CN112063212A

CN112063212A - Application of benzodiazepine indole diketone alkaloid compound in preparation of marine fouling organism control agent

Info

Publication number: CN112063212A
Application number: CN202010791170.1A
Authority: CN
Inventors: 严涛; 曹文浩; 佘志刚; 刘昭明; 林明晴; 林岳光
Original assignee: South China Sea Institute of Oceanology of CAS
Current assignee: South China Sea Institute of Oceanology of CAS
Priority date: 2020-08-07
Filing date: 2020-08-07
Publication date: 2020-12-11
Anticipated expiration: 2040-08-07
Also published as: CN112063212B

Abstract

The invention discloses an application of a benzodiazepine dione alkaloid compound in preparation of a marine fouling organism control agent, wherein the structural formula of the benzodiazepine dione alkaloid compound is shown as a formula (I). The benzodiazepine indole dione alkaloid compound has obvious effect of inhibiting marine organism adhesion when coated on solid surface in low dosage, and thus may be used in preparing marine fouling organism preventing and controlling agent. The marine biofouling inhibitor is a naturally-occurring organic compound, does not pollute the water environment and cause the enrichment of the marine biofouling inhibitor in organisms through food chain transfer, is environment-friendly and high in safety, does not contain heavy metal elements such as copper and tin while effectively inhibiting the attachment of marine organisms, has good social benefits from the aspect of environmental protection, has large popularization and application potentials, and has good application prospects in the prevention and removal of marine biofouling organisms.

Description

Application of benzodiazepine indole diketone alkaloid compound in preparation of marine fouling organism control agent

The technical field is as follows:

the invention belongs to the field of natural products, and particularly relates to an application of a benzodiazepine dione alkaloid compound in preparation of a marine fouling organism control agent.

Background art:

the marine fouling organisms refer to various organisms fixed or inhabiting underwater parts of ships and marine facilities, and the harm caused by the marine fouling organisms is mainly to increase resistance, reduce navigational speed, increase fuel consumption and CO₂Discharging; blocking a seawater pipeline system, changing a metal corrosion process, and initiating local corrosion or perforation corrosion; the dynamic load effect is increased, so that the drift, unbalance and even overturn of the facility are caused; compete with the cultured objects (such as shellfish) for the attachment base and the bait, hinder the growth and development of the cultured objects and reduce the quality of the products.

The antifouling paint is one of the most widely applied technical methods in preventing and removing marine fouling organisms. However, the traditional antifouling paint takes poison release as a main way, inhibits the attachment of fouling organisms through poisoning action, has the defects of high toxicity, short effective period and the like, and the released antifouling agent also has the risk of harming the marine ecological environment, so that the development of a novel, efficient, low-toxicity and environment-friendly antifouling agent becomes a problem and a research hotspot which are urgently needed to be solved at present.

Since marine fouling organisms are composed of animals, plants and microorganisms, the more harmful and difficult to remove species are mainly limy hulls, sessile stemless vines (barnacles) and bivalve mollusks (mussels and oysters). Therefore, screening tests for antifouling compounds have mostly been performed on stemless vines and bivalves.

In tropical coastal waters, balanus reticulatus is a typical representative of sessile tendrils and is an absolutely dominant species in the biofouling organism community; perna viridis is a common bivalve mollusk in the east and south seas, and is also an important fouling organism species attached to ships, navigations (buoys) and aquaculture facilities. Therefore, the verification test of the invention adopts the balanus reticulates and the perna viridis as experimental objects, and the obtained research results have wide representativeness.

Fouling organisms are generally divided into two life stages, namely a planktonic life stage from the development of larva out of an egg membrane to the intermittent exploration of the surface of an object to prepare for attachment and metamorphosis; once the larvae are selected to be in the settlement position, the larvae are attached to the surface of the attachment base and transformed into larvae, and then the larvae are in the fixation or attachment life stage. From an insult point of view, it begins after fixation or attachment to humans. If the settlement and metamorphosis of the larvae can be effectively inhibited, the purpose of preventing and removing can be achieved. Therefore, the invention adopts the larvae of the two types of marine organisms as experimental objects to test the antifouling effect of the compound, and has scientific reasonability and representative significance.

The clarification of the natural chemical antifouling mechanism of marine organisms can provide reference for developing pollution-free antifouling technology, and the marine organisms themselves are important sources of novel antifouling agents. The content of relevant substances in marine organisms is low, and the structure is relatively complex, so that the marine organism is inconvenient to deeply develop and widely apply. Therefore, the natural antifouling agent is searched from terrestrial plants which have large resource quantity and are convenient for planting and cultivation, and the natural antifouling agent is not only a brand new attempt, but also has important theoretical and practical significance.

Benzodiazepine indole dione alkaloid compound named (5aS,13aS,14aS) -5-acetyl-14a- (2-methyl-3-en-2-yl) -5a,13a,14,14a-tetrahydrobenzo [5',6'][1,4]Diazepines [1', 2':1,5]Pyrrole [2,3-b ]]Indole-7,13(5H,12H) -dione, having the name (5aS,13aS,14aS) -5-acetyl-14a- (2-methyllbut-3-en-2-yl) -5a,13a,14,14a-tetrahydrobenzo [5',6'][1,4]diazepino[1',2':1,5]pyrrolo[2,3-b]indole-7,13(5H,12H) -dione, colorless crystal, isolated from an endophytic fungus of the mangrove species Aspergillus terreus H010. The molecular formula is C₂₅H₂₅N₃O₃The molecular weight is 415, and the chemical structural formula is shown as a formula (I).

The structure of the compound has been reported (Rank C, Phipps RK, Harris P, et al 2006.epi-Aszonalenins A, B, and C from Aspergillus novo microorganisms. tetrahedron letters,47(34): 6099-plus 6102), but in the field of marine biofouling organism control, there is no report on the disclosure of its anti-marine biofouling organism attachment activity and its use in the field of biofouling organism control.

The invention content is as follows:

the invention aims to provide application of a benzodiazepine indole diketone alkaloid compound in preparing a marine fouling organism control agent.

The benzodiazepine indole dione alkaloid compound of the present invention may be coated in an amount of 10. mu.g/cm²When compared with the control group, the compound has obvious inhibition effect on the attachment of barnacle larvae (p)<0.01). The preferred coating amount is 10. mu.g/cm². Furthermore, the experiment shows that the coating amount of the benzodiazepine indole diketone alkaloid compound is 1 mu g/cm²In time, the attachment rate of mussel larvae was much lower than that of the control group, and the difference was extremely significant (p)<0.01), indicating that the benzodiazepine dione alkaloid compound is effective in inhibiting the attachment of perna viridis larvae. It is thus seen that benzodiazepine dione alkaloid compounds have a very good inhibitory effect on the adhesion of tendrils and bivalves.

Therefore, the invention provides an application of a benzodiazepine diketone alkaloid compound in preparing a marine fouling organism control agent, wherein the structural formula of the benzodiazepine diketone alkaloid compound is shown as a formula (I):

the marine biofouling organism control agent is preferably a barnacle larva control agent (such as a balanus reticulatus larva control agent) or a mussel larva control agent (such as a perna viridis epidermoid larva control agent).

Compared with the prior art, the invention has the following beneficial effects:

the benzodiazepine indole dione alkaloid compound has obvious effect of inhibiting marine organism adhesion when coated on solid surface in low dosage, and thus may be used in preparing marine fouling organism preventing and controlling agent. The benzodiazepine indole diketone alkaloid compound is a naturally-occurring organic compound, does not pollute the water environment and cause the enrichment of the benzodiazepine indole diketone alkaloid compound in organisms through food chain transfer, is environment-friendly and high in safety, effectively inhibits the attachment of marine organisms, does not contain heavy metal elements such as copper and tin, has good social benefits from the aspect of environmental protection, has large popularization and application potentials, and has good application prospects in the prevention and removal of marine fouling organisms.

The specific implementation mode is as follows:

the following examples are further illustrative of the present invention and are not intended to be limiting thereof.

Example 1:

a quantitative amount of the benzodiazepine dione alkaloid compound was dissolved in chloroform-methanol (3:1/V: V) to give a solution having a concentration of the benzodiazepine dione alkaloid compound of 282.6. mu.g/mL, and 1mL of the solution was added to a 6 cm-diameter petri dish and allowed to uniformly cover the bottom of the dish. After the solvent is completely volatilized, the content of the benzodiazepine dione alkaloid compound coated on the bottom of the culture dish is 10 mu g/cm². 13ml of filtered sterile seawater was added.

Control group: 1ml of solvent (chloroform-methanol (3:1/V: V)) is added to distribute it evenly on the bottom of the culture dish, and 13ml of filtered and sterilized seawater is added after the solution is completely volatilized.

Blank group: 13ml of filtered sterile seawater was added.

Each of the experimental, blank and control groups was provided with 4 replicates. In the adhesion test process, barnacle larvae are randomly distributed to a blank group, a control group and an experimental group, about 30 larvae are placed in each parallel sample, and the sample is placed in a constant-temperature incubator at the temperature of 30 ℃ to be cultured in a dark environment. After 120 hours of culture, the number of attached larvae in each parallel sample was recorded, the attachment rate of larvae (number of attached larvae/number of inserted larvae) in each parallel sample was calculated, the average attachment rate of larvae in each group (i.e., blank group, control group, and experimental group) [ (attachment rate of parallel sample 1 + attachment rate of parallel sample 2 + attachment rate of parallel sample 3 + attachment rate of parallel sample 4)/4 ] was counted, and the significance of differences between groups was examined to confirm the effect of the related compound in inhibiting the attachment of pot larvae. The adhesion rate calculation method of example 2 was also calculated with reference to this method.

Table 1 lists the attachment and mortality rates of the cyprids of the experimental, control and blank groups. It can be seen that, after 120 hours of culture in the incubator, the attachment rate of larvae of the blank group is 74.2%, that of larvae of the control group is 74.5%, and that there is no significant difference (p >0.05) between the attachment rates of larvae of Venus asiatica of the blank group and the control group, indicating that harmful substances affecting the activity of larvae of Venus asiatica are not left after the volatilization of chloroform-methanol as a solvent, and the compound is suitable for dissolving the compound. As for the experimental group treated by the benzodiazepine dione alkaloid compound, the attachment rate of the cyprids is 0, and the difference with the control group is very obvious (p is less than 0.01), which indicates that the compound can effectively inhibit the attachment of cyprids of balanus reticulatus.

Table 1: attachment status of cyprids of balanus reticulates

Group of	Test dose	Adhesion Rate (%)
			Blank group	－	74.2
Control group	－	74.5
			Experimental group	10μg/cm²	0

Example 2

Experimental groups: the benzodiazepine indole dione alkaloid compound was dissolved in chloroform-methanol (3:1/V: V) to prepare a solution in which the concentration of the benzodiazepine indole dione alkaloid compound in the solution was 28.26. mu.g/mL. 1mL of this solution was added to a 6cm diameter dish and allowed to cover the bottom of the dish uniformly. After the solvent is completely volatilized, the content of the benzodiazepine dione alkaloid compound coated on the bottom of the culture dish is 1.0 mu g/cm². 13mL of filtered sterile seawater was added.

Control group: 1mL of solvent (chloroform-methanol (3:1/V: V)) was added to distribute the solution evenly over the bottom of the dish, and 13mL of filtered sterile seawater was added after the solvent had evaporated completely.

Blank group: 13mL of filtered sterile seawater was added.

Determination of the number of larvae: taking water containing perna viridis facial disc larvae from the nursery pond and concentrating by bolting silk. Taking 50mL of the water body after three times, dripping 1-2 drops of formalin solution to kill the larvae, and counting under a microscope to obtain an average value, namely the density of the larvae in the water body.

The experimental group, the blank group and the control group are all provided with 4 parallel samples, a certain amount of water containing perna viridis faceplates larvae is absorbed according to the density of the larvae in the water, and about 30 larvae are added into each sample. Culturing in dark environment in an incubator at a temperature of about 26 ℃. Statistical analysis of the final attachment and death status of each group of larvae was performed after 72 hours of culture.

Table 2 lists the attachment and mortality rates of perna viridis larvae in the experimental, control and blank groups. It can be seen that, after 72 hours of incubation in the incubator, the attachment rate of larvae in the blank group was about 48.2%, that in the control group was about 49.6%, and that there was no significant difference in the attachment rate of larvae in the dough plates between the blank group and the control group (p >0.05), indicating that chloroform-methanol as a solvent was volatilized without leaving harmful substances affecting the activity of larvae in the dough plates, and thus it was suitable for dissolving the compound. The adhesion rate of larvae of the experimental group treated by the benzodiazepine dione alkaloid compound is 0, which is far lower than that of the control group, and the difference is extremely obvious (p is less than 0.01), which indicates that the compound can effectively inhibit the adhesion of larvae of perna viridis.

Table 2: emerald mussel face plate larva attachment condition

Group of	Test dose	Percentage of adhesion%
			Blank group	－	48.2
Control group	－	49.6
			Experimental group	1.0μg/cm²	0

Claims

1. The application of the benzodiazepine diketone alkaloid compound in preparing the marine fouling organism control agent is shown in the formula (I):

2. use according to claim 1, wherein the marine biofouling organism control agent is a barnacle larva control agent.

3. Use according to claim 2, wherein the barnacle larvae control agent is a balanus reticulatus avenae larvae control agent.

4. The use according to claim 1, wherein the marine biofouling organism control agent is a mussel larva control agent.

5. The use as claimed in claim 4, wherein the mussel larvae control agent is emerald mussel faceplates larvae control agent.