Synthesis of bioconjugate sesterterpenoids with phospholipids and polyunsaturated fatty acids

A series of sesterterpenoid bioconjugates with phospholipids and polyunsaturated fatty acids (PUFAs) have been synthesized for biological activity testing as antiproliferative agents in several cancer cell lines. Different substitution analogues of the original lipidic ether edelfosine (1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine) are obtained varying the sesterterpenoid in position 1 or 2 of the glycerol or a phosphocholine or PUFA unit in position 3. Simple bioconjugates of sesterterpenoids and eicosapentaenoic acid (EPA) have been obtained too. All synthetic derivatives were tested against the human tumour cell lines HeLa (cervix) and MCF-7 (breast). Some compounds showed good IC50 (0.3 and 0.2 μM) values against these cell lines.This work was supported by grants from the Spanish Ministry of Economy and Competitiveness (SAF2011-30518 and SAF2014-59716-R). Junta de Castilla y León BIO/SA59/15.Peer reviewe

GENYOi004-A: An induced pluripotent stem cells (iPSCs) line generated from a patient with autism-related ADNP syndrome carrying a pTyr719* mutation

ADNP syndrome is an intellectual disability associated with Autism spectrum disorder caused by mutations in ADNP. We generated an iPSC line from an ADNP syndrome pediatric patient harboring the mutation p.Trp719* (GENYOi004-A). Peripheral blood mononuclear cells were reprogrammed using a non-transmissible form of Sendai viruses expressing the four Yamanaka factors (Oct3/4, SOX2, KLF4 and c-MYC). Characterization of GENYOi004-A included mutation analysis of ADNP by allele-specific PCR, genetic identity by Short Tandem Repeats polymorphism profiling, alkaline phosphatase enzymatic activity, expression of pluripotency-associated factors and pluripotency studies in vivo. GENYOi004-A will be useful to evaluate ADNP syndrome alterations at early developmental stages.This work was supported by the Postdoctoral Subprogramme Juan de la Cierva (JCI_2012_12666) to RM and Ramon y Cajal (RYC-2015-18382) to PJR founded by the Ministry of Economy and Competitiveness; the Instituto de Salud Carlos III-FEDER (CP12/03175 and CPII17/00032) to V.R-M. and (PI12/1598, CPII15/00018 and PI16/01340) to PJR; the Instituto de Investigación Valdecilla (IDIVAL) 2014.041 to JLF-L and DG-L and APG/03 to JLF-L

Cellular and animal models of skin alterations in the autism-related ADNP syndrome

Mutations in ADNP have been recently associated with intellectual disability and autism spectrum disorder. However, the clinical features of patients with this syndrome are not fully identified, and no treatment currently exists for these patients. Here, we extended the ADNP syndrome phenotype describing skin abnormalities in both a patient with ADNP syndrome and an Adnp haploinsufficient mice. The patient displayed thin dermis, hyperkeratotic lesions in periarticular areas and delayed wound healing. Patient-derived skin keratinocytes showed reduced proliferation and increased differentiation. Additionally, detection of cell cycle markers indicated that mutant cells exhibited impaired cell cycle progression. Treatment of ADNP-deficient keratinocytes with the ADNP-derived NAP peptide significantly reduced the expression of differentiation markers. Sonography and immunofluorescence staining of epidermal layers revealed that the dermis was thinner in the patient than in a healthy control. Adnp haploinsufficient mice (Adnp+/-) mimicked the human condition showing reduced dermal thickness. Intranasal administration of NAP significantly increased dermal thickness and normalized the levels of cell cycle and differentiation markers. Our observations provide a novel activity of the autism-linked ADNP in the skin that may serve to define the clinical phenotype of patients with ADNP syndrome and provide an attractive therapeutic option for skin alterations in these patients.This work was supported by grant CI14/09 from Fundacion Instituto de Investigacion Valdecilla to J.L.F.-L., PI14/00900 from Instituto de Salud Carlos III (ISCIII) to A.G., and AMN Foundation and ERA-NET Neuron to I.G. We are grateful to Ana Freije and Laura Ceballos for technical assistance at isolating skin cells, and to Profs. Carmit Levi and Chen Luxenburg and the student Chen Slonimsky for their help in the in vivo experiments. We thank Prof. Joseph Levine for his help with the analysis of the Facebook answers regarding the skin conditions in ADNP children. We also thank Isabel Garcia for her constant support and help to obtain phenotypic information of the skin of patients with ADNP syndrome

Substitution at the indole 3 position yields highly potent indolecombretastatins with reduced susceptibility to MDR resistance

[EN]Resistance to combretastatin A-4 is mediated by metabolic modification of the phenolic hydroxyl and ether groups of the 3-hydroxy-4-methoxyphenyl (B ring). Replacement of the B ring of combretastatin A-4 by a N-methyl-5-indolyl reduces tubulin polymerization inhibition (TPI) and cytotoxicity against human cancer cell lines but cyano, methoxycarbonyl, formyl, and hydroxyiminomethyl substitutions at the indole 3-position restores potent TPI and cytotoxicity against sensitive human cancer cell lines. These highly potent substituted derivatives displayed low nanomolar cytotoxicity against several human cancer cell lines due to tubulin inhibition, as shown by cell cycle analysis, confocal microscopy, and tubulin polymerization inhibitory activity studies and promoted cell killing mediated by caspase-3 activation. Binding at the colchicine site was suggested by molecular modeling studies. Substituted combretastatins displayed higher potencies than the isomeric isocombretastatins and the highest potencies were achieved for the hydroxyiminomethyl (21) and cyano (23) groups, with TPI values in the submicromolar range and cytotoxicities in the subnanomolar range. Dose-response and time-course studies showed that drug concentrations as low as 1 nM (23) or 10 nM (21) led to a complete G2/M cell cycle arrest after 15 h treatment followed by a high apoptosis-like cell § These authors contributed equally to this work. 3 response after 48-72 h treatment. The P-glycoprotein and calcium antagonist verapamil increased 21 and 23 cytotoxicity to IC50 values of 10-10 M, and highly potentiated the cytotoxic activity in 100-fold of the CHO derivative (17), in A549 human non-small cell lung cancer cells. The differences in cytotoxic potency observed between the highly potent cyano (23) and hydroxyiminomethyl (21) groups and other substituents with similar TPI values (17) were very much reduced upon co-treatment with verapamil. A 3,4,5-trimethoxyphenyl ring always afforded more potent derivatives than a 2,3,4-trimethoxyphenyl ring

Identification of new FK866 analogues with potent anticancer activity against pancreatic cancer

11 p.- 2 fig.-4 tab.-8 schem.-1 graph. abst.Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal diseases for which chemotherapy has not been very successful yet. FK866 ((E)-N-(4-(1-benzoylpiperidin-4-yl)butyl)-3-(pyridin-3-yl)acrylamide) is a well-known NAMPT (nicotinamide phosphoribosyltransferase) inhibitor with anti-cancer activities, but it failed in phase II clinical trials. We found that FK866 shows anti-proliferative activity in three PDAC cell lines, as well as in Jurkat T-cell leukemia cells. More than 50 FK866 analogues were synthesized that introduce substituents on the phenyl ring of the piperidine benzamide group of FK866 and exchange its buta-1,4-diyl tether for 1-oxyprop-3-yl, (E)-but-2-en-1,4-diyl and 2- and 3-carbon tethers. The pyridin-3-yl moiety of FK866 was exchanged for chlorinated and fluorinated analogues and for pyrazin-2-yl and pyridazin-4-yl groups. Several compounds showed low nanomolar or sub-nanomolar cell growth inhibitory activity. Our best cell anti-proliferative compounds were the 2,4,6-trimethoxybenzamide analogue of FK866 ((E)-N-(4-(1-(2,4,6-trimethoxybenzoyl)piperidin-4-yl)butyl)-3-(pyridin-3-yl)acrylamide) (9), the 2,6-dimethoxybenzamide (8) and 2-methoxybenzamide (4), which exhibited an IC50 of 0.16 nM, 0.004 nM and 0.08 nM toward PDAC cells, respectively.The research leading to these results has received funding from the European Community's Seventh Framework Programme [FP7-2007-2013] under grant agreement no. 256986 (PANACREAS), from the European Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement no. 813284 (INTEGRATA), by the Spanish Ministry of Science, Innovation, and Universities (SAF2017-89672-R) and by the Associazione Italiana per la Ricerca sul Cancro (AIRC), IG#22098.Peer reviewe

Synthesis of an ent-Halimanolide from ent-Halimic Acid

This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license.-- et al.An efficient synthesis of ent-halimanolide 2 (15,16-epoxy-12-oxo-ent- halima-5(10),13(16),14-trien-18,2β-olide), from ent-halimic acid has been achieved, corroborating the structure of the natural compound and establishing its absolute configuration. © 2008 by the authors; licensee Molecular Diversity Preservation International.The authors also thank the CICYT (CTQ2005-04406) and FSE for financial support and to Junta de Castilla y León for a doctoral fellowship to A.C.A.Peer Reviewe

Synthesis of Isoprenyl Flavonoids: (+)-Denticulaflavonol, Macarangin, and Isomacarangin

A straightforward synthesis of two natural C-alkylflavonoids: macarangin, isomacarangin, and the enantiomer of the natural (-)-denticulaflavonol, has been described. © Georg Thieme Verlag Stuttgart.Peer Reviewe

Expeditious synthesis of nitrogenated spongianes: 4-methyldecarboxyspongolactams

Herein, the synthesis of 4-methyldecarboxyhaumanamide (9) and 4-methyldecarboxyspongolactams A (11) and C (13) is presented. (-)-Sclareol is the starting material and the chloroderivative 7 is the common intermediate. Moreover, this synthesis represents a new strategy for the preparation of pyrrolinones. © 2010 Elsevier Ltd. All rights reserved.The authors thank the Junta de Castilla y León for financial support (GR178 and SA063A07), MICINN (CTQ2009-11557) and for the doctoral fellowships awarded to A.B. and O.B.Peer Reviewe

Hyrtiosanes from labdanes: (-)-Hyrtiosal from sclareol

(-)-Hyrtiosal and its C-16 epimer have been prepared from sclareol in moderate yield. The absolute configuration of natural product (-)-hyrtiosal has being determined.Peer Reviewe

Células primarias de piel como modelo in vitro del Síndrome ADNP: una nueva variante del espectro autista

RESUMEN: Las mutaciones en el gen ADNP (Proteína Neuroprotectora Dependiente de Actividad) se han asociado recientemente con discapacidad intelectual y trastorno del Espectro Autista. Sin embargo, las características clínicas de los pacientes con este síndrome, que se caracteriza por disfunciones multiorgánicas, no están completamente identificadas. Además, actualmente no existe un tratamiento para estos pacientes. En esta Tesis Doctoral hemos ampliado el fenotipo del Síndrome ADNP, describiendo anormalidades tanto en la piel de una paciente como en la piel de ratones haploinsuficientes para dicho gen. La paciente portadora de la mutación patogénica en ADNP presenta una piel delgada, con lesiones hiperqueratósicas en áreas periarticulares. Mediante el aislamiento de células de la piel de dicha paciente, hemos establecido un modelo celular sobre el que poder estudiar las características celulares debidas a la mutación en ADNP, encontrando una menor proliferación celular y un deterioro en la progresión del ciclo celular. El tratamiento de las células portadoras de la mutación con el péptido NAP, el cual deriva de la proteína NAP, mejoro las deficiencias encontradas en estas células. En los ratones haploinsuficientes se observaron resultados similares. Además del modelo celular basado en células de la piel, en esta Tesis también hemos comenzado a desarrollar un modelo celular para el Síndrome ADNP basado en células madre pluripotentes.ABSTRACT: Mutations in ADNP (Activity-Dependent Neuroprotective Protein) have been recently associated with intellectual disability and autism spectrum disorder. However, the clinical features of patients with this syndrome, characterized by multiorganic dysfunctions, are not fully identified, and no treatment currently exists for these patients. In this Thesis we have extended the ADNP syndrome phenotype describing skin abnormalities in both a patient with ADNP syndrome and an Adnp haploinsufficient mice. The patient carrying a pathogenic mutation in ADNP displayed thin skin and hyperkeratotic lesions in periarticular areas. We have established a cellular model based on patient-derived skin cells on which to study the cellular characteristics due to the mutation in ADNP. These cells showed reduced proliferation and exhibited impaired cell cycle progression. Treatment of ADNP-deficient cells with the ADNP-derived NAP peptide improved the deficiencies found in these cells. Adnp haploinsufficient mice mimicked the human conditions. In addition to the cellular model based on skin cells, in the Thesis we have also begun to develop a cellular model for the ADNP syndrome based on pluripotent stem cells