Application of podophyllotoxin in preparation of marine antifouling agent for preventing adhesion of large fouling organisms

Abstract

The invention discloses application of podophyllotoxin in preparation of a marine antifouling agent for preventing adhesion of large fouling organisms. The invention also provides a marine antifouling agent taking podophyllotoxin or the derivative thereof as an effective component. The antifouling agent can optionally contain auxiliary components such as isothiazolinone compounds and the like, and can be prepared into antifouling paint to be applied to the surfaces of marine artificial facilities such as culture netting, ships, wharfs and the like in manners of spraying, dipping and the like. The podophyllotoxin has natural biological activity, an environment-friendly antifouling scheme is provided, the problems that in the existing antifouling technology, an antifouling agent is not high in activity, large in addition amount and poor in environmental friendliness are solved, and the podophyllotoxin is particularly suitable for preventing and treating attachment of large fouling organisms such as perna viridis.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of marine antifouling, and particularly relates to a use of podophyllotoxin in the preparation of a marine antifouling agent for preventing the attachment of large-scale fouling organisms. BACKGROUND

[0002] Marine biofouling refers to the attachment, growth and accumulation of microorganisms, algae and large animals on the surfaces of ships, marine engineering facilities and the like. This phenomenon can cause a series of serious hazards: for the shipping industry, the attachment of organisms on the ship body significantly increases the surface roughness and the sailing resistance, leading to a decrease in sailing speed and a substantial increase in fuel consumption, and indirectly aggravating the emission of greenhouse gases; for marine engineering and fisheries, the fouling organisms increase the structural load of offshore platforms and the like, accelerate metal corrosion, block seawater cooling pipelines, and cause mesh blockage of culture nets and competition for food with cultured organisms; for the marine environment and safety, fouling can interfere with the data accuracy of monitoring instruments, and the attached organisms carried by ocean-going ships can more likely cause the invasion of alien species and disrupt the ecological balance. Therefore, the development of efficient antifouling technology is of great significance to the safe and efficient operation of marine economic activities and the protection of the marine ecological environment.

[0003] Coating antifouling paint is the most economical and effective technical means for preventing and treating marine biofouling at present, and the core lies in the controllable release of antifouling agents. However, the development of mainstream antifouling agents faces severe environmental challenges. Early organic tin antifouling agents, which are highly efficient but highly toxic and have high residues, have been globally banned by the International Convention on the Control of Harmful Anti-Fouling Systems on Ships (AFS Convention) since January 1, 2008 due to their serious harm to the marine environment. Subsequently, cuprous oxide became the main replacement, but the continuous accumulation of copper elements in the marine environment also poses an ecological risk, and some countries and regions have begun to restrict the use of copper-containing antifouling paint.

[0004] Under this background, it has become an urgent technical requirement in the field to develop new environmentally friendly antifouling agents that are efficient, low-toxic and environmentally compatible.

[0005] Podophyllotoxin is a natural product in the rhizome of Podophylloideae plants of Berberidaceae. It can be obtained not only by extraction from the rhizome of Podophylloideae plants of Berberidaceae, but also by plant callus culture or endophytic fungus fermentation. At present, podophyllotoxin and its derivatives (such as etoposide, teniposide, etc.) are mainly used in the fields of anti-tumor, anti-virus and anti-inflammatory drugs, and there is no report on using the biological activity thereof as a marine antifouling agent. SUMMARY

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and provide a new use for podophyllotoxin—its use in the preparation of marine antifouling agents for preventing the attachment of large fouling organisms, thus solving the problems of efficiency and environmental friendliness of marine antifouling technologies in the aforementioned background art.

[0007] The technical solution adopted by the present invention to solve its technical problem is: to provide the use of podophyllotoxin in the preparation of marine antifouling agents that prevent the attachment of large fouling organisms.

[0008] The molecular formula of the podophyllotoxin is C 22 H 22 O8, with a molecular weight of 414.41, has the following structural formula:

[0009]

[0010] In a preferred embodiment of the invention, the marine antifouling agent further includes at least one of isothiazolinone compounds, 2-(p-chlorophenyl)-3-cyano-4-bromo-5-trifluoromethyl-pyrrole, N-2,4,6-trichlorophenylmaleimide and pyridinetriphenylborane.

[0011] In a preferred embodiment of the invention, the large fouling organism is the jade mussel.

[0012] In a preferred embodiment of the invention, the material is applied to the surface of a marine artificial facility, which includes aquaculture nets, aquaculture platforms, docks, ships, buoys, and marine monitoring instruments.

[0013] In a preferred embodiment of the invention, the application method includes spraying, dipping, or coating after being formulated into an antifouling coating.

[0014] Another technical solution adopted by the present invention to solve its technical problem is: to provide a marine antifouling agent that prevents the attachment of large fouling organisms, the active ingredient of which is podophyllotoxin.

[0015] In a preferred embodiment of the invention, the marine antifouling agent further comprises at least one of isothiazolinone compounds, 2-(p-chlorophenyl)-3-cyano-4-bromo-5-trifluoromethyl-pyrrole, N-2,4,6-trichlorophenylmaleimide and pyridinetriphenylborane.

[0016] In a preferred embodiment of the invention, the material is applied to the surface of a marine artificial facility by spraying, impregnation, or formulation into an antifouling coating.

[0017] In a preferred embodiment of the present invention, the antifouling coating further comprises a base material, pigments, additives, and solvents. The base material includes (but is not limited to): acrylic resin, zinc / copper / silicone acrylate resin, chlorinated rubber, chlorinated ether resin, epoxy resin, and polyurethane resin. The pigments include (but are not limited to): lemon yellow, phthalocyanine blue, titanium dioxide, zinc oxide, iron oxide red, and talc. The additives mainly include thixotropic agents, stabilizers, and anti-settling agents. The solvents include (but are not limited to): xylene, cyclohexanone, and n-butanol.

[0018] In a preferred embodiment of the invention, the applied marine artificial facilities include aquaculture nets, aquaculture platforms, docks, ships, buoys, and marine monitoring instruments.

[0019] In a preferred embodiment of the invention, the large fouling organism prevented is the jade mussel.

[0020] Compared with the prior art, this technical solution has the following advantages:

[0021] 1. This invention provides a novel use of the known antitumor / antiviral substance podophyllotoxin in the field of marine antifouling, at extremely low concentrations (EC50) not exceeding 1.0 μg / mL. 50 It can effectively inhibit the attachment of large fouling organisms such as mussels at a concentration of 0.83 μg / mL. This invention's novel application has successfully expanded this substance from the pharmaceutical field to the marine antifouling field, exhibiting a specific activity far exceeding that of conventional antifouling agents, and significantly reducing the dosage and cost.

[0022] 2. This invention differs from its traditional medicinal uses by utilizing podophyllotoxin as a natural and renewable antifouling agent, achieving a cross-domain application from "medicinal" to "antifouling," with significant environmental advantages. Compared to existing synthetic heavy metal antifouling agents (such as cuprous oxide), this shift in application fundamentally avoids heavy metal pollution, providing a more environmentally compatible solution that aligns with the urgent needs of sustainable development.

[0023] 3. This invention utilizes the lipophilic properties of podophyllotoxin, which is poorly soluble in water but readily soluble in organic solvents—a property often considered a challenge in pharmaceutical applications—and turns it into an advantage, making it easy to disperse uniformly in coatings and achieve stable, slow release.

[0024] 4. The raw materials for this invention are widely available and environmentally friendly. They can be prepared in large quantities through fermentation by plant endophytic fungi or culture of plant callus tissue. For the large-scale demand brought about by the new use of podophyllotoxin as an antifouling agent, there are still mature and sustainable supply channels. Compared with some natural antifouling agents, it has certain economic advantages, which provides a guarantee for its promotion and application in the field of marine antifouling. Attached Figure Description

[0025] Figure 1 The results of Example 1 show the inhibitory effect of podophyllotoxin on byssal secretion in jade mussels and its effect on the survival of jade mussels;

[0026] Figure 2 The results of the hanging plate test in Example 2 are shown, where a-control group and b-experimental group;

[0027] Figure 3 The results of the hanging plate test in Example 3 are shown, where a-control group and b-experimental group. Detailed Implementation

[0028] The technical solution of the present invention will be further explained and described below with reference to specific embodiments and accompanying drawings.

[0029] Example 1

[0030] This embodiment provides the use of podophyllotoxin in the preparation of marine antifouling agents to prevent the attachment of large fouling organisms. Specifically, a marine antifouling agent prepared with podophyllotoxin as the active ingredient is applied to large fouling organisms—emerald mussels—collected from the sea area to simulate application on the surface of marine artificial facilities and to test the inhibitory effect of podophyllotoxin on emerald mussels. The specific operation is as follows:

[0031] (1) Acquisition of juvenile jade mussels: Adult jade mussels collected from the sea area were brushed clean and cultured overnight under aeration at 25℃ and salinity 28. Juvenile jade mussels with a size of about 1cm and similar growth conditions were selected, and the byssal threads were cut off with scissors to obtain individual jade mussels. They were then washed with clean seawater for testing.

[0032] (2) Antifouling activity test: Take a clean 24-well plate, add 1990 μL of membrane-filtered seawater (membrane pore size 0.22 μm) to the well, then add 10 μL of the prepared solution (dissolved in DMSO). Set up a membrane-filtered seawater control group (ck1) and a 0.5% DMSO seawater solution control group (ck2). The concentration gradient of the experimental groups is set at 0.5, 1, 5, 10 and 50 μg / mL, with 10 parallel samples in each group. Gently place the selected green mussels into the wells, one per well. Incubate statically in a dark environment at room temperature. After 24 h, remove the green mussels and rinse each well 3 times with clean water. After rinsing, drain the seawater from the wells and add about 3 mL of Coomassie brilliant blue staining solution for 1 h. Observe and record the number of green mussels with blue-stained byssal discs in each well, and record the mortality of the green mussels.

[0033] The results of this embodiment regarding the inhibitory effect of podophyllotoxin on byssal secretion in jade mussels and its impact on the survival of jade mussels are attached. Figure 1 As shown. Its half-inhibitory byssal concentration EC 50The median lethal concentration (LC50) is 0.83 μg / mL. 50 >50 μg / mL, exhibits extremely high inhibitory activity against byssal secretion in green mussels, with low toxicity, demonstrating environmentally friendly characteristics.

[0034] Example 2

[0035] The difference between Example 2 and Example 1 is that in this example, podophyllotoxin is used as an active ingredient in the formulation of the antifouling coating; the specific components of the antifouling coating are:

[0036] 30 parts by weight of zinc acrylate resin

[0037] 12 parts by weight of rosin

[0038] 8 parts by weight of titanium dioxide

[0039] 1 part by weight of organic bentonite

[0040] 23 parts by weight of talc

[0041] 10 parts by weight of podophyllotoxin

[0042] 16 parts by weight of xylene

[0043] The specific preparation method includes the following steps:

[0044] (1) Add xylene to the high-speed dispersion container in proportion, start the high-speed disperser, and stir at a low speed of 400 r / min;

[0045] (2) Slowly add rosin to the material obtained in step (1) and stir at low speed until the rosin is completely dissolved. Then add zinc acrylate resin and continue stirring at low speed until it is completely dissolved.

[0046] (3) Add organic bentonite, titanium dioxide, talc and podophyllotoxin to the material obtained in step (2) in sequence, adjust the speed to 1200 r / min and stir for 10 min;

[0047] (4) Grind the material obtained in step (3) to a fineness of less than 60 μm using a basket mill. After testing the fineness and viscosity, the marine antifouling coating can be obtained.

[0048] The antifouling performance of the marine antifouling coating prepared in this embodiment is tested as follows:

[0049] The shallow-sea siding was tested in the waters off Nan'ao, Guangdong, in accordance with the national standard GB / T 5370-2007. Figure 2 As shown, after 5 months of seabed slab application, the experimental group showed no significant large fouling organisms, while the blank control group had a large amount of fouling organisms attached.

[0050] Example 3

[0051] The difference between Example 3 and Example 2 is that in this example, podophyllotoxin and bromopyrrolidone (i.e., 2-(p-chlorophenyl)-3-cyano-4-bromo-5-trifluoromethyl-pyrrole) are formulated into a marine antifouling agent at a weight ratio of 1:1, and this agent is used as an active ingredient in the formulation of an antifouling coating; the specific components of the antifouling coating are as follows:

[0052] 30 parts by weight of zinc acrylate resin

[0053] 12 parts by weight of rosin

[0054] 8 parts by weight of titanium dioxide

[0055] 1 part by weight of organic bentonite

[0056] 23 parts by weight of talc

[0057] Podophyllotoxin 5 parts by weight

[0058] 5 parts by weight of bromopyrrolidinium

[0059] 16 parts by weight of xylene

[0060] The specific preparation method includes the following steps:

[0061] (1) Add xylene to the high-speed dispersion container in proportion, start the high-speed disperser, and stir at a low speed of 400 r / min;

[0062] (2) Slowly add rosin to the material obtained in step (1) and stir at low speed until the rosin is completely dissolved. Then add zinc acrylate resin and continue stirring at low speed until it is completely dissolved.

[0063] (3) Add organic bentonite, titanium dioxide, talc, podophyllotoxin and bromopyrrolidone to the material obtained in step (2) in sequence, adjust the speed to 1200 r / min and stir for 10 min;

[0064] (4) Grind the material obtained in step (3) to a fineness of less than 60 μm using a basket mill. After testing the fineness and viscosity, the marine antifouling coating can be obtained.

[0065] The antifouling performance of the marine antifouling coating prepared in this embodiment is tested as follows:

[0066] The shallow-sea siding was tested in the waters off Nan'ao, Guangdong, in accordance with the national standard GB / T 5370-2007. Figure 3 As shown, after 12 months of seabed slab application, no large fouling organisms were found in the experimental group, while the blank control group was covered with fouling organisms.

[0067] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of podophyllotoxin in the preparation of marine antifouling agents to prevent the attachment of large fouling organisms.

2. The use according to claim 1, characterized in that: Marine antifouling agents also include at least one of isothiazolinone compounds, 2-(p-chlorophenyl)-3-cyano-4-bromo-5-trifluoromethyl-pyrrole, N-2,4,6-trichlorophenylmaleimide, bromopyrronitrile and pyridinetriphenylborane.

3. The use according to claim 1, characterized in that: The large fouling organisms mentioned are jade mussels, and the half-inhibitory concentration (EC50) of podophyllotoxin or its derivatives in inhibiting byssal secretion in jade mussels is [not specified]. 50 Not higher than 1.0 μg / mL.

4. The use according to claim 1, characterized in that: It is applied to the surface of marine artificial facilities, including aquaculture nets, aquaculture platforms, docks, ships, buoys, and marine monitoring instruments.

5. The use according to claim 4, characterized in that: Application methods include spraying, dipping, or coating after formulation into an antifouling coating.

6. A marine antifouling agent for preventing the attachment of large fouling organisms, characterized in that: Its active ingredient is podophyllotoxin.

7. A marine antifouling agent for preventing the attachment of large fouling organisms according to claim 6, characterized in that: It also includes at least one of isothiazolinone compounds, 2-(p-chlorophenyl)-3-cyano-4-bromo-5-trifluoromethyl-pyrrole, N-2,4,6-trichlorophenylmaleimide and pyridinetriphenylborane.

8. A marine antifouling agent for preventing the attachment of large fouling organisms according to claim 6, characterized in that: It is applied to the surface of marine artificial facilities by spraying, impregnation, or formulation into antifouling coatings.

9. A marine antifouling agent for preventing the attachment of large fouling organisms according to claim 8, characterized in that: The marine artificial facilities used include aquaculture nets, aquaculture platforms, docks, ships, buoys, and marine monitoring instruments.

10. A marine antifouling agent for preventing the attachment of large fouling organisms according to claim 6, characterized in that: The large fouling organisms prevented are jade mussels.

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