Analogue Of Actinomycin Research Articles

Genome Sequencing of Streptomyces griseus SCSIO PteL053, the Producer of 2,2′-Bipyridine and Actinomycin Analogs, and Associated Biosynthetic Gene Cluster Analysis

Marine symbiotic actinomycetes play a key role in drug development and their ecological niches can influence a variety of natural product biosynthesis, providing potential defensive benefits. In this study, we report the whole-genome sequence analysis of marine gastropod mollusk Planaxis sp.-associated Streptomyces griseus SCSIO PteL053, which harbors 28 putative biosynthetic gene clusters (BGCs). Among them, two BGCs encoded by a hybrid non-ribosomal peptide (NRPS)/polyketide (PKS) synthetase and non-ribosomal peptide synthetase (NRPS) are responsible for the synthesis of the known therapeutic metabolites 2,2′-bipyridine and actinomycin analogs, respectively. Detailed bioinformatics analysis revealed the putative BGCs and the functions of the involved genes in the biosynthesis of the known compounds SF2738D (1), SF2738F (2), actinomycin D (3), and Actinomycin Xoβ (4). In the present study, complete-genome sequencing allowed us to rediscover known, clinically useful secondary metabolites in the newly isolated Streptomyces griseus SCSIO PteL053.

Mutasynthesis of Antibacterial Halogenated Actinomycin Analogues.

Through precursor-directed biosynthesis, feeding halogenated (F-, Cl-, Br-, I-) or methoxy-substituted 4-methyl-3-hydroxyanthranilic acid (4-MHA) analogues to the acnGHLM-deleted mutant strain of Streptomyces costaricanus SCSIO ZS0073 led to the production of ten new actinomycin analogues (4-13). Several of the actinomycin congeners displayed impressive antimicrobial activities, with MIC values spanning 0.06-64 μg/mL to clinically derived antibiotic resistant pathogens, including Staphylococcus aureus, Enterococcus faecium, and Candida albicans, with low cytotoxicity.

Efficient, green, and rapid strategy for separating actinomycin D and X2 using supercritical fluid chromatography

Actinomycin D has long been used as a first-line antitumor drug in clinical practice. Actinomycin X2, a new drug lead, is often isolated along with actinomycin D. The minor differences between the two actinomycin analogs pose a daunting challenge in separation. In this study, supercritical fluid chromatography (SFC) was successfully utilized for the purification of actinomycin X2 and actinomycin D from a marine derived Streptomyces sp. DQS-5. After one-step SFC purification, the purities of these two compounds were determined to be 97.3 % and 97.8 %, respectively. This method provides a green alternative for the separation of these pharmacologically important actinomycin antibiotics. This study also demonstrated the development of a simple and rapid method for the separation of natural products based on SFC.

Actinomycin V Inhibits Migration and Invasion via Suppressing Snail/Slug-Mediated Epithelial-Mesenchymal Transition Progression in Human Breast Cancer MDA-MB-231 Cells In Vitro.

Actinomycin V, an analog of actinomycin D produced by the marine-derived actinomycete Streptomyces sp., possessing a 4-ketoproline instead of a 4-proline in actinomycin D. In this study, the involvement of snail/slug-mediated epithelial-mesenchymal transition (EMT) in the anti-migration and -invasion actions of actinomycin V was investigated in human breast cancer MDA-MB-231 cells in vitro. Cell proliferation effect was evaluated by 3-(4,5-Dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay. Wound-healing and Transwell assay were performed to investigate the anti-migration and -invasion effects of actinomycin V. Western blotting was used to detect the expression levels of E-cadherin, N-cadherin, vimentin, snail, slug, zinc finger E-box binding homeobox 1 (ZEB1), and twist proteins and the mRNA levels were detected by rt-PCR. Actinomycin V showed stronger cytotoxic activity than that of actinomycin D. Actinomycin V up-regulated both of the protein and mRNA expression levels of E-cadherin and down-regulated that of N-cadherin and vimentin in the same cells. In this connection, actinomycin V decreased the snail and slug protein expression, and consequently inhibited cells EMT procession. Our results suggest that actinomycin V inhibits EMT-mediated migration and invasion via decreasing snail and slug expression, which exhibits therapeutic potential for the treatment of breast cancer and further toxicity investigation in vivo is needed.

Anti-MRSA actinomycins D1-D4 from the marine sponge-associated Streptomyces sp. LHW52447

Actinomycins D1−D4 (1–4), four new D-type actinomycin analogues, were isolated from the fermentation broth of a strain of marine sponge-associated Streptomyces sp. LHW52447, together with actinomycin D (5). The structures of 1−4 were determined by a combination analysis of HRMS and NMR spectroscopic methods, and their absolute configurations of amino acids were determined by Marfey's analysis. Actinomycins D1 (1) and D2 (2) are the first two naturally occurring actinomycins with incorporation an oxazole unit into the central phenoxazinone chromophore. Both 1 and 2 showed more potent antibacterial activities against three strains of pathogenic methicillin-resistant Staphylococcus aureus (MRSA) with MIC values of 0.125–0.25 μg/mL than those of 3−5 with MIC values of 0.5–1.0 μg/mL, which suggested that the anti-MRSA activity might be enhanced by the incorporation of an additional oxazole unit. In addition, the cytotoxicity evaluation against WI-38 human diploid lung fibroblasts revealed that the incorporation of oxazole unit could decrease the cytotoxicity of actinomycins on human normal cells.

Fluorescence Analysis of 7-Aminoactinomycin–Telomeric Oligonucleotide d[AGGG(TTAGGG)3] Complexes

Interactions of 7-aminoactinomycin D (a fluorescent analogue of actinomycin D, an anticancer antibiotic) and two structural forms of the model guanine-rich telomeric oligonucleotide d[AGGG(TTAGGG)3] have been studied. We have shown that 7-aminoactinomycin D induces fluorescence in two G-quadruplex structures formed with the presence of potassium or sodium ions. The enthalpy of the interaction between the phenoxasone chromophore of the antibiotic and the telomeric oligonucleotide as determined by analysis of excitation spectra is 5.5 kcal/mol. This value differs little from those obtained for complexes with guanine, adenine, or thymine aggregates (6–7 kcal/mol). In the oligonucleotide, the antibiotic is located within dynamic cavities. Therefore, 7-aminoactinomycin D is not released from telomeric structures to the aqueous phase spontaneously or even by photoexcitation, but it is easily released from the surface of aggregates of the respective nucleobases. The entropy term of the interaction energy calculated as a difference between the total energy determined from the binding constant and the enthalpy determined from excitation spectra constitutes approximately 30% for the telomeric oligonucleotide and is virtually null in interactions with nucleobase aggregates.

Synthesis of actinomycin analogs: XXIV. Non-symmetric derivatives of actinocin containing benzo-18-crown-6 moiety and tertiary ammonium group

Non-symmetric derivatives of actinocin containing fragments of benzo-18-crown-6 and dimethylaminopropylamine (with the crown ether group separated from the heterocyclic chromophore by β-alanine moiety) have been prepared as models of actinomycin D.

Antibacterial and Cytotoxic Actinomycins Y6-Y9 and Zp from Streptomyces sp. Strain Gö-GS12.

Four new Y-type actinomycin analogues named Y6-Y9 (1-4) were isolated and characterized from the scale-up fermentation of the Streptomyces sp. strain Gö-GS12, as well as actinomycin Zp (5), which was, for the first time, isolated as a natural product. Structures of the new compounds were elucidated by the cumulative analyses of NMR spectroscopy and HRMS. The 4-hydroxythreonine on the β-ring of 1 uniquely undergoes both a rearrangement by a 2-fold acyl shift and an additional ring closure with the amino group of the phenoxazinone chromophore, and the α-rings of 4 and 5 contain a rare 5-methyl proline. Compounds 2-5 showed potent antibacterial activities against Gram-positive bacteria that correlated with cytotoxicity against representative human cell lines. The combination of a β-ring rearrangement and additional ring closure in 1 rendered this actinomycin significantly less potent relative to the nonrearranged comparator actinomycin Y5 and other actinomycins.

10.1007/s11171-008-2014-y

Using the screening model of hypochromism, we showed that caffeine forms regular clusters consisting of 8–12 molecules. Addition of 7-aminoactinomycin D (7AAMD, a fluorescent analogue of actinomycin D) to the clusters leads to its sorption on the cluster surface. Photoexcitation of 7AAMD leads to its desorption from the surface into the aqueous phase and emission of a quantum. Fluorescence of 7AAMD in the presence of caffeine clusters is quenched by dinitrophenol more weakly than without clusters (the quenching constants are ∼ 85 and ∼280 M−1, respectively) due to decreased steric availability of the antibiotic to the quencher. Addition of 7AAMD-caffeine complexes to DNA leads to a long-wavelength shift in the excitation spectrum and an increase in the fluorescence intensity along with a shift of the fluorescence spectrum to the short-wavelength area. This fact reflects redistribution of the antibiotic from the caffeine surface to the hydrophobic areas inside DNA.

Caffeine clusters as transmitters of actinomycin antibiotics to DNA in solution

Using the screening model of hypochromism, we showed that caffeine forms regular clusters consisting of 8-12 molecules. Addition of 7-aminoactinomycin D (7AAMD, a fluorescent analogue of actinomycin D) to the clusters leads to its sorption on the cluster surface. Photoexcitation of 7AAMD leads to its desorption from the surface into the aqueous phase and emission of a quantum. Fluorescence of 7AAMD in the presence of caffeine clusters is quenched by dinitrophenol more weakly than without clusters (the quenching constants are approximately 85 and approximately 280 M(-1), respectively) due to decreased steric availability of the antibiotic to the quencher. Addition of 7AAMD-caffeine complexes to DNA leads to a long-wavelength shift in the excitation spectrum and an increase in the fluorescence intensity along with a shift of the fluorescence spectrum to the short-wavelength area. This fact reflects redistribution of the antibiotic from the caffeine surface to the hydrophobic areas inside DNA. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2008, vol. 34, no. 2; see also http://www.maik.ru.

Natural and synthetic analogues of actinomycin D as Grb2-SH2 domain blockers

Natural analogues (D, C2, and VII) of actinomycin inhibit Grb2 SH2 domain binding with phosphopeptide-derived from Shc in vitro and in intracellular system. To study structure–activity relationships, 13 actinomycin analogues were synthesized and we found that the inhibition activity depended on the substituents of cyclic peptide groups in actinomycin and two analogues with Tyr residue are the most potent inhibitors with IC 50 value of 0.5 and 0.8 μM, respectively.

A convergent solid-phase synthesis of actinomycin analogues—towards implementation of double-combinatorial chemistry

The actinomycin antibiotics bind to nucleic acids via both intercalation and hydrogen bonding. We found this ‘double-action attack’ mechanism very attractive in our search for a novel class of nucleic acid binders. A highly convergent, solid-phase synthetic strategy has been developed for a class of peptide-aryl-peptide conjugates modeled upon natural actinomycins. The features of this method include the use of Fmoc solid-phase peptide synthesis, side-chain to side-chain cyclization on the solid phase, a chemoselective cleavage step and segment condensation. The synthetic scheme is consistent with the requirements for combinatorial synthesis and furthermore, the final segment condensation allows, for the first time, double-combinatorial chemistry to be performed where two combinatorial libraries can be reacted with each other.

Sequence-specific actinomycin D binding to single-stranded DNA inhibits HIV reverse transcriptase and other polymerases.

Primer extension assays using recombinant templates constructed to contain all 256 possible base quartets in a minimum length sequence were used to examine binding of the anticancer drug actinomycin D to single-stranded DNA. Single-stranded templates were generated by digestion of linearized plasmid with the double-strand-specific T7 gene 6 exonuclease. Actinomycin D formed high-affinity, kinetically stable complexes that paused primer elongation at specific sites by HIV-1 reverse transcriptase, Sequenase (modified T4 DNA polymerase), the Klenow fragment of Escherichia coli DNA polymerase, and Vent (exo-) DNA polymerase. Pauses occurred most commonly near G+C-rich nucleotide clusters, including GpC steps, the preferred sites of double-stranded DNA binding. Complexes were stable for several minutes at temperatures over 50 degrees C as determined by their abilities to pause Vent polymerase at elevated temperatures. Significant variations were noted in pause patterns of different polymerases, demonstrating differential responses of polymerases to a bound actinomycin. Covalent adducts formed on template DNA by a photoaffinity analog of actinomycin D completely stopped primer extension. These results support the possibility that actinomycin D inhibits transcription elongation by complexing single-stranded DNA in the open transcription complex. Single-stranded DNA binding by actinomycin D or analogs may also provide routes for combating HIV or other viruses which replicate through single-stranded intermediates.

Binding of Actinomycin D to the T(G)nT Motif of Double-Stranded DNA: Determination of the Guanine Requirement in Nonclassical, Non-GpC Binding Sites

Strong binding of the antitumor antibiotic actinomycin D to the sequence 5'-TGGGT-3' in double-stranded DNA was recently established by equilibrium binding studies (Bailey et al., 1993). Actinomycin D binding to this -TGGGT- containing sequence was shown to be comparable to that of an -XGCY- containing oligonucleotide (Ka approximately 10(6) M-1). Investigation of -TGGGT- as a high-affinity binding site for actinomycin D follows from our 1989 sequencing study (Rill et al., 1989) in which the photoaffinity analog of actinomycin D (7-azidoactinomycin D) was used to determine DNA base sequence specificities and neighboring base effects. The studies presented here examine the guanine requirements for actinomycin D binding to such nonclassical (non-dGpC) sites by varying the number of central guanine residues in a series of selected duplex oligonucleotides. The central -T(G)nT- motif varies from n equals 1 to 4. Actinomycin D binding to each of these undecamers is characterized and correlated with binding to oligonucleotides of identical length and similar sequences that contain classical dGpC binding sites. Binding affinities of actinomycin D to this series of oligonucleotide duplexes (10 degrees C) can be summarized as -TGGGT- > -TGGT- > TGGGGT- > -TGT. The kinetics of SDS-induced dissociation of actinomycin D from these oligonucleotides reveal single-exponential decays with duration dependent on the sequence at the binding site. With the exception of the -TGGGT- containing oligomer, dissociation times for the T(G)nT duplexes were drastically different and much shorter than times obtained for the dissociation of actinomycin D from oligonucleotides having classical dGpC sites.(ABSTRACT TRUNCATED AT 250 WORDS)

Use of a photoactive derivative of actinomycin to investigate shuffling between binding sites on DNA.

A photoreactive analog of actinomycin (7-azidoactinomycin D) has been used in experiments to probe directly the shuffling hypothesis of Fox and Waring [Fox, K. R., & Waring, M. J. (1984) Nucleic Acids Res. 12, 9271-9285]. According to this theory, actinomycin D molecules initially interact with non-sequence-specific sites on DNA and subsequently "shuffle" along the polymer in a one-dimensional migratory fashion so as to locate their preferred sequence-dependent binding sites. In the study presented here, the drug-DNA complex was allowed to equilibrate (in the dark) for various periods of time, followed by photolysis which renders the complex irreversible and traps the ligand at its instantaneous binding sites. Visualization of the piperidine-labile sites and investigation of how the intensity of reaction at each site changes with time can provide direct confirmation of the shuffling hypothesis. The data reveal that actinomycin D does indeed engage in shuffling along the DNA. After only short equilibration times (20 s) actinomycin D is observed to bind to a variety of sites on the DNA, including many which are not regarded as canonical preferred binding sites at equilibrium. However, after longer periods of equilibration the intensity of reaction is shown to drop as a function of time at nonspecific sites, with corresponding increase at sequence-specific sites. In addition, base sequences which flank the intercalation sites can be seen to play a major role in influencing the binding and sequence specificity of actinomycin.

Synthesis and properties of some peptide analogs of actinomycin D

Analogues of actinomycin D (AMD) were synthesized in which amino acid replacements were made at various sites in the peptide moieties. These include (i) replacement of both N-methylvalines by N-methylleucine, (ii) replacement of both sarcosines by N-[2-(methoxycarbonyl)ethyl]glycine, and (iii) replacement of one or both D-valines by D-threonine. The purpose of replacements ii and iii was to ascertain the effect upon biological activity of introducing a new side chain which could be functionalized to allow the attachment of carrier molecules such as antibodies. NMR data indicated that none of the analogues had solution conformations significantly different from that of AMD. Difference spectra with DNA revealed that replacement i enhanced binding while the other analogues bound less strongly to DNA. All the analogues had lower antimicrobial activities than AMD. In contrast, 5,5'-(MeLeu)2AMD displayed in vitro antitumor activity comparable with that of AMD at approximately 100-fold lower concentrations.

ChemInform Abstract: Synthesis of Actinomycin Analogues. Part 17. Amides of Actinocin with Benzimidazole Fragments.

ChemInformVolume 20, Issue 19 Natural Products ChemInform Abstract: Synthesis of Actinomycin Analogues. Part 17. Amides of Actinocin with Benzimidazole Fragments. I. V. SKLYAROVA, I. V. SKLYAROVA Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorV. A. KUZNETSOV, V. A. KUZNETSOV Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorA. V. GARABADZHIU, A. V. GARABADZHIU Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorE. N. GLIBIN, E. N. GLIBIN Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorO. F. GINZBERG, O. F. GINZBERG Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this author I. V. SKLYAROVA, I. V. SKLYAROVA Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorV. A. KUZNETSOV, V. A. KUZNETSOV Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorA. V. GARABADZHIU, A. V. GARABADZHIU Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorE. N. GLIBIN, E. N. GLIBIN Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorO. F. GINZBERG, O. F. GINZBERG Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this author First published: May 9, 1989 https://doi.org/10.1002/chin.198919309AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume20, Issue19May 9, 1989 RelatedInformation

ChemInform Abstract: Synthesis of Actinomycin Analogues. Part 16. Actinocyl‐bis‐ω‐aminoacyl‐N‐aminoalcohols.

ChemInformVolume 20, Issue 2 Natural Products ChemInform Abstract: Synthesis of Actinomycin Analogues. Part 16. Actinocyl-bis-ω-aminoacyl-N-aminoalcohols. N. G. PLEKHANOVA, N. G. PLEKHANOVA Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorE. B. POPOVA, E. B. POPOVA Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorE. N. GLIBIN, E. N. GLIBIN Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorO. F. GINZBURG, O. F. GINZBURG Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this author N. G. PLEKHANOVA, N. G. PLEKHANOVA Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorE. B. POPOVA, E. B. POPOVA Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorE. N. GLIBIN, E. N. GLIBIN Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this authorO. F. GINZBURG, O. F. GINZBURG Leningradskii tekhnol. inst. im. LensovetaSearch for more papers by this author First published: January 10, 1989 https://doi.org/10.1002/chin.198902297AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume20, Issue2January 10, 1989 RelatedInformation

Effects of actinomycin D analogs on nucleolar phosphoprotein B23 (37,000 daltons/pI 5.1)

Localization of protein B23 in HeLa cells after treatment with actinomycin D and its analogs was studied using indirect immunofluorescence. Bright nucleolar fluorescence was observed in control HeLa cells. After treatment with actinomycin D (250 ng/ml) for 2 hr, a uniform nucleoplasmic fluorescence was observed. Similar results were obtained with the actinomycin analogs, actinomycin Z5 and actinomycin K2T. Only after a much longer incubation (24 hr) with actinomycin 4-4′-gly was nucleoplasmic fluorescence observed. Actinomycin D, actinomycin Z5, and actinomycin K2T inhibited [ 3H]uridine incorporation into the trichloroacetic acid insoluble fraction of HeLa cells with IC 50 values of 9.5 ± 3.2, 59.1 ± 19.6 and 1423.3 ± 212.2 ng/ml respectively. No inhibition of [ 3H]uridine incorporation was observed using actinomycin 4-4′-gly (2000 ng/ml, 2-hr incubation). The order of potency for the loss of nucleolar fluorescence and the concurrent increase in nucleoplasmic fluorescence was actinomycin D > actinomycin Z5 > actinomycin K2T > actinomycin 4-4′-gly, which correlated with the order of their IC 50 values for inhibition of [ 3H]uridine incorporation. Studies of the effects of actinomycin D and its analogs on RNA synthesis and localization of protein B23 indicated that there is a direct relationship between the B23 “translocation” from nucleolus to nucleoplasm and the inhibition of RNA synthesis. At 45–55% inhibition of RNA synthesis, both nuclear and nucleolar B23 immunofluorescence were observed. At 75–85% inhibition, only a uniform nucleoplasmic fluorescence was observed.

"Reverse" and "symmetrical" analogues of actinomycin D: metabolic activation and in vitro and in vivo tumor growth inhibitory activities.

Two new classes of actinomycin D analogues, tetracyclic "reverse" analogues and a tricyclic "symmetrical" analogue of actinomycin D, are reported. These analogues bind to DNA and the binding does not occur by an intercalation mechanism. The analogues inhibit the synthesis of DNA and RNA in P388 tumor cells and the growth of CCRF-CEM cells in vitro at nanomolar concentrations. The tetracyclic "reverse" analogues, which are structurally related to the previously reported actinomycin D oxazolyl analogues, are metabolized in the presence of rat hepatic microsomes and tumor cell homogenates. The metabolism takes place with the loss of the oxazole ring; thus the "reverse" analogues produce a major metabolite which is the "symmetrical" analogue; the actinomycin oxazolyl analogues generate 7-hydroxyactinomycin D. Further, the microsomes activate the analogues to free-radical states which catalyze the production of superoxide as shown by stimulation of epinephrine oxidation and also indicated by electron paramagnetic resonance studies. The "symmetrical" and "reverse" analogues also demonstrate very high activities in these systems. In in vivo studies using P388/S, P388/ADR leukemia, and B16 melanoma in mice, the analogues showed increased activity and superior therapeutic index values, in comparison to actinomycin D.