CO5560591A2 - Vacunas de adn optimizadas por el codon vih-gag - Google Patents

Vacunas de adn optimizadas por el codon vih-gag

Info

Publication number
CO5560591A2
CO5560591A2 CO04025208A CO04025208A CO5560591A2 CO 5560591 A2 CO5560591 A2 CO 5560591A2 CO 04025208 A CO04025208 A CO 04025208A CO 04025208 A CO04025208 A CO 04025208A CO 5560591 A2 CO5560591 A2 CO 5560591A2
Authority
CO
Colombia
Prior art keywords
gag
nef
truncated
optimized codon
hiv
Prior art date
Application number
CO04025208A
Other languages
English (en)
Inventor
Andrew Beaton
Peter Franz Ertl
Gerald Wayne Gough
Andrew Lear
John Philip Tite
Wely Catherine Ann Van
Original Assignee
Glaxo Group Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from PCT/GB2001/004207 external-priority patent/WO2002024225A1/en
Priority claimed from GB0129604A external-priority patent/GB0129604D0/en
Priority claimed from GB0206462A external-priority patent/GB0206462D0/en
Application filed by Glaxo Group Ltd filed Critical Glaxo Group Ltd
Publication of CO5560591A2 publication Critical patent/CO5560591A2/es

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K39/12Viral antigens
    • A61K39/21Retroviridae, e.g. equine infectious anemia virus
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K39/12Viral antigens
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12Antivirals
    • A61P31/14Antivirals for RNA viruses
    • A61P31/18Antivirals for RNA viruses for HIV
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00Drugs for immunological or allergic disorders
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K2039/51Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/53DNA (RNA) vaccination
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K2039/545Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K2039/57Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00Reverse transcribing RNA viruses
    • C12N2740/00011Details
    • C12N2740/10011Retroviridae
    • C12N2740/16011Human Immunodeficiency Virus, HIV
    • C12N2740/16211Human Immunodeficiency Virus, HIV concerning HIV gagpol
    • C12N2740/16222New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00Reverse transcribing RNA viruses
    • C12N2740/00011Details
    • C12N2740/10011Retroviridae
    • C12N2740/16011Human Immunodeficiency Virus, HIV
    • C12N2740/16211Human Immunodeficiency Virus, HIV concerning HIV gagpol
    • C12N2740/16234Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00Reverse transcribing RNA viruses
    • C12N2740/00011Details
    • C12N2740/10011Retroviridae
    • C12N2740/16011Human Immunodeficiency Virus, HIV
    • C12N2740/16311Human Immunodeficiency Virus, HIV concerning HIV regulatory proteins
    • C12N2740/16322New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00Reverse transcribing RNA viruses
    • C12N2740/00011Details
    • C12N2740/10011Retroviridae
    • C12N2740/16011Human Immunodeficiency Virus, HIV
    • C12N2740/16311Human Immunodeficiency Virus, HIV concerning HIV regulatory proteins
    • C12N2740/16334Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Virology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Immunology (AREA)
  • Epidemiology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Communicable Diseases (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Gastroenterology & Hepatology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Hematology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • AIDS & HIV (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Oncology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

1.- Una secuencia de nucleótidos que codifica una proteína gag de VIH-1 o un fragmento de la misma que contiene un epítope gag del mismo y una proteína Nef VIH-1 o un fragmento del mismo que contiene un epitope nef, unida operativamente a un promotor heterólogo. 8.- Una secuencia de nucleótidos seleccionada entre el grupo compuesto por: Gag (p17, p24), Nef truncado Gag (p17, p24), (codón optimizado), Nef (truncado) Gag (p17, p24), RT, Nef (truncado) Gag (p17, p24), codón optimizado, RT, Nef (truncado) Gag (p17, p24) codón optimizado, RT con codón optimizado, Nef truncado RT (codón optimizado), Gag (p17, p24) con codón optimizado, Nef truncado RT (codón optimizado), Nef truncado, gag p17, p24 con codón optimizado.
CO04025208A 2001-09-20 2004-03-17 Vacunas de adn optimizadas por el codon vih-gag CO5560591A2 (es)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
PCT/GB2001/004207 WO2002024225A1 (en) 2000-09-20 2001-09-20 Use of immidazoquinolinamines as adjuvants in dna vaccination
GB0129604A GB0129604D0 (en) 2001-12-11 2001-12-11 Novel compounds
GB0206462A GB0206462D0 (en) 2002-03-19 2002-03-19 Vaccines

Publications (1)

Publication Number Publication Date
CO5560591A2 true CO5560591A2 (es) 2005-09-30

Family

ID=27256076

Family Applications (1)

Application Number Title Priority Date Filing Date
CO04025208A CO5560591A2 (es) 2001-09-20 2004-03-17 Vacunas de adn optimizadas por el codon vih-gag

Country Status (16)

Country Link
US (2) US20070015721A1 (es)
EP (2) EP2322626A1 (es)
JP (1) JP4601956B2 (es)
CN (1) CN100392085C (es)
AR (1) AR036566A1 (es)
AU (1) AU2002362368B2 (es)
BR (1) BR0212619A (es)
CA (1) CA2461056A1 (es)
CO (1) CO5560591A2 (es)
HU (1) HUP0402259A3 (es)
IL (1) IL160809A0 (es)
MX (1) MXPA04002631A (es)
NO (1) NO331826B1 (es)
NZ (1) NZ531814A (es)
PL (1) PL207168B1 (es)
WO (1) WO2003025003A2 (es)

Families Citing this family (29)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2484044A1 (en) * 2002-03-19 2003-10-02 Powdermed Limited Imidazoquinolineamines as adjuvants in hiv dna vaccination
GB0225788D0 (en) 2002-11-05 2002-12-11 Glaxo Group Ltd Vaccine
US8080642B2 (en) * 2003-05-16 2011-12-20 Vical Incorporated Severe acute respiratory syndrome DNA compositions and methods of use
GB2406336A (en) * 2003-09-24 2005-03-30 Oxxon Pharmaccines Ltd HIV Pharmaccines
GB0417494D0 (en) * 2004-08-05 2004-09-08 Glaxosmithkline Biolog Sa Vaccine
DE102004049223A1 (de) * 2004-10-08 2006-04-20 Johannes-Gutenberg-Universität Mainz Zubereitung zum Impfen, Impfverfahren und Verwendung einer Impf-Zubereitung
AU2012201827B2 (en) * 2005-05-12 2014-09-04 Glaxo Group Limited Vaccine composition
TW200716750A (en) * 2005-05-12 2007-05-01 Glaxo Group Ltd Vaccine composition
EP2099486A2 (en) * 2006-11-30 2009-09-16 Government Of The United States Of America, As Represented by the Secretary Codon modified immunogenic compositions and methods of use
GB0700914D0 (en) * 2007-01-18 2007-02-28 Animal Health Inst Polynucleotides and uses thereof
PL2137210T3 (pl) 2007-03-02 2017-06-30 Glaxosmithkline Biologicals Sa Nowy sposób i kompozycje
EP2148697B1 (en) 2007-05-24 2012-10-03 GlaxoSmithKline Biologicals S.A. Lyophilised cpg containing wt-1 composition
WO2009092113A2 (en) * 2008-01-19 2009-07-23 Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Methods and compositions for the delivery of vaccines to disrupted epithelium
EP2485759A4 (en) * 2009-10-09 2013-07-24 Univ New York METHODS, AGENTS AND PEPTIDES FOR INDUCING IMMUNE RESPONSE INNEED IN VACCINATION AGAINST HIV
BRPI0914507A2 (pt) * 2009-12-23 2012-04-10 Fundacao Oswaldo Cruz vacina para lentivìrus baseada em vìrus recombinante vacinal da febre amarela
ES2625406T3 (es) 2010-03-25 2017-07-19 Oregon Health & Science University Glicoproteínas de CMV y vectores recombinantes
LT2569436T (lt) 2010-05-14 2018-03-12 Oregon Health & Science University Rekombinantiniai žcmv ir hcmv vektoriai ir jų panaudojimas
GB201022007D0 (en) 2010-12-24 2011-02-02 Imp Innovations Ltd DNA-sensor
SI2691530T1 (en) 2011-06-10 2018-08-31 Oregon Health & Science University CMV glycoproteins and recombinant vectors
EP2620446A1 (en) 2012-01-27 2013-07-31 Laboratorios Del Dr. Esteve, S.A. Immunogens for HIV vaccination
CA2904001C (en) 2013-03-05 2021-07-13 Oregon Health & Science University Cytomegalovirus vectors enabling control of t cell targeting
US9695171B2 (en) 2013-12-17 2017-07-04 Pfizer Inc. 3,4-disubstituted-1 H-pyrrolo[2,3-b]pyridines and 4,5-disubstituted-7H-pyrrolo[2,3-c]pyridazines as LRRK2 inhibitors
WO2016011293A1 (en) 2014-07-16 2016-01-21 Oregon Health & Science University Human cytomegalovirus comprising exogenous antigens
MA40783A (fr) 2014-10-03 2017-08-08 Los Alamos Nat Security Llc Vaccins contre le vih comprenant un ou plusieurs antigènes episensus de population
CN108064304A (zh) 2015-02-10 2018-05-22 俄勒冈健康与科学大学 可用于产生非典型cd8+ t细胞应答的方法和组合物
RU2722149C1 (ru) 2015-09-14 2020-05-27 Пфайзер Инк. Новые производные имидазо[4,5-c] хинолинов и имидазо[4,5-c][1,5] нафтиридинов в качестве ингибиторов LRRK2
EP3377636A4 (en) 2015-11-20 2019-11-06 Oregon Health & Science University CMV VECTORS WITH MIKRORNA DETECTION ELEMENTS
CN110036112B (zh) 2016-10-18 2024-05-10 俄勒冈健康与科学大学 引发受主要组织相容性复合体e分子限制的t细胞的巨细胞病毒载体
CA3161633A1 (en) 2019-11-14 2021-05-20 Aelix Therapeutics, S.L. Dosage regimens for vaccines

Family Cites Families (29)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4866034A (en) 1982-05-26 1989-09-12 Ribi Immunochem Research Inc. Refined detoxified endotoxin
US4436727A (en) 1982-05-26 1984-03-13 Ribi Immunochem Research, Inc. Refined detoxified endotoxin product
US4877611A (en) 1986-04-15 1989-10-31 Ribi Immunochem Research Inc. Vaccine containing tumor antigens and adjuvants
US4790824A (en) 1987-06-19 1988-12-13 Bioject, Inc. Non-invasive hypodermic injection device
US4912094B1 (en) 1988-06-29 1994-02-15 Ribi Immunochem Research Inc. Modified lipopolysaccharides and process of preparation
US5312335A (en) 1989-11-09 1994-05-17 Bioject Inc. Needleless hypodermic injection device
US5064413A (en) 1989-11-09 1991-11-12 Bioject, Inc. Needleless hypodermic injection device
ES2113371T3 (es) 1989-11-16 1998-05-01 Univ Duke Transformacion de celulas epidermicas de tejidos animales con la ayuda de particulas.
US5697901A (en) 1989-12-14 1997-12-16 Elof Eriksson Gene delivery by microneedle injection
US5383851A (en) 1992-07-24 1995-01-24 Bioject Inc. Needleless hypodermic injection device
US5583035A (en) * 1992-12-07 1996-12-10 Bayer Aktiengesellschaft HIV antisense expression vectors
US5654186A (en) 1993-02-26 1997-08-05 The Picower Institute For Medical Research Blood-borne mesenchymal cells
CN1112943C (zh) 1994-01-21 2003-07-02 粉剂注射疫苗股份有限公司 气体驱动的基因送递装置
ATE420171T1 (de) 1994-07-15 2009-01-15 Univ Iowa Res Found Immunomodulatorische oligonukleotide
US5786464C1 (en) * 1994-09-19 2012-04-24 Gen Hospital Corp Overexpression of mammalian and viral proteins
GB9502879D0 (en) 1995-02-14 1995-04-05 Oxford Biosciences Ltd Particle delivery
ATE309366T1 (de) * 1996-02-22 2005-11-15 Merck & Co Inc Synthetische hiv-gene
US5856462A (en) 1996-09-10 1999-01-05 Hybridon Incorporated Oligonucleotides having modified CpG dinucleosides
AU7332198A (en) * 1997-03-06 1998-09-22 Klaus Uberla Lentivirus based vector and vector system
US5993412A (en) 1997-05-19 1999-11-30 Bioject, Inc. Injection apparatus
GB9727262D0 (en) 1997-12-24 1998-02-25 Smithkline Beecham Biolog Vaccine
US6602705B1 (en) * 1998-12-31 2003-08-05 Chiron Corporation Expression of HIV polypeptides and production of virus-like particles
AU2487300A (en) * 1998-12-31 2000-07-31 Chiron Corporation Polynucleotides encoding antigenic hiv type c polypeptides, polypeptides and uses thereof
ATE391785T1 (de) * 1999-11-16 2008-04-15 Geneart Ag Das genom des hiv-1 intersubtyps (c/b') und seine anwendungen
JP2003516741A (ja) * 1999-12-17 2003-05-20 メルク エンド カムパニー インコーポレーテッド コドン最適化HIV−1Nef及び修飾HIV−1Nefを発現するポリヌクレオチドワクチン
US6656706B2 (en) * 1999-12-23 2003-12-02 The United States Of America As Represented By The Department Of Health And Human Services Molecular clones with mutated HIV gag/pol, SIV gag and SIV env genes
MXPA02006379A (es) * 1999-12-23 2004-06-21 Medical Res Council Mejoriasen o relacionadas con la respuesta inmune a vih.
EP1156112B1 (en) * 2000-05-18 2006-03-01 Geneart GmbH Synthetic gagpol genes and their uses
AU2001284999B2 (en) * 2000-08-14 2006-09-21 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Modifications of HIV Env, Gag and Pol enhance immunogenicity for genetic immunization

Also Published As

Publication number Publication date
WO2003025003A2 (en) 2003-03-27
JP2005511019A (ja) 2005-04-28
CA2461056A1 (en) 2003-03-27
NZ531814A (en) 2005-10-28
NO331826B1 (no) 2012-04-16
HUP0402259A2 (hu) 2005-01-28
EP1427826A2 (en) 2004-06-16
EP2322626A1 (en) 2011-05-18
US20070015721A1 (en) 2007-01-18
PL207168B1 (pl) 2010-11-30
AU2002362368B2 (en) 2006-09-21
CN1606624A (zh) 2005-04-13
WO2003025003A3 (en) 2003-12-04
IL160809A0 (en) 2004-08-31
PL370359A1 (en) 2005-05-16
US20090203144A1 (en) 2009-08-13
JP4601956B2 (ja) 2010-12-22
MXPA04002631A (es) 2004-07-08
AR036566A1 (es) 2004-09-15
CN100392085C (zh) 2008-06-04
NO20041157L (no) 2004-05-19
HUP0402259A3 (en) 2010-01-28
BR0212619A (pt) 2004-08-17
EP1427826B1 (en) 2011-04-20

Similar Documents

Publication Publication Date Title
CO5560591A2 (es) Vacunas de adn optimizadas por el codon vih-gag
MY145614A (en) Vaccine
Silvera et al. Outcome of simian-human immunodeficiency virus strain 89.6 p challenge following vaccination of rhesus macaques with human immunodeficiency virus Tat protein
CO4600706A1 (es) Genes sinteticos del vih
ES2059354T5 (es) Nuevo polipeptido.
ES2150897T3 (es) Procedimiento de fabricacion recombinante de las proteinas derivadas de hiv-2 y cultivo celular que expresa proteinas de hiv-2.
MY136081A (en) Modified hiv - 1gag p17 peptide and immunogenic composition
NO992369D0 (no) Poxvirusbaserte ekspresjonsvektorer inneholdende heterologe innslag avledet fra lentivirus
HK1044778A1 (en) Hiv-peptides, antigens, vaccine compositions, immunoassay kit and a method of detecting antibodies induced by hiv hiv
WO2004050856A3 (en) Polyvalent, primary hiv-1 glycoprotein dna vaccines and vaccination methods
Polacino et al. Protection of macaques against intrarectal infection by a combination immunization regimen with recombinant simian immunodeficiency virus SIVmne gp160 vaccines
ES2069742T3 (es) Proteina de la capsida del calicivirus felino y secuencia nucleotida.
TW200502245A (en) Vaccine
CO5590933A2 (es) Composicion y metodo para el tratamiento de la diabetes
DK0831899T3 (da) Rekombinant koppevirus-calicivirus-præparater og anvendelser
ANDERSON et al. OF GENETICS
US20090076245A1 (en) HIV-1 Subtype Isolate Regulatory/Accessory Genes, and Modification and Derivatives Thereof
MY131949A (en) Vaccines
PE20000759A1 (es) Gen de la citocroma oxidasa de hidroquinona del tipo bd
WO2004027066A3 (fr) Protéine recombinante chimérique et diagnostic in vitro du hiv
DK1511767T3 (da) Oprenset polypeptid, isolerede nukleinsyrer, som koder for polypeptidet, vektorer og anvendelse deraf
GB0325011D0 (en) HIV pharmaccines
MORIMOTO et al. Use of the recombinant chimera proteins, LacZ-Env and Gag-Env, for immunological studies on HIV-1 infection
WO2011113618A1 (en) Immune therapy
Rivera Immunogenic properties of synthetic peptide mixture derived from the hypervariable regions of HIV-1 envelope glycoprotein gp120

Legal Events

Date Code Title Description
FC Application refused