DOCSLIB.ORG

Full Text (PDF)

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Full Text (PDF) Research Article Identification of an Inactivating Cysteine Switch in Protein Kinase CE, a Rational Target for the Design of Protein Kinase CE–Inhibitory Cancer Therapeutics Feng Chu,1 John M. Koomen,2 Ryuji Kobayashi,2 and Catherine A. O’Brian1 Departments of 1Cancer Biology and 2Molecular Pathology, The University of Texas M.D. Anderson Cancer Center, Houston, Texas Abstract sion by selectively inhibiting the protein-tyrosine kinase activity of Critical roles played by some protein kinases in neoplastic BCR-ABL (1). In addition, gefitinib (Iressa) selectively inhibits the transformation and progression provide a rationale for devel- protein-tyrosine kinase activity of the epidermal growth factor oping selective, small-molecule kinase inhibitors as antineo- receptor (EGFR) and is effective against a small fraction of non– plastic drugs. Protein kinase CE (PKCE) is a rational target small cell lung cancers that express various somatic EGFR muta- for cancer therapy, because it is oncogenic and prometastatic tions (3). Both imatinib and gefitinib inhibit their protein kinase in transgenic mouse models. PKCE is activated by sn-1,2- targets by competing with the substrate ATP (1, 2). Similarly, selec- diacylglycerol (DAG). Attempts to develop selective PKCE tive inhibitors have been identified for other protein kinases in inhibitors that block activation by DAG or compete with ATP recent years by screening libraries of chemicals designed to compete have not yet met with success, suggesting a need for new with ATP (5, 6). However, this heavily mined approach has failed to identify selective inhibitors of some protein kinases that are strategies. We previously reported that cystamine and a meta- q q bolic cystine precursor inactivate PKCE in cells in a thiol- rational targets for cancer therapy, such as protein kinase C (PKC ). PKC is a family of 10 isozymes in the AGC group of serine/ reversible manner. In this report, we first determined that q PKCE became resistant to inactivation by disulfides when threonine protein kinases (7, 8). PKC is a rational target for cancer Cys452 was replaced with alanine by site-specific mutagenesis therapy, based on its oncogenic activity in fibroblasts and epithelial of human PKCE or a constitutively active PKCE mutant. These cells (9–11) and prometastatic activity in transgenic mouse models E of chemical carcinogenesis (12, 13). ATP-competitive inhibitors results showed that the disulfides inactivated PKC by thiol- q disulfide exchange, either upon Cys452 S-thiolation or by re- that have been identified for PKC are nonselective; this strategy arrangement to an intra-protein disulfide. Mass spectrometric has also failed to yield selective inhibitors for most of the other PKC isozymes (8, 14). Similarly, allosteric binding site targeting analysis of peptide digests of cystamine-inactivated, carbami- q q domethylated PKCE detected a peptide S-cysteaminylated at has not yielded selective PKC inhibitors. PKC and seven other 452 452 PKC isozymes are activated by phosphatidylserine-dependent Cys , indicating that Cys S-cysteaminylation is a stable 2+ modification. Furthermore, PKCE inactivation by N-ethyl- binding of sn-1,2-diacylglycerol (DAG) to cysteine-rich Zn fingers 452 in the regulatory domain (8). Efforts to develop selective inhibitors maleimide was Cys dependent, providing corroborative evidence that PKCE inhibitors can be designed by targeting of PKC isozymes by targeting the DAG-binding site have not Cys452 with small molecules that stably modify the residue. met with success, and enthusiasm for this strategy has waned as Cys452 is an active site residue that is conserved in only 11 other signaling protein families with homologous DAG-binding human protein kinase genes. Therefore, the PKCE-inactivating sites have been identified (e.g., RAS-GRPs and PKDs; refs. 15, 16). Cys452 switch is a rational target for the design of antineo- In recent years, we have investigated regulatory responses of plastic drugs that selectively inhibit PKCE. (Cancer Res 2005; PKC isozymes to cysteine modification by cystamine and other 65(22): 10478-85) disulfides (17–20). We initially analyzed purified human recombi- nant PKC isozymes and found that structurally diverse disulfides activated PKCy, which is tumor suppressive (21), and inactivated Introduction PKCq,PKCa, and other PKC isozymes to various extents (18, 19). Protein kinases play influential roles in aberrant cell signaling Similarly, treatment of cells with cystamine or a metabolic cystine events involved in neoplastic transformation and malignant prog- precursor activated PKCy and inactivated PKCq in a concentration- ression of epithelial and other types of cells. This is widely recog- dependent and thiol-reversible manner (17, 18). These results iden- nized as a strong rationale for developing selective, small-molecule tified covalent modification of one or more cysteine residues by inhibitors of protein kinases as antineoplastic drugs (1–4). This thiol-disulfide exchange as a novel mode of PKCq inactivation. approach was recently validated as a modality of antineoplastic In this report, we establish that Cys452 is a cysteine switch in therapy by the efficacy of imatinib (Gleevec) against chronic mye- human PKCq (hPKCq) that inactivates the kinase in cells upon q loid leukemia (CML; ref. 1). Imatinib treatment induces CML remis- modification by disulfides or N-ethylmaleimide (NEM). Cys452PKC is an active site residue (22) that is conserved in only 11 of the >400 genes that encode human protein kinases. Further narrowing the q Note: DNA sequence analysis and peptide synthesis were done at M.D. Anderson field, PKCy conserves Cys452PKC but is resistant to inactivation DNA Analysis and Peptide Synthesis Facilities, which are supported in part by NIH by cysteine modification (17–20). Therefore, the design of small- core grant CA16672. Requests for reprints: Catherine A. O’Brian, Department of Cancer Biology, M.D. molecule inhibitors that bind in the active site cavity in an Anderson Cancer Center, 1515 Holcombe Boulevard, Box 173, Houston, TX 77030. orientation favoring reaction with the side chain of Cys452 may Phone: 713-792-7969; E-mail: [email protected]. I2005 American Association for Cancer Research. offer a new avenue for the development of antineoplastic drugs doi:10.1158/0008-5472.CAN-05-1989 that selectively inhibit PKCq. Cancer Res 2005; 65: (22). November 15, 2005 10478 www.aacrjournals.org Downloaded from cancerres.aacrjournals.org on September 23, 2021. © 2005 American Association for Cancer Research. A PKCe-Inactivating Switch Materials and Methods Western blot analysis of FLAG-immunoprecipitated PKCq samples was done using a PKCq mAb from BD Biosciences (San Jose, CA) to detect Cell culture and reagents. COS-7 cells obtained from the American the wt/mut hPKCq species and a PKCq mAb (sc-214) from Santa Cruz Type Culture Collection (Manassas, VA) were maintained in DMEM Biotechnology (Santa Cruz, CA) to detect the mPKCq-CAT species. supplemented with 10% fetal bovine serum and penicillin-streptomycin Enhanced chemiluminescence (Amersham Biosciences, Piscataway, NJ) in humidified air with 5% CO at 37jC. The media, antibiotics, and serum 2 was employed as the detection system. were from Invitrogen (Carlsbad, CA). Protein kinase CE inactivation by N-ethylmaleimide. To measure Plasmids and mutagenesis. Murine PKCq (mPKCq) catalytic domain Cys452-dependent PKCq inactivation in NEM-treated cell lysates, wt hPKCq cDNA encoding a constitutively active PKCq mutant (11) and hPKCq cDNA and C452A hPKCq transfectants were prepared and lysed as described were provided by Dr. I. Bernard Weinstein (Columbia University, New York) in Cellular PKCq Inactivation by Disulfides, except that the cells were and Dr. A.C. Newton (University of California-San Diego), respectively. harvested with Buffer B [0.65 mL/plate; 50 mmol/L HEPES (pH 7.0), Sequence analysis confirmed that the cDNAs encoded the catalytic domain 1 mmol/L EDTA, 1 mmol/L EGTA, 10 Ag/mL leupeptin, 250 Amol/L PMSF, of mPKCq (residues 395-737; Genbank accession no. P16054) and hPKCq 1 mmol/L Na VO , 20 mmol/L NaF, 10 nmol/L microcystin]. The cell (residues 2-737; Genbank accession no. Q02156). 3 4 lysates were briefly centrifuged, assayed for protein content, and nor- To generate cDNAs encoding site-specific mutants of mPKCq and hPKCq malized to 1 mg/mL protein. Next, the lysates were treated with NEM with alanine substituted for cysteine, we used the PCR-based, site-directed for 10 minutes at 30jC; the alkylation reaction was terminated by adding mutagenesis kit QuikChange (Stratagene, La Jolla, CA). pcDNA3-PKCq 2 mmol/L DTT (final concentration) to scavenge unreacted NEM. NEM- expression plasmids served as template DNA and contained the coding induced hPKCq inactivation was measured by immunoprecipitating hPKCq sequence for hPKCq with an NH2-terminal FLAG-epitope tag or the coding sequence for the mPKCq catalytic domain with a COOH-terminal FLAG (wt/mut) with FLAG M2 mAb and analyzing the immunoprecipitated q tag (mPKCq-CAT). Oligonucleotides were commercially synthesized and isozyme as described in Cellular PKC Inactivation by Disulfides, except j purified (Sigma-Genosys, The Woodlands, TX). Mutagenic primers were that the 15-minute incubation at 30 C was omitted. 452 q employed in conjunction with complementary primers to generate To measure Cys -dependent PKC inactivation in NEM-treated cells, q site-specific PKCq mutants. The following are mutagenic primers for wt and C452A hPKC transfectants
Load more

Recommended publications
  • Cysteine, Glutathione, and Thiol Redox Balance in Astrocytes
    antioxidants Review Cysteine, Glutathione, and Thiol Redox Balance in Astrocytes Gethin J. McBean School of Biomolecular and Biomedical Science, Conway Institute, University College Dublin, Dublin, Ireland; [email protected]; Tel.: +353-1-716-6770 Received: 13 July 2017; Accepted: 1 August 2017; Published: 3 August 2017 Abstract: This review discusses the current understanding of cysteine and glutathione redox balance in astrocytes. Particular emphasis is placed on the impact of oxidative stress and astrocyte activation on pathways that provide cysteine as a precursor for glutathione. The effect of the disruption of thiol-containing amino acid metabolism on the antioxidant capacity of astrocytes is also discussed. − Keywords: cysteine; cystine; cysteamine; cystathionine; glutathione; xc cystine-glutamate exchanger; transsulfuration 1. Introduction Thiol groups, whether contained within small molecules, peptides, or proteins, are highly reactive and prone to spontaneous oxidation. Free cysteine readily oxidises to its corresponding disulfide, cystine, that together form the cysteine/cystine redox couple. Similarly, the tripeptide glutathione (γ-glutamyl-cysteinyl-glycine) exists in both reduced (GSH) and oxidised (glutathione disulfide; GSSG) forms, depending on the oxidation state of the sulfur atom on the cysteine residue. In the case of proteins, the free sulfhydryl group on cysteines can adopt a number of oxidation states, ranging from disulfides (–S–S–) and sulfenic acids (–SOOH), which are reversible, to the more oxidised sulfinic (–SOO2H) and sulfonic acids (–SOO3H), which are not. These latter species may arise as a result of chronic and/or severe oxidative stress, and generally indicate a loss of function of irreversibly oxidised proteins. Methionine residues oxidise to the corresponding sulfoxide, which can be rescued enzymatically by methionine sulfoxide reductase [1].
    [Show full text]
  • Pantethine and Cystamine Deplete Cystine from Cystinotic Fibroblasts Via Efflux of Cysteamine-Cysteine Mixed Disulfide
    Pantethine and cystamine deplete cystine from cystinotic fibroblasts via efflux of cysteamine-cysteine mixed disulfide. J D Butler, M Zatz J Clin Invest. 1984;74(2):411-416. https://doi.org/10.1172/JCI111436. Research Article Children suffering from cystinosis, a genetic disease characterized by high levels of lysosomal cystine, are currently being treated with cysteamine to lower the cystine levels in their cells. In fibroblasts from these patients, cysteamine and its disulfide, cystamine, are equally effective in lowering cystine levels. We recently reported that pantethine, a dietary precursor of coenzyme A, depletes cystine from cultured, cystinotic fibroblasts as effectively as cystamine. To determine the mechanism of action of pantethine, and of cystamine, we have compared the fate of [35S]cystine-derived metabolites in the presence and absence of these agents. The results indicate that the ability of pantethine to deplete cystine resides in its being a metabolic precursor of cysteamine. Furthermore, both pantethine and cystamine act by generating the mixed disulfide of cysteamine and cysteine in the lysosomes, which is then rapidly excreted from the cells. The fall in intracellular [35S]cystine caused by these agents was not accompanied by a comparable increase in any intracellular metabolite; rather, it could be accounted for by the appearance of mixed disulfide in the medium. There was no accumulation of mixed disulfide in the cells. Radioactivity in cytoplasmic glutathione was, however, increased by cystamine or pantethine. Thus, cysteamine (formed intracellularly in these experiments) undergoes thiol-disulfide exchange with cystine in the lysosomes, producing cysteamine-cysteine mixed disulfide and free cysteine, which enter the […] Find the latest version: https://jci.me/111436/pdf Pantethine and Cystamine Deplete Cystine from Cystinotic Fibroblasts via Efflux of Cysteamine-Cysteine Mixed Disulfide Jean DeB.
    [Show full text]
  • Inhibition of Melanin Synthesis by Cystamine in Human Melanoma Cells
    CORE Metadata, citation and similar papers at core.ac.uk Provided by Elsevier - Publisher Connector Inhibition of Melanin Synthesis by Cystamine in Human Melanoma Cells Ling Qiu, Mei Zhang, Rick A. Sturm,* Brooke Gardiner,* Ian Tonks, Graham Kay, and Peter G. Parsons Queensland Cancer Fund Laboratories, Queensland Institute of Medical Research and University of Queensland Joint Experimental Oncology Program, Herston, Queensland, Australia; *Center for Molecular and Cell Biology, University of Queensland, Australia In studies to determine whether pigmentation can be of MM96L and HeLa cells. Cystamine treatment regulated physiologically by thiols, human melanoma lowered the degree of cross-linking of the pigment- cells (MM418c5)and melanocytes were found to ation antigen gp75/TRP-1 in MM418c5 cells. become depigmented when cultured continuously in Tyrosinase protein and mRNA levels in MM418c5 50 mM cystamine. Cystamine was depleted from the cells were not affected by cystamine. The results culture medium and the treatment was nontoxic and show that cystamine at a concentration close to reversible. Cysteamine, dithiothreitol, and phenyl- physiologic levels has multiple effects on the melano- thiourea were less effective, and glutathione, genic pathway. In amelanotic cells, tyrosinase has a cysteine, and cystine were inactive. Tyrosinase (dopa short half-life and is readily inhibited by cystamine/ oxidase)activity was not greatly affected except for cysteamine whereas tyrosinase in the more mature induction of a lag period. In contrast, tyrosinase melanosomes of the pigmented cell appears to be activity in an amelanotic melanoma cell line less accessible to proteolytic and thiol attack. (MM96L)was rapidly inhibited without consumption Inhibition of melanin synthesis in the latter cell type of cystamine/cysteamine, in association with the may arise to a signi®cant degree from reduction of generation of free thiol in the culture medium, and cystamine to cysteamine, which sequesters quinones.
    [Show full text]
  • Subject Index to Volume 73 JANUARY-DE(MEMBER 1976
    Subject Index to Volume 73 JANUARY-DE(MEMBER 1976 Introduction The terms in the Subject Index for Vol ume 73, January-December 1976, of the PROCEEDINGSOF THE NATIONALACA DEMY OF SCIENCESUSA were chosen mainly from titles and key terms of artic] es. The index terms are alphabetized by computer; numbers, conformational pref ixes, Greek letters, hyphens, and spaces between words are disregarded in alphabe tization. After each index term is printed the title of the article (or a suitable modi fication of the title) and the appropriate page number. Titles are listed in alphabe tical order under the index terms. Classifications (e.g., Physics, Mather latics) under which papers have been published are used as index terms only if it seemed this would be helpful. Papers that are concerned in some major way with methodology are indexed under "Methodology" as well as under more sj ,ecific terms. Papers relevant to human diseases are indexed under "Diseases of I iuman beings" (with some subclassifica- tions) as well as under more specific tern is. Corrections to papers in which errors occurred are indexed under the term "Coi rection" as well as under the index terms selected for the paper itself. Organisms are indexed by their scientific names when scientific names were provided in the pape rs; suitable cross-references are provided. Genes are listed together as, for example ,"Gene, lac." Because the PROCEEDINGSurges auth ors to follow the tentative rules and rec- ommendations of the nomenclature cor amissions (e.g., for biochemistry, those proposed by the International Union ol 'Pure and Applied Chemistry and the Commission on Biochemical Nomenclatu re), an effort has been made to construct an index that conforms with this policy.
    [Show full text]
  • Cellular Thiol Pools Are Responsible for Sequestration of Cytotoxic Reactive Aldehydes: Central Role of Free Cysteine and Cysteamine
    BRAIN RESEARCH 1158 (2007) 158– 163 available at www.sciencedirect.com www.elsevier.com/locate/brainres Research Report Cellular thiol pools are responsible for sequestration of cytotoxic reactive aldehydes: Central role of free cysteine and cysteamine Paul L. Wood⁎, M. Amin Khan, Joseph R. Moskal The Falk Center for Molecular Therapeutics, Department of Biomedical Engineering, McCormick School of Engineering and Applied Sciences, Northwestern University, 1801 Maple Ave., Suite 4306, Evanston, IL 60201, USA ARTICLE INFO ABSTRACT Article history: Cellular thiol pools have been shown to be important in the regulation of the redox status Accepted 4 May 2007 of cells, providing a large antioxidant pool consisting of free thiols, thiols bound in the Available online 10 May 2007 disulfide form and thiols bound to proteins. However, experimental studies with the thiol cysteamine and its disulfide cystamine have demonstrated dramatic cytoprotection in Keywords: experimental models where antioxidants provide only minor protection. These data Cystamine suggest that an alternate action of thiols is important in their cytoprotective actions. A Cysteamine common feature of the in vitro and in vivo models, where these thiol agents demonstrate Cysteine cytoprotection, is the generation of cytotoxic aldehydes. We therefore studied the actions Pantethine of cystamine, cysteamine and several reference thiol agents as cytoprotectants against Thiol cell death induced by increased “aldehyde load”. We found that all the thiol agents Aldehyde sequestration examined provided dramatic protection against aldehyde-induced cell death in SN56 Acrolein cholinergic neurons, under conditions in which acrolein induced 100% cell death. 3-Aminopropanal With regard to mechanism of action, the reference thiols cysteine, N-acetylcysteine, Neuroprotection 2-mercaptoethanesulfonic acid, mercapto-propionyglycine, and cysteamine can directly SN56 cholinergic neuron sequester aldehydes.
    [Show full text]
  • TB 2-F-Di.Indd
    22825 DAVIS DRIVE USE OF 2-F-dI TO PRODUCE N2-MODIFIED dG DERIVATIVES STERLING, VIRGINIA 20164 Introduction FIGURE 1: CONVERSION OF 2-F-dI TO 2-AMINO DERIVATIVES To our list of Convertible Nucleosides1, we are adding 2- PHONE NO2 F-dI-CE Phosphoramidite (1). 2-Fluoro-2’-deoxyInosine 703-437-6191 (Figure 1) can be converted to 2-substituted dG derivatives by reaction with a primary amine, which displaces the O 800-327-GLEN 2-5 fluorine atom. The timing of the conversion step is N N a little tricky because small alkyl primary amines are FAX capable of doing the conversion while also cleaving and F N N 703-435-9774 deprotecting the oligonucleotide. For example, reaction DMTO with ethylamine would convert 2-F-dI to N2-ethyl-dG O INTERNET but would simultaneously cleave and deprotect the oligonucleotide. Although that may be interesting in its OP N(iPr) WWW.GLENRES.COM own right, we have chosen to focus on larger primary 2 amines in our development work. For example, treatment O CNEt of the oligonucleotide (while still fully protected on the (1) 2-F-dI-CE Phosphoramidite synthesis column) with dansyl cadaverine converts the nucleoside to an N2-dansyl-dG derivative, as shown in Figure 2. Following removal of the O6-protecting 2-F-dI Conversion group with DBU, further conventional deprotection of the oligonucleotide leads to the final product. The 2-F-dI is a convertible nucleoside that allows synthesis product oligonucleotide now has a fluorescent tag which, of oligos containing an N2 substituted deoxy-guanosine.
    [Show full text]
  • Www .Alfa.Com
    Bio 2013-14 Alfa Aesar North America Alfa Aesar Korea Uni-Onward (International Sales Headquarters) 101-3701, Lotte Castle President 3F-2 93 Wenhau 1st Rd, Sec 1, 26 Parkridge Road O-Dong Linkou Shiang 244, Taipei County Ward Hill, MA 01835 USA 467, Gongduk-Dong, Mapo-Gu Taiwan Tel: 1-800-343-0660 or 1-978-521-6300 Seoul, 121-805, Korea Tel: 886-2-2600-0611 Fax: 1-978-521-6350 Tel: +82-2-3140-6000 Fax: 886-2-2600-0654 Email: [email protected] Fax: +82-2-3140-6002 Email: [email protected] Email: [email protected] Alfa Aesar United Kingdom Echo Chemical Co. Ltd Shore Road Alfa Aesar India 16, Gongyeh Rd, Lu-Chu Li Port of Heysham Industrial Park (Johnson Matthey Chemicals India Toufen, 351, Miaoli Heysham LA3 2XY Pvt. Ltd.) Taiwan England Kandlakoya Village Tel: 866-37-629988 Bio Chemicals for Life Tel: 0800-801812 or +44 (0)1524 850506 Medchal Mandal Email: [email protected] www.alfa.com Fax: +44 (0)1524 850608 R R District Email: [email protected] Hyderabad - 501401 Andhra Pradesh, India Including: Alfa Aesar Germany Tel: +91 40 6730 1234 Postbox 11 07 65 Fax: +91 40 6730 1230 Amino Acids and Derivatives 76057 Karlsruhe Email: [email protected] Buffers Germany Tel: 800 4566 4566 or Distributed By: Click Chemistry Reagents +49 (0)721 84007 280 Electrophoresis Reagents Fax: +49 (0)721 84007 300 Hydrus Chemical Inc. Email: [email protected] Uchikanda 3-Chome, Chiyoda-Ku Signal Transduction Reagents Tokyo 101-0047 Western Blot and ELISA Reagents Alfa Aesar France Japan 2 allée d’Oslo Tel: 03(3258)5031 ...and much more 67300 Schiltigheim Fax: 03(3258)6535 France Email: [email protected] Tel: 0800 03 51 47 or +33 (0)3 8862 2690 Fax: 0800 10 20 67 or OOO “REAKOR” +33 (0)3 8862 6864 Nagorny Proezd, 7 Email: [email protected] 117 105 Moscow Russia Alfa Aesar China Tel: +7 495 640 3427 Room 1509 Fax: +7 495 640 3427 ext 6 CBD International Building Email: [email protected] No.
    [Show full text]
  • Therapeutic Effects of Cystamine in a Murine Model of Huntington's Disease
    The Journal of Neuroscience, October 15, 2002, 22(20):8942–8950 Therapeutic Effects of Cystamine in a Murine Model of Huntington’s Disease Alpaslan Dedeoglu,1,2 James K. Kubilus,1,2 Thomas M. Jeitner,2,4 Samantha A. Matson,2 Misha Bogdanov,3,5 Neil W. Kowall,1,2 Wayne R. Matson,3 Arthur J. L. Cooper,4,5,6 Rajiv R. Ratan,7 M. Flint Beal,5* Steven M. Hersch,8* and Robert J. Ferrante1,2 1Geriatric Research Education and Clinical Center, Bedford Veterans Affairs Medical Center, Bedford, Massachusetts 01730, 2Neurology, Pathology, and Psychiatry Departments, Boston University School of Medicine, Boston, Massachusetts 02118, 3ESA Laboratories, Inc., Chelmsford, Massachusetts 01824, Departments of 4Biochemistry and 5Neurology and Neuroscience, Weill Medical College of Cornell University, Presbyterian Hospital, New York, New York 10021, 6Burke Medical Research Institute, White Plains, New York 10605, 7Department of Neurology and Program in Neuroscience, Harvard Medical School and The Beth Israel-Deaconess Medical Center, Boston, Massachusetts 02115, and 8Center for Aging, Genetics, and Neurodegeneration, Neurology Service, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02129 The precise cause of neuronal death in Huntington’s disease marker of Tgase activity, was markedly elevated in the neocor- (HD) is unknown. Proteolytic products of the huntingtin protein tex and caudate nucleus in HD patients. Both Tgase and GGEL can contribute to toxic cellular aggregates that may be formed immunoreactivities colocalized to huntingtin aggregates. Cys- in part by tissue transglutaminase (Tgase). Tgase activity is tamine treatment normalized transglutaminase and GGEL lev- increased in HD brain. Treatment in R6/2 transgenic HD mice, els in R6/2 mice.
    [Show full text]
  • Challenges for Cysteamine Stabilization, Quantification, and Biological Effects Improvement Carla Atallah, Catherine Charcosset, Hélène Greige-Gerges
    Challenges for cysteamine stabilization, quantification, and biological effects improvement Carla Atallah, Catherine Charcosset, Hélène Greige-Gerges To cite this version: Carla Atallah, Catherine Charcosset, Hélène Greige-Gerges. Challenges for cysteamine stabilization, quantification, and biological effects improvement. Journal of Pharmaceutical Analysis, Elsevier B.V. on behalf of Xi’an Jiaotong University, In press, 10.1016/j.jpha.2020.03.007. hal-03028849 HAL Id: hal-03028849 https://hal.archives-ouvertes.fr/hal-03028849 Submitted on 27 Nov 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Journal of Pharmaceutical Analysis xxx (xxxx) xxx Contents lists available at ScienceDirect Journal of Pharmaceutical Analysis journal homepage: www.elsevier.com/locate/jpa Review paper Challenges for cysteamine stabilization, quantification, and biological effects improvement * Carla Atallah a, b, Catherine Charcosset b,Hel ene Greige-Gerges a, a Bioactive Molecules Research Laboratory, Doctoral School of Sciences and Technologies, Faculty of Sciences, Lebanese University, Lebanon b Laboratory of Automatic Control, Chemical and Pharmaceutical Engineering, University Claude Bernard Lyon 1, France article info abstract Article history: The aminothiol cysteamine, derived from coenzyme A degradation in mammalian cells, presents several Received 22 October 2019 biological applications.
    [Show full text]
  • Redox-Sensitive Linear and Cross-Linked Cystamine-Based Polymers for Colon-Targeted Drug Delivery: Design, Synthesis, and Characterisation
    pharmaceutics Article Redox-Sensitive Linear and Cross-Linked Cystamine-Based Polymers for Colon-Targeted Drug Delivery: Design, Synthesis, and Characterisation Yoke Mooi Ng 1, Siti Nur Aishah Mat Yusuf 1,2, Hock Ing Chiu 1 and Vuanghao Lim 1,* 1 Integrative Medicine Cluster, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Kepala Batas 13200, Penang, Malaysia; [email protected] (Y.M.N.); [email protected] (S.N.A.M.Y.); [email protected] (H.I.C.) 2 Department of Chemical Engineering Technology, Faculty of Engineering Technology, UniCITI Alam Campus, Universiti Malaysia Perlis, Padang Besar 02100, Perlis, Malaysia * Correspondence: [email protected]; Tel.: +604-5622427 Received: 14 February 2020; Accepted: 23 April 2020; Published: 18 May 2020 Abstract: Cystamine-based polymers may help to achieve controlled and targeted drug delivery to the colon due to their susceptibility to breakage of the disulfide linkage in the low redox potential environment of the colon. In this study, two linear cystamine-based polymers with similar repeating units (LP1 and LP2) and a cross-linked cystamine-based polymer (BP) were synthesised and their kinetics and the various physical conditions underlying cystamine-based polymerisation were evaluated. In brief, N1,N6-bis(2-(tritylthio)ethyl)adipamide (2) was synthesised from the reaction of triphenylmethanol and cysteamine. Next, the trityl group of 2 was removed with trifluoroacetic acid and triethylsilane before proceeding to oxidative polymerisation of the end product, N1,N6-bis(2-mercaptoethyl)adipamide (3) to LP1. The Schotten-Bauman reaction was applied to synthesise LP2 and BP from the reaction of cystamine with adipoyl chloride or trimesoyl chloride.
    [Show full text]
  • Enterically Coated Cysteamine, Cystamine And
    (19) TZZ__¥T (11) EP 1 919 458 B3 (12) NEW EUROPEAN PATENT SPECIFICATION After limitation procedure (B3-1) (45) Mention of the grant of the patent: (51) Int Cl.: 11.07.2012 Bulletin 2012/28 A61K 31/145 (2006.01) A61K 9/32 (2006.01) A61P 43/00 (2006.01) (45) Date of publication and mention of the limitation decision: (86) International application number: B3-1 11.01.2017 Bulletin 2017/02 PCT/US2007/002325 (21) Application number: 07762690.1 (87) International publication number: WO 2007/089670 (09.08.2007 Gazette 2007/32) (22) Date of filing: 26.01.2007 (54) ENTERICALLY COATED CYSTEAMINE, CYSTAMINE AND DERIVATIVES THEREOF MAGENSAFTRESISTENT BESCHICHTETES CYSTAMIN, CYSTAMIN UND DERIVATE DAVON CYSTEAMINE A ENROBAGE ENTERIQUE, CYSTAMINE ET LEURS DERIVES (84) Designated Contracting States: (74) Representative: Hill, Christopher Michael et al AT BE BG CH CY CZ DE DK EE ES FI FR GB GR Page White & Farrer HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI Bedford House SK TR John Street London, WC1N 2BF (GB) (30) Priority: 27.01.2006 US 762715 P (56) References cited: (43) Date of publication of application: WO-A1-03/070020 WO-A1-2005/063226 14.05.2008 Bulletin 2008/20 US-A- 5 668 117 US-A1- 2005 245 433 US-B1- 6 331 316 (60) Divisional application: 12169866.6 / 2 535 044 • BUTLER DEB J ET AL: "PANTETHINE AND CYSTAMINE DEPLETE CYSTINE FROM (73) Proprietor: The Regents of the University of CYSTINOTIC FIBROBLASTS VIA EFFLUX OF California CYSTEAMINE-CYSTEINE MIXED DISULFIDE" Oakland, CA 94607 (US) JOURNAL OF CLINICAL INVESTIGATION, AMERICAN SOCIETY FOR CLINICAL (72) Inventors: INVESTIGATION, US LNKD- • DOHIL, Ranjan DOI:10.1172/JCI111436, vol.
    [Show full text]
  • Toxicological Profile for Carbon Tetrachloride
    TOXICOLOGICAL PROFILE FOR CARBON TETRACHLORIDE U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service Agency for Toxic Substances and Disease Registry August 2005 CARBON TETRACHLORIDE ii DISCLAIMER The use of company or product name(s) is for identification only and does not imply endorsement by the Agency for Toxic Substances and Disease Registry. CARBON TETRACHLORIDE iii UPDATE STATEMENT A Toxicological Profile for Carbon Tetrachloride, Draft for Public Comment was released in September 2003. This edition supersedes any previously released draft or final profile. Toxicological profiles are revised and republished as necessary. For information regarding the update status of previously released profiles, contact ATSDR at: Agency for Toxic Substances and Disease Registry Division of Toxicology/Toxicology Information Branch 1600 Clifton Road NE Mailstop F-32 Atlanta, Georgia 30333 CARBON TETRACHLORIDE vii QUICK REFERENCE FOR HEALTH CARE PROVIDERS Toxicological Profiles are a unique compilation of toxicological information on a given hazardous substance. Each profile reflects a comprehensive and extensive evaluation, summary, and interpretation of available toxicologic and epidemiologic information on a substance. Health care providers treating patients potentially exposed to hazardous substances will find the following information helpful for fast answers to often-asked questions. Primary Chapters/Sections of Interest Chapter 1: Public Health Statement: The Public Health Statement can be a useful tool for educating patients about possible exposure to a hazardous substance. It explains a substance’s relevant toxicologic properties in a nontechnical, question-and-answer format, and it includes a review of the general health effects observed following exposure. Chapter 2: Relevance to Public Health: The Relevance to Public Health Section evaluates, interprets, and assesses the significance of toxicity data to human health.
    [Show full text]