DOCSLIB.ORG

(12) Patent Application Publication (10) Pub. No.: US 2007/0191272 A1 Stemmer Et Al

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

US 200701.91272A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2007/0191272 A1 Stemmer et al. (43) Pub. Date: Aug. 16, 2007 (54) PROTEINACEOUS PHARMACEUTICALS Publication Classification AND USES THEREOF (76) Inventors: Willem P.C. Stemmer, Los Gatos, CA (51) Int. Cl. (US); Volker Schellenberger, Palo A6II 38/16 (2006.01) Alto, CA (US); Martin Bader, C40B 40/08 (2006.01) Mountain View, CA (US); Michael C40B 40/10 (2006.01) Scholle, Mountain View, CA (US) C07K I4/47 (2006.01) (52) U.S. Cl. ................. 514/12: 435/7.1: 435/6; 530/324 Correspondence Address: WILSON SONSN GOODRCH & ROSAT 650 PAGE MILL ROAD (57) ABSTRACT PALO ALTO, CA 94304-1050 (US) (21) Appl. No.: 11/528,927 The present invention provides cysteine-containing scaf folds and/or proteins, expression vectors, host cell and (22) Filed: Sep. 27, 2006 display systems harboring and/or expressing such cysteine containing products. The present invention also provides Related U.S. Application Data methods of designing libraries of Such products, methods of (60) Provisional application No. 60/721,270, filed on Sep. screening Such libraries to yield entities exhibiting binding 27, 2005. Provisional application No. 60/721,188, specificities towards a target molecule. Further provided by filed on Sep. 27, 2005. Provisional application No. the invention are pharmaceutical compositions comprising 60/743,622, filed on Mar. 21, 2006. the cysteine-containing products of the present invention. Patent Application Publication Aug. 16, 2007 Sheet 1 of 46 US 2007/0191272 A1 Takara togra: Patent Application Publication Aug. 16, 2007 Sheet 2 of 46 US 2007/0191272 A1 FIG. 7 was 8Aas Furse actics in proteirss is awarage proteir Awsages lossei Patent Application Publication Aug. 16, 2007 Sheet 3 of 46 US 2007/0191272 A1 F.G. 14 P P LECHNOQSSQPPTTKTCPGETNCYKKVWRDHRGTIERGCGCPTVKPG IKLNCCTTDKCN 3 15 25 T5 36 35 45 45 55 55 6 N FIG. 18 LicYNCPNPTADCKTAVNCSSDFDACLITKAGLovnkcwKFEHCNFNDvTTRLRENELT. = . 1. 15 2b. 25 3. 4. s B top YYCCKKDLCN FNEQLEN 6 I 7 Patent Application Publication Aug. 16, 2007 Sheet 5 of 46 US 2007/0191272 A1 FIG. 24 L.was essIS FIG. 26 aessA Ex e--- A. sists is S. assistasiaataasaia8 was w ws seas a Patent Application Publication Aug. 16, 2007 Sheet 6 of 46 US 2007/0191272 A1 FG. 30 f |AH T.HH Patent Application Publication Aug. 16, 2007 Sheet 7 of 46 US 2007/0191272 A1 FIG. 36 Cysteine i. 2. & S 6 Bgk vgRDWFKETAgrRAKSL TSQKYRAN-SA Shk RSIDTIFKSRS----T SMKYRISFg : stecrisp 182frRENKFTSNTMVQQSSEQD-NYMKTN-rais-FioNKii & s 22 PST PKRNNDFSKSLAKKSKQT-EWIKKK-g SEN I pseudecin PNYNNDFSS SLAKKSKQT-EWIKKK-stra -FSHN HTX PixONDVYNgPDLKKQVGGGH-PIMK-D-S-STS-KSLTEK Eara sess & is a 's so is is a a 4 a 4: a is 8 s Patent Application Publication Aug. 16, 2007 Sheet 8 of 46 US 2007/0191272 A1 FIG. 41 n AS YKs CRyaw- seekyll'sww. T Eastess-------------------------------------------------------------r SS5's SSA ESS as 1-5 2-436 till s Yellvil tWe a WBALLIEUlstAAAAA Patent Application Publication Aug. 16, 2007 Sheet 9 of 46 US 2007/0191272 A1 rakhtawirraat A FAA 'reaweerses rwpackaw Patent Application Publication Aug. 16, 2007 Sheet 10 of 46 US 2007/0191272 A1 's JLC : SufusAAF Aki| Patent Application Publication Aug. 16, 2007 Sheet 11 of 46 US 2007/0191272 A1 Patent Application Publication Aug. 16, 2007 Sheet 12 of 46 US 2007/0191272 A1 FIG. 67 s d AR al Patent Application Publication Aug. 16, 2007 Sheet 13 of 46 US 2007/0191272 A1 ALAEGQS CGVYTERCAQGLRCL PRODEEKPLHALLHGRGVCLNEKS 13 2O 25 3O 3S 4O 45 SO S5. Active site:- V BS Patent Application Publication Aug. 16, 2007 Sheet 14 of 46 US 2007/0191272 A1 FIG. 81 FIG. 83 FIG. 84 AscNGvcs PFEMPP cors ACRCI Pvg Lv I GYCRNP sc 3. lb. 5 2b 25 35 FG. 85 Patent Application Publication Aug. 16, 2007 Sheet 15 of 46 US 2007/0191272 A1 aws. Yearsaw FG. 94 A w a - - - - - - - - - - - - - - - a a is a error Vs Arka ce-Yee is escalarate criss E away pms WEochy s y s ss s dis Patent Application Publication Aug. 16, 2007 Sheet 16 of 46 US 2007/0191272 A1 FIG. 95 saww.saw-sursres Eks. TrAccor cpcrks ppg.cgcroy Rekcts AckscLc Rs procRico or cyps 2 3. 4. s al s FIG. 99 - P LRoc LP copogkorcFGP s cccbecace vocale Aircea is NYL ps Pcces. Geep cosgesR 25. - - - - - 3. a. s s 37 75 s s Patent Application Publication Aug. 16, 2007 Sheet 20 of 46 US 2007/0191272 A1 36 kektCanast tunnthdlicts yistclvksg ggCdnrtCan aptntinda CeayftgnnC 421 itksgggovtnttcaaitle aacvknSSgstcfwdtaSSSckdktcvnapatnthdlcq 481 pfintctwins tsagcvektcensivlaicid kdtsSraciw kgkcykkocylassattha 54. FIG. 113 .2a. leaglessells Elsiegels FIG. 114 elesslyas ' aasassesses O ars s y ps s sease . s FIG. 116 MGRSNSLRLL FLFCLLLALS TAASRRLKROCGCSNFCNCO COPVFFAOS 50 LPACSCQQAPICQPQCPRAE INSDCSATCv RACIPSCSKSTGNTFACSTT 100 CESTCDKTCA SAAQQAMSHIQVSPPNPQPLVPIAAAPTVD DSCQNVCONV 150 CQGACVSQNSPPAVCQQTCR QSCQFGCATN EQLPTTSSTS TNAPTIKITL 200 NINDAYFDSN CAPKCTOSCHSOCISOGNPAASCSNSCNTE CSDKCSTRPV 250 QAovoaiaPQ QVQITIPQTC QSRCENRCLSTCTASQPSVCAPSCSTACQL 300 SCDSGAATAPQSTNPTPVE WRALCTPTCMPQCLPSCTATTTTTTTOTPY 350 Patent Application Publication Aug. 16, 2007 Sheet 21 of 46 US 2007/0191272 A1 FIG. 117 N-cap Library I Library II F.G. 118 F.G. 119 1H Second cross-over First cross-over Patent Application Publication Aug. 16, 2007 Sheet 22 of 46 US 2007/0191272 A1 FIG. 120 10 2O 3O 40 50 library VESCEQYTSCGECLGSGDPH-CGWCVLENRCTRRSDCQRAEEPNRWASSISQCVKL 1ss GPGCRHFLTCSMCLRAPRFMGCGWC-GGVCSRQHEC-DGGWWQDS-CPPP. 10 20 30 40 FIG. 121 FIG. 122 | || is a s ass series Patent Application Publication Aug. 16, 2007 Sheet 23 of 46 US 2007/0191272 A1 FIG. 124 Cysteine-Rich Repeat Proteins Natural repeat Protein Repeat domain Repeat domain Non-Repeat domain Repeat contain N-termis linker . N-tettina Cap module -optional, depending Repeat on bonding pattem often 1-2 fewer Cys Modeo unit (optional)E, since it does not link to previous module. Spiral AA backbone library 1 Library 2 library 3 Binding Binding Binding Site 1 Site 2 Site 3 Repeat Protein Affinity Maturation: Different Walking Processes Radotte Select, Select, AdditionModule "S" "S" Repeat Protein --- . Select, Select, Module "S" i Randgnize next Replacement Randotzed Selected Randomized Unselected Selected Randonized Unselected Modules Modules Modules Modules Modules Modules Modules FIG 125 Granulin Repeat Proteins Repeat unit: - - - 3C6C5CC8CC6CCSCC5C6C2 CCXCCX or N- N-1 V-1 CX5CCXC on 1-3 2-5 4-7 6-9 8-11 1 0-12 XCCXCC, where Highest disulfide density in database X = 5,6,7,8 Repeat unit F 4. Cys Nat Struct Biol. 1996 Sep;3(9):747-52. The hairpin stack fold, a novel protein architecture for a new family of protein growth factors granuins have shot antibaralled beta-sheets that may impose sourie degree of sequence conservation Hrabal FR, Chen Z, Jarnes S. Bennett HP, Nif. and prevent full randomization as it may be necessary to use amino acids that are capable of beta-aheet formation. Natural granuins have some length variation in the loops which may of ?tay not be irthportant for folding. One solution is to define a short corsensus replaat motifie (CCs) arwd another solution is to define a long repeat consensus motif that retains the naturallength variation (ie coscC8ccescC5cc5). Patent Application Publication Aug. 16, 2007 Sheet 24 of 46 US 2007/0191272 A1 FIG, 126. Re peat Proteins Approach 1 Rarizatio: -DegreeSelectingth dependent on (get it ofstructural positions variation) App roach 2 Sirrultaneous Stepwise New Synthetic lib alonization Cycle 1 diverse Sequences Cycle 3 Randomization requires carefit balancing of the length AA arc degree AAposition of randomization. The actual numbers depend on how sensitive the structure is to perturtation. synthetic mutagenesis is preferred because it is well-controlled. However, Cycle 4 its difficus to synthetically re-mutagenize an area that was synthetically mutagenized before, the reason is that one needs to re-mutagenize a whole pool of cores, each having a different sequence, which requires sequencing all of the dones and dividing the pool into families of related cores. Resynthesis of a family allows retention of a family-specific sequence motif, but any detailbeyond the nati is destroyed. Thus, approaches that re-mutagenizes sequences blindly, without requiring sequence information, are much preferred. Cwcle 5 NA shufflic. error-prone PCR and Tutator strains art such big approachers. y the best corportise is to synthetically mutageize each past of a sequerce only once, Multiple cycles of Litagenesis are generatyroxuired to accumulate sufficiant binding activity, but each cycle targets a different part of the protein aequence. it is possible to use recombination to permutate previously selected sequences. Other approaches: D However, binding proteirs derived fram random sequence tend to bind to different epitopes, belong to different sequence families and differ in their register, F.Cistricts avaney of approaches. one can randomize only one face of a protein instead of the whole surface. ots can randomize alterating blocks of 2,368,739 or 1 artino acids, eiths successively or simultaneously. Antifreeze Protein-derived Repeat Protein FIG. 127 Natural repeat proteins are imperfect repeats but comparison of the repeats can be used to define a consensus sequence that can be repeated as rany tungna sequence: times as desired. he diagrams below show how two different solutions can - - be derived from the same original sequence. Both likely work but one may have - - - advantages. g QCTGGADCTSCTGACTGCGNCPNAV.......... at Unit S - Patent Application Publication Aug. 16, 2007 Sheet 25 of 46 US 2007/0191272 A1 | FIG. 128 Simple Designs for Spiral Repeat Protein Scaffolds Disulfide Bond. Examples of +3: Types Öxcxcke - Antifreeze Protein -3 XCXCXCX - Granulin On Oen More different disulfidelinkages result in less efficient folding. The simplest linkages Are preferred, such as the +3 patten shown. -3-4-4 to Affinity Maturation of Repeat Proteins Randomization: -Sequence t length get structural variation) -degree dependent on is of positions -Examples below do not use recombination Approach 3 Approach 4 Approach 5 Approach 6 Cycle 1 E.
Recommended publications
  • Examples of Successful Protein Expression with SUMO Reference Protein Type Family Kda System (Pubmed ID)

    Examples of Successful Protein Expression with SUMO Reference Protein Type Family Kda System (Pubmed ID)

    Examples of Successful Protein Expression with SUMO Reference Protein Type Family kDa System (PubMed ID) 23 (FGF23), human Growth factor FGF superfamily ~26 E. coli 22249723 SARS coronavirus (SARS-CoV) membrane 3C-like (3CL) protease Viral membrane protein protein 33.8 E. coli 16211506 5′nucleotidase-related apyrase (5′Nuc) Saliva protein (apyrase) 5′nucleotidase-related proteins 65 E. coli 20351782 Acetyl-CoA carboxylase 1 (ACC1) Cytosolic enzyme Family of five biotin-dependent carboxylases ~7 E. coli 22123817 Acetyl-CoA carboxylase 2 (ACC2) BCCP domain Cytosolic enzyme Family of five biotin-dependent carboxylases ~7 E. coli 22123817 Actinohivin (AH) Lectin Anti-HIV lectin of CBM family 13 12.5 E. coli DTIC Allium sativum leaf agglutinin (ASAL) Sugar-binding protein Mannose-binding lectins 25 E. coli 20100526 Extracellular matrix Anosmin protein Marix protein 100 Mammalian 22898776 Antibacterial peptide CM4 (ABP-CM4) Antibacterial peptide Cecropin family of antimicrobial peptides 3.8 E. coli 19582446 peptide from centipede venoms of Scolopendra Antimicrobial peptide scolopin 1 (AMP-scolopin 1) small cationic peptide subspinipes mutilans 2.6 E. coli 24145284 Antitumor-analgesic Antitumor-analgesic peptide (AGAP) peptide Multifunction scorpion peptide 7 E. coli 20945481 Anti-VEGF165 single-chain variable fragment (scFv) Antibody Small antibody-engineered antibody 30 E. coli 18795288 APRIL TNF receptor ligand tumor necrosis factor (TNF) ligand 16 E. coli 24412409 APRIL (A proliferation-inducing ligand, also named TALL- Type II transmembrane 2, TRDL-1 and TNFSF-13a) protein Tumor necrosis factor (TNF) family 27.51 E. coli 22387304 Aprotinin/Basic pancreatic trypsin inhibitor (BPTI) Inhibitor Kunitz-type inhibitor 6.5 E.
  • The Unique Cysteine Knot Regulates the Pleotropic Hormone Leptin

    The Unique Cysteine Knot Regulates the Pleotropic Hormone Leptin

    The Unique Cysteine Knot Regulates the Pleotropic Hormone Leptin Ellinor Haglund1, Joanna I. Sułkowska1, Zhao He2, Gen-Sheng Feng2, Patricia A. Jennings1*, Jose´ N. Onuchic3* 1 Department of Chemistry and Biochemistry and Center for theoretical Biological Physics (CTBP), University of California San Diego, La Jolla, California, United States of America, 2 Department of Pathology; School of Medicine and Molecular Biology Section, Division of Biological Sciences, University of California San Diego, La Jolla, California, United States of America, 3 Center for Theoretical Biological physics and Department of Physics and Astronomy, Chemistry, and Biochemistry and Cell Biology, Rice University, Houston, Texas, United States of America Abstract Leptin plays a key role in regulating energy intake/expenditure, metabolism and hypertension. It folds into a four-helix bundle that binds to the extracellular receptor to initiate signaling. Our work on leptin revealed a hidden complexity in the formation of a previously un-described, cysteine-knotted topology in leptin. We hypothesized that this unique topology could offer new mechanisms in regulating the protein activity. A combination of in silico simulation and in vitro experiments was used to probe the role of the knotted topology introduced by the disulphide-bridge on leptin folding and function. Our results surprisingly show that the free energy landscape is conserved between knotted and unknotted protein, however the additional complexity added by the knot formation is structurally important. Native state analyses led to the discovery that the disulphide-bond plays an important role in receptor binding and thus mediate biological activity by local motions on distal receptor-binding sites, far removed from the disulphide-bridge.
  • Animal Venom Derived Toxins Are Novel Analgesics for Treatment Of

    Animal Venom Derived Toxins Are Novel Analgesics for Treatment Of

    Short Communication iMedPub Journals 2018 www.imedpub.com Journal of Molecular Sciences Vol.2 No.1:6 Animal Venom Derived Toxins are Novel Upadhyay RK* Analgesics for Treatment of Arthritis Department of Zoology, DDU Gorakhpur University, Gorakhpur, UP, India Abstract *Corresponding authors: Ravi Kant Upadhyay Present review article explains use of animal venom derived toxins as analgesics of the treatment of chronic pain and inflammation occurs in arthritis. It is a [email protected] progressive degenerative joint disease that put major impact on joint function and quality of life. Patients face prolonged inappropriate inflammatory responses and bone erosion. Longer persistent chronic pain is a complex and debilitating Department of Zoology, DDU Gorakhpur condition associated with a large personal, mental, physical and socioeconomic University, Gorakhpur, UttarPradesh, India. burden. However, for mitigation of inflammation and sever pain in joints synthetic analgesics are used to provide quick relief from pain but they impose many long Tel: 9838448495 term side effects. Venom toxins showed high affinity to voltage gated channels, and pain receptors. These are strong inhibitors of ion channels which enable them as potential therapeutic agents for the treatment of pain. Present article Citation: Upadhyay RK (2018) Animal Venom emphasizes development of a new class of analgesic agents in form of venom Derived Toxins are Novel Analgesics for derived toxins for the treatment of arthritis. Treatment of Arthritis. J Mol Sci. Vol.2 No.1:6 Keywords: Analgesics; Venom toxins; Ion channels; Channel inhibitors; Pain; Inflammation Received: February 04, 2018; Accepted: March 12, 2018; Published: March 19, 2018 Introduction such as the back, spine, and pelvis.
  • Anti-Inflammatory and Immune Regulatory Actions of Naja Naja

    Anti-Inflammatory and Immune Regulatory Actions of Naja Naja

    toxins Review Anti-Inflammatory and Immune Regulatory Actions of Naja naja atra Venom Shu-Zhi Wang 1,2 and Zheng-Hong Qin 3,* 1 Institute of Pharmacy and Pharmacology, University of South China, Hengyang 421001, China; [email protected] 2 Hunan Province Cooperative Innovation Center for Molecular Target New Drug Study, University of South China, Hengyang 421001, China 3 Department of Pharmacology and Laboratory of Aging and Nervous Diseases, Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, College of Pharmaceutical Science, Soochow University, Suzhou 215123, China * Correspondence: [email protected]; Tel./Fax: +86-512-65882071 Received: 23 December 2017; Accepted: 24 February 2018; Published: 28 February 2018 Abstract: Naja naja atra venom (NNAV) is composed of various proteins, peptides, and enzymes with different biological and pharmacological functions. A number of previous studies have reported that NNAV exerts potent analgesic effects on various animal models of pain. The clinical studies using whole venom or active components have confirmed that NNAV is an effective and safe medicine for treatment of chronic pain. Furthermore, recent studies have demonstrated that NNAV has anti-inflammatory and immune regulatory actions in vitro and in vivo. In this review article, we summarize recent studies of NNAV and its components on inflammation and immunity. The main new findings in NNAV research show that it may enhance innate and humoral immune responses while suppressing T lymphocytes-mediated cellular immunity, thus suggesting that NNAV and its active components may have therapeutic values in the treatment of inflammatory and autoimmune diseases.
  • A Functional Nav1.7-Navab Chimera with a Reconstituted High-Affinity Protx-II Binding Site S

    A Functional Nav1.7-Navab Chimera with a Reconstituted High-Affinity Protx-II Binding Site S

    Supplemental material to this article can be found at: http://molpharm.aspetjournals.org/content/suppl/2017/06/23/mol.117.108712.DC1 1521-0111/92/3/310–317$25.00 https://doi.org/10.1124/mol.117.108712 MOLECULAR PHARMACOLOGY Mol Pharmacol 92:310–317, September 2017 Copyright ª 2017 by The American Society for Pharmacology and Experimental Therapeutics A Functional NaV1.7-NaVAb Chimera with a Reconstituted High-Affinity ProTx-II Binding Site s Ramkumar Rajamani, Sophie Wu, Iyoncy Rodrigo, Mian Gao, Simon Low, Lisa Megson, David Wensel, Rick L. Pieschl, Debra J. Post-Munson, John Watson, David R. Langley, Michael K. Ahlijanian, Linda J. Bristow, and James Herrington Molecular Discovery Technologies, Wallingford, Connecticut, Princeton, New Jersey, and Waltham, Massachusetts (R.R., S.W., I.R., M.G., S.L., L.M., D.W., D.R.L.); Discovery Biology (R.L.P., D.J.P.-M., M.K.A., L.J.B., J.H.) and Lead Discovery and Optimization (J.W.), Bristol-Myers Squibb Company, Wallingford, Connecticut Downloaded from Received March 6, 2017; accepted June 14, 2017 ABSTRACT The NaV1.7 voltage-gated sodium channel is implicated in part of the voltage sensor domain 2 (VSD2) of NaV1.7. Importantly, human pain perception by genetics. Rare gain of function this chimera, DII S1–S4, forms functional sodium channels and is molpharm.aspetjournals.org mutations in NaV1.7 lead to spontaneous pain in humans whereas potently inhibited by the NaV1.7 VSD2 targeted peptide toxin loss of function mutations results in congenital insensitivity to pain. ProTx-II. Further, we show by [125I]ProTx-II binding and surface Hence, agents that specifically modulate the function of NaV1.7 plasmon resonance that the purified DII S1–S4 protein retains high have the potential to yield novel therapeutics to treat pain.
  • Chemical Synthesis and NMR Solution Structure of Conotoxin GXIA from Conus Geographus

    Chemical Synthesis and NMR Solution Structure of Conotoxin GXIA from Conus Geographus

    marine drugs Article Chemical Synthesis and NMR Solution Structure of Conotoxin GXIA from Conus geographus David A. Armstrong 1, Ai-Hua Jin 2, Nayara Braga Emidio 2 , Richard J. Lewis 2 , Paul F. Alewood 2 and K. Johan Rosengren 1,* 1 School of Biomedical Sciences, Faculty of Medicine, The University of Queensland, Brisbane, QLD 4072, Australia; [email protected] 2 Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD 4072, Australia; [email protected] (A.-H.J.); [email protected] (N.B.E.); [email protected] (R.J.L.); [email protected] (P.F.A.) * Correspondence: [email protected] Abstract: Conotoxins are disulfide-rich peptides found in the venom of cone snails. Due to their exquisite potency and high selectivity for a wide range of voltage and ligand gated ion channels they are attractive drug leads in neuropharmacology. Recently, cone snails were found to have the capability to rapidly switch between venom types with different proteome profiles in response to predatory or defensive stimuli. A novel conotoxin, GXIA (original name G117), belonging to the I3-subfamily was identified as the major component of the predatory venom of piscivorous Conus geographus. Using 2D solution NMR spectroscopy techniques, we resolved the 3D structure for GXIA, the first structure reported for the I3-subfamily and framework XI family. The 32 amino acid peptide is comprised of eight cysteine residues with the resultant disulfide connectivity forming an ICK+1 motif. With a triple stranded β-sheet, the GXIA backbone shows striking similarity to Citation: Armstrong, D.A.; Jin, A.-H.; several tarantula toxins targeting the voltage sensor of voltage gated potassium and sodium channels.
  • Rope Parasite” the Rope Parasite Parasites: Nearly Every Au�S�C Child I Ever Treated Proved to Carry a Significant Parasite Burden

    Rope Parasite” the Rope Parasite Parasites: Nearly Every AuSC Child I Ever Treated Proved to Carry a Significant Parasite Burden

    Au#sm: 2015 Dietrich Klinghardt MD, PhD Infec4ons and Infestaons Chronic Infecons, Infesta#ons and ASD Infec4ons affect us in 3 ways: 1. Immune reac,on against the microbes or their metabolic products Treatment: low dose immunotherapy (LDI, LDA, EPD) 2. Effects of their secreted endo- and exotoxins and metabolic waste Treatment: colon hydrotherapy, sauna, intes4nal binders (Enterosgel, MicroSilica, chlorella, zeolite), detoxificaon with herbs and medical drugs, ac4vaon of detox pathways by solving underlying blocKages (methylaon, etc.) 3. Compe,,on for our micronutrients Treatment: decrease microbial load, consider vitamin/mineral protocol Lyme, Toxins and Epigene#cs • In 2000 I examined 10 au4s4c children with no Known history of Lyme disease (age 3-10), with the IgeneX Western Blot test – aer successful treatment. 5 children were IgM posi4ve, 3 children IgG, 2 children were negave. That is 80% of the children had clinical Lyme disease, none the history of a 4cK bite! • Why is it taking so long for au4sm-literate prac44oners to embrace the fact, that many au4s4c children have contracted Lyme or several co-infec4ons in the womb from an oVen asymptomac mother? Why not become Lyme literate also? • Infec4ons can be treated without the use of an4bio4cs, using liposomal ozonated essen4al oils, herbs, ozone, Rife devices, PEMF, colloidal silver, regular s.c injecons of artesunate, the Klinghardt co-infec4on cocKtail and more. • Symptomac infec4ons and infestaons are almost always the result of a high body burden of glyphosate, mercury and aluminum - against the bacKdrop of epigene4c injuries (epimutaons) suffered in the womb or from our ancestors( trauma, vaccine adjuvants, worK place related lead, aluminum, herbicides etc., electromagne4c radiaon exposures etc.) • Most symptoms are caused by a confused upregulated immune system (molecular mimicry) Toxins from a toxic environment enter our system through damaged boundaries and membranes (gut barrier, blood brain barrier, damaged endothelium, etc.).
  • Pain Research Product Guide | Edition 2

    Pain Research Product Guide | Edition 2

    Pain Research Product Guide | Edition 2 Chili plant Capsicum annuum A source of Capsaicin Contents by Research Area: • Nociception • Ion Channels • G-Protein-Coupled Receptors • Intracellular Signaling Tocris Product Guide Series Pain Research Contents Page Nociception 3 Ion Channels 4 G-Protein-Coupled Receptors 12 Intracellular Signaling 18 List of Acronyms 21 Related Literature 22 Pain Research Products 23 Further Reading 34 Introduction Pain is a major public health problem with studies suggesting one fifth of the general population in both the USA and Europe are affected by long term pain. The International Association for the Study of Pain (IASP) defines pain as ‘an unpleasant sensory and emotional experience associated with actual or potential tissue damage, or described in terms of such damage’. Management of chronic pain in the clinic has seen only limited progress in recent decades. Treatment of pain has been reliant on, and is still dominated by two classical medications: opioids and non-steroidal anti-inflammatory drugs (NSAIDs). However, side effects such as dependence associated with opioids and gastric ulceration associated with NSAIDs demonstrates the need for new drug targets and novel compounds that will bring in a new era of pain therapeutics. Pain has been classified into three major types: nociceptive pain, inflammatory pain and neuropathic or pathological pain. Nociceptive pain involves the transduction of painful stimuli by peripheral sensory nerve fibers called nociceptors. Neuropathic pain results from damage or disease affecting the sensory system, and inflammatory pain represents the immunological response to injury through inflammatory mediators that contribute to pain. Our latest pain research guide focuses on nociception and the transduction of pain to the spinal cord, examining some of the main classical targets as well as emerging pain targets.
  • Amphiphilic B-Sheet Cobra Cardiotoxin Targets Mitochondria and Disrupts Its Network

    Amphiphilic B-Sheet Cobra Cardiotoxin Targets Mitochondria and Disrupts Its Network

    CORE Metadata, citation and similar papers at core.ac.uk Provided by Elsevier - Publisher Connector FEBS 29626 FEBS Letters 579 (2005) 3169–3174 Amphiphilic b-sheet cobra cardiotoxin targets mitochondria and disrupts its network Chia-Hui Wang, Wen-guey Wu* Department of Life Sciences, Institute of Bioinformatics and Structural Biology, National Tsinghua University, 101, Section 2, Kuang Fu Road, Hsinchu 30013, Taiwan Received 29 March 2005; revised 2 May 2005; accepted 2 May 2005 Available online 23 May 2005 Edited by Maurice Montal brane is likely to be involved in the CTX-induced perturbation Abstract Recent advance in understanding the role of toxin proteins in controlling cell death has revealed that pro-apoptotic of cytosolic calcium homeostasis and hypercontracture in rat viral proteins targeting mitochondria contain amphiphilic a-heli- ventricular myocytes [5,10]; however, evidence indicating ces with pore-forming properties. Herein, we describe that the that CTX may target intracellular organelle within the cell is pore-forming amphiphilic b-sheet cardiotoxins (or cytotoxins, lacking. CTXs) from Taiwan cobra (Naja atra) also target mitochondrial A large number of protein toxins from bacteria and virus membrane after internalization and act synergistically with have been extensively studied in order to understand their CTX-induced cytosolic calcium increase to disrupt mitochondria mechanism of action as well as of intracellular membrane network. It is suggested that CTX-induced fragmentation of trafficking [11–13]. For instance, cholera toxin and related mitochondria play a role in controlling CTX-induced necrosis AB5-subunit bacterial toxin have been shown to follow a of myocytes and cause severe tissue necrosis in the victims.
  • Product Update Price List Winter 2014 / Spring 2015 (£)

    Product Update Price List Winter 2014 / Spring 2015 (£)

    Product update Price list winter 2014 / Spring 2015 (£) Say to affordable and trusted life science tools! • Agonists & antagonists • Fluorescent tools • Dyes & stains • Activators & inhibitors • Peptides & proteins • Antibodies hellobio•com Contents G protein coupled receptors 3 Glutamate 3 Group I (mGlu1, mGlu5) receptors 3 Group II (mGlu2, mGlu3) receptors 3 Group I & II receptors 3 Group III (mGlu4, mGlu6, mGlu7, mGlu8) receptors 4 mGlu – non-selective 4 GABAB 4 Adrenoceptors 4 Other receptors 5 Ligand Gated ion channels 5 Ionotropic glutamate receptors 5 NMDA 5 AMPA 6 Kainate 7 Glutamate – non-selective 7 GABAA 7 Voltage-gated ion channels 8 Calcium Channels 8 Potassium Channels 9 Sodium Channels 10 TRP 11 Other Ion channels 12 Transporters 12 GABA 12 Glutamate 12 Other 12 Enzymes 13 Kinase 13 Phosphatase 14 Hydrolase 14 Synthase 14 Other 14 Signaling pathways & processes 15 Proteins 15 Dyes & stains 15 G protein coupled receptors Cat no. Product name Overview Purity Pack sizes and prices Glutamate: Group I (mGlu1, mGlu5) receptors Agonists & activators HB0048 (S)-3-Hydroxyphenylglycine mGlu1 agonist >99% 10mg £112 50mg £447 HB0193 CHPG Sodium salt Water soluble, selective mGlu5 agonist >99% 10mg £59 50mg £237 HB0026 (R,S)-3,5-DHPG Selective mGlu1 / mGlu5 agonist >99% 10mg £70 50mg £282 HB0045 (S)-3,5-DHPG Selective group I mGlu receptor agonist >98% 1mg £42 5mg £83 10mg £124 HB0589 S-Sulfo-L-cysteine sodium salt mGlu1α / mGlu5a agonist 10mg £95 50mg £381 Antagonists HB0049 (S)-4-Carboxyphenylglycine Competitive, selective group 1
  • (73) Assignee If Its of the Sawengy

    (73) Assignee If Its of the Sawengy

    US 2008O131500A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2008/0131500 A1 CHANG (43) Pub. Date: Jun. 5, 2008 (54) METHODS AND COMPOSITIONS FOR AOIP I/00 (2006.01) RAPID INACTIVATION OF PROTEINS A6II 47/02 (2006.01) A 6LX 39/395 (2006.01) (75) Inventor: ROWEN.J.-Y. CHANG, Houston, C07K I4/00 (2006.01) TX (US) AOIN 25/00 (2006.01) (52) U.S. Cl. ......... 424/451; 530/410; 530/408; 530/402; Correspondence Address: 530/409:514/769; 424/130.1; 424/45 FULBRIGHT & UAWORSK L.L.P. 600 CONGRESSAVE., SUITE 2400 (57) ABSTRACT AUSTIN, TX 78701 Disclosed are methods of inactivating a protein, Such as leavi disulfide bond of tein, that invol tacti (73) Assignee niversityif of Texasits Systemof the ideSAWENGy ion, wherein the pH of the composition is about 10.0 to about 14.0. Also disclosed are methods of treating or prevent (21) Appl. No.: 11/566,595 ing a disease in a subject, such as a toxin-related disease or a 1-1. prion-related disease, that involve contacting a subject with a (22) Filed: Dec. 4, 2006 pharmaceutically effective amount of a reducing agent, a O O denaturant, and a hydroxide ion, wherein the pH of the com Publication Classification position is about 9.0 to about 14.0. Also disclosed are com (51) Int. Cl. positions that include a reducing agent, a denaturant, and a A6 IK 9/48 (2006.01) hydroxide ion, wherein the pH of the composition is about 9.0 A6IP 25/28 (2006.01) to about 14.0.
  • Antitumoral Activity of Snake Venom Proteins: New Trends in Cancer Therapy

    Antitumoral Activity of Snake Venom Proteins: New Trends in Cancer Therapy

    Hindawi Publishing Corporation BioMed Research International Volume 2014, Article ID 203639, 19 pages http://dx.doi.org/10.1155/2014/203639 Review Article Antitumoral Activity of Snake Venom Proteins: New Trends in Cancer Therapy Leonardo A. Calderon,1 Juliana C. Sobrinho,1 Kayena D. Zaqueo,1 Andrea A. de Moura,1 Amy N. Grabner,1 Maurício V. Mazzi,2 Silvana Marcussi,3 Auro Nomizo,4 CarlaF.C.Fernandes,1 Juliana P. Zuliani,1 Bruna M. A. Carvalho,5 Saulo L. da Silva,5 Rodrigo G. Stábeli,1 and Andreimar M. Soares1 1 Centro de Estudos de Biomoleculas´ Aplicadas aSa` ude,´ CEBio, Fundac¸ao˜ Oswaldo Cruz, Fiocruz Rondoniaˆ e Departamento de Medicina, Universidade Federal de Rondonia,ˆ UNIR, Porto Velho, RO, Brazil 2 Fundac¸ao˜ Herm´ınio Ometto, UNIARARAS, Nucleo´ de Cienciasˆ da Saude-NUCISA,´ 13607-339 Araras, SP, Brazil 3 Departamento de Qu´ımica, Universidade Federal de Lavras, UFLA, 37200-000 Lavras, MG, Brazil 4 Departamento de Analises´ Cl´ınicas, Toxicologicas´ e Bromatologicas,´ Faculdade de Cienciasˆ Farmaceuticasˆ de Ribeirao˜ Preto, Universidade de Sao˜ Paulo, USP, Ribeirao˜ Preto, SP, Brazil 5 Departamento de Qu´ımica, Biotecnologia e Engenharia de Bioprocessos, Universidade Federal de Sao˜ Joao˜ del Rei, UFSJ, Campus Alto paraopeba, Ouro Branco, MG, Brazil Correspondence should be addressed to Andreimar M. Soares; [email protected] Received 20 September 2013; Revised 7 December 2013; Accepted 8 December 2013; Published 13 February 2014 Academic Editor: Fernando Albericio Copyright © 2014 Leonardo A. Calderon et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.