Epothilones as lead structures for the synthesis-based discovery of new chemotypes for microtubule stabilization

Epothilones are macrocyclic bacterial natural products with potent microtubule-stabilizing and antiproliferative activity. They have served as successful lead structures for the development of several clinical candidates for anticancer therapy. However, the structural diversity of this group of clinical compounds is rather limited, as their structures show little divergence from the original natural product leads. Our own research has explored the question of whether epothilones can serve as a basis for the development of new structural scaffolds, or chemotypes, for microtubule stabilization that might serve as a basis for the discovery of new generations of anticancer drugs. We have elaborated a series of epothilone-derived macrolactones whose overall structural features significantly deviate from those of the natural epothilone scaffold and thus define new structural families of microtubule-stabilizing agents. Key elements of our hypermodification strategy are the change of the natural epoxide geometry from cis to trans, the incorporation of a conformationally constrained side chain, the removal of the C3-hydroxyl group, and the replacement of C12 with nitrogen. So far, this approach has yielded analogs 30 and 40 that are the most advanced, the most rigorously modified, structures, both of which are potent antiproliferative agents with low nanomolar activity against several human cancer cell lines in vitro. The synthesis was achieved through a macrolactone-based strategy or a high-yielding RCM reaction. The 12-aza-epothilone ("azathilone" 40) may be considered a "non-natural" natural product that still retains most of the overall structural characteristics of a true natural product but is structurally unique, because it lies outside of the general scope of Nature's biosynthetic machinery for polyketide synthesis. Like natural epothilones, both 30 and 40 promote tubulin polymerization in vitro and at the cellular level induce cell cycle arrest in mitosis. These facts indicate that cancer cell growth inhibition by these compounds is based on the same mechanistic underpinnings as those for natural epothilones. Interestingly, the 9,10-dehydro analog of 40 is significantly less active than the saturated parent compound, which is contrary to observations for natural epothilones B or D. This may point to differences in the bioactive conformations of N-acyl-12-aza-epothilones like 40 and natural epothilones. In light of their distinct structural features, combined with an epothilone-like (and taxol-like) in vitro biological profile, 30 and 40 can be considered as representative examples of new chemotypes for microtubule stabilization. As such, they may offer the same potential for pharmacological differentiation from the original epothilone leads as various newly discovered microtubule-stabilizing natural products with macrolactone structures, such as laulimalide, peloruside, or dictyostatin.     

Natural products as leads for anticancer drug discovery: discovery of new chemotypes of microtubule stabilizers through reengineering of the epothilone scaffold

Epothilones are bacterial macrolides with potent microtubule-stabilizing and antiproliferative activity, which have served as successful lead structures for the discovery of several clinical candidates for cancer treatment. Overall, seven epothilone-type agents have been advanced to clinical evaluation in humans so far and one of these has been approved by the FDA in 2007 for clinical use in breast cancer patients. Notwithstanding these impressive numbers, however, the structural diversity represented by the collection of epothilone analogs that have been (or still are) investigated clinically is rather limited and their individual structures show little divergence from the original natural product leads. In contrast, we have elaborated a series of epothilone-derived macro-lactones, whose overall structural features significantly deviate from those of the natural epothilone scaffold and thus define new structural families of microtubule-stabilizing agents. Key elements of our hypermodification strategy are the change of the natural epoxide geometry from cis to trans, the incorporation of conformationally constrained side chains, the removal of the C(3)-hydroxyl group, and the replacement of C(12) with nitrogen. The latter modification leads to aza-macrolides that may be described as 'non-natural natural products'.     

Translating Nature's Library: The Bryostatins and Function-Oriented Synthesis

We review in part our computational, design, synthesis, and biological studies on a remarkable class of compounds and their designed analogs that have led to preclinical candidates for the treatment of cancer, a first-in-class approach to Alzheimer's disease, and a promising strategy to eradicate HIV/AIDS. Because these leads target, in part, protein kinase C (PKC) isozymes, they have therapeutic potential even beyond this striking set of therapeutic indications. This program has given rise to new synthetic methodology and represents an increasingly important direction of synthesis focused on achieving function through synthesis-informed design (function-oriented synthesis).     

Regioselective Cobalt‐Catalysed Hydrovinylation for the Synthesis of Non‐Conjugated Enones and 1,4‐Diketones

The highly regioselective cobalt‐catalysed 1,4‐hydrovinylation of terminal alkenes with 2‐trimethylsilyloxy‐1,3‐butadiene generates in a stereospecific fashion unsaturated E‐configured silyl enol ether intermediates that are suitable for diastereoselective Mukaiyama‐aldol reactions with bulky aliphatic aldehydes. The acidic hydrolysis of the enol ethers to γ,δ‐unsaturated ketones followed by ozonolysis can be used for the synthesis of various 1,4‐diketones and polycarbonyl derivatives. The 1,4‐diketones and polycarbonyl derivatives were successfully tested for the synthesis of some mono‐ and bis‐pyrrole derivatives. The γ,δ‐unsaturated ketones are useful building blocks (e.g., in natural product synthesis) and can be generated in a one‐pot procedure.     

阿魏酸衍生物的设计、合成及除草活性

[目的]以天然产物阿魏酸为先导设计合成具有较强除草活性的衍生物.[方法]运用农药分子合理设计原理设计合成阿魏酸酰胺类似物,采用小杯法测定除草活性.[结果]合成了11个新型的阿魏酸衍生物;小杯法测定显示,在400 mg/L质量浓度下,化合物8f和8k的除草活性较好;半抑制浓度测定结果表明:8f对马唐根和茎的IC50值为分...     

Deletion of the C26 Methyl Substituent from the Bryostatin Analogue Merle 23 Has Negligible Impact on Its Biological Profile and Potency

Important strides are being made in understanding the effects of structural features of bryostatin 1, a candidate therapeutic agent for cancer and dementia, in conferring its potency toward protein kinase C and the unique spectrum of biological responses that it induces. A critical pharmacophoric element in bryostatin 1 is the secondary hydroxy group at the C26 position, with a corresponding primary hydroxy group playing an analogous role in binding of phorbol esters to protein kinase C. Herein, we describe the synthesis of a bryostatin homologue in which the C26 hydroxy group is primary, as it is in the phorbol esters, and show that its biological activity is almost indistinguishable from that of the corresponding compound with a secondary hydroxy group.     

Heterogeneous-Backbone Foldamer Mimics of a Computationally Designed,Disulfide-Rich Miniprotein

Disulfide-rich peptides have found widespread use in the development of bioactive agents; however, low proteolytic stability and the difficulty of exerting synthetic control over chain topology present barriers to their application in some systems. Herein, we report a method that enables the creation of artificial backbone (“foldamer”) mimics of compact, disulfide-rich tertiary folds. Systematic replacement of a subset of natural α-residues with various artificial building blocks in the context of a computationally designed prototype sequence leads to “heterogeneous-backbone” variants that undergo clean oxidative folding, adopt tertiary structures indistinguishable from that of the prototype, and enjoy proteolytic protection beyond that inherent to the topologically constrained scaffold. Collectively, these results demonstrate systematic backbone substitution to be a viable method to engineer the properties of disulfide-rich sequences and expands the repertoire of protein mimicry by foldamers to an exciting new structural class.     

Phospholipases: Occurrence and production in microorganisms,assay for high-throughput screening,and gene discovery from natural and man-made diversity

Various kinds of phospholipids have wide industrial applications such as in food and nutraceuticals, cosmetics, agricultural products, and pharmaceuticals. The demand for reliable biocatalysts for the production of phospholipid products, such as phospholipases A₁, A₂, C, and D, has steadily increased over the past several decades. A large number of microbial phospholipases have been isolated and characterized, and the increasing availability of these enzymes could eventually lead to the sustained development of phospholipid-related biotechnology. Although a number of reactions have been performed using phospholipases, a reliable and efficient supply of superior phospholipases in quantity is still a challenge for their practical application. High-throughput functional assay methods for phospholipases should be devised to develop superior new species from the huge diversity of phospholipases. Recent biotechnological advances in the discovery of new phospholipase genes from natural sources, such as extremophiles of phospholipases by protein engineering, such as directed evolution, can provide valuable means of rapidly developing practical uses of phospholipases for various applications.     

Carbon nanotubes: synthesis,integration, and properties

Synthesis of carbon nanotubes by chemical vapor deposition over patterned catalyst arrays leads to nanotubes grown from specific sites on surfaces. The growth directions of the nanotubes can be controlled by van der Waals self-assembly forces and applied electric fields. The patterned growth approach is feasible with discrete catalytic nanoparticles and scalable on large wafers for massive arrays of novel nanowires. Controlled synthesis of nanotubes opens up exciting opportunities in nanoscience and nanotechnology, including electrical, mechanical, and electromechanical properties and devices, chemical functionalization, surface chemistry and photochemistry, molecular sensors, and interfacing with soft biological systems.     

Isolation,identification, synthesis,and bioassay of the pheromone of the comstock mealybug and some analogs

The sex pheromone of the Comstock mealybug,Pseudococcus comstocki (Kuwana) was isolated from volatiles trapped from air passing over virgin female insects. Combined gas chromatography, mass and infrared spectroscopy, and microreactions indicated that the structure was 2,6-dimethyl-1,5-heptadien-3-ol acetate. This was confirmed by synthesis; several analogs were also prepared. The natural and synthetic pheromone caused similar trap capture of male insects. Synthetic analogs were significantly less attractive in field tests and the addition of minor amounts of the corresponding alcohol to the pheromone appeared to enhance trap capture, but the results were not statistically conclusive.     

Selective C−H Functionalizations by Electrochemical Reactions with Palladium Catalysts

Efficient, selective, transition-metal-catalyzed C−H functionalizations have been widely studied and are recognized as atom- and step-economical tools in organic synthesis. During the past two decades, a variety of catalytic reactions involving C−H bond cleavage have been developed. In this review, we briefly survey studies dealing with transition-metal-catalyzed efficient and selective C−H functionalization by electrochemical reactions.     

Synthesis and evaluation of 1-amino-6-halo-β-carbolines as antimalarial and antiprion agents

Malaria is one of the world's most devastating parasitic diseases, causing almost one million deaths each year. Growing resistance to classical antimalarial drugs, such as chloroquine, necessitates the discovery of new therapeutic agents for successful control of this global disease. Here, we report the synthesis of some 6-halo-β-carbolines as analogues of the potent antimalarial natural product, manzamine A, retaining its heteroaromatic core whilst providing compounds with much improved synthetic accessibility. Two compounds displayed superior activity to chloroquine itself against a resistant Plasmodium falciparum strain, identifying them as promising leads for future development. Furthermore, in line with previous reports of similarities in antimalarial and antiprion effects of aminoaryl-based antimalarial agents, the 1-amino-β-carboline libraries were also found to possess significant bioactivity against a prion-infected cell line.     

Signaling effects of demethylasterriquinone B1, a selective insulin receptor modulator

A possible breakthrough in the treatment of diabetes was made with the discovery that a fungal natural product, demethylasterriquinone B1 (DAQ B1), is an orally active, small-molecule mimic of insulin. Subsequent work has shown that the glucose-lowering effects of DAQ B1 are not accompanied by enhanced vascular proliferation, which is a side effect of chronic insulin administration that can lead to arteriosclerosis. Our recent short and modular total synthesis of DAQ B1 could be readily modified to create congeners and afforded ample supplies of the natural product, which permitted intracellular signal transduction of DAQ B1 to be examined. The activities of DAQ B1 and over a dozen related structures were studied for insulin receptor (IR) and insulin receptor substrate-1 phosphorylation. Examination of the effect of DAQ B1 on kinases downstream of the IR in insulin signal transduction showed selective activation of Akt kinase (a metabolic effect) but not of extracellular-regulated kinase (a proliferative effect). The influence of DAQ B1 on gene expression (determined by a microarray study) was also divergent from that of insulin, which activates both proliferative and metabolic pathways. The action of DAQ B1 as a selective insulin receptor modulator can be accounted for by its ability to selectively activate one kinase among the many emanating from insulin receptor autophosphorylation and its reduced effects on gene expression.     

以天然产物为先导化合物开发的农药品种(Ⅰ)--杀菌剂

研究天然产物的重点不是直接利用它，而是以其为先导化舍物开发活性更优的天然产物替代品。只有如此，才能保护有限的资源，更好的为人类持续发展服务。在农药创制研究中以天然产物为先导化合物进行研究、开发新农药品种的方法仍将是一种有效的方法；因为以具有活性的天然产物为先导化合物不仅可以更快、更经济的发现活性更优的类似物，而且因其内在的性能是产品更易符合环境保护与持续发展之需要。本文概述了以天然产物为先导化合物进行的新农药创制研究以及开发的农药品种，其中杀菌剂包括乙蒜素、稻瘟灵、恶霉灵、肉桂酸衍生物、吡咯类化合物、甲氧基丙烯酸酯类等。旨在吸取他人成功经验，利用其研究之成果研制我们自己的新农药品种。     

Design,Synthesis and Structure–Activity Relationships Studies on the D Ring of the Natural Product Triptolide

Triptolide is a diterpene triepoxide natural product isolated from Tripterygium wilfordii Hook F, a traditional Chinese medicinal herb. Triptolide has previously been shown to possess antitumor, anti‐inflammatory, immunosuppressive, and antifertility activities. Earlier reports suggested that the five‐membered unsaturated lactone ring (D ring) is essential for potent cytotoxicity, however, to the best of our knowledge, systematic structure–activity relationship studies have not yet been reported. Here, four types of D ring‐modified triptolide analogues were designed, synthesized and evaluated against human ovarian (SKOV‐3) and prostate (PC‐3) carcinoma cell lines. The results suggest that the D ring is essential to potency, however it can be modified, for example to C18 hydrogen bond acceptor and/or donor furan ring analogues, without complete loss of cytotoxic activity. Interestingly, evaluation of the key series of C19 analogues showed that this site is exquisitely sensitive to polarity. Together, these results will guide further optimization of this natural product lead compound for the development of potent and potentially clinically useful triptolide analogues.     

The effects of dehydroepiandrosterone on carcinogenesis, obesity, the immune system, and aging

With the passage of the U.S. Dietary Supplement Health and Education Act of 1994, dehydroepiandrosterone (DHEA, 5-androsten-3β-ol-17-one) has become widely available, and a large and growing market has developed for this “fountain of youth”. DHEA has been shown to have significant beneficial effects in animals, which may lead to clinical uses in man. Historically, the U.S. Food and Drug Administration removed DHEA from the over-the-counter market in 1985 because there was no support for the health claims that were made for this product. Almost all of the biological data was on animals and there was a lack of demonstrated efficacy in humans. Recently there have been a number of small clinical trials in humans but the results have not been as positive as in the animal tests. This review will be restricted to the effects of DHEA on carcinogenesis, obesity, the immune system, and aging. Four hypotheses have been proposed to explain the underlying biochemical mechanism(s) by which DHEA exerts its beneficial properties. The first is based on the inhibitory effect of DHEA on mammalian glucose-6-phosphate dehydrogenase. This mechanism can explain the antiinitiation and antipromotion steps in some cases of carcinogenesis. The second biochemical mechanism involves the induction of peroxisomes and peroxisome-associated enzymes. The third explanation is that DHEA works in a similar fashion to the known anticarcinogenic action of food restriction. An antiglucocorticoid mechanism has also been suggested. A hypothesis for the increase followed by the decrease in the levels of DHEA with age is proposed. A number of new synthetic DHEA analogs have been synthesized and tested. They offer the best hope for the development of a clinically useful drug based on the properties of DHEA.     

Design and synthesis of 1,4-bis[4-(1,1-dicyanovinyl)styryl]-2,5-bis(alkoxy)benzenes as red organic electroluminescent PPV analogs

A series of 1,4-distyrylbenzene derivatives containing 1,1-dicyanovinyl moieties were designed as poly-p-phenylenevinylene analogs to obtain the red electroluminescence assisted by the computational results. Based on the subsequent synthesis and optical studies, the present work indicated that introduction of 1,1-dicyano group as electron receptor in PPV skeleton could shift the electroluminescence significantly to the red region. The present work has shown a successful strategy to tune the emission of PPV analogs based on the theoretical design and organic synthesis.     

Fluorescent Nucleoside Analogs: Probes for Investigating Nucleic Acid Structure and Function

Base-modified fluorescent nucleoside analog probes have been very valuable in the study of nucleic acid structure and function. Many of them structurally resemble natural bases, and also display useful properties, such as large Stokes shifts and sensitivity to microenvironment changes. Therefore, unlike traditional fluorescence probes, which mostly report global changes, nucleoside analogs, when incorporated into oligonucleotides, can photophysically report changes that occur around the site of interest, at the nucleotide level. In this review, we provide an overview of various strategies that have been employed to design base-modified fluorescent nucleoside analogs. Then we review recent developments and applications of new generation fluorescent nucleoside analogs with a particular focus on the synthesis, photophysical characterizations and applications of heterobicycle-conjugated pyrimidine nucleoside analogs that have been developed by our group. These analogs, which have a minimal effect on the structures of the oligonucleotides into which they are incorporated, show emission in the visible region and excellent fluorescence solvatochromism. Notably, unlike the majority of fluorescent nucleoside analogs developed so far, these analogs retain their fluorescence efficiency when incorporated into oligonucleotides. We anticipate that these nucleoside analogs, with such photophysical properties, would be useful in designing robust biophysical assays to study nucleic acids.     

Preparation and Dissolution Profiles of the Amorphous,Dihydrated Crystalline,and Anhydrous Crystalline Forms of Paclitaxel

The selection of pharmaceutical polymorphisms in the final production step is very important in terms of product recovery, properties, and storage. The amorphous, dihydrated crystalline, and anhydrous crystalline forms of paclitaxel were prepared using precipitation, spray drying, and colloid formation methods. These methods were found to be highly efficient and convenient, giving high recovery, short processing time, and good stability, as compared with conventional methods such as freeze drying, evaporation, recrystallization, and melting. The polymorphic natures of the resulting paclitaxel samples were confirmed by XRPD, IR, TGA, DSC, and SEM. The dissolution rates of the paclitaxel samples were studied in pharmaceutical solvents, which included cotton seed oil, corn oil, tricaprylin, and tributyrin. For each solvent, all of the amorphous paclitaxel samples showed much higher dissolution rates than the dihydrated crystalline, anhydrous crystalline, and commercial forms, and can be used for clinical applications that demand improvements in drug delivery.