Synthesis and in vitro Bioactivity of Polyunsaturated Fatty Acid Conjugates of Combretastatin A-4

Combretastatin A-4 (CA-4) (1) is a plant-derived anticancer agent binding to the tubulin colchicine site. Polyunsaturated fatty acids (PUFA) are readily taken up by cancer cells and have been used to improve cell targeting. In the present study, four CA-4-PUFA conjugates were synthesized by coupling combretastatin A-4 (1) with several polyunsaturated fatty acids. The conjugates (2a-d) were characterized using spectroscopic methods. Their cytotoxicity was evaluated against human breast cancer cells (MCF-7) and the inhibition of tubulin polymerization was determined in vitro. All conjugates influenced tubulin polymerization with the arachidonic acid conjugate (2c) displaying cytotoxicity similar in potency to the natural product CA-4 (1)

Caffeine affects the biological responses of human hematopoietic cells of myeloid lineage via downregulation of the mTOR pathway and xanthine oxidase activity

Correction of human myeloid cell function is crucial for the prevention of inflammatory and allergic reactions as well as leukaemia progression. Caffeine, a naturally occurring food component, is known to display anti-inflammatory effects which have previously been ascribed largely to its inhibitory actions on phosphodiesterase. However, more recent studies suggest an additional role in affecting the activity of the mammalian target of rapamycin (mTOR), a master regulator of myeloid cell translational pathways, although detailed molecular events underlying its mode of action have not been elucidated. Here, we report the cellular uptake of caffeine, without metabolisation, by healthy and malignant hematopoietic myeloid cells including monocytes, basophils and primary acute myeloid leukaemia mononuclear blasts. Unmodified caffeine downregulated mTOR signalling, which affected glycolysis and the release of pro-inflammatory/pro-angiogenic cytokines as well as other inflammatory mediators. In monocytes, the effects of caffeine were potentiated by its ability to inhibit xanthine oxidase, an enzyme which plays a central role in human purine catabolism by generating uric acid. In basophils, caffeine also increased intracellular cyclic adenosine monophosphate (cAMP) levels which further enhanced its inhibitory action on mTOR. These results demonstrate an important mode of pharmacological action of caffeine with potentially wide-ranging therapeutic impact for treating non-infectious disorders of the human immune system, where it could be applied directly to inflammatory cells

The Azinomycins, Discovery, Synthesis and DNA binding Studies

Nature is a rich source of antitumour agents that can act as a paradigm for the development of synthetic agents that retain the biological activity of the natural product but are targeted to the tumor, have increased bioavailability or are simply prodrug forms that are inactive until administered. Examples of potent antitumour antibiotics that interact with DNA include CC-1065 and the duocarmycins, ecteinascidin 743 and rebeccamycin. The azinomycins A (1) and B (2) (Fig. 24.1) and the truncated analogue 3 represent novel structures that have been extensively explored from a synthetic viewpoint, bot for which studies of structure-activity relationships from a DNA binding and biological viewpoint have been few and widely spaced chronologically. In this chapter we aim to bring together the details of the isolation, synthesis, structural, and DNA-binding studies of the azinomycins, to generate a starting point from which to begin to understand the properties of these natural products that could lead to new, therapeutically relevant compounds

Synthesis and in vitro Bioactivity of Polyunsaturated Fatty Acid Conjugates of Combretastatin A-4

Combretastatin A-4 (CA-4) (1) is a plant-derived anticancer agent binding to the tubulin colchicine site. Polyunsaturated fatty acids (PUFA) are readily taken up by cancer cells and have been used to improve cell targeting. In the present study, four CA-4-PUFA conjugates were synthesized by coupling combretastatin A-4 (1) with several polyunsaturated fatty acids. The conjugates (2a-d) were characterized using spectroscopic methods. Their cytotoxicity was evaluated against human breast cancer cells (MCF-7) and the inhibition of tubulin polymerization was determined in vitro. All conjugates influenced tubulin polymerization with the arachidonic acid conjugate (2c) displaying cytotoxicity similar in potency to the natural product CA-4 (1)

Antitumour antibiotics with potent activity against multidrug resistant (MDR) Staphylococcus aureus: a new approach to targeting resistant bacteria.

As hospital reports of strains of resistant bacteria are continuing to increase, a new approach is required for the identification of small molecules with antibacterial activity. Natural products that bind covalently to their biological target have been largely unexplored, although in the field of cancer chemotherapy, such molecules have been shown to counter resistance developed through efflux mechanisms. The azinomycins are potent antitumour agents that alkylate DNA and one of the natural products, compound 1, is a mono-alkylator that has been reported to retain potent antitumour activity. All four diastereomers of 1 were synthesized via a route involving late stage introduction of the epoxide stereocentre and separation of the resulting compounds. A non-alkylating analogue and a potential alkylator that cannot intercalate were also made. All four diastereomers are potent antibacterial agents in cell lines containing efflux-based resistance mechanisms. MIC values in the range of 0.25-1.0 microg/ml were observed. Comparison with the antitumour activity of the compounds suggests that the antibacterial activity stems from a similar mechanism of action involving DNA alkylation. As the ultimate molecular target of the azinomycins is unknown, bacterial strains may represent an interesting route for the discovery of the downstream mechanisms affected by DNA alkylation

Truncated azinomycin analogues intercalate into DNA.

NoThe design and synthesis of a potentially more therapeutically-viable azinomycin analogue 4 based upon 3 has been completed. It involved coupling of a piperidine mustard to the acid chloride of the azinomycin chromophore. Both the designed azinomycin analogue 4 and the natural product 3 bind to DNA and cause unwinding, supporting an intercalative mode of binding. Graphical abstract A designed analogue of the left half of azinomycin has been synthesized and unwinds supercoiled DNA

Design and synthesis of a DNA-crosslinking azinomycin analogue

NoThe azinomycins are potent antitumour antibiotics that are able to crosslink DNA, but are relatively unstable and unlikely to progress as therapeutic candidates. A prototype analogue 4 with more clinical potential has been designed and synthesised and incorporates the epoxide function of the azinomycins and a nitrogen mustard. Two further analogues 5 and 6 that can alkylate DNA but cannot crosslink the duplex have also been synthesised. Compound 4 crosslinks DNA efficiently at nM concentrations. Compounds 4¿6 were submitted to the NCI 60 cell line screen and have similar antitumour activity, although 4 is slightly less active than the non-crosslinking compounds. These observations will be important in the design of further azinomycin analogues with antitumour activity