Exporting Metal-Carbene Chemistry to Live Mammalian Cells: Copper-Catalyzed Intracellular Synthesis of Quinoxalines Enabled by N-H Carbene Insertions

Implementing catalytic organometallic transformations in living settings can offer unprecedented opportunities in chemical biology and medicine. Unfortunately, the number of biocompatible reactions so far discovered is very limited, and essentially restricted to uncaging processes. Here, we demonstrate the viability of performing metal carbene transfer reactions in live mammalian cells. In particular, we show that copper (II) catalysts can promote the intracellular annulation of alpha-keto diazocarbenes with ortho-amino arylamines, in a process that is initiated by an N-H carbene insertion. The potential of this transformation is underscored by the in cellulo synthesis of a product that alters mitochondrial functions, and by demonstrating cell selective biological responses using targeted copper catalysts. Considering the wide reactivity spectrum of metal carbenes, this work opens the door to significantly expanding the repertoire of life-compatible abiotic reactionsThis work has received financial support from Spanish grants (PID2019-108624RB-I00, RTI2018-093813-J-I00 and ORFEO-CINQA network CTQ2016-81797-REDC), the Consellería de Cultura, Educación e Ordenación Universitaria (2015-CP082, ED431C-2017/19 and Centro Singular de Investigación de Galicia accreditation 2019–2022, ED431G 2019/03), the European Union (European Regional Development Fund-ERDF corresponding to the multiannual financial framework 2014–2020), and the European Research Council (Advanced Grant No. 340055). S.G. thanks the European Union (European Regional Development Fund, Interreg V-A POCTEP España-Portugal programme, 2iqbioneuro project) and M.T.G. thanks the financial support from the Agencia Estatal de Investigación (RTI2018-093813-J-I00)S

Intracellular Ruthenium‐Promoted (2+2+2) Cycloadditions

This is the peer reviewed version of the following article: J. Miguel-Ávila, M. Tomás-Gamasa, J. L. Mascareñas, Angew. Chem. Int. Ed. 2020, 59, 17628, which has been published in final form at https://doi.org/10.1002/anie.202006689. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived VersionsMetal‐mediated intracellular reactions are becoming invaluable tools in chemical and cell biology, and hold promise for strongly impacting the field of biomedicine. Most of the reactions reported so far involve either uncaging or redox processes. Demonstrated here for the first time is the viability of performing multicomponent alkyne cycloaromatizations inside live mammalian cells using ruthenium catalysts. Both fully intramolecular and intermolecular cycloadditions of diynes with alkynes are feasible, the latter providing an intracellular synthesis of appealing anthraquinones. The power of the approach is further demonstrated by generating anthraquinone AIEgens (AIE=aggregation induced emission) that otherwise do not go inside cells, and by modifying the intracellular distribution of the products by simply varying the type of ruthenium complexThis work has received financial support from the Spanish Government (SAF2016‐76689‐R, ORFEO‐CINQA network CTQ2016‐81797‐REDC) the Consellería de Cultura, Educación e Ordenación Universitaria (2015‐CP082, ED431C‐2017/19 and Centro Singular de Investigación de Galicia Accreditation 2019–2022, ED431G 2019/03), the European Union (European Regional Development Fund‐ERDF corresponding to the multiannual financial framework 2014–2020), and the European Research Council (Advanced Grant No. 340055). J.M.Á. thanks the Ministerio de Educación, Cultura y Deporte for the FPU fellowship (FPU16/00711) and M.T.G. thanks the financial support from the Agencia Estatal de Investigación (RTI2018‐093813‐J‐I00)S

Synthesis of gamma-labeled nucleoside 5 `-triphosphates using click chemistry

Real-time enzymatic studies are gaining importance as their chemical and technical instrumentation improves. Here we report the efficient synthesis of gamma-alkyne modified triphosphate amidates that are converted into a variety of gamma-fluorophore labeled triphosphates by Cu(I) catalyzed alkyne/azide click reactions. The synthesized triphosphates are incorporated into DNA by DNA polymerases

Synthesis and properties of a Cu(ii) complexing pyrazole ligandoside in DNA

The development of metal base pairs is of immense importance for the construction of DNA nanostructures. Here we report the synthesis of a biaryl pyrazole-phenol nucleoside that forms in DNA a stable self-pair upon complexation of a Cu(ii) ion. A sequence with five consecutive pyrazole nucleotides allows the complexation of five Cu(ii) ions in a row

Transition metal catalysis in the mitochondria of living cells

The development of transition metal catalysts capable of promoting non-natural transformations within living cells can open significant new avenues in chemical and cell biology. Unfortunately, the complexity of the cell makes it extremely difficult to translate standard organometallic chemistry to living environments. Therefore, progress in this field has been very slow, and many challenges, including the possibility of localizing active metal catalysts into specific subcellular sites or organelles, remain to be addressed. Herein, we report a designed ruthenium complex that accumulates preferentially inside the mitochondria of mammalian cells, while keeping its ability to react with exogenous substrates in a bioorthogonal way. Importantly, we show that the subcellular catalytic activity can be used for the confined release of fluorophores, and even allows selective functional alterations in the mitochondria by the localized transformation of inert precursors into uncouplers of the membrane potentialWe are thankful for the support given by the Spanish grant SAF2013-41943-R, the Xunta de Galicia (GRC2013-041 and 2015-CP082), the ERDF and the European Research Council (Advanced Grant No. 340055). M.T.G. thanks the Ministerio de Economía y Competitividad for the Postdoctoral fellowship. M.M.C. thanks Ministerio de Economía y Competitividad for the Juan de la Cierva-Incorporacio´n fellowship (IJCI-2014-19326).S

Ruthenium-Catalyzed Redox Isomerizations inside Living Cells

Tailored ruthenium(IV) complexes can catalyze the isomerization of allylic alcohols into saturated carbonyl derivatives under physiologically relevant conditions, and even inside living mammalian cells. The reaction, which involves ruthenium-hydride intermediates, is bioorthogonal and biocompatible, and can be used for the “in cellulo” generation of fluorescent and bioactive probes. Overall, our research reveals a novel metal-based tool for cellular intervention, and comes to further demonstrate the compatibility of organometallic mechanisms with the complex environment of cellsThis work has received financial support from the Spanish Government (SAF2016-76689-R, Orfeo-cinqa network CTQ2016-81797-REDC), the Consellería de Cultura, Educación e Ordenación Universitaria (2015-CP082, ED431C 2017/19), Centro Singular de Investigación de Galicia accreditation 2016–2019, ED431G/09), the European Union (European Regional Development Fund-ERDF), and the European Research Council (Advanced Grant No. 340055)S

Kebijakan Penanggulangan Bencana Berbasis Komunitas: Kampung Siaga Bencana dan Desa/kelurahan Tangguh Bencana

Bencana alam sering terjadi di Indonesia, Kementerian Sosial membuat kebijakan programkampung siaga bencana dan Badan Nasional Penanggulangan Bencana membuat kebijakan programdesa/kelurahan tangguh bencana. Keduanya, merupakan kebijakan pemerintah dalam penanggulanganbencana berbasis komunitas. Sehingga terkesan terjadi tumpang tindih program. Oleh karena itu penelitianini membandingkan kebijakan program kampung siaga bencana dan desa tangguh bencana dilihat darilembaga pembuat kebijakan, tujuan, konsep desa/kelurahan dan kampung, organisasi pelaksana, pelaksana,mitra organisasi, konteks ekologikal, protokol intervensi, populasi target.Hasil penelitian menunjukkan berbeda dengan Badan Nasional Penanggulangan Bencana,Kementerian Sosial RI tidak hanya sebagai pembuat kebijakan akan tetapi juga melaksanakan fasilitasilangsung pembentukan kelembagaan kampung siaga bencana. Konsep kampung pada kampung siagabencana cenderung pada merek program bukan kampung sebagai wilayah sedangkan pada desa/kelurahan merupakan konsep kewilayahan desa/kelurahan itu sendiri. Tujuan dari kampung siaga bencanacenderung lebih kompleks yaitu memberikan pemahaman dan kesadaran masyarakat, membentuk jejaringdan memperkuat interaksi sosial, mengorganisasikan, menjamin kesinambungan, mengoptimalkan potensidan sumber daya sedangkan pada desa/kelurahan tangguh bencana lebih cenderung sebagai upayapeningkatan penanggulangan berbasis komunita

Intramolecular C-C Coupling Reactions of Alkynyl, Vinylidene, and Alkenylphosphane Ligands in Rhodium(III) Complexes

Electrophilic attack with methyl trifluoromethanesulfonate or tetrafluoroboric acid, to new alkynyl rhodium complexes containing alkenylphosphanes, leads to butenynyl coupling products or to the unprecedented rhodaphosphacycle complexes Rh(¿5-C5Me5){¿4-(P, C, C, C)-iPr2PCH2C(=CH2)C(CH2R)C=C(R)}]BF4] (R = Ph (11a), p-tol (11b)). These complexes 11a, b can be explained as a result of the coupling of three organic fragments in the molecule, the alkynyl, the vinylidene, generated in situ by reaction with HBF4 (A), and the C-C double bond from the alkenylphosphane. DFT computational studies on the formation of complex 11a suggest the 2 + 2] intramolecular cycloaddition between the double bond of the allylphosphane and the Ca-Cß of the vinylidene A as the most plausible pathway for this reaction

DNA based multi-copper ions assembly using combined pyrazole and salen ligandosides

The DNA structure is an ideal building block for the construction of functional nano-objects. In this direction, metal coordinating base pairs (ligandosides) are an appealing tool for the future specific functionalization of such nano-objects. We present here a study, in which we combine the metal ion coordinating pyrazole ligandoside with the interstrand crosslinking salen ligandoside system. We show that both ligandosides, when combined, are able to create stable multi-copper ion complexing DNA double helix structures in a cooperative fashion