SYNTHESIS, CHARACTERIZATION, ANTICANCER ACTIVITY AND REACTIVITY TOWARD BIOLOGIC

合成、表征、抗癌活性和生物反应性

• 批准号: 7381556
• 负责人: CESAR LOZANO
• 金额: $ 8.48万
• 依托单位: UNIVERSITY OF PUERTO RICO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7381556
• 关键词: SYNTHESIS; CHARACTERIZATION; ANTICANCER; ACTIVITY; REACTIVITY

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The major objective of this proposal is to search for new platinum drugs with improved properties compared to currently used metal based anticancer agents such as cisplatin, carboplatin, etc..., and to discover the molecular basis of their cytotoxicity by studying their reactivity toward biological important molecules (DNA, oligonucleotides) of some of the promising discovered drugs. In order to achieve these goals we plan to determine the better conditions for the synthesis and characterization of two series of platinum(II) complexes. 1.- anionic trihalo complexes of platinum(II) in which either a styrylbenzoheterazole or amide derivatives of important biological active carboxylic acids will be used as ligands. These complexes were designed based on the reported anticancer activity of previously synthesized tribromo anionic complexes of styrylbenzoheterazole which demonstrated a different pattern of cross-linking compared to cisplatin. Although no toxicity studies have been performed on these complexes, we expect to decrease any possible unwanted toxicity by substitution of the bromide ligands by chloride bound to the platinum(II) center on other series of compounds. To study the effect of the ligand on these complexes, the new styrylbenzoheterazole ligands to be used will be modified by placing other substituents (electrondonating and/or electronwithdrawing) on either the benzazolo and/or the styryl moieties, and also by changing the heteroatom (NH, S, O) on the benzazolo part of the ligands. Other ligands to be used for the anionic trihalo complexes include amides derived from reported bioactive carboxylic acids, such as 2-phenylacetic acid, phenylbutyric acid, branched and unbranched saturated fatty acids and unsaturated and polyunsaturated fatty acids. 2.- Neutral platinum(II) complexes in which the one or both of the chloride leaving groups in the parent drugs are substituted by the biological active carboxylic acids mentioned above. The parent drugs involves the use of cisplatin, cis-Pt(DACH)CI2, and cis-Pt(en)CI2, where DACH stands for 1,2-diaminocyclohexane and en for ethylenediamine. The use of these ligands are expected to increase the lipophyllicity and membrane permeability of the new complexes and probably to induce new ways of interactions with the main targets for bioactivity. Also the presence of the carboxylate groups are expected to produce a multifunctional compound in such a way that is known that the leaving groups are realized by the platinum complexes before it is bound to DNA. So, the freed carboxylates should exert their own bioactivity on the cells. All of these compounds will be purified and characterized by elemental analysis, UV-Vis spectroscopy, 1H, 3C, 195Pt and two dimension NMR experiments. They will be also crystallized to obtain crystal good enough for X-Ray Diffraction studies. The biological testing against cancer cell lines will be performed at the Biotesting Laboratory at UPR, Rio Piedras Campus. The long term objectives of the proposed projects is to study the interaction of promising complexes with important biological molecules, such as DNA, which is the main target of known platinum complexes. These interactions will be followed by standard procedures andby NMR spectroscopy and HPLC, and their products characterized. In order to obtain information on the preferred sites of binding of the new complexes, the platinated DNA will be enzymatically digested and their adducts separated chromatographically and identified by 1H NMR spectroscopy.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。该提案的主要目的是寻找与目前使用的基于金属的抗癌剂如顺铂、卡铂等相比具有改进性质的新铂药物。并通过研究它们对一些有希望的已发现药物的生物学重要分子（DNA、寡核苷酸）的反应性来发现它们的细胞毒性的分子基础。为了实现这些目标，我们计划确定两个系列的铂（II）配合物的合成和表征的更好的条件。 1.-铂（II）的阴离子三卤代络合物，其中苯乙烯基苯并杂四唑或重要的生物活性羧酸的酰胺衍生物将用作配体。这些复合物的设计基于先前合成的苯乙烯基苯并杂四唑的三溴阴离子复合物的抗癌活性，所述苯乙烯基苯并杂四唑的三溴阴离子复合物与顺铂相比表现出不同的交联模式。虽然没有毒性研究已经进行了这些配合物，我们希望减少任何可能的不必要的毒性，通过取代溴代配体的氯化物结合到铂（II）中心的其他系列的化合物。为了研究配体对这些配合物的影响，将通过在苯并唑和/或苯乙烯基部分上放置其他取代基（给电子和/或吸电子）以及通过改变配体的苯并唑部分上的杂原子（NH，S，O）来修饰待使用的新的苯乙烯基苯并杂唑配体。用于阴离子三卤配合物的其他配体包括衍生自已报道的生物活性羧酸的酰胺，例如2-苯乙酸、苯丁酸、支化和非支化饱和脂肪酸以及不饱和和多不饱和脂肪酸。 2.-中性铂（II）络合物，其中母体药物中的一个或两个氯离去基团被上述生物活性羧酸取代。母体药物涉及使用顺铂、顺式-Pt（DACH）Cl 2和顺式-Pt（en）Cl 2，其中DACH代表1，2-二氨基环己烷，en代表乙二胺。这些配体的使用预计将增加新的复合物的脂溶性和膜渗透性，并可能诱导与生物活性的主要靶标相互作用的新方式。此外，羧酸根基团的存在预期以已知的方式产生多官能化合物，即离去基团在其与DNA结合之前通过铂络合物实现。因此，游离的羧酸盐应该对细胞发挥其自身的生物活性。所有这些化合物均经过元素分析、紫外-可见光谱、~ 1H、~ 3C、~（195）Pt和二维核磁共振实验进行了纯化和表征。它们也将被结晶以获得足够好的晶体用于X射线衍射研究。针对癌细胞系的生物学试验将在Rio Piedras Campus的UPR生物试验实验室进行。 拟议项目的长期目标是研究有前途的配合物与重要生物分子的相互作用，如DNA，这是已知铂配合物的主要目标。这些相互作用将遵循标准程序和核磁共振光谱和高效液相色谱法，其产品的特点。为了获得关于新复合物的优选结合位点的信息，铂化DNA将被酶促消化，并且它们的加合物通过色谱法分离并通过1H NMR光谱法鉴定。