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1.
ACS Omega ; 7(38): 33963-33984, 2022 Sep 27.
Article in English | MEDLINE | ID: mdl-36188331

ABSTRACT

Addressing the growing burden of cancer and the shortcomings of chemotherapy in cancer treatment are the current research goals. Research to overcome the limitations of curcumin and to improve its anticancer activity via its heterocycle-fused monocarbonyl analogues (MACs) has immense potential. In this study, 32 asymmetric MACs fused with 1-aryl-1H-pyrazole (7a-10h) were synthesized and characterized to develop new curcumin analogues. Subsequently, via initial screening for cytotoxic activity, nine compounds exhibited potential growth inhibition against MDA-MB-231 (IC50 2.43-7.84 µM) and HepG2 (IC50 4.98-14.65 µM), in which seven compounds showing higher selectivities on two cancer cell lines than the noncancerous LLC-PK1 were selected for cell-free in vitro screening for effects on microtubule assembly activity. Among those, compounds 7d, 7h, and 10c showed effective inhibitions of microtubule assembly at 20.0 µM (40.76-52.03%), indicating that they could act as microtubule-destabilizing agents. From the screening results, three most potential compounds, 7d, 7h, and 10c, were selected for further evaluation of cellular effects on breast cancer MDA-MB-231 cells. The apoptosis-inducing study indicated that these three compounds could cause morphological changes at 1.0 µM and could enhance caspase-3 activity (1.33-1.57 times) at 10.0 µM in MDA-MB-231 cells, confirming their apoptosis-inducing activities. Additionally, in cell cycle analysis, compounds 7d and 7h at 2.5 µM and 10c at 5.0 µM also arrested MDA-MB-231 cells in the G2/M phase. Finally, the results from in silico studies revealed that the predicted absorption, distribution, metabolism, excretion, and the toxicity (ADMET) profile of the most potent MACs might have several advantages in addition to potential disadvantages, and compound 7h could bind into (ΔG -10.08 kcal·mol-1) and access wider space at the colchicine-binding site (CBS) than that of colchicine or nocodazole via molecular docking studies. In conclusion, our study serves as a basis for the design of promising synthetic compounds as anticancer agents in the future.

2.
ESMO Open ; 6(3): 100134, 2021 06.
Article in English | MEDLINE | ID: mdl-33984676

ABSTRACT

BACKGROUND: The impact of the first coronavirus disease 2019 (COVID-19) wave on cancer patient management was measured within the nationwide network of the Unicancer comprehensive cancer centers in France. PATIENTS AND METHODS: The number of patients diagnosed and treated within 17 of the 18 Unicancer centers was collected in 2020 and compared with that during the same periods between 2016 and 2019. Unicancer centers treat close to 20% of cancer patients in France yearly. The reduction in the number of patients attending the Unicancer centers was analyzed per regions and cancer types. The impact of delayed care on cancer-related deaths was calculated based on different hypotheses. RESULTS: A 6.8% decrease in patients managed within Unicancer in the first 7 months of 2020 versus 2019 was observed. This reduction reached 21% during April and May, and was not compensated in June and July, nor later until November 2020. This reduction was observed only for newly diagnosed patients, while the clinical activity for previously diagnosed patients increased by 4% similar to previous years. The reduction was more pronounced in women, in breast and prostate cancers, and for patients without metastasis. Using an estimated hazard ratio of 1.06 per month of delay in diagnosis and treatment of new patients, we calculated that the delays observed in the 5-month period from March to July 2020 may result in an excess mortality due to cancer of 1000-6000 patients in coming years. CONCLUSIONS: In this study, the delays in cancer patient management were observed only for newly diagnosed patients, more frequently in women, for breast cancer, prostate cancer, and nonmetastatic cancers. These delays may result is an excess risk of cancer-related deaths in the coming years.


Subject(s)
COVID-19 , Neoplasms/complications , COVID-19/complications , Female , France , Humans , Male , SARS-CoV-2
3.
ACS Omega ; 5(25): 15229-15239, 2020 Jun 30.
Article in English | MEDLINE | ID: mdl-32637796

ABSTRACT

In this study, cellulose extracted from straw was modified using N(4)-morpholinothiosemicarbazide to generate a novel adsorbent as a chelate-complex-based material. The effects of pH, time, temperature, and mass ratios of KIO4: cellulose on the yield of the oxidation were analyzed using iodometric titration and photometric methods. The accuracy and precision of the above two methods were evaluated using Student and Fisher statistical distribution. The structure of the material was characterized by Fourier-transform infrared spectroscopy, thermogravimetric analysis, X-ray diffraction, scanning electron microscopy, and Brunauer-Emmett-Teller surface area analysis. The kinetic order of Ni(II) adsorption was dependent on the concentration of Ni(II). The surface response design enabled to optimize the condition for Ni(II) adsorption at 58 °C, pH of 4.98, within 106 min. The maximum Ni(II) adsorption capacity was 90 mg g-1. This kind of adsorbent can be reused at least five times without a significant decrease in its adsorption efficiency.

4.
J Expo Sci Environ Epidemiol ; 30(4): 689-697, 2020 07.
Article in English | MEDLINE | ID: mdl-32355212

ABSTRACT

The literature shows associations between maternal exposures to PM2.5 and adverse pregnancy outcomes. There are few data from Latin America. We have examined PM2.5 and pregnancy outcomes in Lima. The study included 123,034 births from 2012 to 2016, at three public hospitals. We used estimated daily PM2.5 from a newly created model developed using ground measurements, satellite data, and a chemical transport model. Exposure was assigned based on district of residence (n = 39). Linear and logistic regression analyzes were used to estimate the associations between air pollution exposure and pregnancy outcomes. Increased exposure to PM2.5 during the entire pregnancy and in the first trimester was inversely associated with birth weight. We found a decrease of 8.13 g (-14.0; -1.84) overall and 18.6 g (-24.4, -12.8) in the first trimester, for an interquartile range (IQR) increase (9.2 µg/m3) in PM2.5. PM2.5 exposure was positively associated with low birth weight at term (TLBW) during entire pregnancy (OR: 1.11; 95% CI: 1.03-1.20), and at the first (OR: 1.11; 95% CI: 1.03-1.20), second (OR: 1.09; 95% CI: 1.01-1.17), and third trimester (OR: 1.10; 95% CI: 1.02-1.18) per IQR (9.2 µg/m3) increase. Higher exposure to PM2.5 was also associated with increased risk of small for gestational age (SGA). There were no statistically significant associations between PM2.5 exposure and preterm births (PTB). Exposure to higher concentrations of PM2.5 in Lima may decrease birth weight and increase the frequency of TLBW and SGA. Our study was inconsistent with the literature in finding no associations with preterm birth.


Subject(s)
Maternal Exposure/statistics & numerical data , Particulate Matter/analysis , Pregnancy Outcome/epidemiology , Adult , Air Pollutants/analysis , Air Pollution/adverse effects , Birth Weight , Female , Humans , Infant, Low Birth Weight , Infant, Newborn , Logistic Models , Maternal Exposure/adverse effects , Peru/epidemiology , Pregnancy , Pregnancy Trimester, Third , Premature Birth
5.
J Expo Sci Environ Epidemiol ; 30(4): 680-688, 2020 07.
Article in English | MEDLINE | ID: mdl-31745179

ABSTRACT

INTRODUCTION: There have been no time-series studies of air pollution in Peru. Here we evaluate the effect of ambient PM2.5 on emergency room (ER) visits in Lima. METHODS: We estimated daily PM2.5 levels at a 1 km2 resolution during 2010-2016 using ground measurements, satellite data, and chemical transport model simulations. Population-weighted average daily PM2.5 levels were calculated for each district in Lima (n = 40), and assigned to patients based on residence. ER visits for respiratory and circulatory diseases were gathered from nine large public hospitals. Poisson regression was used to estimate the rate ratio for daily ER visits with change in daily PM2.5, controlling for meteorology, time trends, and district. RESULTS: For each interquartile range (IQR) increase in PM2.5, respiratory disease ER visits increased 4% (95% CI: 0-5%), stroke visits 10% (3-18%), and ischemic heart disease visits (adults, 18-64 years) 11% (-1, 24%). Districts with higher poverty showed significantly stronger associations of PM2.5 and respiratory disease ER visits than districts with lower poverty. Effects were diminished 24-42% using Lima-wide instead of district-specific PM2.5 levels. CONCLUSIONS: Short-term exposure to ambient PM2.5 is associated with increases in ER visits in Lima for respiratory diseases and stroke, and among middle-aged adults, ischemic heart disease.


Subject(s)
Air Pollution/statistics & numerical data , Cardiovascular Diseases/epidemiology , Environmental Exposure/statistics & numerical data , Respiratory Tract Diseases/epidemiology , Adult , Aged , Air Pollutants/analysis , Air Pollution/analysis , Emergency Service, Hospital/statistics & numerical data , Environmental Exposure/analysis , Female , Humans , Male , Meteorology , Middle Aged , Particulate Matter/analysis , Peru/epidemiology , Poverty , Stroke , Time
6.
Nat Med ; 25(7): 1131-1142, 2019 07.
Article in English | MEDLINE | ID: mdl-31263285

ABSTRACT

Huntington's disease (HD) is a dominantly inherited neurodegenerative disorder caused by a CAG trinucleotide expansion in the huntingtin gene (HTT), which codes for the pathologic mutant HTT (mHTT) protein. Since normal HTT is thought to be important for brain function, we engineered zinc finger protein transcription factors (ZFP-TFs) to target the pathogenic CAG repeat and selectively lower mHTT as a therapeutic strategy. Using patient-derived fibroblasts and neurons, we demonstrate that ZFP-TFs selectively repress >99% of HD-causing alleles over a wide dose range while preserving expression of >86% of normal alleles. Other CAG-containing genes are minimally affected, and virally delivered ZFP-TFs are active and well tolerated in HD neurons beyond 100 days in culture and for at least nine months in the mouse brain. Using three HD mouse models, we demonstrate improvements in a range of molecular, histopathological, electrophysiological and functional endpoints. Our findings support the continued development of an allele-selective ZFP-TF for the treatment of HD.


Subject(s)
Alleles , Huntingtin Protein/genetics , Huntington Disease/therapy , Mutation , Transcription, Genetic , Zinc Fingers , Animals , Cells, Cultured , Disease Models, Animal , Female , Humans , Huntington Disease/genetics , Male , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Neuroprotection , Trinucleotide Repeats
7.
Cancer Radiother ; 18(5-6): 458-60, 2014 Oct.
Article in French | MEDLINE | ID: mdl-25195115

ABSTRACT

In 2013, about 6000 patients were treated with brachytherapy, the number diminishing by 2.6% per year since 2008. Prostate, breast and gynecological cancers are the most common types of cancers. Since 2008, the number of brachytherapy facilities has decreased by 18%. In medicoeconomic terms, brachytherapy faces many problems: the coding system is outdated; brachytherapy treatments cost as much as internal radiation; fees do not cover costs; since iridium wire has disappeared from the market, the technique will be transferred to more expensive high-speed or pulse dose rates. The French financing grid based on the national study of costs lags behind changes in such treatments and in the best of cases, hospitals resorting to alternatives such as in-hospital brachytherapy are funded at 46% of their additional costs. Brachytherapy is a reference technique. With intense pressure on hospital pricing, financing brachytherapy facilities will become even more problematic as a consequence of the disappearance of iridium 192 wires. The case of brachytherapy illustrates the limits of the French financing system and raises serious doubts as to its responsiveness.


Subject(s)
Brachytherapy/instrumentation , Iridium Radioisotopes/administration & dosage , Brachytherapy/economics , Brachytherapy/methods , Brachytherapy/statistics & numerical data , Brachytherapy/trends , Cancer Care Facilities/economics , Commerce , Equipment Design , Equipment and Supplies/supply & distribution , France , Hospital Costs , Humans , Neoplasms/economics , Neoplasms/radiotherapy , Radiotherapy Dosage , Technology, High-Cost/economics , Universal Health Insurance/economics , Vocabulary, Controlled
8.
Mol Immunol ; 59(1): 79-90, 2014 May.
Article in English | MEDLINE | ID: mdl-24491490

ABSTRACT

We have designed a 39 amino acid peptide mimic of the conformation-dependent main immunogenic region (MIR) of the Torpedo acetylcholine receptor (TAChR) that joins three discontinuous segments of the Torpedo α-subunit, α(1-12), α(65-79), and α(110 - 115) with two GS linkers: This 39MIR-mimic was expressed in E. coli as a fusion protein with an intein-chitin-binding domain (IChBD) to permit affinity collection on chitin beads. Six MIR-directed monoclonal antibodies (mAbs) bind to this complex and five agonist/antagonist site directed mAbs do not. The complex of MIR-directed mAb-132A with 39MIR has a Kd of (2.11±0.11)×10(-10)M, which is smaller than (7.13±1.20)×10(-10)M for the complex of mAb-132A with α(1-161) and about the same as 3.4×10(-10)M for that of mAb-132A with TAChR. Additionally, the 39MIR-IChBD adsorbs all MIR-directed antibodies (Abs) from an experimental autoimmune myasthenia gravis (EAMG) rat serum. Hence, the 39MIR-mimic has the potential to inactivate or remove pathogenic Torpedo MIR-directed Abs from EAMG sera and to direct a magic bullet to the memory B-cells that produce those pathogenic Abs. The hope is to use this as a guide to produce a mimic of the human MIR on the way to an antigen specific therapeutic agent to treat MG.


Subject(s)
Fish Proteins/immunology , Peptides/immunology , Receptors, Cholinergic/immunology , Torpedo/immunology , Amino Acid Sequence , Animals , Antibodies, Monoclonal/immunology , Antibody Affinity/immunology , Base Sequence , Binding Sites/genetics , Binding Sites/immunology , Blotting, Western , Drug Design , Fish Proteins/chemistry , Fish Proteins/genetics , Immune Sera/immunology , Models, Molecular , Molecular Mimicry , Molecular Sequence Data , Myasthenia Gravis/blood , Myasthenia Gravis/immunology , Peptides/chemistry , Peptides/genetics , Protein Binding/immunology , Protein Structure, Tertiary , Rats , Receptors, Cholinergic/chemistry , Receptors, Cholinergic/genetics , Torpedo/genetics
9.
Neurobiol Dis ; 62: 381-6, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24148854

ABSTRACT

Parkinson's disease associated mutations in leucine rich repeat kinase 2 (LRRK2) impair mitochondrial function and increase the vulnerability of induced pluripotent stem cell (iPSC)-derived neural cells from patients to oxidative stress. Since mitochondrial DNA (mtDNA) damage can compromise mitochondrial function, we examined whether LRRK2 mutations can induce damage to the mitochondrial genome. We found greater levels of mtDNA damage in iPSC-derived neural cells from patients carrying homozygous or heterozygous LRRK2 G2019S mutations, or at-risk individuals carrying the heterozygous LRRK2 R1441C mutation, than in cells from unrelated healthy subjects who do not carry LRRK2 mutations. After zinc finger nuclease-mediated repair of the LRRK2 G2019S mutation in iPSCs, mtDNA damage was no longer detected in differentiated neuroprogenitor and neural cells. Our results unambiguously link LRRK2 mutations to mtDNA damage and validate a new cellular phenotype that can be used for examining pathogenic mechanisms and screening therapeutic strategies.


Subject(s)
DNA Damage , DNA, Mitochondrial/metabolism , Neural Stem Cells/metabolism , Parkinson Disease/genetics , Parkinson Disease/metabolism , Protein Serine-Threonine Kinases/genetics , Targeted Gene Repair , Adult , Aged , DNA Repair , DNA, Mitochondrial/genetics , Female , Humans , Induced Pluripotent Stem Cells/metabolism , Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 , Male , Middle Aged , Mutation , Zinc Fingers
10.
Mol Immunol ; 58(1): 116-31, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24333757

ABSTRACT

To develop antigen-specific immunotherapies for autoimmune diseases, knowledge of the molecular structure of targeted immunological hotspots will guide the production of reagents to inhibit and halt production of antigen specific attack agents. To this end we have identified three noncontiguous segments of the Torpedo nicotinic acetylcholine receptor (AChR) α-subunit that contribute to the conformationally sensitive immunological hotspot on the AChR termed the main immunogenic region (MIR): α(1-12), α(65-79), and α(110-115). This region is the target of greater than 50% of the anti-AChR Abs in serum from patients with myasthenia gravis (MG) and animals with experimental autoimmune myasthenia gravis (EAMG). Many monoclonal antibodies (mAbs) raised in one species against an electric organ AChR cross react with the neuromuscular AChR MIR in several species. Probing the Torpedo AChR α-subunit with mAb 132A, a disease inducing anti-MIR mAb raised against the Torpedo AChR, we have determined that two of the three MIR segments, α(1-12) and α(65-79), form a complex providing the signature components recognized by mAb 132A. These two segments straddle a third, α(110-115), that seems not to contribute specific side chains for 132A recognition, but is necessary for optimum antibody binding. This third segment appears to form a foundation upon which the three-dimensional 132A epitope is anchored.


Subject(s)
Myasthenia Gravis/immunology , Receptors, Nicotinic/chemistry , Amino Acid Sequence , Animals , Antibodies, Monoclonal/immunology , Binding Sites, Antibody/immunology , Green Fluorescent Proteins/genetics , Humans , Molecular Sequence Data , Myasthenia Gravis/blood , Peptide Fragments/immunology , Protein Binding/immunology , Protein Conformation , Protein Structure, Tertiary , Receptors, Nicotinic/immunology , Recombinant Fusion Proteins/genetics , Sequence Analysis, Protein , Torpedo
11.
Environ Pollut ; 166: 172-81, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22512998

ABSTRACT

Polycyclic aromatic hydrocarbon (PAH) extracts of fine particles (PM(2.5)) collected from combustion of seven wood species and briquettes were tested for mutagenic activities using Ames test with Salmonella typhimurium TA98 and TA100. The woods were Pinus pinaster (maritime pine), Eucalyptus globulus (eucalypt), Quercus suber (cork oak), Acacia longifolia (golden wattle), Quercus faginea (Portuguese oak), Olea europea (olive), and Quercus ilex rotundifolia (Holm oak). Burning experiments were done using woodstove and fireplace, hot start and cold start conditions. A mutagenic response was recorded for all species except golden wattle, maritime pine, and briquettes. The mutagenic extracts were not correlated with high emission factors of carcinogenic PAHs. These extracts were obtained both from two burning appliances and start-up conditions. However, fireplace seemed to favour the occurrence of mutagenic emissions. The negative result recorded for golden wattle was interesting, in an ecological point of view, since after confirmation, this invasive species, can be recommended for domestic use.


Subject(s)
Aerosols/toxicity , Air Pollutants/toxicity , Mutagens/toxicity , Polycyclic Aromatic Hydrocarbons/toxicity , Aerosols/analysis , Air Pollutants/analysis , Eucalyptus , Fires , Incineration , Mutagenicity Tests , Mutagens/analysis , Particulate Matter/analysis , Particulate Matter/toxicity , Pinus , Polycyclic Aromatic Hydrocarbons/analysis , Portugal , Salmonella typhimurium/drug effects , Wood
12.
Chem Biol Drug Des ; 79(2): 157-65, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22107736

ABSTRACT

HIV-1 integrase (IN) is a validated therapeutic target for the treatment of AIDS. However, the emergence of resistance to raltegravir, the sole marketed FDA-approved IN inhibitor, emphasizes the need to develop second-generation inhibitors that retain efficacy against clinically relevant IN mutants. We report herein bicyclic hydroxy-1H-pyrrolopyridine-triones as a new family of HIV-1 integrase inhibitors that were efficiently prepared using a key 'Pummerer cyclization deprotonation cycloaddition' cascade of imidosulfoxides. In in vitro HIV-1 integrase assays, the analogs showed low micromolar inhibitory potencies with selectivity for strand transfer reactions as compared with 3'-processing inhibition. A representative inhibitor (5e) retained most of its inhibitory potency against the three major raltegravir-resistant IN mutant enzymes, G140S/Q148H, Y143R, and N155H. In antiviral assays employing viral vectors coding these IN mutants, compound 5e was approximately 200- and 20-fold less affected than raltegravir against the G140S/Q148H and Y143R mutations, respectively. Against the N155H mutation, 5e was approximately 10-fold less affected than raltegravir. Thus, our new compounds represent a novel structural class that may be further developed to overcome resistance to raltegravir, particularly in the case of the G140S/Q148H mutations.


Subject(s)
Bridged Bicyclo Compounds/chemistry , HIV Integrase Inhibitors/chemistry , HIV-1 , Imides/chemistry , Pyridines/chemistry , Pyridones/chemistry , Amino Acid Substitution , Binding Sites , Bridged Bicyclo Compounds/chemical synthesis , Bridged Bicyclo Compounds/pharmacology , Cell Line, Tumor , Computer Simulation , Drug Resistance, Viral/drug effects , HIV Integrase/chemistry , HIV Integrase/genetics , HIV Integrase/metabolism , HIV Integrase Inhibitors/chemical synthesis , HIV Integrase Inhibitors/pharmacology , HIV-1/drug effects , HIV-1/enzymology , Humans , Imides/chemical synthesis , Imides/pharmacology , Mutation , Pyridones/chemical synthesis , Pyridones/pharmacology , Pyrrolidinones/pharmacology , Raltegravir Potassium
13.
Eur J Clin Nutr ; 66(1): 75-82, 2012 Jan.
Article in English | MEDLINE | ID: mdl-21811293

ABSTRACT

BACKGROUND/OBJECTIVES: The effect of vegan diet on bone loss has not been studied. The aim of this study was to examine the association between veganism and bone loss in postmenopausal women. SUBJECTS/METHODS: The study was designed as a prospective longitudinal investigation with 210 women, including 105 vegans and 105 omnivores. Femoral neck (FN) bone mineral density (BMD) was measured in 2008 and 2010 by dual-energy X-ray absorptiometry (Hologic QDR4500). The incidence of vertebral fracture was ascertained by X-ray report. Serum levels of C-terminal telopeptide of type I collagen (ßCTX) and N-terminal propeptide of type I procollagen (PINP) were measured by Roche Elecsys assays. Serum concentration of 25-hydroxyvitamin D and parathyroid hormone were measured by electrochemiluminescence. RESULTS: Among the 210 women who initially participated in the study in 2008, 181 women had completed the study and 29 women were lost to follow-up. The rate of loss in FN BMD was -1.91±3.45%/year in omnivores and -0.86±3.81%/year (P=0.08) in vegans. Lower body weight, higher intakes of animal protein and lipid, and corticosteroid use were associated with greater rate of bone loss. The 2-year incidence of fracture was 5.7% (n=5/88) in vegans, which was not significantly different from omnivores (5.4%, n=6/93). There were no significant differences in ßCTX and PINP between vegans and omnivores. The prevalence of vitamin D insufficiency in vegans was higher than in omnivores (73% versus 46%; P=0.0003). CONCLUSIONS: Vegan diet did not have adverse effect on bone loss and fracture. Corticosteroid use and high intakes of animal protein and animal lipid were negatively associated with bone loss.


Subject(s)
Bone Density , Collagen Type I/blood , Diet, Vegetarian , Fractures, Bone , Osteoporosis, Postmenopausal/etiology , Peptides/blood , Vitamin D/analogs & derivatives , Absorptiometry, Photon , Adrenal Cortex Hormones/adverse effects , Aged , Asian People , Body Weight , Bone Resorption/blood , Bone Resorption/etiology , Diet, Vegetarian/adverse effects , Dietary Fats/adverse effects , Dietary Proteins/adverse effects , Feeding Behavior , Female , Humans , Longitudinal Studies , Lost to Follow-Up , Middle Aged , Osteoporosis, Postmenopausal/blood , Prevalence , Prospective Studies , Vietnam/epidemiology , Vitamin D/blood , Vitamin D Deficiency/blood , Vitamin D Deficiency/epidemiology
14.
Cell ; 146(2): 318-31, 2011 Jul 22.
Article in English | MEDLINE | ID: mdl-21757228

ABSTRACT

Patient-specific induced pluripotent stem cells (iPSCs) derived from somatic cells provide a unique tool for the study of human disease, as well as a promising source for cell replacement therapies. One crucial limitation has been the inability to perform experiments under genetically defined conditions. This is particularly relevant for late age onset disorders in which in vitro phenotypes are predicted to be subtle and susceptible to significant effects of genetic background variations. By combining zinc finger nuclease (ZFN)-mediated genome editing and iPSC technology, we provide a generally applicable solution to this problem, generating sets of isogenic disease and control human pluripotent stem cells that differ exclusively at either of two susceptibility variants for Parkinson's disease by modifying the underlying point mutations in the α-synuclein gene. The robust capability to genetically correct disease-causing point mutations in patient-derived hiPSCs represents significant progress for basic biomedical research and an advance toward hiPSC-based cell replacement therapies.


Subject(s)
Parkinson Disease/pathology , Pluripotent Stem Cells , Point Mutation , Cell Line , Embryonic Stem Cells , Genetic Engineering , Genome-Wide Association Study , Humans , Mutagenesis , Oligonucleotides/metabolism , alpha-Synuclein/genetics
15.
Bioorg Med Chem Lett ; 21(10): 2986-90, 2011 May 15.
Article in English | MEDLINE | ID: mdl-21493066

ABSTRACT

New tricyclic HIV-1 integrase (IN) inhibitors were prepared that combined structural features of bicyclic pyrimidinones with recently disclosed 4,5-dihydroxy-1H-isoindole-1,3(2H)-diones. This combination resulted in the introduction of a nitrogen into the aryl ring and the addition of a fused third ring to our previously described inhibitors. The resulting analogues showed low micromolar inhibitory potency in in vitro HIV-1 integrase assays, with good selectivity for strand transfer relative to 3'-processing.


Subject(s)
HIV Integrase Inhibitors/chemistry , HIV Integrase Inhibitors/chemical synthesis , HIV-1/enzymology , Pyrimidinones/chemical synthesis , Biological Assay , Cells, Cultured , Cyclization , HIV Integrase/metabolism , HIV Integrase Inhibitors/pharmacology , HIV-1/drug effects , Humans , Hydroxylation , Inhibitory Concentration 50 , Molecular Structure , Pyrimidinones/chemistry , Pyrimidinones/pharmacology , Structure-Activity Relationship
16.
Antimicrob Agents Chemother ; 55(5): 2379-89, 2011 May.
Article in English | MEDLINE | ID: mdl-21343443

ABSTRACT

It is important to develop new anti-HIV drugs that are effective against the existing drug-resistant mutants. Because the excision mechanism is an important pathway for resistance to nucleoside analogs, we are preparing analogs that retain a 3'-OH and can be extended after they are incorporated by the viral reverse transcriptase. We show that 4'-C-alkyl-deoxyadenosine (4'-C-alkyl-dA) compounds can be phosphorylated in cultured cells and can inhibit the replication of HIV-1 vectors: 4'-C-methyl- and 4'-C-ethyl-dA show both efficacy and selectivity against HIV-1. The compounds are also effective against viruses that replicate using reverse transcriptases (RTs) that carry nucleoside reverse transcriptase inhibitor resistance mutations, with the exception of the M184V mutant. Analysis of viral DNA synthesis in infected cells showed that viral DNA synthesis is blocked by the incorporation of either 4'-C-methyl- or 4'-C-ethyl-2'-deoxyadenosine. In vitro experiments with purified HIV-1 RT showed that 4'-C-methyl-2'-dATP can compete with dATP and that incorporation of the analog causes pausing in DNA synthesis. The 4'-C-ethyl compound also competes with dATP and shows a differential ability to block DNA synthesis on RNA and DNA templates. Experiments that measure the ability of the compounds to block DNA synthesis in infected cells suggest that this differential block to DNA synthesis also occurs in infected cells.


Subject(s)
Anti-HIV Agents/pharmacology , DNA Replication/drug effects , Deoxyadenosines/pharmacology , HIV-1/drug effects , Anti-HIV Agents/chemistry , Cell Line , Deoxyadenosines/chemistry , HIV Reverse Transcriptase/genetics , HIV Reverse Transcriptase/metabolism , HIV-1/enzymology , Humans , Magnetic Resonance Spectroscopy , Polymerase Chain Reaction
17.
J Med Chem ; 52(17): 5356-64, 2009 Sep 10.
Article in English | MEDLINE | ID: mdl-19678643

ABSTRACT

A major pathway for HIV-1 resistance to nucleoside reverse transcriptase inhibitors (NRTIs) involves reverse transcriptase (RT) mutations that enhance ATP-dependent pyrophosphorolysis, which excises NRTIs from the end of viral DNA. We analyzed novel NRTIs for their ability to inhibit DNA synthesis of excision-proficient HIV-1 RT mutants. D-carba T is a carbocyclic nucleoside that has a 3' hydroxyl on the pseudosugar. The 3' hydroxyl group allows RT to incorporate additional dNTPs, which should protect D-carba TMP from excision. D-carba T can be converted to the triphosphate form by host cell kinases with moderate efficiency. D-carba T-TP is efficiently incorporated by HIV-1 RT; however, the next dNTP is added slowly to a D-carba TMP at the primer terminus. D-carba T effectively inhibits viral vectors that replicate using NRTI-resistant HIV-1 RTs, and there is no obvious toxicity in cultured cells. NRTIs based on the carbocyclic pseudosugar may offer an effective approach for the treatment of HIV-1 infections.


Subject(s)
Anti-HIV Agents/pharmacology , HIV Reverse Transcriptase/antagonists & inhibitors , HIV-1/drug effects , HIV-1/enzymology , Pyrimidine Nucleosides/pharmacology , Reverse Transcriptase Inhibitors/pharmacology , Thymidine/analogs & derivatives , Anti-HIV Agents/adverse effects , Anti-HIV Agents/chemistry , Base Sequence , Cell Line, Tumor , HIV-1/physiology , Humans , Models, Molecular , Molecular Conformation , Pyrimidine Nucleosides/adverse effects , Pyrimidine Nucleosides/chemistry , Reverse Transcriptase Inhibitors/adverse effects , Reverse Transcriptase Inhibitors/chemistry , Thymidine/adverse effects , Thymidine/chemistry , Thymidine/pharmacology , Virus Replication/drug effects
18.
ChemMedChem ; 4(8): 1354-63, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19533724

ABSTRACT

The syntheses of new conformationally locked North- and South-bicyclo[3.1.0]hexene nucleosides is reported. The North analogues were synthesized by a convergent approach from the known (1S,2R,5R)-5-[(tert-butyldiphenylsilyloxy)methyl]bicyclo[3.1.0]hex-3-en-2-ol by Mitsunobu coupling with the nucleobases. The South analogues were synthesized from their bicyclo[3.1.0]hexane nucleoside precursors by the selective protection of the primary hydroxy group, conversion of the secondary alcohol into a good leaving group, and base-catalyzed elimination to generate the olefin. The transformation of a bicyclo[3.1.0]hexane nucleoside into a bicyclo[3.1.0]hexene nucleoside flattens the five-membered ring of the bicyclic system and rescues anti-HIV activity for North-D4T, North-D4A, and South-D4C. The relationship between planarity and the anti/syn disposition of the nucleobase that is favored by a particular pseudosugar platform are proposed as key parameters in controlling biological activity.


Subject(s)
Anti-HIV Agents/chemistry , Bridged Bicyclo Compounds/chemistry , Nucleosides/chemistry , Anti-HIV Agents/chemical synthesis , Anti-HIV Agents/pharmacology , Bridged Bicyclo Compounds/chemical synthesis , Bridged Bicyclo Compounds/pharmacology , Cell Line, Tumor , Crystallography, X-Ray , HIV Reverse Transcriptase/metabolism , Humans , Molecular Conformation , Nucleosides/chemical synthesis , Nucleosides/pharmacology
19.
Bioorg Med Chem Lett ; 19(10): 2714-7, 2009 May 15.
Article in English | MEDLINE | ID: mdl-19364649

ABSTRACT

Using 2,3-dihydro-6,7-dihydroxy-1H-isoindol-1-one and 4,5-dihydroxy-1H-isoindole-1,3(2H)-dione based HIV-1 integrase inhibitors as display platforms, we undertook a thorough examination of the effects of modifying the halogen substituents on a key benzyl ring that is hypothesized to bind in a hydrophobic pocket of the integrase.DNA complex. Data from this study suggest that in general dihalo-substituted analogues have higher potency than monohalo-substituted compounds, but that further addition of halogens is not beneficial.


Subject(s)
HIV Integrase Inhibitors/chemistry , HIV Integrase/metabolism , Halogens/chemistry , Isoindoles/chemistry , Cells, Cultured , HIV Integrase/chemistry , HIV Integrase Inhibitors/chemical synthesis , HIV Integrase Inhibitors/pharmacology , Halogens/chemical synthesis , Humans , Isoindoles/chemical synthesis , Isoindoles/pharmacology , Structure-Activity Relationship
20.
Nucleic Acids Symp Ser (Oxf) ; (52): 623-4, 2008.
Article in English | MEDLINE | ID: mdl-18776534

ABSTRACT

The conformationally locked carbocyclic nucleoside phosphonates 2 and 2' and key intermediates for the synthesis of 3 and 3' were prepared from a chiral cyclopentene derivative and epicholorohydrine, respectively. The structure of the nucleoside precursor 6 was confirmed by X-ray crystallography. These carbocyclic nucleoside phosphonates were designed to probe their binding interactions at the active site of HIV-1-RT.


Subject(s)
Adenosine/analogs & derivatives , Anti-HIV Agents/chemical synthesis , HIV Reverse Transcriptase/drug effects , Organophosphonates/chemical synthesis , Reverse Transcriptase Inhibitors/chemical synthesis , Adenosine/chemical synthesis , Adenosine/chemistry , Anti-HIV Agents/chemistry , Catalytic Domain , Organophosphonates/chemistry , Reverse Transcriptase Inhibitors/chemistry
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