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  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/02—Immunomodulators
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2319/00—Fusion polypeptide
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  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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  • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
  • C12N2710/00011—Details
  • C12N2710/10011—Adenoviridae
  • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
  • C12N2710/10322—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
  • C12N2710/00011—Details
  • C12N2710/14011—Baculoviridae
  • C12N2710/14111—Nucleopolyhedrovirus, e.g. autographa californica nucleopolyhedrovirus
  • C12N2710/14141—Use of virus, viral particle or viral elements as a vector
  • C12N2710/14143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector

Definitions

• this novel vector is capable of efficient transduction of primary tumor cells.
• Two t o three orders of magnitude of increased gene transfer to ovarian cancer cells was demonstrated by the vector, suggesting that recombinant adenoviruses containing fibers with an incorporated RGD peptide may be of great utility for treatment of neoplasms characterized by deficiency of the primary Ad5 receptor.
• Figure 6 shows an adenovirus binding assay.
• D ata normalized for protein concentration is shown. Background luciferase activity in mock-infected cells is also displayed (open box). Each po int represents the mean ⁇ standard deviation of three determinations. D ata from two representative samples, ( Figure 19A) and ( Figure 19B), are shown .
• 1 5 fiber knob allowed the virus to utilize the RGD/integrin interactions as a n alternative infection pathway, dramatically improved the ability of th e virus to transduce several types of cells, which normally are refractory t o Ad infection.
• this viral vector was employed as a means for efficient gene transfer to primary ovarian cancer cells.
• the recombinant adenovirus vector containing fibers with RGD-motif in the HI loop was capable of dramatically augmenting gene delivery to target cells via the a CAR-independent cell entry mechanism.
• a composition of matter comprising a modified adenoviral vector containing fiber gene variants .
• a "DNA molecule” refers to the polymeric form of deoxyribonucleotides (adenine, guanine, thymine, or cytosine) in its either single stranded form, or a double-stranded helix. This term refers only to the primary and secondary structure of the molecule, and does not limit it to any particular tertiary forms. Thus, this term includes double-stranded DNA found, inter alia, in linear DNA molecules (e.g., restriction fragments), viruses, plasmids, and chromosomes. In discussing the structure herein according to the normal convention of giving only the sequence in the 5' to 3' direction along the nontranscribed strand of DNA (i.e., the strand having a sequence homologous to the mRNA).
• a DNA "coding sequence” is a double-stranded DNA sequence which is transcribed and translated into a polypeptide in vivo when placed under the control of appropriate regulatory sequences. The boundaries of the coding sequence are determined by a start codon at th e 5' (amino) terminus and a translation stop codon at the 3' (carboxyl) terminus.
• a coding sequence can include, but is not limited to, prokaryotic sequences, cDNA from eukaryotic mRNA, genomic DNA sequences from eukaryotic (e.g., mammalian) DNA, and even synthetic DNA sequences. A polyadenylation signal and transcription termination sequence will usually be located 3' to the coding sequence.
• Recombinant DNA technology refers to techniques for uniting two heterologous DNA molecules, usually as a result of in vitro ligation of DNAs from different organisms. Recombinant DNA molecules are commonly produced by experiments in genetic engineering. Synonymous terms include “gene splicing", “molecular cloning” a n d “genetic engineering”. The product of these manipulations results in a “recombinant” or “recombinant molecule”.
• Labels include, for example, fluorescein, rhodamine, auramine, Texas Red, AMCA blue and Lucifer Yellow.
• a particular detecting material is anti-rabbit antibody prepared in goats and conjugated with fluorescein through an isothiocyanate. Proteins can also be labeled with a radioactive element or with a n enzyme. The radioactive label can be detected by any of the currently available counting procedures.
• the preferred isotope may be selected from 3 H, 14 C, 32 P, 35 S, 36 C1, S IQ, 57 Co, 58Co, 59p e , 9 ⁇ , i25 I; i3i I? and 186 Re .
• Anti-fiber monoclonal antibodies 4D2 (19) and 1D6.14 ( 14) were generated at the University of Alabama at Birmingham Hybridoma Core Facility.
• Anti-FLAG monoclonal antibody M2 and M2-affinity gel were purchased from Scientific Imaging Systems (Eastman Kodak, New Haven, Conn.).
• Anti- ⁇ v ⁇ 3 integrin monoclonal antibody LM609 and anti- ⁇ v ⁇ 5 integrin monoclonal antibody P1F6 were purchased from Chemicon (Chemicon, Temecula, Cal.) and Gibco-BRL (Gibco BRL, Gaithersburg, Md.), respectively.
• Anti-CAR monoclonal antibody RmcB was obtained from R. W. Finberg (Dana-Farber Cancer Institute, Harvard Medical School, Boston, Mass.).
• the supernatant containing unbound material, the wash, and the eluate were then employed t o detect the presence of the virus. For this, aliquots of these fractions were treated for 1 hour at 37°C with sodium dodecyl sulfate, EDTA, a n d proteinase K at final concentrations of 1%, 10 mM and 100 ⁇ g/ml, respectively. The samples were analyzed by agarose gel electrophoresis t o detect viral DNA.
• Recombinant fibers were recovered from the lysates of baculovirus-infected insect cells with Ni-NTA-Sepharose designed for purification of the six-His-tagged proteins.
• the yield of purified fibers was in the range of 10 ⁇ g of protein per 10" infected cells. Analysis by SDS- polyacrylamide gel electrophoresis of both recombinant proteins showed
• the RGD-4C containing fiber protein, fiber-RGD was expressed in a baculovirus expression system in order to characterize th e
• the virus was derived by the method described by Chartier e t al. To simplify the downstream gene transfer assays, an expression cassette containing the firefly luciferase gene driven by cytomegalovirus promoter was introduced in place of El region of the adenoviral genome .
• the genome of the new virus designated Ad51ucRGD was generated in E. coli via a two step protocol utilizing homologous DNA recombination between the plasmid pVK50 and fragments of DNA isolated from two shuttle vectors, pNEB.PK.FfflRGD and pACCMV.Luc ⁇ PC, which contain th e fiber gene and the luciferase expression cassette flanked with adenoviral DNA sequences, respectively.

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• 1999
  • 1999-02-05 IL IL13768199A patent/IL137681A0/xx unknown
  • 1999-02-05 EP EP99906761A patent/EP1053013A4/en not_active Withdrawn
  • 1999-02-05 CN CN99804659A patent/CN1304316A/zh active Pending
  • 1999-02-05 KR KR1020007008593A patent/KR20010034487A/ko not_active Withdrawn
  • 1999-02-05 WO PCT/US1999/002549 patent/WO1999039734A1/en not_active Ceased
  • 1999-02-05 US US09/245,603 patent/US7297542B2/en not_active Expired - Fee Related
  • 1999-02-05 JP JP2000530231A patent/JP2002507391A/ja active Pending
  • 1999-02-05 AU AU26595/99A patent/AU744252B2/en not_active Ceased
  • 1999-02-05 BR BR9908613-1A patent/BR9908613A/pt not_active IP Right Cessation
  • 1999-02-05 CA CA002327545A patent/CA2327545A1/en not_active Abandoned
• 2000
  • 2000-08-04 NO NO20003956A patent/NO20003956L/no not_active Application Discontinuation

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