5-Iodotubercidin (NSC-113939)   中文名称: 5-碘代杀结核菌素--庄

Adenosine kinase (IC50 = 26 nM)

5-Iodotubercidin (NSC 113939) 的 IC50 值分别为 0.4、3.5、5-10、5-10、10.9 和 27.7 μM，并抑制 CK1、胰岛素受体酪氨酸激酶、磷酸化酶激酶、PKA、CK2 和 PKC[ 1]。 5-碘结核菌素 (20 μM) 导致 ATP 浓度显着下降，而 AMP 浓度相应略有增加。 5. 碘结核菌素降低脂肪酸和胆固醇的合成速率以及ACC的活性。 5-碘结核菌素显着降低细胞内丙二酰辅酶A的浓度，这与碘结核菌素介导的ACC抑制作用一致[4]。

---

酶抑制的SAR。[2]
表1中列出的化合物被评估为重组人AK的抑制剂，IC50是通过稍微修改的文献方法通过放射化学测定法测定的。34酶抑制研究的目的是评估结核菌素C4-、C5-和C5'-位置的修饰对AKI活性的影响。最初的努力集中在C5位置，因为用碘基团替代会将适度的AK底物tubercidin（KM=10μM）转化为强效的AK抑制剂（1a/5-Iodotubercidin，IC50=0.026μM）。结果表明，吡咯并嘧啶核苷C5位的卤素原子如Cl、Br或I对强AKI活性至关重要。此外，抑制效力随着卤素原子尺寸和电子密度的增加而增加。I的大原子尺寸似乎提供了与酶活性位点的疏水相互作用。为了研究疏水性和供电子基团对AKI活性的影响，在C5位引入了CH3和SCH3等基团。然而，这些化合物（11a，b）被发现是酶的不良抑制剂。11a所示的AKI效力的损失可能是由于与任何卤素原子相比，CH3基团的尺寸较小，只能提供微弱的疏水相互作用。尽管SCH3是一个富含电子的基团，但11b的效力降低似乎是由于该基团在活性位点的拟合不良。因此，1a/5-Iodotubercidin，c，d表现出的强效AKI活性似乎是由于C5取代基的亲脂性和电负性的共同作用。

---

添加微摩尔浓度的腺苷衍生物5-Iodotubercidin（Itu）通过引起磷酸化酶失活和糖原合酶激活来启动分离肝细胞中的糖原合成[Flückiger Isler和Walter（1993）Biochem.J.29285-91]。我们在这里报告说，Itu还可以拮抗饱和浓度的胰高血糖素和加压素对这些酶的影响。Itu诱导的糖原合酶激活不能用磷酸化酶a（糖原相关合酶磷酸酶的强效抑制剂）的去除来解释。当在纯化的酶上进行测试时，Itu不影响主要Ser/Thr特异性蛋白磷酸酶（PP-1、PP-2A、PP-2B和PP-2C）的活性，但它抑制了各种Ser/Thr特异性蛋白激酶以及胰岛素受体的酪氨酸激酶活性（IC50在10-15微M ATP下为0.4至28微M）。结核菌素不影响肝细胞中的糖原合酶或磷酸化酶，作为蛋白激酶抑制剂的效力要低300倍。酪蛋白激酶-1和蛋白激酶A抑制的动力学分析表明，Itu对ATP起竞争性抑制剂的作用，对蛋白质底物起混合型抑制剂的作用。我们提出Itu通过抑制磷酸化酶激酶和各种糖原合酶激酶来灭活磷酸化酶并激活糖原合酶。与Itu在体外的广泛特异性一致，该化合物降低了完整肝细胞中许多磷酸多肽的磷酸化水平。我们的数据表明，Itu的至少一些生物学效应可以通过抑制蛋白激酶来解释。1.

---

尽管5-Iodotubercidin被广泛用作腺苷激酶抑制剂，从而干扰腺苷和腺嘌呤核苷酸的代谢，但已经提出了不同的作用机制。大鼠肝细胞与碘胸苷的孵育对脂质代谢产生了重要影响。（i） 乙酰辅酶A羧化酶和脂肪酸从头合成都被5-碘酸杆菌素平行抑制，脂肪酸合酶的活性没有变化。对这两种活性的抑制显示出对碘酸浓度的类似依赖性，并且伴随着细胞内丙二酰辅酶a浓度的类似降低（约60%）。（ii）碘酸杆菌素刺激棕榈酸氧化，但辛酸氧化不受影响。然而，这种效应可归因于丙二酰辅酶A浓度的降低以及随之而来的肉碱棕榈酰基转移酶I抑制的缓解，因为在用洋地黄皂苷渗透的细胞中测定时，发现这种酶的活性没有改变。（iii）碘酸也能从头抑制胆固醇合成。因此，结果表明，碘胸苷以脂肪生成为代价增加了肝脏的脂肪酸氧化活性，我们认为这些对脂肪酸代谢的影响是通过抑制乙酰辅酶a羧化酶介导的，这可能是由于AMP/ATP比值增加了两倍多，同时刺激了AMP活化的蛋白激酶[3]。

在 AKI 局部给予梨状皮层后，5-碘结核菌素（1 mL/kg，腹腔注射）已被证明可有效对抗荷包牡丹碱诱发的癫痫发作[2]。

---

SAR的抗癫痫活性。[2]
在标准最大电击发作（MES）模型中评估了几种AKI（表2）。使用5-Iodotubercidin/5-碘代结核菌素（1a）观察到的抗惊厥作用与在将AKI局部施用到prepiriform皮层后观察到的针对荷包牡丹碱诱导的癫痫发作的活性一致。20最有效的化合物1b的ED50为0.3 mg/kg，而其他化合物的效力在4.2-7.1 mg/kg的范围内。体内抗惊厥活性的等级顺序与体外酶抑制试验中的效力顺序没有很强的相关性，这表明分子取代基的变化会导致药代动力学特性的实质性变化，如清除率、脑渗透或胞内酶的可及性。例如，化合物15a、b、1b、c的C5'-氨基类似物在体内显示出较低的效力，尽管对酶的效力更大；这可能是C5'-氨基类似物渗透血脑屏障减少的结果，这些类似物的疏水性低于其相应的5'-脱氧类似物。这些化合物也可能通过可能影响体内活性的其他机制发挥作用。这些机制可能与腺苷有关，因为这些化合物的体内活性被腺苷受体拮抗剂的给药所拮抗。39由于这些化合物对受体几乎没有亲和力，因此排除了A1或A2腺苷受体上直接激动剂活性的参与。15同样，这些化合物对腺苷脱氨酶（ADA）或腺苷单磷酸脱氨酶没有可检测的活性，对与转运机制相关的硝基苄硫肌苷结合位点几乎没有亲和力。15然而，这些化合物可能会调节未定义的转运系统，这可能会影响细胞外腺苷的积累。

酶活性测定 [2]
AK活性是在类似于Yamada等人的程序的放射化学分析中测量的，但略有修改。最终反应体积为100μL，含有70 mM Tris-leate（pH 7.0）、0.1%（w/v）牛血清白蛋白、1.0 mM MgCl2、1.0 mM ATP、1.0μM[U-14C]腺苷（400−600 mCi/mmol；Moravek Biochemicals，股份有限公司）和各种抑制剂浓度。抑制剂制备为DMSO中的10mM储备溶液。测定中DMSO的最终浓度为5%（v/v）。使用0.001至10.0μM的11种不同浓度的试验溶液来确定酶抑制的剂量反应曲线。通过加入适量纯化的人重组AK开始反应，并在37°C下孵育20分钟。通过加入强效AKI GP3269.40终止反应。将每种反应的30μL等分试样点在DEAE纤维素滤纸上（切成约1×1 cm的正方形），风干30分钟。然后将干燥的过滤器在去离子水中洗涤3分钟，以去除残留的[U-14C]腺苷，用乙醇冲洗并在90°C下干燥20分钟。使用Beckman LS3801闪烁计数器在5.5 mL Ready Safe液体闪烁混合物中计数滤纸。对照AK活性由在5%DMSO存在下形成的[14C]AMP的量测定。抑制50%AK活性所需的抑制剂浓度（IC50）由抑制剂浓度与对照酶活性百分比（%）的图以图形方式确定。结果如表1所示。

---

生化分析[3]
在与脂肪酸合酶反应相结合的测定中，测定了洋地黄皂苷透性肝细胞中的ACC活性。为了测量酶活性，将100μL肝细胞悬浮液加入到100μL预热的含洋地黄皂苷的测定培养基中，反应2分钟，完全按照所述进行。如前所述，在洋地黄皂苷渗透的肝细胞中监测脂肪酸合酶（FAS）活性。将肝细胞悬浮液（100μL）加入100μL含洋地黄皂苷的测定培养基中，反应4分钟。在洋地黄皂苷渗透的肝细胞中测定CPT-I活性，即放射性标记的L-肉碱在棕榈酰肉碱中的十四烷基甘氨酸酯（TDGA）敏感掺入。简而言之，在有或没有5μM TDGA（CPT-I的强效特异性抑制剂）的情况下，将肝细胞预孵育20分钟。将两组肝细胞孵育的等分试样（100μL）加入100μL预热的含洋地黄皂苷的检测培养基中，反应进行40秒。 通过放射酶法测定中和的高氯酸细胞提取物中丙二酰辅酶a的细胞内水平。胆固醇从头合成的速率被确定为[1- ]用轻质石油醚将乙酸盐提取成不皂化的甾醇。按照Gualix等人的描述，通过HPLC测定中和的高氯酸细胞提取物中的核苷酸水平。蛋白质采用Lowry等人的方法测定。

MES Seizure Assay. [2]
Male SA rats (100−150 ) were maintained on a 12:12 light:dark cycle in temperature-controlled facilities with free access to food and water. One hour prior to seizure testing, the animals were injected intraperitoneally (1 mL/kg) with DMSO vehicle or with test compound dissolved in DMSO. At the time of the test, an electrolyte solution (2% lidocaine in 0.9% sodium chloride) was applied to the eyes. Maximal electroshock seizures were induced by administering a 60-Hz current of 150 mA for 0.2 s via corneal electrodes, using a Wahlquist Model H stimulator.41 The endpoint measured was suppression of hindlimb tonic extension (HTE) and expressed as percentage of animals in which the response was inhibited. At this supramaximal stimulation level, virtually 100% of control (vehicle-treated) animals showed HTE. ED50 values were calculated from a dose−response curve using probit analysis.42 The N for the screening doses was 6−8; dose−response determinations were conducted with at least 5 animals/dose. [2]

---

Dissolved in saline; 1, 2.5 and 5 mg/kg; i.p. injection

Male Mongolian gerbils

[1]. Identification of the glycogenic compound 5-iodotubercidin as a general protein kinase inhibitor. Biochem J. 1994 Apr 1;299 (Pt 1):123-8.

[2]. Adenosine kinase inhibitors. 1. Synthesis, enzyme inhibition, and antiseizure activity of 5-iodotubercidin analogues. J Med Chem. 2000 Jul 27;43(15):2883-93.

[3]. Effects of 5-iodotubercidin on hepatic fatty acid metabolism mediated by the inhibition of acetyl-CoA carboxylase. Biochem Pharmacol. 2002 Jun 1;63(11):1997-2000.

[4]. A small-molecule inhibitor of Haspin alters the kinetochore functions of Aurora B. J Cell Biol. 2012 Oct 15;199(2):269-84.

[5]. Adenosine Kinase Inhibition Protects against Cranial Radiation-Induced Cognitive Dysfunction. Front Mol Neurosci. 2016 Jun 3;9:42.

5-iodotubercidin is an organoiodine compound. It is functionally related to a tubercidin.

---

In summary, we have explored the effect of various substitutions at the C4-, C5-, and C5‘-positions of tubercidin on AK inhibition and have described new AKIs with increased potency. We have also demonstrated anticonvulsant activity of several of these compounds against MES induced seizures in rats. It is clear from these studies that a halogen atom such as Cl, Br, or I at the C5-position and either a Cl, NH2, or SCH3 group at the C4-position of the pyrrolo[2,3-d]pyrimidine ring system are essential for tubercidin analogues to potently inhibit AK. The C5‘-position appears to accommodate both hydrophobic and hydrophilic groups, with the amino group at this position resulting in three of the most potent AK inhibitors described to date (15a,b and 16). In vivo, the potencies of these compounds were similar to that of 5-Iodotubercidin, and further modifications will be required to enhance in vivo properties. The ability of potent AK inhibitors to inhibit maximal seizures induced by electroshock indicates that these compounds are highly efficacious as anticonvulsants and suggests that inhibition of AK is a viable alternative to adenosine receptor agonists as a purinergic approach to inhibition of seizure activity.[2]

---

aken together, our results allow us to conclude that Itu markedly affects hepatic fatty acid metabolism by stimulating mitochondrial fatty acid oxidation at the expense of lipogenesis. We have identified acetyl-CoA carboxylase as the target enzyme for these effects of Itu. Moreover, these data afford indirect evidence which can be taken to suggest a mechanism responsible for this cellular response, based on the increase of AMP/ATP ratio and the concomitant stimulation of the AMPK. Clearly, further research is required to elucidate fully this suggested mechanism of action of Itu, and studies investigating this issue are in progress. In addition, these studies may contribute to a better understanding of this widely used inhibitor of adenosine kinase. Interestingly, Itu has been shown to inhibit partially purified AMPK, but only in the absence of AMP, therefore, not likely in isolated hepatocytes. [3]

---

By phosphorylating Thr3 of histone H3, Haspin promotes centromeric recruitment of the chromosome passenger complex (CPC) during mitosis. Aurora B kinase, a CPC subunit, sustains chromosome bi-orientation and the spindle assembly checkpoint (SAC). Here, we characterize the small molecule 5-iodotubercidin (5-ITu) as a potent Haspin inhibitor. In vitro, 5-ITu potently inhibited Haspin but not Aurora B. Consistently, 5-ITu counteracted the centromeric localization of the CPC without affecting the bulk of Aurora B activity in HeLa cells. Mislocalization of Aurora B correlated with dephosphorylation of CENP-A and Hec1 and SAC override at high nocodazole concentrations. 5-ITu also impaired kinetochore recruitment of Bub1 and BubR1 kinases, and this effect was reversed by concomitant inhibition of phosphatase activity. Forcing localization of Aurora B to centromeres in 5-ITu also restored Bub1 and BubR1 localization but failed to rescue the SAC override. This result suggests that a target of 5-ITu, possibly Haspin itself, may further contribute to SAC signaling downstream of Aurora B.[4]

---

Clinical radiation therapy for the treatment of CNS cancers leads to unintended and debilitating impairments in cognition. Radiation-induced cognitive dysfunction is long lasting; however, the underlying molecular and cellular mechanisms are still not well established. Since ionizing radiation causes microglial and astroglial activation, we hypothesized that maladaptive changes in astrocyte function might be implicated in radiation-induced cognitive dysfunction. Among other gliotransmitters, astrocytes control the availability of adenosine, an endogenous neuroprotectant and modulator of cognition, via metabolic clearance through adenosine kinase (ADK). Adult rats exposed to cranial irradiation (10 Gy) showed significant declines in performance of hippocampal-dependent cognitive function tasks [novel place recognition, novel object recognition (NOR), and contextual fear conditioning (FC)] 1 month after exposure to ionizing radiation using a clinically relevant regimen. Irradiated rats spent less time exploring a novel place or object. Cranial irradiation also led to reduction in freezing behavior compared to controls in the FC task. Importantly, immunohistochemical analyses of irradiated brains showed significant elevation of ADK immunoreactivity in the hippocampus that was related to astrogliosis and increased expression of glial fibrillary acidic protein (GFAP). Conversely, rats treated with the ADK inhibitor 5-iodotubercidin (5-ITU, 3.1 mg/kg, i.p., for 6 days) prior to cranial irradiation showed significantly improved behavioral performance in all cognitive tasks 1 month post exposure. Treatment with 5-ITU attenuated radiation-induced astrogliosis and elevated ADK immunoreactivity in the hippocampus. These results confirm an astrocyte-mediated mechanism where preservation of extracellular adenosine can exert neuroprotection against radiation-induced pathology. These innovative findings link radiation-induced changes in cognition and CNS functionality to altered purine metabolism and astrogliosis, thereby linking the importance of adenosine homeostasis in the brain to radiation injury.[5]

C11H13IN4O4

392.15

391.998

24386-93-4

97297

White to gray solid powder

2.5±0.1 g/cm3

701.5±60.0 °C at 760 mmHg

216-217ºC dec.

378.0±32.9 °C

0.0±2.3 mmHg at 25°C

1.919

0.91

126.65

4

7

2

20

365

4

IC1C2=C(N([H])[H])N=C([H])N=C2N(C=1[H])[C@@]1([H])[C@@]([H])([C@@]([H])([C@@]([H])(C([H])([H])O[H])O1)O[H])O[H]

WHSIXKUPQCKWBY-IOSLPCCCSA-N

InChI=1S/C11H13IN4O4/c12-4-1-16(10-6(4)9(13)14-3-15-10)11-8(19)7(18)5(2-17)20-11/h1,3,5,7-8,11,17-19H,2H2,(H2,13,14,15)/t5-,7-,8-,11-/m1/s1

(2R,3R,4S,5R)-2-(4-amino-5-iodopyrrolo[2,3-d]pyrimidin-7-yl)-5-(hydroxymethyl)oxolane-3,4-diol

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)

配方 1 中的溶解度: ≥ 2.5 mg/mL (6.38 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

---

配方 2 中的溶解度: ≥ 2.5 mg/mL (6.38 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

---

View More

配方 3 中的溶解度: ≥ 2.5 mg/mL (6.38 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液添加到 900 μL 玉米油中并混合均匀。

---

配方 4 中的溶解度: ≥ 2.5 mg/mL (6.38 mM) (饱和度未知) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

---

配方 5 中的溶解度: ≥ 2.5 mg/mL (6.38 mM) (饱和度未知) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

---

配方 6 中的溶解度: ≥ 0.5 mg/mL (1.28 mM) (饱和度未知) in 1% DMSO 99% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

---

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。