CN103265550B - Alkaloid compounds and preparation method thereof and the application in the anti-marine biofouling coating of preparation and antitumor drug - Google Patents
Alkaloid compounds and preparation method thereof and the application in the anti-marine biofouling coating of preparation and antitumor drug Download PDFInfo
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Abstract
The invention discloses alkaloid compounds and preparation method thereof and the application in the anti-marine biofouling coating of preparation and antitumor drug.Its structure of alkaloid compound as shown in formula I, wherein compound 1:R
1=H, R
2=OH, R
3=Cl; Compound 2:R
1=OH, R
2=H, R
3=H.Compound 1 and compound 2 are from fungi Aspergillus westerdijkiae DFFSCS013(CCTCC M2013078) preparative separation obtains fermented liquid.Alkaloid compound of the present invention has anti-ocean Macro-fouling Organisms grass tongue worm and kentrogon attachment activity, and the growth of the various tumor cell strains such as malignant myeloid cell lines K562 can be suppressed, therefore in the anti-marine biofouling coating of preparation and antitumor drug, there is good application prospect.
formula I is compound 1:R wherein
1=H, R
2=OH, R
3=Cl; Compound 2:R
1=OH, R
2=H, R
3=H.
Description
Technical field:
The invention belongs to biological technical field, be specifically related to alkaloid compounds and preparation method thereof and the application in the anti-marine biofouling coating of preparation and antitumor drug.
Background technology:
Fungi can produce the secondary metabolite of the various structures type comprising alkaloids, peptide class, polyketone class, steroidal and terpene etc., wherein much has antitumor, antibacterium, the biological activity such as antimycotic, antiviral, pest-resistant.Alkaloid is the alkaline organic compound that in natural product, a class is nitrogenous, and great majority have complicated ring texture, is one of effective constituent important in natural drug.There are many alkaloids for clinical example at present, as the camptothecine in camplotheca acuminata and the vincristine(VCR) in Vinca are used for antitumor etc.In addition, marine biofouling become people make full use of oceanic resources one of the matter of utmost importance that will urgently solve.Along with the poisonous paint such as TBT are disabled in the whole world, develop efficient, nontoxic, eco-friendly ocean novel antifouling agent demand urgent, the research of this respect is dashed forward important ecological significance and economic implications.Marine Natural Product Antifoulants agent belongs to nontoxic (or low toxicity) stain control agent, easy degraded, and do not endanger halobiontic life, be conducive to keeping ecological balance, therefore, develop Marine Natural Product Antifoulants agent and become one of important channel obtaining high effect nontoxic stain control agent.
Summary of the invention:
First object of the present invention is to provide a class and has antitumor and alkaloid compound that is anti-fouling activity.
The present invention is by multiple column chromatography and one dimension, two dimensional NMR wave spectrum, be separated from the fermented liquid of fungi Aspergillus westerdijkiae DFFSCS013 and obtain the new alkaloid compound of a class, this alkaloid compound has the cytotoxic activity suppressing multiple growth of cancer cells, can be used for preparing antitumor drug; And also have anti-marine biofouling attachment activity, can be used for preparing anti-marine biofouling coating, thus achieve object of the present invention.
Alkaloid compound of the present invention, its structure is as shown in formula I:
Formula I
Compound 1:R
1=H, R
2=OH, R
3=Cl; Compound 2:R
1=OH, R
2=H, R
3=H.
Second object of the present invention is to provide the preparation method of the alkaloid compound as shown in formula I.
The preparation method of the alkaloid compound as shown in formula I of the present invention, is characterized in that, comprise the following steps:
(1) fermented liquid of fungi Aspergillus westerdijkiae DFFSCS013 is prepared;
(2) fermented liquid ethyl acetate, methylene dichloride or chloroform solvent extraction step (1) obtained, concentrates and obtains ethyl acetate extract, dichloromethane extract or chloroform extract;
(3) by the ethyl acetate extract described in step (2), dichloromethane extract or chloroform extract are through normal pressure silica gel column chromatography, with chloroform-methanol, chloroform-acetone, chlorofonn-ethylacetate, sherwood oil-acetone or petroleum ether-ethyl acetate solvent systems are eluent, gradient elution is carried out from volume ratio 100:0 to 0:100, follow the trail of with thin-layer chromatography and merge component, by the component can launched with the chloroform-methanol solvent systems in volume ratio 95:5 to 90:10 scope on thin-layer chromatography, crude product is obtained through gel filtration chromatography, purified, obtain the alkaloid compound 1 and 2 as shown in formula I.
Fungi Aspergillus westerdijkiae DFFSCS013 can be inoculated in the applicable substratum of aspergillus fungi by the fermented liquid of the fungi Aspergillus westerdijkiae DFFSCS013 described in step (1), obtained under common fermentation condition.Fungi Aspergillus westerdijkiae DFFSCS013 is inoculated in PDA nutrient agar by preferred preparation method, cultivate 3 days for 26 DEG C, obtain cultivating the flat board having bacterial classification, then strain inoculation in flat board is entered in PDB liquid nutrient medium, cultivate 2 days in rotating speed 200rpm shaking table, temperature 26 DEG C, obtain seed liquor, then seed liquor is inoculated in rice solid medium, in temperature 26 DEG C of quiescent culture 26 days, obtain fermented liquid.The composition of above-mentioned PDA nutrient agar is potato 200g, glucose 20g, sea salt 30g, agar 20g and 1L water; The composition of PDB liquid nutrient medium is potato 200g, glucose 20g, sea salt 30g and 1L water; The composition of rice solid medium is rice 400g, yeast extract 2g, glucose 2g, sea salt 18g and water 600mL.
The most handy ethyl acetate of extraction described in step (2), the described concentrated method of routine that can adopt is as concentrating under reduced pressure.
Purifying described in step (3) can adopt chromatographic column to be separated or recrystallization.
Found by anti-ocean Macro-fouling Organisms grass tongue worm larva and the test of kentrogon attachment activity, in formula I of the present invention, compound 1 and compound 2 suppress the EC that careless tongue worm larva adheres to
50value is respectively 13.51 μ g/mL and 28.12 μ g/mL, and suppresses the EC of kentrogon attachment
50value is less than 25 μ g/mL, when measured maximum concentration 200 μ g/mL, no toxicity is not shown to larva, there is good anti-marine biofouling active, can for the preparation of anti-marine biofouling coating, as it infiltrated alone or in combination or being spread in film forming natural resin, polyethylene vinyl acetate multipolymer and other hydrolyzable, in the polymkeric substance such as solvable or insoluble resin, make antifouling paint, effective constituent to the surface that antifouling paint can discharge q.s reaches antifouling effect, and these compounds are active skull cap components, and it is not all soluble in water, very easily be dissolved in chloroform, the low polar organic solvent such as ethyl acetate, there is good lipophilicity, therefore can be applied to alone or in combination prepares in marine antifoulant, have a good application prospect.
By anti tumor activity in vitro screening experiment, result shows: the compound 1 and 2 shown in formula I of the present invention suppresses the IC of human leukemia cell line K562 growth
50value is respectively 44.2 and 52.8 μMs, and has the effect that certain suppression malignant melanoma cell strain A375, lung cancer cell types, cervical cancer cell lines HeLa, breast cancer cell line mcf-7, laryngeal cancer cell strain Hep-2, hepatoma H22 cells, colon-cancer cell strain SW-620 grow.
Therefore, the 3rd object of the present invention is to provide the alkaloid compound shown in formula I or the application of its salt in the anti-marine biofouling coating of preparation and antitumor drug.
Described antitumor drug is preferably human body leukemia medicament.
The present invention is from fungi Aspergillus westerdijkiae DFFSCS013(CCTCC M2013078) preparative separation obtains alkaloid compounds-compound 1 and compound 2 fermented liquid.Compound 1 and compound 2 have anti-ocean Macro-fouling Organisms grass tongue worm and kentrogon attachment activity, and the growth of the various tumor cell strains such as malignant myeloid cell lines K562 can be suppressed, therefore have a good application prospect in the anti-marine biofouling coating of preparation and antitumor drug.
Fungi Aspergillus westerdijkiae DFFSCS013 of the present invention is preserved in China typical culture collection center (CCTCC), address on March 12nd, 2013: China, Wuhan, Wuhan University, preserving number CCTCC M2013078.
accompanying drawing illustrates:
Fig. 1 is the NOESY coherent signal of compound 1 and 2, wherein 1 representation compound 1,2 representation compound 2.
Embodiment:
Following examples further illustrate of the present invention, instead of limitation of the present invention.
Embodiment 1: the preparation of alkaloid compound 1 and 2
PDA nutrient agar is like this preparation: by 200 grams of potatoes, 20 grams of glucose, 30 grams of sea salt, and 20 grams of agar mixing, add 1 premium on currency and fully dissolve, 121 DEG C of high pressure steam sterilizations 25 minutes.
PDB liquid nutrient medium was like this preparation: by 200 grams of potatoes, 20 grams of glucose, 30 grams of sea salt mixing, adds after 1 premium on currency fully dissolves, then is sub-packed in the Erlenmeyer flask of 500mL, every bottle of about 150mL, 121 DEG C of high pressure steam sterilizations 25 minutes.
Rice solid medium was like this preparation: by 400 grams of rice, 2 grams of yeast extracts, 2 grams of glucose, 18 grams of sea salt mixing, adds after 600 ml waters fully dissolve, loads in the Erlenmeyer flask of 5000mL, 121 DEG C of high pressure steam sterilizations 25 minutes.
With aseptic bamboo let by fungi Aspergillus westerdijkiae DFFSCS013 strain inoculation on PDA nutrient agar, cultivate 3 days for 26 DEG C, obtain cultivating the flat board having bacterial classification, then from flat board, choose about 3 microlitre strain inoculation with aseptic bamboo let to enter and be equipped with in the Erlenmeyer flask of PDB liquid nutrient medium containing 150mL is above-mentioned, cultivate 2 days in shaking table (rotating speed 200rpm) temperature 26 DEG C, obtain seed liquor, 50mL seed liquor is inoculated again in the triangular flask of above-mentioned every bottle of rice solid medium, after 26 days, fermented liquid is obtained in temperature 26 DEG C of quiescent culture.
3 times are extracted by cultivating through rice solid medium (altogether 2kg rice) the fermented liquid acetone-water (volume ratio 80:20) obtained, then this extracting solution is evaporated to without acetone residue, by the equal-volume extraction into ethyl acetate 3 times of the aqueous phase after concentrated, concentrating under reduced pressure obtains ethyl acetate extract 30g.After ethyl acetate extract purification on normal-phase silica gel (200-300 order) dry method mixes sample, load glass chromatography column (being about 500g containing 200-300 object purification on normal-phase silica gel), carry out Column at Normal Temperature chromatography, using chloroform-methanol as eluent, gradient elution is carried out, according to thin-layer chromatography (GF from volume ratio 100:0 to 0:100
254silica-gel plate) situation merges each cut, and reclaim eluting solvent, evaporate to dryness cut methyl alcohol shifts.The component launched using the chloroform-methanol solvent systems in volume ratio 95:5 to 90:10 scope on thin-layer chromatography is through gel filtration chromatography (diameter 18mm, column length 1600mm, gel is sephedex LH-20, moving phase is the chloroform-methanol of volume ratio 1:1) be separated, collect cut, separate out crude product, crude product adopts high performance liquid phase half preparation, and (determined wavelength is 254nm, flow velocity is 3mL/min, chromatographic column is Phenomenex Gemini 5 μ C18110A column (250 × 10mm, 5 μm), the methanol-water of moving phase to be volume ratio be 65:35) when appearance time is 25 and 33 minutes, separation and purification obtains compound 1 and 2 respectively.
Wherein the structure elucidation of compound 1 is as follows:
Compound 1 is white solid, and its high resolution mass spectrum (HRESIMS) is at m/z498.1778 [M+H]
+place provides quasi-molecular ion peak, and isotopic peak m/z500.1762 [M+H+2]
+be 1:3 with the relative abundance of quasi-molecular ion peak, show in molecule, containing 1 chlorine atom, in conjunction with NMR spectroscopic data (Table1), to learn that the molecular formula of compound 1 is C
26h
28clN
3o
5.
1there are 4 methyl [δ in HNMR spectrum display molecule
h0.70 (3H, s), 0.78 (3H, s), 1.42 (6H, s)], 4 methylene radical [δ
h1.77 (1H, dd, J=9.5,12.5Hz), 1.84 (2H, overlapped), 1.96 (1H, dd, J=10.0,12.5Hz), 2.03 (1H, m), 2.54 (1H, overlapped), 3.41 (2H, t, J=6.0Hz)], 1 High-Field methyne [δ
h3.76 (1H, t, J=9.5Hz)], the methyne [δ of 1 oxidation
h5.19 (1H, d, J=7.5Hz)], 3 alkene hydrogen [δ
h5.84 (1H, d, J=10.0Hz), 6.54 (1H, d, J=10.0Hz), 6.91 (1H, s)], 1 hydroxyl hydrogen [δ
h5.65 (1H, d, J=8.0Hz)] and 2 amino hydrogen [δ
h8.01 (1H, s), 10.81 (1H, s)].
13c NMR and DEPT composes (Table2) and shows to there are 26 carbon in molecule, comprises 4 methyl (δ
c18.9,22.5,27.5,27.5), 4 methylene radical (δ
c24.2,28.8,29.5,43.2), 1 High-Field methyne (δ
c54.9), the methyne (δ of 1 oxidation
c72.9), 5 quaternary carbon (δ
c43.1,65.3,68.4,69.0,77.1), 8 olefinic carbon (δ
c105.3,112.0,116.2,122.1,125.2,130.8,138.1,147.7) and 3 carbonyl (δ
c168.6,172.5,177.4).Above-mentioned data show that compound 1 and known compound sclerotiamide have similar structures [Whyte, A.C.; Gloer, J.B.; Wicklow, D.T.; Dowd, P.F.J Nat Prod.1996,59,1093-1095.], compared with sclerotiamide, compound 1
1h NMR has lacked an aromatic signal in composing, its
13c NMR and DEPT has lacked a fragrant methine signals in composing but has added 1 fragrant quaternary carbon signal, the difference of further its molecular formula of contrast, can infer that difference unique in compound 1 and the two dimensional structure of sclerotiamide is that 1 aromatic of sclerotiamide is replaced by chlorine atom.Due to H-4(δ in hydrogen spectrum
h6.91, s) for unimodal, and H-4 and C-3/C-5/C-6/C-8 is correlated with in HMBC spectrum (Figure1), infers that the position of substitution of chlorine atom is C-5 thus.Thus, the qualification of the two dimensional structure of compound 1 as shown in formula I, wherein all carbon, hydrogen signal also all pass through HSQC, HMBC and
1h-
1h COSY spectrum belongs to.
The steric configuration of compound 1 is identified by NOESY spectrum (Fig. 1), H-4, H-10 and CH
3nOE coherent signal between-24 shows H-10 and CH
3-24 is beta comfiguration, and H-10 and CH
3-24 are positioned at the surface of pentamethylene ring and towards H-4, the relative configuration of deducibility C-3 is as shown in formula I thus; Meanwhile, H-19 and NH-21/CH
3-23 have NOE is correlated with, and shows H-19 and CH
3-23 is α configuration.It can thus be appreciated that, the relative configuration of compound 1 is identical with sclerotiamide, again because the absolute configuration of sclerotiamide is identified by the method contrasted with analog paraherquamide, therefore the absolute configuration of compound 1 is accredited as (3R too, 10S, 11R, 17S, 19S).The concrete structure of compound 1 as shown in formula I, wherein R
1=H, R
2=OH, R
3=C1.
NMR data (the 500and125MH of table 1. compound 1 and 2
z, respectively, in DMSO-d6, δ ppm)
asignals overlapping
Compound 2 is white solid, and its high resolution mass spectrum (HRESIMS) is at m/z464.2166 [M+H]
+place provides quasi-molecular ion peak, in conjunction with NMR spectroscopic data (Table1), learns that compound 2 has the molecular formula C identical with sclerotiamide
26h
29n
3o
5.Compound 2
1h and
13c NMR modal data is close with sclerotiamide ten points, except 10-CH(δ
c77.5, δ
h4.62) and C-3(δ
c65.7) in chemical shift, there is notable difference.Further to compound 2
1h NMR,
13c NMR, HSQC and HMBC data analysis, find that compound 2 has identical two dimensional structure with sclerotiamide, but its steric configuration there are differences [1.Whyte, A.C.; Gloer, J.B.; Wicklow, D.T.; Dowd, P.F.J Nat Prod.1996,59,1093-1095.].The relative configuration of compound 2 is identified, 10-OH and H-4/H-5/CH by NOESY spectrum (Fig. 1)
3the NOE coherent signal of-24 shows OH-10 and CH
3-24 is beta comfiguration, and H-10, H-19, NH-21 and CH
3nOE coherent signal between-23 shows H-10, H-19 and CH
3-23 is α configuration.Above-mentioned deduction shows that compound 2 is the epimer of sclerotiamide, the relative configuration at the two C-10 place is just in time contrary, but the configuration of all the other hand-type carbon is completely the same, because the absolute configuration of sclerotiamide is determined, therefore, the absolute configuration of compound 2 is accredited as (3R, 10R, 11R, 17S, 19S).The concrete structure of compound 2 as shown in formula I, wherein R
1=OH, R
2=H, R
3=H.
Formula I.
Embodiment 2: the anti-careless tongue worm larva attachment activity test of the alkaloid compound shown in formula (I)
The attachment inhibition test of one of the marine fouling organism model that active testing model adopts current laboratory the most frequently used multicell grass tongue worm larva (Bugula neritina).The multicell of the maturation collected grass tongue worm is placed in the seawater with 0.22 μm of membrane filtration, with light stimulation, collects the larva that ripe multicell grass tongue worm produces through light stimulation.Larva is placed in the watch-glass filled with the seawater of 0.22 μm of membrane filtration.Activity test is done with the larva that careless tongue worm produces through illumination.24 hole polystyrene plates are adopted to measure the anti-larva attachment activity of compound.Formula (I) compound (compound 1 or compound 2) is dissolved in DMSO respectively, is diluted to different strength solution (60 – 140 μ g/mL) as test fluid with sterile filtration seawater.Add 1mL test fluid and 20 ± 2 ripe careless tongue worm larvas in each hole, each concentration establishes 5 to repeat samples, and sterile filtration seawater does blank.24 porocyte culture plates are placed in 28 DEG C of incubators to place 3 hours.Add up the larva number of attachment under the microscope, carry out statistical study with spss11.0 software, calculate EC
50value.
In experimental result display type (I), compound 1 and 2 suppresses the EC of careless tongue worm larva attachment
50value is respectively 13.52 μ g/mL and 28.12 μ g/mL, and the alkaloid compound shown in formula (I) has anti-careless tongue worm larva attachment activity thus, can for the preparation of in anti-marine biofouling coating.
Embodiment 3: the anti-kentrogon attachment activity test of the alkaloid compound shown in formula (I)
Active testing model is the attachment inhibition test of one of the marine fouling organism model adopting current laboratory the most frequently used line barnacle worm cyprids (Barnacles larvae).The nauplius larva of barnacle for food, is cultivated the cypris larva stage with Chaetoceros gracilis (Chaetoceros gracilis Schutt) by adult barnacle in 0.22 μm of filtering sea, every milliliter of 2 larvas, culture temperature 28 DEG C.Activity test is carried out with the Venus stage juvenile produced.24 hole polystyrene plates are adopted to measure the anti-larva attachment activity of compound.Formula (1) compound (compound 1 or compound 2) is dissolved in DMSO respectively, is diluted to different strength solution (60 – 140 μ g/mL) as test fluid with sterile filtration seawater.Add 1mL test fluid and 20 ± 2 ripe kentrogons in each hole, each concentration establishes 5 to repeat samples, and sterile filtration seawater does blank.24 porocyte culture plates are placed in 30 DEG C of incubators to place 24 hours.Add up the larva number of attachment under the microscope.
Experimental result shows, and the inhibiting rate that in formula (I), compound 1 and 2 suppresses kentrogon to adhere to when starting point concentration 25 μ g/mL is respectively 70% and 64%, its EC
50value should be less than 25 μ g/mL(due to the limited not test further of sample size), when measured maximum concentration 200 μ g/mL, no toxicity is not shown to larva, alkaloid compound shown in formula (I) has anti-kentrogon attachment activity thus, can for the preparation of in anti-marine biofouling coating.
Embodiment 4: the alkaloid compound anti tumor activity in vitro test shown in formula (I)
Collect the malignant myeloid cell lines K562 of logarithmic phase, malignant melanoma cell strain A375, lung cancer cell types, cervical cancer cell lines HeLa, breast cancer cell line mcf-7, laryngeal cancer cell strain Hep-2, hepatoma H22 cells and colon-cancer cell strain SW-620 respectively, it is made to suspend with 10% serum 1640 substratum, be inoculated in 96 well culture plates, every porocyte number is 5000/80 μ L, is placed in 5%CO
2incubator 37 DEG C cultivation.With 10% serum 1640 substratum, compound 1 and 2 in formula (I) being diluted to concentration is 3.125 μ g/mL, 6.25 μ g/mL, 12.5 μ g/mL, 25 μ g/mL, 50 μ g/mL, 100 μ g/mL, the experimental solutions of 200 μ g/mL.It is 3.125 μ g/mL that next day adds concentration respectively in the culture plate of different experimental group, 6.25 μ g/mL, 12.5 μ g/mL, 25 μ g/mL, 50 μ g/mL, the experimental solutions 20 μ L of 100 μ g/mL, 200 μ g/mL, and make the final concentration in every hole reach test (experimental group concentration adopts two-fold dilution).10% serum 1640 substratum of equivalent is added in addition in negative control group.After 48h, inhale the nutrient solution abandoned in experimental group and control group, every hole adds MTT20 μ L (2.5mg/mL), continue to cultivate 4h, every hole adds DMSO100 μ L termination reaction again, place 20min, detect the absorbance A value of each hole at 570nm place by microplate reader for 37 DEG C, calculate inhibitory rate of cell growth.Cell growth rate=(experimental group OD ÷ control group OD) × 100%.
Experimental result shows, and the compound 1 and 2 shown in formula I of the present invention suppresses the IC of human leukemia cell line K562 growth
50value is respectively 44.2 and 52.8 μMs, and have the effect that certain suppression malignant melanoma cell strain A375, lung cancer cell types, cervical cancer cell lines HeLa, breast cancer cell line mcf-7, laryngeal cancer cell strain Hep-2, hepatoma H22 cells, colon-cancer cell strain SW-620 grow, IC
50value is 80 ~ 100 μMs.Therefore the alkaloid compound shown in formula of the present invention (I) can for the preparation of antitumor drug.
Claims (6)
1. the alkaloid compound shown in formula I:
Compound 1:R
1=H, R
2=OH, R
3=Cl; Compound 2:R
1=OH, R
2=H, R
3=H.
2. a preparation method for alkaloid compound according to claim 1, is characterized in that, comprises the following steps:
(1) fermented liquid of fungi Aspergillus westerdijkiae DFFSCS013 is prepared;
(2) fermented liquid ethyl acetate, methylene dichloride or chloroform solvent extraction step (1) obtained, concentrates and obtains ethyl acetate extract, dichloromethane extract or chloroform extract;
(3) by the ethyl acetate extract described in step (2), dichloromethane extract or chloroform extract are through normal pressure silica gel column chromatography, with chloroform-methanol, chloroform-acetone, chlorofonn-ethylacetate, sherwood oil-acetone or petroleum ether-ethyl acetate solvent systems are eluent, gradient elution is carried out from volume ratio 100:0 to 0:100, follow the trail of with thin-layer chromatography and merge component, by the component can launched with the chloroform-methanol solvent systems in volume ratio 95:5 to 90:10 scope on thin-layer chromatography, crude product is obtained through gel filtration chromatography, purified, obtain compound 1 and 2 according to claim 1,
Fungi Aspergillus westerdijkiae DFFSCS013 is inoculated in PDA nutrient agar by the preparation method of the fermented liquid of the fungi Aspergillus westerdijkiae DFFSCS013 described in step (1), cultivate 3 days for 26 DEG C, obtain cultivating the flat board having bacterial classification, then by the strain inoculation on flat board in PDB liquid nutrient medium, cultivate 2 days in rotating speed 200rpm shaking table, temperature 26 DEG C, obtain seed liquor, again seed liquor is inoculated in rice solid medium, in room temperature quiescent culture 26 days, obtain fermented liquid; The formula of described rice solid medium is: add rice 400g, yeast extract 2g, glucose 2g and sea salt 18g in every 600mL water.
3. the preparation method of alkaloid compound according to claim 2, is characterized in that, the extraction ethyl acetate described in step (2), described concentrated employing concentrating under reduced pressure; Purifying described in step (3) adopts chromatographic column to be separated or recrystallization.
4. alkaloid compound according to claim 1 or the application of its salt in preparation anti-marine fouling organism attachment coating.
5. the application in antitumor drug prepared by alkaloid compound according to claim 1 or its salt.
6. application according to claim 5, is characterized in that, described antitumor drug is human body leukemia medicament.
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| CN111234586B (en) * | 2020-02-24 | 2021-01-19 | 中国科学院南海海洋研究所 | Application of pyrazinoquinazolinetrione alkaloid compound in preparation of marine fouling organism control agent |
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