Anti-biofilm compounds derived from marine sponges

Bacterial biofilms are surface-attached communities of microorganisms that are protected by an extracellular matrix of biomolecules. In the biofilm state, bacteria are significantly more resistant to external assault, including attack by antibiotics. In their native environment, bacterial biofilms underpin costly biofouling that wreaks havoc on shipping, utilities, and offshore industry. Within a host environment, they are insensitive to antiseptics and basic host immune responses. It is estimated that up to 80% of all microbial infections are biofilm-based. Biofilm infections of indwelling medical devices are of particular concern, since once the device is colonized, infection is almost impossible to eliminate. Given the prominence of biofilms in infectious diseases, there is a notable effort towards developing small, synthetically available molecules that will modulate bacterial biofilm development and maintenance. Here, we highlight the development of small molecules that inhibit and/or disperse bacterial biofilms specifically through non-microbicidal mechanisms. Importantly, we discuss several sets of compounds derived from marine sponges that we are developing in our labs to address the persistent biofilm problem. We will discuss: discovery/synthesis of natural products and their analogues-including our marine sponge-derived compounds and initial adjuvant activity and toxicological screening of our novel anti-biofilm compounds.     

Polyketides from a Marine-Derived Fungus Xylariaceae sp.

Eighteen polyketides (1–18) including six citrinin derivatives, two phenol derivatives, one cyclopentenone, two naphthol derivatives, and seven tetralone derivatives were isolated from the culture broth of a marine-derived fungal strain Xylariaceae sp. SCSGAF0086. Five of these compounds (1, 2, 8, 9, and 10) were new, and their structures were determined by spectroscopic methods. Compounds 4, 6, 7, and 17 showed enzyme-inhibitory activities towards several tested enzymes, and 6 and 7 showed strong antifouling activity against Bugula neritina larvae settlement. This is the first time that the antifouling and enzyme-inhibitory activities of these compounds has been reported.     

Adverse Effect of Antifouling Compounds on the Reproductive Mechanisms of the Ascidian Ciona intestinalis

Fertilization and embryo development that occur in sea water are sensitive to xenobiotics from anthropogenic sources. In this work, we evaluated the influence of two antifouling biocides, tributyltin (TBT) and diuron, on the reproductive mechanisms of the marine invertebrate Ciona intestinalis. By using electrophysiological techniques, we examined the impact of these compounds on the electrical properties of the mature oocytes and of events occurring at fertilization. With different toxicity assays, we studied the effect of the two biocides on the gametes by evaluating fertilization rate and embryo development. Results show that sodium (Na⁺) currents were significantly reduced by either of the two biocides, whereas conductance was significantly increased. The fertilization current frequency and amplitude, fertilization rate and larval development were affected only by TBT. This study suggests that: (i) the two biocides affect either the electrical properties of the oocyte plasma membrane and the reproductive success representing a risk factor for the survival of the species exposed to environmental pollution; (ii) the ascidian Ciona intestinalis may represent a good model organism to test toxicity of marine pollutants. Possible mechanisms of action of the two biocides are discussed.     

大连海区海藻和无脊椎动物附生细菌对养殖网笼污损细菌的拮抗活性

以从养殖网笼上分离的6株污损细菌为指示菌,采用双层琼脂扩散法对从藻类表面分离出的192株和从无脊椎动物表面分离出的127株附生细菌进行筛选,从中获得有拮抗活性的菌株分别为60株和49株,占其测试菌株的31.3%和38.6%。从不同海藻和无脊椎动物附生细菌中可以筛选出不同得率的活性菌株,其抗菌谱和活性强度各不相同,将抗菌谱和活性强度两个参数结合,设置活性指数Index(I),经计算可知,P31、HE426和AU4111的活性最高,其I值(≥2.00)最大。研究结果表明,海藻和无脊椎动物附生细菌可能帮助宿主阻止污损细菌等污损生物的附着。     

Territrem and Butyrolactone Derivatives from a Marine-Derived Fungus Aspergillus Terreus

Seventeen lactones including eight territrem derivatives (1–8) and nine butyrolactone derivatives (9–17) were isolated from a marine-derived fungus Aspergillus terreus SCSGAF0162 under solid-state fermentation of rice. Compounds 1–3 and 9–10 were new, and their structures were elucidated by spectroscopic analysis. The acetylcholinesterase inhibitory activity and antiviral activity of compounds 1–17 were evaluated. Among them, compounds 1 and 2 showed strong inhibitory activity against acetylcholinesterase with IC₅₀ values of 4.2 ± 0.6, 4.5 ± 0.6 nM, respectively. This is the first time it has been reported that 3, 6, 10, 12 had evident antiviral activity towards HSV-1 with IC₅₀ values of 16.4 ± 0.6, 6.34 ± 0.4, 21.8 ± 0.8 and 28.9 ± 0.8 μg·mL⁻¹, respectively. Antifouling bioassay tests showed that compounds 1, 11, 12, 15 had potent antifouling activity with EC₅₀ values of 12.9 ± 0.5, 22.1 ± 0.8, 7.4 ± 0.6, 16.1 ± 0.6 μg·mL⁻¹ toward barnacle Balanus amphitrite larvae, respectively.     

船舶防污涂料与防污剂的研究进展

概括介绍了海洋生物的附着过程及其影响因素,讨论了各类船舶防污涂料及其防污剂的最新研究进展,指出了新型低无毒防污涂料的发展趋势。     

Bioactive Phenylalanine Derivatives and Cytochalasins from the Soft Coral-Derived Fungus,Aspergillus elegans

One new phenylalanine derivative 4′-OMe-asperphenamate (1), along with one known phenylalanine derivative (2) and two new cytochalasins, aspochalasin A1 (3) and cytochalasin Z24 (4), as well as eight known cytochalasin analogues (5–12) were isolated from the fermentation broth of Aspergillus elegans ZJ-2008010, a fungus obtained from a soft coral Sarcophyton sp. collected from the South China Sea. Their structures and the relative configurations were elucidated using comprehensive spectroscopic methods. The absolute configuration of 1 was determined by chemical synthesis and Marfey’s method. All isolated metabolites (1–12) were evaluated for their antifouling and antibacterial activities. Cytochalasins 5, 6, 8 and 9 showed strong antifouling activity against the larval settlement of the barnacle Balanus amphitrite, with the EC₅₀ values ranging from 6.2 to 37 μM. This is the first report of antifouling activity for this class of metabolites. Additionally, 8 exhibited a broad spectrum of antibacterial activity, especially against four pathogenic bacteria Staphylococcus albus, S. aureus, Escherichia coli and Bacillus cereus.     

Bioactive Bromotyrosine Derivatives from the Pacific Marine Sponge Suberea clavata (Pulitzer-Finali, 1982)

Chemical investigation of the South-Pacific marine sponge Suberea clavata led to the isolation of eight new bromotyrosine metabolites named subereins 1–8 (2–9) along with twelve known co-isolated congeners. The detailed configuration determination of the first representative major compound of this family 11-epi-fistularin-3 (11R,17S) (1) is described. Their chemical characterization was achieved by HRMS and integrated 1D and 2D NMR (nuclear magnetic resonance) spectroscopic studies and extensive comparison with literature data. For the first time, a complete assignment of the absolute configurations for stereogenic centers C-11/17 of the known members (11R,17S) 11-epi-fistularin-3 (1) and 17-deoxyfistularin-3 (10) was determined by a combination of chemical modifications, Mosher’s technology, and ECD spectroscopy. Consequently, the absolute configurations of all our new isolated compounds 2–9 were determined by the combination of NMR, Mosher’s method, ECD comparison, and chemical modifications. Interestingly, compounds 2–7 were obtained by chemical transformation of the major compound 11-epi-fistularin-3 (1). Evaluation for acetylcholinesterase inhibition (AChE), DNA methyltransferase 1 (DNMT1) modulating activity and antifouling activities using marine bacterial strains are also presented.     

空心莲子草中抗污损活性皂苷成分的研究

海洋污损生物危害水产养殖,为发现抗污损的天然资源,对入侵植物空心莲子草(Alternantheraphiloxeroides)进行了抗污损活性化学成分研究。分离鉴定了3个皂苷类化合物,分别为3-O-β-D-吡喃葡萄糖醛酸基齐墩果酸-28-O-β-D-吡喃葡萄糖酯(1)、3-O-β-D-吡喃葡萄糖醛酸甲酯基齐墩果酸-28-O-β-D-吡喃葡萄糖酯(2)、3-O-β-D-吡喃葡萄糖醛酸基-30-去甲基-12,20(29)-二烯齐墩果酸-28-O-β-D-吡喃葡萄糖酯(3),其中化合物2为首次从该植物中分离得到。初步的抗污损结果表明,皂苷3具有明显的杀灭污损生物藤壶的作用,活性与阳性对照CuSO4相近,但是对卤虫的毒性则远低于相同剂量的CuSO4;部分皂苷混合物也具有一定的选择性杀灭藤壶的作用。     

海水网箱网衣碱性硅酸盐防污涂料的初步研究

本文以碱性硅酸盐为主防污剂，开发了一种防污期长、无毒、环保且高效的海水网箱网衣防污涂料：涂有该种防污涂料的试验网衣于2002年4月开始进行浅海挂网试验．为期七个月；每月一次定期对试验网片进行附着情况检查、拍照及称重。结果证明．与对照组相比，涂有硅酸盐防污涂料的网衣材料的网片增重及被附着的面积百分比均有大幅度的降低；实拍照片也证明涂有防污涂料的网片具有极佳的防污效果：根据试验结果，讨论了影响防污涂料防污效果的诸多因素。