mitochondrial pyruvate oxidation.

6. Use of the dihydroorotate dehydrogenase inhibitor according to claim 1 in the preparation of an insect growth regulator, characterized in that: The dihydroorotate dehydrogenase inhibitor regulates mitochondrial pyruvate oxidation by inhibiting the de novo synthesis of pyrimidine.

7. Use of the dihydroorotate dehydrogenase inhibitor according to any one of claims 3 to 6 in the preparation of an insect growth regulator, characterized in that: The insects include agricultural pests.

8. Use of the dihydroorotate dehydrogenase inhibitor according to claim 7 in the preparation of an insect growth regulator, characterized in that: The agricultural pests include cotton bollworm, diamondback moth, fall armyworm, beet armyworm and Spodoptera litura.

9. An insect growth regulator, characterized in that: The invention comprises the dihydroorotate dehydrogenase inhibitor according to any one of claims 1 or 2, and further comprises an auxiliary material acceptable in pesticides.

10. Use of the insect growth regulator as claimed in claim 9 in agricultural pest control.

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