Dendritic Pyridine-Imine Copper Complexes as Metallo-Drugs.

Since the discovery of cisplatin in the 1960s, the search for metallo-drugs that are more efficient than platinum complexes with negligible side effects has attracted much interest. Among the other metals that have been examined for potential applications as anticancer agents is copper. The interest in copper was recently boosted by the discovery of cuproptosis, a recently evidenced form of cell death mediated by copper. However, copper is also known to induce the proliferation of cancer cells. In view of these contradictory results, there is a need to find the most suitable copper chelators, among which Schiff-based derivatives offer a wide range of possibilities. Gathering several metal complexes in a single, larger entity may provide enhanced properties. Among the nanometric objects suitable for such purpose are dendrimers, precisely engineered hyperbranched macromolecules, which are outstanding candidates for improving therapy and diagnosis. In this review article, we present an overview of the use of a particular Schiff base, namely pyridine-imine, linked to the surface of dendrimers, suitable for complexing copper, and the use of such dendrimer complexes in biology, in particular against cancers.

Phenyl dialkynylcarbinols, a Bioinspired Series of Synthetic Antitumor Acetylenic Lipids.

A series of 25 chiral anti-cancer lipidic alkynylcarbinols (LACs) were devised by introducing an (hetero)aromatic ring between the aliphatic chain and the dialkynylcarbinol warhead. The resulting phenyl-dialkynylcarbinols (PACs) exhibit enhanced stability, while retaining cytotoxicity against HCT116 and U2OS cell lines with IC50 down to 40 nM for resolved eutomers. A clickable probe was used to confirm the PAC prodrug behavior: upon enantiospecific bio-oxidation of the carbinol by the HSD17B11 short-chain dehydrogenase/reductase (SDR), the resulting ynones covalently modify cellular proteins, leading to endoplasmic reticulum stress, ubiquitin-proteasome system inhibition, and apoptosis. Insights into the design of LAC prodrugs specifically bioactivated by HSD17B11 vs its paralogue HSD17B13 were obtained. The HSD17B11/HSD17B13-dependent cytotoxicity of PACs was exploited to develop a cellular assay to identify specific inhibitors of these enzymes. A docking study was performed with the HSD17B11 AlphaFold model, providing a molecular basis of the SDR substrates mimicry by PACs. The safety profile of a representative PAC was established in mice.

BODIPY-Ethynylestradiol molecular rotors as fluorescent viscosity probes in endoplasmic reticulum.

Due to their capability for sensing changes in viscosity, fluorescent molecular rotors (FMRs) have emerged as potential tools to develop several promising viscosity probes; most of them, however, localize non-selectively within cells, precluding changes in the viscosity of specific cellular microdomains to be studied by these means. Following previous reports on enhanced fluorophore uptake efficiency and selectivity by incorporation of biological submolecular fragments, here we report two potential BODIPY FMRs based on an ethynylestradiol spindle, a non-cytotoxic semisynthetic estrogen well recognized by human cells. A critical evaluation of the potential of these fluorophores for being employed as FMRs is presented, including the photophysical characterization of the probes, SXRD studies and TD-DFT computations, as well as confocal microscopy imaging in MCF-7 (breast cancer) cells.

SDR enzymes oxidize specific lipidic alkynylcarbinols into cytotoxic protein-reactive species.

Hundreds of cytotoxic natural or synthetic lipidic compounds contain chiral alkynylcarbinol motifs, but the mechanism of action of those potential therapeutic agents remains unknown. Using a genetic screen in haploid human cells, we discovered that the enantiospecific cytotoxicity of numerous terminal alkynylcarbinols, including the highly cytotoxic dialkynylcarbinols, involves a bioactivation by HSD17B11, a short-chain dehydrogenase/reductase (SDR) known to oxidize the C-17 carbinol center of androstan-3-alpha,17-beta-diol to the corresponding ketone. A similar oxidation of dialkynylcarbinols generates dialkynylketones, that we characterize as highly protein-reactive electrophiles. We established that, once bioactivated in cells, the dialkynylcarbinols covalently modify several proteins involved in protein-quality control mechanisms, resulting in their lipoxidation on cysteines and lysines through Michael addition. For some proteins, this triggers their association to cellular membranes and results in endoplasmic reticulum stress, unfolded protein response activation, ubiquitin-proteasome system inhibition and cell death by apoptosis. Finally, as a proof-of-concept, we show that generic lipidic alkynylcarbinols can be devised to be bioactivated by other SDRs, including human RDH11 and HPGD/15-PGDH. Given that the SDR superfamily is one of the largest and most ubiquitous, this unique cytotoxic mechanism-of-action could be widely exploited to treat diseases, in particular cancer, through the design of tailored prodrugs.

Methinylogation Approach in Chiral Pharmacophore Design: from Alkynyl- to Allenyl-carbinol Warheads against Tumor Cells.

Extension of a structure-activity relationship study of the antitumor cytotoxicity of lipidic dialkynylcarbinols (DACs) is envisaged by formal methinylogation of one of the ethyndiyl moieties of the DAC warhead into the corresponding allenylalkynylcarbinol (AllAC) counterpart. External AllACs were directly obtained by methinylation of the parent DACs with formaldehyde in either the racemic or scalemic series. Isomers containing external progargyl and propynyl motifs were also prepared. Internal AllACs were obtained as racemic statistical mixtures of stereoisomers in two steps from the key C5 -DAC rac-TIPS-C≡C-CH(OH)-C≡CH and aldehydes. Kinetic resolution of the (S)-C5 -DAC in 97 % ee and (R)-C5 -DAC in 99 % ee was achieved by sequential lipase-mediated acetylation/hydrolysis using the Candida antartica lipase (Novozyme 435). The four internal AllAC stereoisomers were prepared by asymmetric methinylation with (R)- or (S)-diphenylprolinol as chiral auxiliary. Cytotoxicity assays on HCT116 cancer cells showed that the most active (eutomeric) external or internal AllAC exhibits an S configuration, a fatty chain length of n=12, and a 50 % inhibitory concentration IC50 ≈1.0 µm.

Skeletal Optimization of Cytotoxic Lipidic Dialkynylcarbinols.

In line with a recent study of the pharmacological potential of bioinspired synthetic acetylenic lipids, after identification of the terminal dialkynylcarbinol (DAC) and butadiynyl alkynylcarbinol (BAC) moieties as functional antitumor pharmacophoric units, this work specifically addresses the issue of carbon backbone length. A systematic variation of the aliphatic chain length was thus carried out in both the DAC and BAC series. The critical impact of the length of the lipidic skeleton was first confirmed in the racemic series, with the highest cytotoxic activity observed for C17 to C18 backbones. Enantiomerically enriched samples were prepared by asymmetric synthesis of the optimal C18 DAC and C17 BAC derivatives. Samples with upgraded enantiomeric purity were alternatively produced by enzymatic kinetic resolution. Eutomers possessing the S configuration displayed cytotoxicity IC50 values as low as 15 nm against HCT116 cancer cells, the highest level of activity reached to date in this series.

Steric/π-Electronic Insulation of the carbo-Benzene Ring: Dramatic Effects of tert-Butyl versus Phenyl Crowns on Geometric, Chromophoric, Redox, and Magnetic Properties.

Hexa-tert-butyl-carbo-benzene (C18 tBu6 ) and three phenylated counterparts (C18 tBum Ph6-m ; m=4, 2) have been synthesized. The peralkylated version (m=6) provides experimental access to intrinsic features of the insulated C18 core independently from the influence of π-conjugated substituent. Over the series, structural, spectroscopic, and electrochemical properties are compared with those of the hexaphenylated reference (m=0). Anchoring tBu substituents at the C18 macrocycle is shown to enhance stability and solubility, and to dramatically modify UV/Vis absorption and redox properties. Whereas all carbo-benzenes reported previously were obtained as dark-reddish/greenish solids, crystals and solutions of C18 tBu6 happen to be yellow (λmax =379 vs. 472 nm for C18 Ph6 ). In comparison to C18 Ph6 , the reduction of C18 tBu6 remains reversible, but occurs at twice as high an absolute potential (E1/2 =-1.36 vs. -0.72 V). Systematic XRD analyses and DFT calculations show that the C18 ring symmetry is the nearest to D6h for m=6, which indicates a maximum geometric aromaticity. According to calculated nucleus-independent chemical shifts (NICS), the macrocyclic magnetic aromaticity is also maximum for C18 tBu6 : NICS(0)=-17.2 ppm versus (-18.0±0.1) ppm for the theoretical references C18 H6 and C18 F6 , and -13.5 ppm for C18 Ph6 . Accurate correlations of NICS(0) with experimentally recorded or calculated maximum UV/Vis absorption wavelengths, λmax , and chemical hardness, η=ELUMO -EHOMO , are evidenced.

Fluorophore-tagged pharmacophores for antitumor cytotoxicity: Modified chiral lipidic dialkynylcarbinols for cell imaging.

Chiral lipidic dialkynylcarbinols (DACs), recently highlighted as antitumoral pharmacophores, have been conjugated to difluoroboron-dipyrromethene (bodipy), 7-hydroxy-coumarine, and 7-nitro-benzoxadiazole (NBD) fluorophore motifs through triazole clips. The labeled lipids preserve cytotoxic activity against HCT116 cells, and fluorescence microscopy of the stained cells showed clear signals in the intra-cellular membrane system. While the bodipy conjugate also labels lipid droplets very brightly, as expected, the coumarine and NBD probes appear as promising specific tools for the identification of the intra-cellular targets of DACs' cytotoxicity.

Carbo-benzene's aromaticity, before and beyond: a tribute to Nozoe.

Beyond rigorous inspection of the literature, selected comments and drawings from the Nozoe Autograph Books show how concepts and targets of the chemistry of today already existed 40 years ago. This is illustrated for the chemistry of carbo-mers, which was actually unknowingly inspired by chemists of the golden age. After a personal travel through time and pages by the last author of this essay, who meets autographs by people who have been important in his career, in particular on the occasions of Nozoe's visits to France or past ISNA events, unpublished results in non-benzenoid chemistry are dedicated to Nozoe's memory. Experimental advances are first disclosed on the comparative chemistry of the three ortho, meta and para isomers of tetraphenyl-carbo-benzene C18 H2 Ph4 , the known ortho isomer serving as a reference. The two CH vertices of these chromophores make their [6]pericyclyne precursors quite sensitive and their yields and purifications challenging to optimize. The meta isomer was, however, isolated in 18% yield, and described by NMR and UV-vis absorption spectroscopy and crystallography. The para isomer was characterized both in pure form and in a 1:1 mixture with a single isomer of its HCl adduct, including by X-ray diffraction analysis of a co-crystal showing that the interior of the C18 ring of the HCl adduct is filled by the Cl atom. Theoretical advances are then offered regarding the "aromatic character" of a triple bond, by showing that the topological acyclic reference of acetylene is the Möbius-twisted cyclobutadiene. Acetylenoids are finally proposed as a novel class of non-benzenoid aromatics.

Chiral alkynylcarbinols from marine sponges: asymmetric synthesis and biological relevance.

Covering: up to March 2014. Previous review on the topic: B. W. Gung, C. R. Chim., 2009, 12, 489-505. Chiral α-functional lipidic propargylic alcohols extracted from marine sponges, in particular of the pacific genus Petrosia, constitute a class of acetylenic natural products exhibiting remarkable in vitro biological activities, especially anti-tumoral cytotoxicity. These properties, associated to functionalities that are uncommon among natural products, have prompted recent projects on asymmetric total synthesis. On the basis of a three-sector structural typology, three main sub-types of secondary alkynylcarbinols (with either alkyl, alkenyl, or alkynyl as the second substituent) can be identified as the minimal pharmacophoric units. Selected natural products containing these functionalities have been targeted using previously known or on purpose-designed procedures, where the stereo-determining step can be: (i) a C-C bond forming reaction (e.g. the Zn-mediated addition of alkynyl nucleophiles to aldehydes in the presence of chiral aminoalcohols), (ii) a functional layout (e.g. the asymmetric organo- or metallo-catalytic reduction of ynones), or (iii) an enantiomeric resolution (e.g. a lipase-mediated kinetic resolution via acetylation). The promising medicinal importance of these targets is finally surveyed, and future investigation prospects are proposed, such as: (i) further total synthesis of known or future extraction products; (ii) the synthesis of non-natural analogues, with simpler lipophilic environments of the alkynylcarbinol-based pharmacophoric units; (iii) the variation and optimization of both the pharmacophoric units and their lipophilic environment; and (iv) investigations into the biological mode of action of these unique structures.

Functional carbo-butadienes: nonaromatic conjugation effects through a 14-carbon, 24-π-electron backbone.

A systematic study of carbo-butadiene motifs not embedded in an aromatic carbo-benzene ring is described. Dibutatrienylacetylene (DBA) targets R(1) C(R)CCC(Ph)C≡CC(Ph)CCC(R)R(2) are devised, in which R is C≡CSiiPr3 and R(1) and R(2) are R, H, or 4-X-C6 H4 , with the latter including three known representatives (X: H, NMe2 , or NH2 ). The synthesis method is based on the SnCl2 -mediated reduction of pentaynediols prepared by early or late divergent strategies; the latter allows access to a OMe-NO2 push-pull diaryl-DBA. If R(1) and R(2) are H, an over-reduced dialkynylbutatriene (DAB) with two allenyl caps was isolated instead of the unsubstituted DBA. If R(1) =R(2) =R, the tetraalkynyl-DBA target was obtained, along with an over-reduced DBA product with a 12-membered 1,2-alkylidene-1H2 ,2H2 -carbo-cyclobutadiene ring. X-ray crystallography shows that all of the acyclic DBAs adopt a planar trans-transoid-trans configuration. The maximum UV/Vis absorption wavelength is found to vary consistently with the overall π-conjugation extent and, more intriguingly, with the π-donor character of the aryl X substituents, which varies consistently with the first (reversible) reduction potential and first (irreversible) oxidation peak, as determined by voltammetry.