ATP synthase; Antibiotic

从海洋链霉菌B8496和B8739发酵液中分离得到4种抗生素，分别为帕霉素、低霉素A、低霉素B和青孢素。生物测定显示，这些化合物都以剂量和时间依赖的方式损害了葡萄假单胞游动孢子的运动性并导致随后的裂解。帕霉素的运动抑制和裂解活性最强(IC50为0.1 μ mL(-1))，其次是低霉素B (IC50为0.15和0.2 μ mL(-1))和低霉素F (IC50为0.3和0.5 μ mL(-1))。寡霉素A和棘孢素对游动孢子也有运动抑制作用，IC50值分别为3.0和10.0 μ mL(-1)。这是首次报道这些抗生素对植物致病性peronosporomyter的游动孢子的运动抑制和裂解活性。所有分离化合物的结构都是根据详细的光谱分析确定的。[2]

游动孢子的产生及生物测定[2]
为分离葡萄侵染叶片的葡萄浆原菌孢子囊。在含1.5%水琼脂的培养皿中，在25°C和95%相对湿度条件下，在葡萄幼苗叶片下表面连续培养该菌株(Islam and Tiedemann 2011;Islam, Tiedemann and Laatsch 2011)。培养第6天，用微型真空吸尘器将含有柠檬形孢子囊的孢子囊收集到埃本多夫小瓶中。将新鲜收获的孢子囊与孢子囊分离，用尼龙筛(50目)过滤，蒸馏水洗涤2次，室温(23℃)消毒后的自来水(3 × 104个/ mL)黑暗培养6 h，释放运动孢子。游动孢子在灭菌水中保持运动长达12小时，并用于运动生物测定(Islam和Tahara 2001)。首先用二甲基亚砜(DMSO)配制抗生素原液，然后用蒸馏水稀释。游动孢子悬浮液中DMSO的最终浓度从未超过1% (v/v)，这一条件不影响游动孢子的运动。化合物对游动孢子的活力和活力的影响采用先前描述的方案进行了分析(Islam等，2005;Abdalla等人2011;Islam, Tiedemann and Laatsch 2011)。在光镜下观察游动孢子的运动。如前所述，对游动孢子的运动性和裂解的时间变化进行了量化(Islam, Tiedemann和Laatsch 2011)。每个治疗重复三次。计算各处理染病孢子的平均值%±SE(标准误差)，并通过单、双向方差分析(ANOVA)和t检验确定统计学显著性。采用双因素方差分析(见表S1，支持信息)检验抗生素在不同治疗时间和剂量下的运动抑制和溶解活性差异的显著性。以P < 0.05为差异有统计学意义。

疥癣素酯(SEs)是最近从黄斑地衣(Xanthoparmelia scabrosa)中分离到的一种具有活性二硫键的epipolythiiodioxopiperazine (ETP)类次生代谢产物。集落形成试验表明，这些毒素在纳摩尔浓度下对人类肿瘤细胞系有活性。ETP类毒素的其他成员，如胶质毒素，已被证明可诱导细胞凋亡，尽管ETP毒素的细胞靶点目前尚不清楚。ETP毒素已被证明通过与敏感的半胱氨酸残基相互作用抑制多种酶。通过JC-1染色，我们发现典型的醋酸丁酸皂素酯诱导早期线粒体膜超极化，并伴有细胞凋亡。该毒素降低完整细胞中的ATP并抑制渗透细胞中ATP的合成速率。与已知的ATP合成酶抑制剂寡霉素B的作用比较，ATP合成酶作为皂素酯诱导细胞死亡的早期靶点是一致的[1]。

Production of zoospores and bioassay[2]
For the isolation of sporangia of Plasmopara viticola, infected leaves of grapevine. The strain was maintained by continuous culturing on the lower surface of young grapevine leaves on Petri dishes containing 1.5% water agar at 25°C and 95% relative humidity (Islam and Tiedemann 2011; Islam, Tiedemann and Laatsch 2011). On the sixth day of cultivation, the sporangiophores containing the lemon-shaped sporangia were harvested into an Eppendorf vial by a micro-vacuum cleaner. The freshly harvested sporangia were separated from sporangiophores by filtration through a nylon sieve (50 mesh), washed twice with distilled water and then incubated in sterilized tap water (3 × 104 sporangia per mL) in the dark for 6 h at room temperature (23°C) to release zoospores. The zoospores remained motile up to 12 h in sterilized water, and they were used for the motility bioassay (Islam and Tahara 2001). Stock solutions of the antibiotics were first prepared in dimethyl sulfoxide (DMSO) and then diluted with distilled water. The final concentration of DMSO in the zoospore suspension never exceeded 1% (v/v), a condition that does not affect zoospore motility. The effects of compounds on motility and viability of zoospores were assayed using protocols described earlier (Islam et al.2005; Abdalla et al.2011; Islam, Tiedemann and Laatsch 2011). Motility of the zoospores was observed under a light microscope at 100-fold magnification. Quantification of time-course changes of motility and lysis of the zoospores were carried out as described earlier (Islam, Tiedemann and Laatsch 2011). Each treatment was replicated three times. The mean value % ±SE (standard error) of the affected spores in each treatment was calculated and the statistical significance was determined by one- and two-way Analysis of Varience (ANOVA) and t-test. Two-way ANOVA (see Table S1, Supporting Information) was used to test the significance of differences in motility inhibition and lytic activities of the antibiotics at different times and doses of treatment. The difference is considered statistically significant with a value of P < 0.05.

mouse LD50 intraperitoneal 2900 ug/kg

[1]. Evidence that the lichen-derived scabrosin esters target mitochondrial ATP synthase in P388D1 cells. Toxicol Appl Pharmacol. 2003 Aug 1;190(3):232-40.

[2]. Oligomycins and pamamycin homologs impair motility and induce lysis of zoospores of the grapevine downy mildew pathogen, Plasmopara viticola. FEMS Microbiol Lett. 2016 Aug;363(16).

Oligomycin B is an oligomycin with formula C45H72O12 that is oligomycin A in which the spirocyclic ring bearing the 2-hydroxypropyl substituent has been substituted by an oxo group at the carbon which is directly attached to the spirocentre. It is a nonselective inhibitor of the mitochondrial F1F0 ATP synthase. It has a role as an EC 3.6.3.14 (H(+)-transporting two-sector ATPase) inhibitor. It is an oligomycin, a triketone and a pentol.
Oligomycin A, 28-oxo- has been reported in Apis cerana with data available.

C45H72O12

805.04598

804.502

11050-94-5

Oligomycin A;579-13-5;Oligomycin;1404-19-9;Oligomycin D;1404-59-7;Oligomycin C;11052-72-5

76958645

Typically exists as White to off-white solid at room temperature

1.2±0.1 g/cm3

911.4±65.0 °C at 760 mmHg

160-161℃

260.3±27.8 °C

0.0±0.6 mmHg at 25°C

1.547

5.5

197.12

5

12

3

57

1470

18

CC[C@@H]1C=CC=CC[C@H](C)[C@H]([C@@](C)(C(=O)[C@H](C)[C@H]([C@H](C)C(=O)[C@H](C)[C@H]([C@H](C)C=CC(=O)O[C@H]2[C@@H](C)[C@@H](CC1)O[C@]3([C@@H]2C)C(=O)C[C@@H](C)[C@@H](C[C@H](C)O)O3)O)O)O)O

QPRQJOHKNJIMGN-WVUAJZTGSA-N

InChI=1S/C45H72O12/c1-12-33-17-15-13-14-16-25(3)42(52)44(11,54)43(53)31(9)40(51)30(8)39(50)29(7)38(49)24(2)18-21-37(48)55-41-28(6)34(20-19-33)56-45(32(41)10)36(47)22-26(4)35(57-45)23-27(5)46/h13-15,17-18,21,24-35,38,40-42,46,49,51-52,54H,12,16,19-20,22-23H2,1-11H3/b14-13+,17-15+,21-18+/t24-,25+,26-,27+,28+,29-,30-,31-,32-,33-,34-,35-,38+,40+,41+,42-,44+,45+/m0/s1

(1R,4E,5'S,6S,6'S,7R,8S,10R,11R,12S,14R,15S,16R,18E,20E,22R,25S,27S,28S,29R)-22-ethyl-7,11,14,15-tetrahydroxy-6'-[(2R)-2-hydroxypropyl]-5',6,8,10,12,14,16,28,29-nonamethylspiro[2,26-dioxabicyclo[23.3.1]nonacosa-4,18,20-triene-27,2'-oxane]-3,3',9,13-tetrone

Oligomycin B; 11050-94-5; Oligomycin A, 28-oxo-; 387OE420H7; EINECS 234-275-5; BRN 5705502; (1R,4E,5'S,6S,6'S,7R,8S,10R,11R,12S,14R,15S,16R,18E,20E,22R,25S,27S,28S,29R)-22-ethyl-7,11,14,15-tetrahydroxy-6'-[(2R)-2-hydroxypropyl]-5',6,8,10,12,14,16,28,29-nonamethyl-5',6'-dihydro-3H,9H,13H-spiro[2,26-dioxabicyclo[23.3.1]nonacosa-4,18,20-triene-27,2'-pyran]-3,3',9,13(4'H)-tetrone; 28-OXOOLIGOMYCIN A;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

---

注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

View More

注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

---

口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

View More

口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

---

注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

---

请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。