Alkylating Activity Research Articles

Kinetic studies on the tert-butylation of toluene over H-BEA zeolite

The highly selective synthesis of 4- tert-butyltoluene from toluene with tert-butyl alcohol over zeolite catalysts was performed in the liquid phase. The order of activity for alkylation over zeolite catalysts was as follows: H-BEA > H-Y > H-MOR > H-ZSM-5. The main reaction products observed were 4- tert-butyltoluene and 3- tert-butyltoluene. 2- tert-Butyltoluene was completely absent in the product. A kinetic study of the reaction of toluene with tert-butyl alcohol over pure H-BEA (Si/Al = 25) was performed. Experimental runs were conducted at 160, 170, 180 and 190 °C and reaction times of up to 240 min. Based on the results obtained, a simplified kinetic model was developed. The kinetics could be described by the pseudo first-order model.

When alcohol is the answer: Trapping, identifying and quantifying simple alkylating species in aqueous environments

Alkylating agents are a significant class of environmental carcinogens as well as commonly used anticancer therapeutics. Traditional alkylating activity assays have utilized the colorimetric reagent 4-(4-nitrobenzyl)pyridine (4NBP). However, 4NBP based assays have a relatively low sensitivity towards harder, more oxophilic alkylating species and are not well suited for the identification of the trapped alkyl moiety due to adduct instability. Herein we describe a method using water as the trapping agent which permits the trapping of simple alkylating electrophiles with a comparatively wide range of softness/hardness and permits the identification of donated simple alkyl moieties.

Shape selectivity of beta and MCM-49 zeolites in liquid-phase alkylation of benzene with ethylene

NH3, pyridine and 2,4,6-trimethyl-pyridine were selected as the base probes to characterize the different pore acidity of H-beta and H-MCM-49 zeolites, revealing the essence in their catalytic performance from the view of shape selectivity in different pore systems. Based on the size of base probes, reactants, product, transition states and pore channels, the difference in catalytic performance over H-beta and H-MCM-49 was investigated. Easy diffusion of reactants into and long diffusion distance for ethylbenzene escaping out of confined space within beta are the main reason for superior activity and poorer ethylbenzene (EB) selectivity. As for H-MCM-49, acid sites located in 10 MR channels contributed little to the formation of EB due to transition state selectivity; Acid sites located in 12 MR cups on the outer surface with little restriction on the diffusion of reactant/product toward/from the active centers were the major active centers for EB formation, which could be detected by 2,4,6-trimethyl-pyridine; There was diffusion limitation on reactant/product in 12 MR supercages through 10 MR windows to influence activity/selectivity. The enhancement in activity and selectivity in liquid-phase alkylation of benzene with ethylene over MWW zeolites should be mainly focused on transforming confined 12 MR supercages into 12 MR cups with improved openness.

Comparative activity of aryl, alkyl, and cycloalkyl halides in the suzuki reaction catalyzed with acyclic diaminocarbene complex of palladium

Relative activity of halogenated arenes, alkanes, and alkanes in the Suzuki reaction catalyzed with acyclic diaminocarbene complex of palladium has been investigated. Under all the investigated conditions, 4-iodoanisole has been more active than the alkyl halides. The reaction with 4-methyl-1-(chloromethyl)benzene has afforded the target 4-methyl-1-(phenylmethyl)benzene along with significant amount of by-products; other alkyl and cycloalkyl halides do not participate into the cross-coupling reaction. Ethanol has been found the most suitable solvent for the reaction. The reaction in acetonitrile provides noticeable yield of the products only in the presence of polyethylene glycol and water.

Selective Targeting of the KRAS Codon 12 Mutation Sequence by Pyrrole-Imidazole Polyamide seco-CBI Conjugates.

Mutation of KRAS is a key step in many cancers. Mutations occur most frequently at codon 12, but the targeting of KRAS is notoriously difficult. We recently demonstrated selective reduction in the volume of tumors harboring the KRAS codon 12 mutation in a mouse model by using an alkylating hairpin N-methylpyrrole-N-methylimidazole polyamide seco-1,2,9,9a-tetrahydrocyclopropa[1,2-c]benz[1,2-e]indol-4-one conjugate (conjugate 4) designed to target the KRAS codon 12 mutation sequence. Herein, we have compared the alkylating activity of 4 against three other conjugates that were also designed to target the KRAS codon 12 mutation sequence. Conjugate 4 displayed greater affinity for the G12D mutation sequence than for the G12V sequence. A computer-minimized model suggested that conjugate 4 could bind more efficiently to the G12D match sequence than to a one-base-pair mismatch sequence. Conjugate 4 was modified for next-generation sequencing. Bind-n-Seq analysis supported the evidence showing that conjugate 4 could target the G12D mutation sequence with exceptionally high affinity and the G12V mutation sequence with much higher affinity than that for the wild-type sequence.

Synthesis and Catalytic Performance of Hierarchically Porous MIL-100(Fe)@polyHIPE Hybrid Membranes.

Metal-organic frameworks (MOFs) nanoparticles in combination with a nonionic surfactant (Pluronic L-121) are used to stabilize dicyclopentadiene (DCPD)-in-water high internal phase emulsions (HIPEs). The resulting HIPEs containing the MIL-100(Fe) nanoparticles (MIL: Materials of Institut Lavoisier) at the interface between the oil- and the water-phases are then cured, and 100 μm thick, fully open, hierarchically porous hybrid membranes are obtained. The properties of the MIL-100(Fe)@pDCPD polyHIPE membranes are characterized and it is found that up to 14 wt% of the MIL-100(Fe) nanoparticles are incorporated in the hybrid material resulting in an increase of the microporosity up to 130 m(2) g(-1). Hybrid membranes show an appealing catalytic activity in Friedel-Crafts alkylation in a batch mode as well as in a flow-through mode, thereby demonstrating the preserved accessibility of Lewis acidic sites in the MOF nanostructures.

Impregnation of tungstophosphoric acid on poly(methacrylamide-co-methyl methacrylate) and its acid catalytic activity in TMB alkylation

A poly(methacrylamide-co-methylmethacrylate) (abbreviated PMAA-MMA) polymer support was studied for supporting a heteropolyacid (tungstophosphoric acid, H3PW12O40) with its surface positively charged in the polymerization step. PMAA-MMA supports could be obtained in a porous form by eliminating template reagent molecules (benzylmalonic acid) combined with properly selected monomer (methacrylamide). The amount of amine groups in PMAA-MMA directly determined the amount of H3PW12O40 impregnated, because the amine groups induced a positive charge on the PMAA-MMA surface. Finally, H3PW12O40/PMAA-MMA showed better acid catalytic activities than unsupported H3PW12O40 in alkylation of 1,3,5-trimethylbenzene with cyclohexene, which confirmed that PMAA-MMA supported H3PW12O40 effectively.

A further catalysis mechanism study on Amberlyst 35 resins application in alkylation desulfurization of gasoline

Olefinic alkylation of thiophenic sulfurs (OATS) technology can be handled under mild conditions without any hydrogen consumption, which is a promising way to produce clean gasoline. However, the side reaction in this desulfurization process would lead to significant levels of coke. To maintain the high activity for alkylation of sulfurs, the selectivity of OATS catalyst must be improved by reducing side reactions. In this paper, the catalytic mechanism of macroporous sulfonic resins Amberlyst 35 (A35) in the OATS process was further investigated by the calculation of density functional theory (DFT), to understand the reaction path of different reactant at molecular level. The calculated results indicated that the beginning of main and side reactions were both from a stable alkoxide intermediate, which was the protonation product of adsorbed olefin on the catalyst. Compared with alkenes, thiophenic compounds were more inclined to be coadsorbed on the alkoxide intermediate for further reaction, and the alkylation rate of sulfurs with alkenes was faster than the self-dimerization of alkenes. Moreover, the calculated results also indicated that the alkylation of thiophenic sulfurs as main reaction was exothermic while the dimerization of alkenes as side reaction was endothermic over A35. Additionally, the conversion curves of different reactants over A35 and the related kinetics at different temperature were also studied by experimental methods. The obtained experimental results could be used to verify the reliability of relevant theoretical calculations. Based on the differences in the reaction mechanisms obtained by the theoretical and experimental studies, two measures were proposed, which would be useful to reduce side reactions to a lower extent. The study would be beneficial to the further industrial application of A35 in the alkylation desulfurization of gasoline.

Effects of alkylation and immunopotentiation against Ehrlich ascites murine carcinoma in vivo using novel tetra-O-acetate haloacetamido carbohydrate analogs

Tetra-O-acetate haloacetamido carbohydrate analogs (Tet-OAHCs) are novel alkylating agents that appear to have alkylating activity at the plasma membrane, specificity against neoplastic cells, and may potentiate host leukocyte influx. This study sought to characterize the chemical attributes and in vivo activity of Tet-OAHCs. Four Tet-OAHCs were assessed for their partition coefficient and alkylating activity to determine cellular environments where adduct formation would be favorable. In vitro, IC50 values of all four Tet-OAHCs were determined against Ehrlich ascites murine carcinoma, as well as two leukemias (U937 human monocytic leukemia and L1210 murine lymphoid leukemia) to assess their cytotoxicity in multiple neoplastic cell lines. In vivo, B6D2F1 and CD2F1 mice were challenged i.p. with Ehrlich ascites carcinoma prior to, or after being treated with a single dose of one of the analogs. Finally, a quantitative comparison of host leukocyte influx between Tet-OAHCs and other alkylating agents was performed to confirm previous in vivo observations that the tetra-O-acetate carbohydrate moiety is important for inducing a host leukocyte response in murine models.The results can be summarized as follows: 1) Tet-OAHCs appear to demonstrate high alkylating activity in amphiphilic environments. 2) All four congeners have comparable in vitro cytotoxicities against the neoplastic cell lines examined. 3) The analogs demonstrate marked in vivo activity in both B6D2F1 and CD2F1 mice challenged with a lethal dose of Ehrlich ascites carcinoma, and frequently produce long term survival at 60 days, which is not observed in simple halo derivatives or two currently approved antineoplastic agents (daunorubicin and mechlorethamine). These effects are observed when the agents are administered either before or after the tumor challenge. 4) The carbohydrate moiety appears to be important for potentiating host leukocyte influx, as Tet-OAHCs, but not other alkylating agents demonstrated such activity in vivo.

Pharmacokinetic characterization of BMS-936561, an anti-CD70 antibody-drug conjugate, in preclinical animal species and prediction of its pharmacokinetics in humans.

CD70 is a tumor necrosis factor (TNF)-like type II integral membrane protein that is transiently expressed on activated T- and B-lymphocytes. Aberrant expression of CD70 was identified in both solid tumors and haematologic malignancies. BMS-936561 (αCD70_MED-A) is an antibody-drug conjugate composed of a fully human anti-CD70 monoclonal antibody (αCD70) conjugated with a duocarmycin derivative, MED-A, through a maleimide-containing citrulline-valine dipeptide linker. MED-A is a carbamate prodrug that is activated by carboxylesterase to its active form, MED-B, to exert its DNA alkylation activity. In vitro serum stability studies suggested the efficiencies of hydrolyzing the carbamate-protecting group in αCD70_MED-A followed a rank order of mouse>rat > >monkey>dog~human. Pharmacokinetics of αCD70_MED-A was evaluated in mice, monkeys, and dogs after single intravenous doses. In mice, αCD70_MED-A was cleared rapidly, with no detectable exposures after 15 min following dosing. In contrast, αCD70_MED-A was much more stable in monkeys and dogs. The clearance of αCD70_MED-A in monkeys was 58 mL/d/kg, ~2-fold faster than that in dogs (31 mL/d/kg). The human PK profiles of the total αCD70 and αCD70_MED-A were predicted using allometrically scaled monkeys PK parameters of αCD70 and the carbamate hydrolysis rate constant estimated in dogs. Comparing the predicted and observed human PK from the phase I study, the dose-normalized concentration-time profiles of αCD70_MED-A and the total αCD70 were largely within the 5(th)-95(th) percentile of the predicted profiles.

Multicomponent synthesis and antimicrobial activity of alkyl 4-arylamino-1,2,6-triaryl-1,2,5,6-tetrahydropyridine-3-carboxylates

Reaction of arylamines with aromatic aldehydes and β-ketoesters in the presence of bismuth nitrate or crystalline iodine as a catalyst afforded to alkyl 4-arylamino-1,2,6-triaryl-1,2,5,6-tetrahydropyridine-3-carboxy-lates. Antimicrobial activity of the obtained compounds was studied.

The slow dissociation rate of K-1602 contributes to the enhanced inhibitory activity of this novel alkyl–aryl-bearing fluoroketolide

Ketolides belong to the latest generation of macrolides and are not only effective against macrolide susceptible bacterial strains but also against some macrolide resistant strains. Here we present data providing insights into the mechanism of action of K-1602, a novel alkyl–aryl-bearing fluoroketolide. According to our data, the K-1602 interacts with the ribosome as a one-step slow binding inhibitor, displaying an association rate constant equal to 0.28 × 104 M−1 s−1 and a dissociation rate constant equal to 0.0025 min−1. Both constants contribute to produce an overall inhibition constant Ki equal to 1.49 × 10−8 M, which correlates very well with the superior activity of this compound when compared with many other ketolides or fluoroketolides.

EPI-001 is a selective peroxisome proliferator-activated receptor-gamma modulator with inhibitory effects on androgen receptor expression and activity in prostate cancer.

The androgen receptor (AR) is a driver of prostate cancer (PCa) cell growth and disease progression. Therapies for advanced PCa exploit AR dependence by blocking the production or action of androgens, but these interventions inevitably fail via multiple mechanisms including mutation or deletion of the AR ligand binding domain (LBD). Thus, the development of new inhibitors which act through non-LBD interfaces is an unmet clinical need. EPI-001 is a bisphenol A-derived compound shown to bind covalently and inhibit the AR NH2-terminal domain (NTD). Here, we demonstrate that EPI-001 has general thiol alkylating activity, resulting in multilevel inhibitory effects on AR in PCa cell lines and tissues. At least one secondary mechanism of action associated with AR inhibition was found to be selective modulation of peroxisome proliferator activated receptor-gamma (PPARγ). These multi-level effects of EPI-001 resulted in inhibition of transcriptional activation units (TAUs) 1 and 5 of the AR NTD, and reduced AR expression. EPI-001 inhibited growth of AR-positive and AR-negative PCa cell lines, with the highest sensitivity observed in LNCaP cells. Overall, this study provides new mechanistic insights to the chemical biology of EPI-001, and raises key issues regarding the use of covalent inhibitors of the intrinsically unstructured AR NTD.

Bis-3-chloropiperidines containing bridging lysine linkers: Influence of side chain structure on DNA alkylating activity

A series of bis-3-chloropiperidines containing lysine linkers was synthesised as DNA alkylating model compounds by using a bidirectional synthetic strategy. These novel piperidine mustard based agents have been evaluated for their alkylating properties towards nucleic acids and were shown to alkylate and cleave DNA with strong preference for guanine residues. Our studies reveal that the introduction of aromatic groups in the side chain of the lysine linker has an impact on DNA alkylating activity. Analysis by ESI mass spectrometry enabled the verification of the reactive aziridinium ion formation. Overall, the results confirm our recently proposed reaction mechanism of bis-3-chloropiperidines.

Cytotoxic activity of triazole-containing alkyl β-D-glucopyranosides on a human T-cell leukemia cell line.

BackgroundSimple glycoside surfactants represent a class of chemicals that are produced from renewable raw materials. They are considered to be environmentally safe and, therefore, are increasingly used as pharmaceuticals, detergents, and personal care products. Although they display low to moderate toxicity in cells in culture, the underlying mechanisms of surfactant-mediated cytotoxicity are poorly investigated.ResultsWe synthesized a series of triazole-linked (fluoro)alkyl β-glucopyranosides using the copper-catalyzed azide-alkyne reaction, one of many popular “click” reactions that enable efficient preparation of structurally diverse compounds, and investigate the toxicity of this novel class of surfactant in the Jurkat cell line. Similar to other carbohydrate surfactants, the cytotoxicity of the triazole-linked alkyl β-glucopyranosides was low, with IC50 values decreasing from 1198 to 24 μM as the hydrophobic tail length increased from 8 to 16 carbons. The two alkyl β-glucopyranosides with the longest hydrophobic tails caused apoptosis by mechanisms involving mitochondrial depolarization and caspase-3 activation.ConclusionsTriazole-linked, glucose-based surfactants 4a-g and other carbohydrate surfactants may cause apoptosis, and not necrosis, at low micromolar concentrations via induction of the intrinsic apoptotic cascade; however, additional studies are needed to fully explore the molecular mechanisms of their toxicity.Graphical Triazole-linked, glucose-based surfactants cause apoptosis, and not necrosis, at low micromolar concentrations via induction of the intrinsic apoptotic cascade.Electronic supplementary materialThe online version of this article (doi:10.1186/s13065-014-0072-1) contains supplementary material, which is available to authorized users.

Positive effect of secondary structure creation in mordenites on alkylation of benzene with 1-tetradecene

The liquid-phase alkylation of benzene with 1-tetradecene was realized over dealuminated mordenites with different mesoporous surface areas and three different levels of acidity. The mordenite catalysts were characterized by N2-sorption, FTIR pyridine adsorption, and TPDA. The number of Brønsted acid sites decreases; however, it represents a higher proportion of the total acidity. Alkylation results demonstrated that the mordenite activity in alkylation accelerated with increasing mesoporous surface area in spite of the lower total acidity. This fact indicates that the accessibility of acid sites and the internal mass transport in mordenite channels play a significant role in liquid-phase alkylation of aromatics with 1-alkenes.

Enhancing the anti-inflammatory activity of chalcones by tuning the Michael acceptor site.

Inflammatory signaling pathways orchestrate the cellular response to infection and injury. These pathways are known to be modulated by compounds that alkylate cysteinyl thiols. One class of phytochemicals with strong thiol alkylating activity is the chalcones. In this study we tested fourteen chalcone derivatives, α-X-substituted 2',3,4,4'-tetramethoxychalcones (α-X-TMCs, X = H, F, Cl, Br, I, CN, Me, p-NO2-C6H4, Ph, p-OMe-C6H4, NO2, CF3, COOEt, COOH), for their ability to modulate inflammatory responses, as monitored by their influence on heme oxygenase-1 (HO-1) activity, inducible nitric oxide synthase (iNOS) activity, and cytokine expression levels. We confirmed that the transcriptional activity of Nrf2 was activated by α-X-TMCs while for NF-κB it was inhibited. For most α-X-TMCs, anti-inflammatory activity was positively correlated with thiol alkylating activity, i.e. stronger electrophiles (X = CF3, Br and Cl) being more potent. Notably, this correlation did not hold true for the strongest electrophiles (X = CN and NO2) which were found to be ineffective as anti-inflammatory compounds. These results emphasize the idea that chemical fine-tuning of electrophilicity is needed to achieve and optimize desired therapeutic effects.

In-Vitro Actvity of Pixantrone (PIX) in Plasma Cell Leukaemia (PCL) and Early Clinical Activity in a Pixantrone–Pomalidomide–Dexamethasone (PiPoD) Combination

IntroductionPlasma cell leukemia (PCL) is a leukemic variant of Myeloma arising either de novo or from clinically pre-existent Multiple Myeloma (MM). Treatment options are limited and no drug has ever been explicitly registered in this indication. Furthermore, there is no generally accepted standard of care, although most haematologists would, depending on previous treatments, use combinations of MM drugs and chemotherapy as induction therapy.Pixantrone (PIX) is an aza-anthracenedione which has both alkylating and intercalating activity as well as a unique method of action the induction of cell death through mitotic perturbations and subsequent aberrant cell divisions. PIX is registered in both the US and EC for the treatment of relapsed or refractory aggressive NHL that has failed to respond to at least two previous regimens. Our group has previously shown significant in-vitro activity of PIX in both MM cell lines and primary patient materials*.MethodsPeripheral blood mononuclear cells from patient samples containing > 80% plasma cells at high leucocyte counts were isolated by density gradient using Ficoll and incubated for 24 and 48h with various pharmacokinetically realistic concentrations of PIX ranging from 0.01 – 5.00 µM at 37°C in a humidified air atmosphere. Apoptosis was determined by annexin/propidium iodide staining of the whole population (Pt. 1) and 7-AAD staining of the CD38 high/CD45 low cells on a FACSCanto using Diva software (BD).CasesPt 1.was a 63 year old female diagnosed MAR 2012 with an IgG lambda ISS stage I, Durie & Salmon stage III A MM. Cytogenetics showed a t(11;14). 1st line treatment consisted of VTD -> ASCT with a resulting PR. Post ASCT the pt. was stable on Rd – maintenance until JAN 13 when a leukemic transformation and a light chain escape phenomenon occurred. Cytogenetics showed clonal evolution with acquisition of additional aberrations (+18, amp 1p36, amp 11q22.3). From APR 2013 – JUL 2013 the pt. could be stabilized with a Bortezomib-liposomal Anthracyclin-Dexamethason combination. On progression she opted for palliative care and succumbed to PCL in AUG 2013. Her PCL cells were evaluated in-vitro.Pt. 2was a 51 years old female diagnosed with FLC kappa ISS stage III, Durie & Salmon stage III A MM in NOV 2011. In JUN 2014 the pt. presented with a 2nd relapse having been pre-treated with VTD-ASCT in 1st line, and Rd in 2nd line. Out of PR under continuous Rd for 9 months she developed a rapidly progressive PCL with a complex aberrant karyotype and massively elevated sFLC kappa. The patient was severely anaemic and thrombopenic from presentation onwards. Her PCL cells were evaluated in-vitro while treatment was initiated with the Pi-Po-D regime:Table 1Pi-Po-D regimeDRUGAPPLICATIONDOSAGESCHEDULEREPETITIONPixantronei.v.50mg/m²d1-8-15d28Pomalidomidep.o.4mgd1-21d28Dexamethasonp.o.40mgd1-4, 8,15d28In-Vitro-ResultsPt. 1 showed a significant induction of apoptosis concomitantly with a reduction of living cells after an incubation period of 24h starting with a PIX concentration of 2.50 µM and with 1.0 µM at 48h (Table 2). Similar results were obtained with the cells of Pt. 2, despite methodological problems however, plasma cells proofed not to be as sensitive as the whole cell population.Abstract 5732. Table 2In-Vitro activity of Pixantrone in pt. 1Pixantrone Concentration (µM)Living cells after 24 h (%)Apoptotic cells after 24h (%)Living cells after 48h (%)Apoptotic cells after 48h (%)0.0095.8504.1597.3502.650.0196.1503.8596.4503.550.0595.0504.9596.7503.250.1093.3506.6595.7004.300.2590.7509.2593.7006.300.5085.4014.6084.2515.751.0079.9520.0561.1038.902.5031.3068.7006.6593.355.0000.1599.8505.0099.70Clinical course with Pi-Po-D in PCLPt. 2 received the Pi-Po-D regime as no suitable clinical alternative was available and in-vitro data were promising. We noticed a sharp decline of sFLC (> 30%/3weeks) and peripheral plasma cell counts within cycle I as early sign of clinical activity. Unfortunately the pt. had a severe fall while being grade IV CTC thrombopenic and died from unstoppable intracranial haemorrhage.DiscussionPixantrone shows promising in-vitro activity in PCL and MM* at pharmacologically realistic concentrations and first signs of clinical activity in combination therapy. In our opinion PIX deserves further pre-clinical and clinical evaluation in MM and PCL.*J Clin Oncol 32, 2014 (suppl; abstr e19569) [Display omitted] DisclosuresOff Label Use: Use of Pixantrone in Plasma Cell Leukaemia.

The Alkylating Histone Deacetylase Inhibitor Fusion Molecule Edo-S101 Displays Full Bi-Functional Properties in Preclinical Models of Hematological Malignancies

BackgroundAlkylating histone deacetylase inhibitors (HDACi) enhance the anticancer efficacy of alkylators by increasing chromatin accessibility and also down regulating DNA repair. EDO-S101 is a first-in-class fusion molecule that combines DNA damaging effect of bendamustine with the pan-HDACi vorinostat.ObjectivesTo study the bi-functional properties of EDO-S101 as an alkylating agent and a pan-HDACi in various in vitro and in vivo xenograft models of hematological malignancies.MethodsIn vitro inhibition of HDAC Class I and II enzymes by EDO-S101 and vorinostat was tested using an recombinant human enzymatic assay (BPS Bioscience, Enzo Life Science) and in vivo in rat peripheral blood mononuclear cells (PBMCs). The degree of inhibition was measured 1 hour following a single dose of 10–50 mg/kg i.v. and duration of inhibition over 24 hours after a single i.v. dose of EDO-S101 of 25 mg/kg. HDAC inhibition, alkylation and apoptotic activity were evaluated in vitro in myeloid (HL60 AML cell line) and lymphoid cell lines, including Daudi Burkitt’s lymphoma (BL) and a panel of 6 MM cell lines (MM1S, MM1R, RPMI-8226, RPMI-LR5, U266, U266-LR7). In vivo intra-tumor effects were analyzed after short courses of treatment with EDO-S101 in MM1S human plasmacytoma (PC) and BL xenograft models. Changes in pathway activation, protein expression and activities influencing the cell cycle were measured by Western blot and immunohistochemistry. Anti-tumor activity in vitro was measured by MTT and in vivo using a caliper to assess tumor size at regular intervals.ResultsIn vitro, EDO-S101’s pan-HDACi activity, at nanomolar concentrations in Class I and II recombinant enzymes, was similar to vorinostat. In vivo, in intact rat PBMCs, HDAC inhibition was maximal at 1 hour after a single dose of 10 mg/kg i.v.–the dose where antitumor activity starts. HDAC inhibition did not increase with doses up to 50 mg/kg, recovery began within 3 hours and was nearly complete at 16 hours.In the AML HL60 cell line in vitro, hyperacetylation of lysine residues K9, K14, K23 and K56 on histone 3 was found after exposure to 2–4 µM of EDO-S101. Histone 3 and 4 hyperacetylation was also demonstrated in MM cell lines at 1–5 µM concentrations. In xenograft models of human plasmacytoma and BL, EDO-S101 induced histone 3 hyperacetylation, indicating an HDACi effect in vivo.Alkylating activity was demonstrated in vitro in HL60 and MM cell lines by DNA cross-linking and double strand break formation in the comet assay by immunofluorescence. In vivo, in xenograft models of human plasmacytoma (60 mg/kg d 1, 8, 15) and BL (40 and 80mg/kg d1) exposure to EDO-S101 caused a strong DNA-repair response shown by activation of pH2AX and p53 (PC and BL) followed by an increase of DNA damage check point proteins pCHK1 (PC) and even more prominent pCHK2 (PC and BL). The kinetics of this effect, studied in vivo in BL tumors, showed that the pH2AX response fell at Day 8 after dosing while the p53 response lasted, particularly in the group treated with 80mg/kg. In Daudi-bearing mice tumors, p-ATR was completely suppressed at Day 8 after treatment, which was not clear in the PC tumors.EDO-S101 triggered apoptosis in vitro and in vivo, resulting in strong antitumor activity in HL60, Daudi and the panel of six MM cell lines. Initial in vitro experiments in HL60 cells showed an activation of the intrinsic pathway of apoptosis with cleavage of caspases 3, 9 and PARP and a marked reduction of anti-apoptotic proteins XIAP and Mcl-1. In the MM cell line, MM1S activation of the intrinsic and extrinsic pathways of apoptosis (C 8, 9, 3, 7 and PARP cleavage) was seen with a loss of mitochondrial membrane potential by DiOC6.Tumors of human plasmacytoma and BL in vivo were rapidly shrinking or completely eradicated after i.v. administration of EDO-S101. A decrease in proliferation (Ki67) and slight PARP cleavage was found in the tumor tissue (PC), and evidence of activation of apoptosis by cleavage of caspases 7 and 9 at Day 4 and caspase 8 and PARP at Day 8 after treatment in BL tumors. The level of caspase 3, different to MM, remained unchanged. Importantly, EDO-S101 induced a rapid and dose-dependent strong decrease of XIAP and Mcl-1 which lasted until Day 8.ConclusionsThis study demonstrates the bi-functional mechanism of ED0-S101 in both myeloid and lymphoid hematological malignancies. The data support the clinical investigation of EDO-S101 in treating hematological malignancies. DisclosuresOcio:Mundipharma: Honoraria, Research Funding. Mehrling:Mundipharma: Employment.

Novel magnetic carbon based solid acid for alkylation of benzene and dodecene

Novel magnetic carbon based solid acid has been synthesized through the hydrothermal carbonization of glucose with magnetic cores and sulfonation of carbonaceous intermediate with hydroxyethylsulfonic acid. The carbon based solid acid owned the core–shell structure and high BET surface, which provided the easily accessible acid sites on the shell. The magnetic solid acid exhibited high activities for the hydrophobic alkylation of benzene and dodecene with complete conversion of dodecene, which was difficult for the traditional carbon based acids for low hydrophilic surface. The simple magnetic recovery process and high stability added the advantages of the solid acid. The high activities, reusability and simple recovery process were the key properties of the solid acid, which hold great potential for green chemical processes.