Application of phenothiazine derivatives in the preparation of drugs or preparations for inhibiting the growth of cyanobacteria

Abstract

The present invention discloses the use of phenothiazine derivatives in the preparation of drugs or preparations for inhibiting the growth of cyanobacteria, wherein the phenothiazine derivatives have a general structure as shown in formula (1), and R1 is selected from F, Cl, Br, I, CF3, CH2F, CHF2, C m H 2m+1 , OC m H 2m+1 , S.C. m H 2m+1 、N(C m H 2m+1 )2, R2 is selected from N(C n H 2n+1 )2.OC n H 2n+1 , SC n H 2n+1 etc., m and n are selected from integers of 0-12. The present invention also provides a cyanobacteria inhibitor, which comprises one or more of the phenothiazine derivatives, and the concentration of the phenothiazine derivatives is 0.01-80 μg / mL. The phenothiazine derivatives in the present invention have a significant inhibitory effect on cyanobacteria, and the inhibitory effect can reach more than 95%. Under the same conditions, they have good selectivity for green algae, and the phenothiazine derivatives are used to inhibit the growth of cyanobacteria. The effect is significant.

Description

Technical Field

[0001] The invention belongs to the field of resource and environmental technology, and particularly relates to the application of phenothiazine derivatives in preparing medicines or preparations for inhibiting the growth of cyanobacteria. Background Art

[0002] In recent years, harmful algal blooms (HABs) have occurred frequently due to the increase in eutrophication of water bodies and have been exacerbated by global warming. Nutrients such as nitrogen and phosphorus from domestic sewage, industrial and agricultural runoff flow into water bodies in large quantities, causing excessive growth of algal cells. Environmental factors (such as light intensity, temperature and pH) can also affect harmful algal blooms. Rising temperatures caused by climate change can further accelerate the growth of algae. The rapid growth of cyanobacteria poses a huge risk to the aquatic environment and the health of aquatic organisms and humans.

[0003] Algal cells usually grow on the water surface, reducing the light transmittance of the water body, thereby inhibiting the photosynthesis of aquatic plants, reducing the oxygen content of the water body, and hindering the normal growth of aquatic organisms. When harmful algal blooms break out in raw water, the extremely high density of algal cells will greatly affect the efficiency of the water treatment process and seriously affect the operation of the water plant and the safety of water supply. In addition, the release of cyanobacterial metabolites into water bodies, such as odorous substances and cyanobacterial toxins, especially algal toxins, may have irreversible adverse effects on aquatic organisms and humans. Microcystins are common, toxic and widespread algal toxins that cannot be ideally removed by water plants due to their stable and complex circulation structure. Lian et al.'s research shows that if people are exposed to food or water contaminated with cyanobacterial toxins, microcystins can endanger the human kidneys, liver and nervous system, and in severe cases may cause death.

[0004] Faced with the severe situation of HABs pollution, we first need to effectively reduce the algae content in polluted water bodies. The existing algae inhibition methods can be divided into three categories according to their means: physical, biological, and chemical. Among them: physical means mainly remove harmful algae in water bodies directly through mechanical salvage, flocculation sedimentation, filtration, etc. In the flocculation sedimentation method, the use of modified clay to flocculate and precipitate HAB cells is considered to be a promising strategy. Biological means mainly use biological manipulation to inhibit the excessive reproduction of algae. Fish, shrimps, crabs and other organisms in water bodies mostly feed on algae. The introduction of such species in captivity can effectively prevent the formation of algal blooms. In addition, some heterotrophic bacteria also have a certain inhibitory effect on the growth of algae. Chemical means mainly remove algae cells in water bodies by adding chemical reagents with strong oxidizing properties, allelopathic effects or other functions to water bodies containing large amounts of harmful algae or where HABs have occurred. Chemical reagents have the advantages of rapid effect and good selectivity in algae removal. Chemical methods are now more commonly used in the control of algal blooms and red tides. Commonly used chemical algaecides are divided into two categories: the first category is inorganic algaecides such as copper sulfate; the second category is organic algaecides, such as herbicides.

[0005] In summary, physical algae inhibition methods are costly and difficult to popularize; biological methods have long-term effects and are relatively low in cost, but are slow to take effect, and improper use will affect the lake ecosystem; most of the chemical algaecides currently used have some disadvantages, such as poor selectivity, easy resistance, low efficiency, and great environmental pollution. Based on this, the present invention was completed. Summary of the invention

[0006] The purpose of the present invention is to provide the use of phenothiazine derivatives in the preparation of drugs or preparations for inhibiting the growth of cyanobacteria. The phenothiazine derivatives have an inhibitory effect on cyanobacteria and solve the problems of poor selectivity and low efficiency of existing chemical algaecides.

[0007] To achieve the above object, the present invention adopts the following technical solutions:

[0008] The present invention provides the use of phenothiazine derivatives in the preparation of drugs or preparations for inhibiting the growth of cyanobacteria, wherein the phenothiazine derivatives have a general structure as shown in the following formula (1):

[0009]

[0010] In formula (1), p is selected from an integer of 0-12;

[0011] R1 is selected from F, Cl, Br, I, CF3, CH2F, CHF2, C m H 2m+1 , OC m H 2m+1 , SC m H 2m+1 、N(C m H 2m+1 )2, m is selected from an integer of 0-12;

[0012] R2 is selected from N(C n H 2n+1 )2.OC n H 2n+1 , SC n H 2n+1 ,

[0013]

[0014] n is selected from integers of 0-12.

[0015] Preferably, in formula (1), p is an integer selected from 1-5.

[0016] Preferably, in formula (1), R1 is selected from one or more of Cl, CF3, CHF2, SCH3, OCH3, OC2H5 or N(CH3)2.

[0017] Preferably, in formula (1), R2 is selected from N(CH3)2,

[0018]

[0019] Preferably, the phenothiazine derivative is selected from any of the following compounds:

[0020]

[0021] Preferably, the cyanobacteria is Synechocystis pcc6803.

[0022] Preferably, the drug or preparation contains the phenothiazine derivative as an active ingredient, and its effective dose is 0.01-80 μg / mL.

[0023] More preferably, the effective dose of the phenothiazine derivative is 5 μg / mL.

[0024] The present invention also provides a cyanobacteria inhibitor, which comprises one or more of the phenothiazine derivatives, and the concentration of the phenothiazine derivatives is 0.01-80 μg / mL, preferably 5 μg / mL.

[0025] Compared with the prior art, the present invention has the following beneficial effects: the phenothiazine derivatives as shown in the chemical formula (1) of the present invention have a selective inhibitory effect on cyanobacteria, and the inhibitory effect can reach more than 95%. Under the same conditions, they have good selectivity for green algae (using Chlorella pyrenoidosa as a model), especially having no obvious killing effect on Chlorella pyrenoidosa. Such phenothiazine derivatives can be used to prepare drugs or preparations for inhibiting the growth of cyanobacteria. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is the general structure diagram of phenothiazine derivatives.

[0027] Figure 2 : (a) Algae killing pictures of different concentrations of Compound A1 in the examples; (b) Inhibition rate of cyanobacteria treated with different concentrations of Compound A1 for 12 h, where the light intensity was 100 μmol·m -2 s -1 , temperature was 27°C, initial cell density OD 730 nm=1.5.

[0028] Figure 3 : (a) Pictures of algae killing by compound A2 at different concentrations in the examples; (b) Inhibition rate of cyanobacteria treated with compound A2 at different concentrations for 12 h, where the light intensity was 100 μmol·m -2 s -1 , temperature was 27°C, initial cell density OD 730nm=1.5.

[0029] Figure 4 : (a) Pictures of algae killing by compound A3 at different concentrations in the examples; (b) Inhibition rate of cyanobacteria treated with compound A3 at different concentrations for 12 h, where the light intensity was 100 μmol·m -2 s -1 , temperature was 27°C, initial cell density OD 730 nm=1.5.

[0030] Figure 5 : (a) Pictures of algae killing by compound A4 at different concentrations in the examples; (b) Inhibition rate of cyanobacteria treated with compound A4 at different concentrations for 12 h, where the light intensity was 100 μmol·m -2 s -1 , temperature was 27°C, initial cell density OD 730 nm=1.5.

[0031] Figure 6 : (a) Algae killing pictures of compound A5 at different concentrations in the examples; (b) Inhibition rate of cyanobacteria treated with compound A5 at different concentrations for 12 h, where the light intensity was 100 μmol·m -2 s -1 , temperature was 27-28℃, initial cell density OD 730 nm=1.5.

[0032] Figure 7 : Inhibition rate of Chlorella pyrenoidosa treated with different concentrations of Compound A2 for 12 h, where the light intensity was 100 μmol·m -2 s -1 , the temperature is 27-28℃.

[0033] Figure 8 : Inhibition rate of cyanobacteria treated with different concentrations of compound A2 for 12 h in the embodiment, where the light intensity was 100 μmol·m -2 s -1 , the temperature is 27-28℃.

[0034] Fig. 9 : Effect of 5 μg / mL Compound A2 on initial cell density OD in Example 730 nm=0.5 The inhibition rate of cyanobacteria, where the light intensity is 40 μmol·m -2 s -1 , the temperature is 27℃.

[0035] Fig.10 : Effect of 5 μg / mL compound A2 on initial cell density OD in the example 730nm=1.0 The inhibition rate of cyanobacteria, where the light intensity is 40 μmol·m -2 s -1 , the temperature is 27℃.

[0036] Fig.11 : Effect of 5 μg / mL compound A2 on initial cell density OD in the example 730 nm=1.5 The inhibition rate of cyanobacteria, where the light intensity is 40 μmol·m -2 s -1 , the temperature is 27-28℃. DETAILED DESCRIPTION

[0037] In order to make the purpose, technical solution and advantages of the embodiment of the present invention clearer, the technical solution of the embodiment of the present invention will be clearly and completely described below in conjunction with the accompanying drawings of the embodiment of the present invention. Obviously, the described embodiment is a part of the embodiment of the present invention, not all of the embodiments. Based on the described embodiment of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0038] The sources of compounds A1-A9 used in the following examples are from Chaudhari, A., Pramanik, C., Patil, P., Vedpathak, H., Bandishti, M., Tripathy, NK, Gurjar, MK, J. Heterocycl. Chem. 2022, 59(6), 1054.

[0039] Example 1

[0040] 1. Algae Cultivation

[0041] 1.1 Cultivation method of Chlorella pyrenoidosa

[0042] 1.1.1 Preparation of stock solution STOCK I:

[0043] Prepare 1000mL of stock solution STOCKⅠ (Hurner's trace elements): Add NH4Cl (15.0g), CaCl2 (1.5g), MgSO4 (3.2g), and deionized water (fixed to 1000mL) into a 1L beaker and fix to 1000mL. Stir and mix well, transfer to a glass bottle, seal it, sterilize it at high temperature and high pressure, and place it at room temperature.

[0044] 1.1.2 Preparation of stock solution II:

[0045] Preparation of STOCKⅡ: Add K2HPO4 (28.8g), KH2PO4 (14.4g), and deionized water (diluted to 100mL) into a 500mL beaker and dilute to 100mL. Stir and mix well, transfer to a glass bottle, seal it, sterilize it at high temperature and high pressure, and store it in a refrigerator at 4℃.

[0046] 1.1.3 Preparation of stock solution III:

[0047] Preparation of STOCK III: Dissolve ZnSO4·7H2O (15.0 g) in deionized water (50 mL), H3BO3 (5.7 g) in deionized water (100 mL), MnCl2·4H2O (2.53 g) in deionized water (25 mL), Co(NO3)2·6H2O (0.9845 g) in deionized water (25 mL), CuSO4·5H2O (0.785 g) in deionized water (25 mL), Na2MoO4·2H2O (0.75375 g) in deionized water (25 mL), FeSO4·7H2O (2.495 g) in deionized water (25 mL), and disodium EDTA. Salt (25g) was dissolved in deionized water (125mL). STOCKⅢ was prepared strictly in the above order. They were dissolved in turn and then poured into a 500mL beaker. The large beaker was placed in a microwave oven and heated several times until boiling (multiple heating ensures uniform heating inside). Then EDTA was poured in several times to fully dissolve it. Pour slowly to prevent boiling and splashing. The beaker was then placed in a 70℃ water bath and the pH was adjusted to 6.7 with KOH solution. After the pH adjustment, the solution was greenish. Then the solution was fixed to 500mL with water for the second time. Finally, a ventilation tube was added and the solution was oxidized in an incubator for two weeks until the solution turned dark purple. Finally, it was sterilized by filtration and stored in a refrigerator at 4℃.

[0048] 1.1.4 Preparation of TAP medium:

[0049] After the three mother solutions were prepared, the components STOCKⅠ (125mL), STOCKⅡ (625μL), STOCKⅢ (5mL), glacial acetic acid (5mL), Tris-base (12.1g), and deionized water (to 5000mL) were added to a 5L beaker. Secondary water was added to make the volume to 5L, pH was adjusted to 7.3, and sterilized at high temperature and high pressure and stored at room temperature. The operation of adding mother solutions was carried out in a clean bench.

[0050] 1.1.5 Cultivation of Chlorella pyrenoidosa:

[0051] The research object was Chlorella pyrenoidosa. All algae strains were grown in TAP medium with pH = 7.3, with an inoculation of 1% and in a light incubator at 25°C with a concentration of 40 μE·m-2 s -1 The light intensity was continuously illuminated and aerated for 36 h. After the culture was completed, the cell concentration was counted with a cell counter. 7 -2.2×10 7 / mL.

[0052] 1.2 Cyanobacteria culture

[0053] 1.2.1 Preparation of A5 stock solution:

[0054] A5 1L formula: Add H3BO3 (2.86g), MnCI2·4H2O (1.81g), ZnSO4·7H2O (0.222g), Na2MoO4·2H2O (0.39g), CuSO4·5H2O (0.079g), Co(NO3)2·6H2O (0.0494g) to a 1L beaker and dilute to 1L. Stir to mix well, transfer to a glass bottle, seal it, sterilize it at high temperature and high pressure, and store it at room temperature.

[0055] 1.2.3 Preparation of BGⅡ medium:

[0056] Prepare BGⅡ1L formula: add NaNO3 (1.5g), K2HPO4·3H2O (0.04g), MgSO4·7H2O (0.075g), CaCl2·2H2O (0.036g), citric acid (0.006g), ferric citrate ammonium (0.006g), EDTA·Na2 (0.001g), Na2CO3 (0.02g), A5 (1mL), and deionized water (1000mL) into a 1L beaker and dilute to 1L, adjust pH to 8, stir to mix, transfer to a glass bottle, seal, sterilize at high temperature and high pressure, and store at room temperature.

[0057] The cyanobacterial strain used in the following examples is Synechocystis pcc6803. All algal cells were cultured in BGⅡ medium with pH=8.0 and 5 mM Tris-HCl. The culture temperature was maintained at about 30°C and the light intensity was 40 μE·m -2 s -1 The algae cells were cultured under continuous illumination by LED lights and sterile air containing 2% (v / v) CO2. 730 It can be used when nm=1.5.

[0058] 2. Treatment of Cyanobacterial Cells

[0059] After the cyanobacteria or Chlorella vulgaris was cultured, 10 mL of the culture and different concentrations of phenothiazine derivatives were transferred to each 60 mL hydrogen production glass tube. The mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix. Then, it was placed in a room with a light intensity of 100 μmol·m -2 s -1 , leave it at a temperature of 27-28℃ for several hours.

[0060] 3. Inhibitory effects of phenothiazine derivatives on Chlorella pyrenoidosa

[0061] Chlorella vulgaris was cultured to different cell concentrations (5×10^6 / mL-2×10^7 / mL), 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, and 0-800 μg of phenothiazine derivatives ( Figure 1 ), a solution with a concentration of 0-80 μg / mL was obtained and transferred into the corresponding hydrogen production glass tube. The mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, calculate the chlorophyll a concentration, and calculate the inhibition rate of protein nucleus Chlorella.

[0062] 4. Inhibitory effects of phenothiazine derivatives on cyanobacteria

[0063] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10, 20, and 30 mL of algae solution were centrifuged at 4000 r / min, the supernatant was discarded, and 30 mL of fresh culture medium was added. After suspension was shaken on a vortex shaker, the cells were transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=0.5, 1.0, 1.5, 0-800 μg of phenothiazine derivative ( Figure 1 ), a solution with a concentration of 0-80 μg / mL was obtained and transferred into the corresponding hydrogen production glass tube. The mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix. Then, it was placed in a room with a light intensity of 100 μmol·m -2 s -1, and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, calculate the chlorophyll a concentration, and calculate the inhibition rate of cyanobacteria.

[0064] Example 2

[0065] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm = 0.5 solution, 0, 50, 100, 200, 400, 800 μg of compound A1 were dissolved in 100 μL of DMSO respectively. The solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL were obtained and transferred into the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube. Then add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, put it in a 4°C refrigerator and let it stand for 12h, then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 20%, 18%, 30%, 61%, 92% ( Figure 2 ).

[0066] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A1 was dissolved in 100 μL of DMSO to obtain 0, 5, 10, 20, 40, 80 μg / mL concentration solutions, which were transferred to the corresponding hydrogen production glass tubes, the mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s-1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 9%, 8%, 6%, 6%, and 3%.

[0067] Example 3

[0068] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm = 0.5 solution. Dissolve 0, 50, 100, 200, 400, 800 μg of compound A2 in 100 μL of DMSO respectively. The solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL were obtained and transferred into the corresponding hydrogen production glass tubes. The tubes were sealed with rubber stoppers and slightly shaken to mix. The tubes were then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae liquid from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube. Then add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 23%, 21%, 30%, 76%, 82% ( Figure 3 ).

[0069] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, and 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube. 0, 50, 100, 200, 400, and 800 μg of compound A2 were dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL, which were then transferred to the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and gently shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 5%, 2%, 3%, 3%, and 1%.

[0070] Example 4

[0071] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm = 0.5 solution. 0, 50, 100, 200, 400, 800 μg of compound A3 were dissolved in 100 μL of DMSO respectively. The solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL were obtained and transferred into the corresponding hydrogen production glass tubes. The tube mouths were sealed with rubber stoppers and slightly shaken to mix. Then, the tubes were placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae liquid from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 22%, 28%, 35%, 70%, 80% ( Figure 4).

[0072] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A3 was dissolved in 100 μL of DMSO to obtain 0, 5, 10, 20, 40, 80 μg / mL concentration solutions, which were transferred to the corresponding hydrogen production glass tubes, the mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and put it in a 4℃ refrigerator for 12h. Then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, calculate the chlorophyll a concentration, and calculate the inhibition rate of protein nucleus Chlorella to be 0, 8%, 5%, 6%, 7%, and 4%.

[0073] Example 5

[0074] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm = 0.5 solution. 0, 50, 100, 200, 400, 800 μg of compound A4 were dissolved in 100 μL of DMSO respectively. The solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL were obtained and transferred into the corresponding hydrogen production glass tubes. The tubes were sealed with rubber stoppers and slightly shaken to mix. The tubes were then placed under a light intensity of 100 μmol·m -2 s -1, and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae liquid from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 18%, 21%, 30%, 60%, 82% ( Figure 5 ).

[0075] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A4 was dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, 80 μg / mL, and transferred to the corresponding hydrogen production glass tube, the mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 9%, 11%, 12%, 8%, and 7%.

[0076] Example 6

[0077] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm = 0.5 solution, 0, 50, 100, 200, 400, 800 μg of compound A5 were dissolved in 100 μL of DMSO respectively. The solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL were obtained and transferred into the corresponding hydrogen production glass tubes. The tube mouths were sealed with rubber stoppers and slightly shaken to mix. Then, the tubes were placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27°C. After standing, extract 5mL of algae liquid from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 18%, 21%, 28%, 78%, 83% ( Figure 6 ).

[0078] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A6 was dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, 80 μg / mL, and transferred to the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 9%, 11%, 12%, 8%, and 7%.

[0079] Example 7

[0080] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730nm = 0.5 solution, 0, 50, 100, 200, 400, 800 μg of compound A6 were dissolved in 100 μL of DMSO respectively. The solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL were obtained and transferred into the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, put it in a 4°C refrigerator and let it stand for 12h, then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 22%, 23%, 35%, 62%, and 85%.

[0081] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A6 was dissolved in 100 μL of DMSO to obtain 0, 5, 10, 20, 40, 80 μg / mL concentration solutions, which were transferred to the corresponding hydrogen production glass tubes, the mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 10%, 11%, 10%, 9%, and 6%.

[0082] Example 8

[0083] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD730 nm = 0.5 solution, 0, 50, 100, 200, 400, 800 μg of compound A7 were dissolved in 100 μL of DMSO respectively. The solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL were obtained and transferred into the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, put it in a 4°C refrigerator and let it stand for 12h, then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 18%, 23%, 45%, 62%, and 79%.

[0084] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A7 was dissolved in 100 μL of DMSO to obtain 0, 5, 10, 20, 40, 80 μg / mL concentration solutions, which were transferred to the corresponding hydrogen production glass tubes, the mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 15%, 13%, 10%, 9%, and 6%.

[0085] Example 9

[0086] Cyanobacteria were cultured to a cell density of OD 730After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm = 0.5 solution, 0, 50, 100, 200, 400, 800 μg of compound A8 were dissolved in 100 μL of DMSO respectively. The solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL were obtained and transferred into the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, put it in a 4°C refrigerator and let it stand for 12h, then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 22%, 23%, 35%, 62%, and 85%.

[0087] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A8 was dissolved in 100 μL of DMSO to obtain 0, 5, 10, 20, 40, 80 μg / mL concentration solutions, which were transferred to the corresponding hydrogen production glass tubes, the mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 10%, 10%, 13%, 9%, and 7%.

[0088] Example 10

[0089] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm = 0.5 solution, 0, 50, 100, 200, 400, 800 μg of compound A9 were dissolved in 100 μL of DMSO respectively. The solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL were obtained and transferred into the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, put it in a 4°C refrigerator and let it stand for 12h, then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 18%, 23%, 36%, 62%, and 85%.

[0090] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A9 was dissolved in 100 μL of DMSO to obtain 0, 5, 10, 20, 40, 80 μg / mL concentration solutions, which were transferred to the corresponding hydrogen production glass tubes, the mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 8%, 16%, 13%, 9%, and 17%.

[0091] Embodiment 11

[0092] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, take 20 mL of algae solution and centrifuge at 4000 r / min. After discarding the supernatant, add 30 mL of fresh culture medium, vortex and suspend on a vortex shaker, and transfer to a 60 mL glass tube to obtain the cell density OD 730 nm=1.0. 0, 50, 100, 200, 400, 800 μg of compound A1 were dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL, which were then transferred into the corresponding hydrogen production glass tubes. The tubes were sealed with rubber stoppers and gently shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 18%, 16%, 32%, 59%, and 90%.

[0093] Chlorella vulgaris was cultured to a cell concentration of 1×10^7 / mL, and 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube. 0, 50, 100, 200, 400, and 800 μg of compound A1 were dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL, which were transferred to the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 10%, 7%, 5%, 6%, and 4%.

[0094] Example 12

[0095] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, take 20 mL of algae solution and centrifuge at 4000 r / min. After discarding the supernatant, add 30 mL of fresh culture medium, vortex and suspend on a vortex shaker, and transfer to a 60 mL glass tube to obtain the cell density OD 730 nm=1.0, 0, 50, 100, 200, 400, 800 μg of compound A2 were dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, 80 μg / mL, and transferred into the corresponding hydrogen production glass tubes. The tube mouths were sealed with rubber stoppers and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 20%, 25%, 34%, 62%, and 84%.

[0096] Cultivate the pyrenoid algae to a cell concentration of 1×10^7 / mL, and transfer 10 mL of the culture to each 60 mL hydrogen production glass tube. Dissolve 0, 50, 100, 200, 400, and 800 μg of compound A2 in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL, and transfer them to the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 6%, 4%, 5%, 5%, and 2%.

[0097] Example 13

[0098] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, take 20 mL of algae solution and centrifuge at 4000 r / min. After discarding the supernatant, add 30 mL of fresh culture medium, vortex and suspend on a vortex shaker, and transfer to a 60 mL glass tube to obtain the cell density OD 730 nm=1.0. Dissolve 0, 50, 100, 200, 400, and 800 μg of compound A3 in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL, and transfer them into the corresponding hydrogen production glass tubes. Seal the tube mouth with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 23%, 20%, 33%, 79%, and 83%.

[0099] Cultivate the pyrenoid algae to a cell concentration of 1×10^7 / mL, and transfer 10 mL of the culture to each 60 mL hydrogen production glass tube. Dissolve 0, 50, 100, 200, 400, and 800 μg of compound A3 in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL, and transfer them to the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 9%, 6%, 5%, 8%, and 4%.

[0100] Embodiment 14

[0101] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, take 20 mL of algae solution and centrifuge at 4000 r / min. After discarding the supernatant, add 30 mL of fresh culture medium, vortex and suspend on a vortex shaker, and transfer to a 60 mL glass tube to obtain the cell density OD 730 nm=1.0, 0, 50, 100, 200, 400, 800 μg of compound A4 were dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, 80 μg / mL, and transferred into the corresponding hydrogen production glass tubes. The tubes were sealed with rubber stoppers and gently shaken to mix, and then placed under a light intensity of 100 μmol·m -2 .s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 16%, 23%, 30%, 78%, and 81%.

[0102] Cultivate Chlorella vulgaris to a cell concentration of 5×10^6 / mL, and transfer 10mL of the culture to each 60mL hydrogen production glass tube. Dissolve 0, 50, 100, 200, 400, and 800μg of compound A4 in 100μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80μg / mL, and transfer them to the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 13%, 12%, 10%, 8%, and 7%.

[0103] Embodiment 15

[0104] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, take 20 mL of algae solution and centrifuge at 4000 r / min. After discarding the supernatant, add 30 mL of fresh culture medium, vortex and suspend on a vortex shaker, and transfer to a 60 mL glass tube to obtain the cell density OD 730 nm=1.0. 0, 50, 100, 200, 400, 800 μg of compound A5 were dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, 80 μg / mL, and transferred into the corresponding hydrogen production glass tubes. The tube mouths were sealed with rubber stoppers and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 20%, 21%, 28%, 75%, and 81%.

[0105] Cultivate Chlorella vulgaris to a cell concentration of 1×10^7 / mL, and transfer 10mL of the culture to each 60mL hydrogen production glass tube. Dissolve 0, 50, 100, 200, 400, and 800μg of compound A5 in 100μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80μg / mL, and transfer them to the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 10%, 11%, 8%, 8%, and 7%.

[0106] Example 16

[0107] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=0.5, 0, 50, 100, 200, 400, 800 μg of compound A6 were dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, 80 μg / mL, which were transferred into the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and gently shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, put it in a 4°C refrigerator and let it stand for 12h, then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 22%, 23%, 35%, 62%, and 85%.

[0108] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A6 was dissolved in 100 μL of DMSO to obtain 0, 5, 10, 20, 40, 80 μg / mL concentration solutions, which were transferred to the corresponding hydrogen production glass tubes, the mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 10%, 11%, 10%, 9%, and 6%.

[0109] Embodiment 17

[0110] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=0.5, 0, 50, 100, 200, 400, 800 μg of compound A7 were dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, 80 μg / mL, which were transferred into the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and gently shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, put it in a 4°C refrigerator and let it stand for 12h, then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 18%, 23%, 45%, 62%, and 79%.

[0111] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A7 was dissolved in 100 μL of DMSO to obtain 0, 5, 10, 20, 40, 80 μg / mL concentration solutions, which were transferred to the corresponding hydrogen production glass tubes, the mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 15%, 13%, 10%, 9%, and 6%.

[0112] Embodiment 18

[0113] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=0.5, 0, 50, 100, 200, 400, 800 μg of compound A8 were dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, 80 μg / mL, which were transferred into the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and gently shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, put it in a 4°C refrigerator and let it stand for 12h, then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 22%, 23%, 35%, 62%, and 85%.

[0114] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A8 was dissolved in 100 μL of DMSO to obtain 0, 5, 10, 20, 40, 80 μg / mL concentration solutions, which were transferred to the corresponding hydrogen production glass tubes, the mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 10%, 10%, 13%, 9%, and 7%.

[0115] Embodiment 19

[0116] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=0.5, 0, 50, 100, 200, 400, 800 μg of compound A9 were dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, 80 μg / mL, which were transferred into the corresponding hydrogen production glass tubes. The mouth of the hydrogen production tube was sealed with a rubber stopper and gently shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube, centrifuge at 4000r / min for 10min, remove the supernatant, and shake on a vortex shaker until there is no algae cell attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, put it in a 4°C refrigerator and let it stand for 12h, then centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 18%, 23%, 36%, 62%, and 83%.

[0117] Chlorella vulgaris was cultured to a cell concentration of 5×10^6 / mL, 10 mL of the culture was transferred to each 60 mL hydrogen production glass tube, 0, 50, 100, 200, 400, 800 μg of compound A9 was dissolved in 100 μL of DMSO to obtain 0, 5, 10, 20, 40, 80 μg / mL concentration solutions, which were transferred to the corresponding hydrogen production glass tubes, the mouth of the hydrogen production tube was sealed with a rubber stopper and slightly shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 8%, 16%, 13%, 9%, and 16%.

[0118] Embodiment 20

[0119] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 30 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, and 30 mL of fresh culture medium was added. After suspension was shaken on a vortex shaker, the suspension was transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=1.5. Dissolve 0, 50, 100, 200, 400, and 800 μg of compound A1 in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL, and transfer them into the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 21%, 19%, 31%, 62%, and 93%.

[0120] Cultivate Chlorella vulgaris to a cell concentration of 2×10^7 / mL, and transfer 10mL of the culture to each 60mL hydrogen production glass tube. Dissolve 0, 50, 100, 200, 400, and 800μg of compound A1 in 100μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80μg / mL, and transfer them to the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100μmol·m -2 s -1, and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 8%, 7%, 5%, 5%, and 2%.

[0121] Embodiment 21

[0122] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 30 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, and 30 mL of fresh culture medium was added. After suspension was shaken on a vortex shaker, the suspension was transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=1.5, 0, 50, 100, 200, 400, 800 μg of compound A2 were dissolved in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, 80 μg / mL, and transferred into the corresponding hydrogen production glass tubes. The hydrogen production tube was sealed with a rubber stopper and gently shaken to mix, and then placed under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 19%, 25%, 33%, 62%, and 83%.

[0123] Cultivate Chlorella vulgaris to a cell concentration of 2×10^7 / mL, and transfer 10mL of the culture to each 60mL hydrogen production glass tube. Dissolve 0, 50, 100, 200, 400, and 800μg of compound A2 in 100μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80μg / mL, and transfer them to the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100μmol·m -2 s-1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 6%, 3%, 5%, 7%, and 1%.

[0124] Embodiment 22

[0125] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 30 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, and 30 mL of fresh culture medium was added. After suspension was shaken on a vortex shaker, the suspension was transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=1.5. Dissolve 0, 50, 100, 200, 400, and 800 μg of compound A3 in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL, and transfer them into the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 26%, 21%, 33%, 79%, and 81%.

[0126] Cultivate Chlorella vulgaris to a cell concentration of 2×10^7 / mL, and transfer 10mL of the culture to each 60mL hydrogen production glass tube. Dissolve 0, 50, 100, 200, 400, and 800μg of compound A3 in 100μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80μg / mL, and transfer them to the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100μmol·m -2s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 7%, 4%, 5%, 6%, and 4%.

[0127] Embodiment 23

[0128] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 30 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, and 30 mL of fresh culture medium was added. After suspension was shaken on a vortex shaker, the suspension was transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=1.5. Dissolve 0, 50, 100, 200, 400, and 800 μg of phenothiazine compound A2 in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL, and transfer them into the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100 μmol·m -2 s -1 , at a temperature of 27-28°C. Let stand for 12 hours. After standing, extract 5mL of algae liquid from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. After 12h, centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, -2%, 3%, 18%, 39%, 48% ( Figure 7 ).

[0129] Cultivate Chlorella vulgaris to a cell concentration of 5×10^7 / mL, and transfer 10mL of the culture to each 60mL hydrogen production glass tube. Dissolve 0, 50, 100, 200, 400, and 800μg of phenothiazine compound A2 in 100μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80μg / mL, and transfer them to the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100μmol·m -2 s -1 , at a temperature of 27-28°C. Let stand for 12 hours. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. After 12h, centrifuge at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 20%, 15%, 50%, 75%, 80% ( Figure 8 ).

[0130] Embodiment 24

[0131] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 30 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, and 30 mL of fresh culture medium was added. After suspension was shaken on a vortex shaker, the suspension was transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=1.5. Dissolve 0, 50, 100, 200, 400, and 800 μg of compound A5 in 100 μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80 μg / mL, and transfer them into the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100 μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28℃. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4℃ refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0, 19%, 22%, 29%, 79%, and 82%.

[0132] Cultivate Chlorella vulgaris to a cell concentration of 5×10^6 / mL, and transfer 10mL of the culture to each 60mL hydrogen production glass tube. Dissolve 0, 50, 100, 200, 400, and 800μg of compound A5 in 100μL of DMSO to obtain solutions with concentrations of 0, 5, 10, 20, 40, and 80μg / mL, and transfer them to the corresponding hydrogen production glass tubes. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix, then place it under a light intensity of 100μmol·m -2 s -1 , and let it stand for 12 hours at a temperature of 27-28°C. After standing, extract 5mL of algae solution from each tube and centrifuge at 4000r / min for 10min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 5mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator and let it stand for 12h. Then centrifuge it at 4000r / min for 10min, detect the absorbance value of the supernatant at 665nm, and calculate the chlorophyll a concentration. The inhibition rate of protein nucleus Chlorella is calculated to be 0, 8%, 10%, 11%, 7%, and 6%.

[0133] Embodiment 25

[0134] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, 10 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, 30 mL of fresh culture medium was added, and the suspension was shaken on a vortex shaker and transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=0.5 solution, dissolve 50 μg of compound A2 in 100 μL of DMSO, i.e., the concentration is 5 μg / mL, and transfer it into a glass tube. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix it, then place it in a glass tube with a light intensity of 100 μmol·m -2 s -1, at a temperature of 27-28°C. On days 0, 1, 3, 5, and 7, take photos and extract 3 mL of algae solution, and centrifuge at 4000 r / min for 10 min. After removing the supernatant, shake on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 3 mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator for 12 hours. Then centrifuge at 4000 r / min for 10 minutes, detect the absorbance value of the supernatant at 665 nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0% on the 0th day, 13% on the 1st day, 48% on the 3rd day, 48% on the 5th day, and 37% on the 7th day. It can be seen that the concentration of 5 μg / mL compound A2 has the best effect on inhibiting cyanobacteria on the third day ( Fig. 9 ).

[0135] Embodiment 26

[0136] Cyanobacteria were cultured to a cell density of OD 730 After nm=1.5, take 20 mL of algae solution and centrifuge at 4000 r / min. After discarding the supernatant, add 30 mL of fresh culture medium, vortex and suspend on a vortex shaker, and transfer to a 60 mL glass tube to obtain the cell density OD 730 nm=1. Dissolve 50 μg of compound A2 in 100 μL of DMSO, i.e., the concentration is 5 μg / mL, and transfer it into a glass tube. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix it. Then place it in a glass tube with a light intensity of 100 μmol·m -2 s -1 , at a temperature of 27-28°C. On days 0, 1, 3, 5, and 7, take photos and extract 3 mL of algae solution, and centrifuge at 4000 r / min for 10 min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 3 mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator for 12 hours. Then centrifuge it at 4000 r / min for 10 minutes, detect the absorbance value of the supernatant at 665 nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0% on the 0th day, 19% on the 1st day, 23% on the 3rd day, 22% on the 5th day, and 22% on the 7th day. It can be seen that the concentration of 5 μg / mL compound A2 has the best effect on inhibiting cyanobacteria on the third day ( Fig.10 ).

[0137] Embodiment 27

[0138] Cyanobacteria were cultured to a cell density of OD 730After nm=1.5, 30 mL of algae solution was centrifuged at 4000 r / min, the supernatant was discarded, and 30 mL of fresh culture medium was added. After suspension was shaken on a vortex shaker, the suspension was transferred into a 60 mL glass tube to obtain the cell density OD 730 nm=1.5. Dissolve 50 μg of compound A2 in 100 μL of DMSO, i.e., the concentration is 5 μg / mL, and transfer it into a glass tube. Seal the mouth of the hydrogen production tube with a rubber stopper and shake it slightly to mix it, then place it in a glass tube with a light intensity of 100 μmol·m -2 s -1 , at a temperature of 27-28°C. On days 0, 1, 3, 5, and 7, take photos and extract 3 mL of algae solution, and centrifuge at 4000 r / min for 10 min. After removing the supernatant, shake it on a vortex shaker until there are no algae cells attached to the wall of the centrifuge tube, add 3 mL of methanol, continue to shake on the vortex shaker for 2 minutes, and then put it in a 4°C refrigerator for 12 hours. Then centrifuge it at 4000 r / min for 10 minutes, detect the absorbance value of the supernatant at 665 nm, and calculate the chlorophyll a concentration. The inhibition rate of cyanobacteria is calculated to be 0% on the 0th day, 9% on the 1st day, 21% on the 3rd day, 16% on the 5th day, and 13% on the 7th day. It can be seen that the concentration of 5 μg / mL compound A2 has the best effect on inhibiting cyanobacteria on the third day ( Fig.11 ).

[0139] It can be seen from the above examples that the phenothiazine derivatives of the present invention have a significant inhibitory effect on cyanobacteria, and the inhibitory effect can reach more than 95%. Under the same conditions, they have no obvious killing effect on Chlorella pyrenoidosa, and have good selectivity for green algae (using Chlorella pyrenoidosa as a model).

[0140] The above description of the embodiments is to facilitate the understanding and use of the invention by those skilled in the art. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative work. Therefore, the present invention is not limited to the above embodiments, and improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the present invention should be within the scope of protection of the present invention.

Claims

1. Use of a phenothiazine derivative in the preparation of a drug or preparation for inhibiting the growth of cyanobacteria, wherein the phenothiazine derivative is selected from any of the following compounds:

2. The use according to claim 1, characterized in that: The cyanobacteria is Synechocystis pcc6803.

3. The use according to claim 1, characterized in that: The medicine or preparation uses the phenothiazine derivative as an active ingredient, and its effective dosage is 0.01-80 μg / mL.

4. The use according to claim 1, characterized in that: The effective dose of the phenothiazine derivative is 5 μg / mL.

5. A cyanobacteria inhibitor, characterized in that: The method comprises one or more phenothiazine derivatives as claimed in claim 1, wherein the concentration of the phenothiazine derivatives is 0.01-80 μg / mL.

6. The cyanobacteria inhibitor according to claim 5, characterized in that The concentration of the phenothiazine derivative was 5 μg / mL.

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