43 results about "Lethal factor" patented technology

The present invention provides compounds that efficiently and specifically inhibit lethal factor (LF) protease activity of anthrax toxin.

The invention is related to polynucleotide-based anthrax vaccines. In particular, the invention is plasmids operably encoding Bacillus anthracis antigens, in which the naturally-occurring coding regions for the B. anthracis antigens have been modified for improved translation in human or other mammalian cells through codon optimization. In certain embodiments, the coding regions are also modified so as to remove potential N-linked glycosylation sites. B. anthracis antigens which are useful in the invention include, but are not limited to protective antigen (PA), lethal factor (LF), and fragments, variants or derivatives of either of these antigens. The invention is further directed to methods to induce an immune response to B. anthracis in a mammal, for example, a human, comprising delivering a plasmid encoding a codon-optimized B. anthracis antigen as described above. The invention is also directed to pharmaceutical compositions comprising plasmids encoding a codon-optimized B. anthracis antigen as described above, and further comprising adjuvants, excipients, or immune modulators.

Recombinant immunogenic compositions and methods for protecting against lethal infections from Bacillus anthracis having a variant of recombinant Bacillus anthracis protective antigen (rPA) and a variant of recombinant Bacillus anthracis lethal factor (rLF). These proteins may be expressed separately or as a fusion protein. The recombinant proteins are produced in an avirulent strain of Bacillus anthracis that overproduces the desired antigens. The compositions and methods induce the animal host to produce antibodies against a virulent strain of Bacillus anthracis.

The present invention provides a protease biosensor that can be used to detect the presence of the lethal factor protease from Bacillus anthracis, as well as methods for using the protease biosensor.

The present invention is directed to a method for delivering exogenous proteins to the cytosol, by binding a target antigen (such as a protein) to a transport factor that contains a fragment of a bipartite protein exotoxin, but not the corresponding protective antigen. Preferably, the target antigen is fused to the transport factor. Preferred transport factors include the protective antigen binding domain of lethal factor (LFn) from B. anthracis, consisting of amino acids 1-255, preferably a fragment of at least 80 amino acids that shows at least 80% homology to LFn, and a fragment of about 105 amino acids from the carboxy portion that does not bind PA. The target antigen can include any molecule for which it would be desirable to elicit a CMI response, including viral antigens and tumor antigens.

This invention relates to compounds of formula (I), and a method for treating anthrax or inhibiting lethal factor by administrating a composition containing a compound of formula (I) and a pharmaceutically acceptable carrier. This invention further relates to the use of the compounds of formula (I) to treat other conditions related to an anthrax infection.

Methods for inhibiting metalloprotease activity; treating a pathological condition influenced by the action of MMP; treating a pathological condition influenced by cyclooxygenases, endonucleases, metallopepitidases and 5-epoxigenase; treating a pathological condition for which the presence of a metal ion is required; inhibiting the lethal factor produced by toxigenic strains of anthrax; inhibiting activities of fungi, bacteria or plants that utilize zinc-dependent methionine synthetase; inhibiting the activity of a zinc-dependent enzyme in prokaryotic systems; and inhibiting the activity of zinc-dependent enzymes; and compositions thereof, are disclosed.

The invention provides compounds that can efficiently and specifically inhibit bacterial toxins, such as inhibit the lethal factor (LF) protease activity of anthrax toxin and / or botulinum neurotoxin type A. The invention also provides methods for inhibiting proteases, such as lethal factor protease, as well as methods for treating bacterial infections, such as anthrax and botulinum.

Provided are oligonucleotides for isolating human antibody cDNAs from cells or cell lines, such as hybridomas. The invention also provides cDNAs that encode at least one provided CDR of a heavy chain or a light chain of a human monoclonal antibody that binds to B. anthracis protective antigen; and cDNAs that encode at least one provided CDR of a heavy chain or a light chain of a human monoclonal antibody that binds to B. anthracis lethal factor. The invention further provides expression vectors that contain one or more cDNAs isolated according to the methods of the invention, host cells expressing one or more inventive cDNAs, and transgenic plants and animals that express one or more inventive cDNAs. In certain embodiments of the invention the expression system is a plant-based expression system. The invention further provides antibody compositions comprising one or more antibodies produced by expressing a cDNA isolated according to the methods of the invention in a suitable expression system. Additionally encompassed in the invention are kits containing one or more of provided compositions, as well as methods of production and use of provided compositions.

Disclosed herein is a pharmacophore model for inhibiting anthrax lethal factor protease activity which comprises a first aromatic center A, a second aromatic center B, a first polar center C, a second polar center D, a third polar center E, and a neutral linker F. In some embodiments, the distance between the first aromatic center A and the neutral linker F is about 4.7 to about 6.7 Å, preferably about 5.7 Å. In some embodiments, the distance between the neutral linker F and the second aromatic center B is about 3.4 to about 4.4 Å, preferably about 3.9 Å. In some embodiments, the distance between first aromatic center A and the first polar center C is about 5.5 to about 7.5 Å, preferably about 6.5 Å. In some embodiments, the distance between the first aromatic center A and the second polar center D is about 4.6 to about 6.6 Å, preferably about 5.6 Å. In some embodiments, the distance between the second aromatic center B and the second polar center D is about 3.6 to about 4.6 Å, preferably about 4.1 Å. In some embodiments, the distance between the second aromatic center B and the third polar center E is about 2.6 to about 3.6 Å, preferably about 3.1 Å. In some embodiments, the distance between the first polar center C and the third polar center E is about 15.6 to about 19.6 Å, preferably about 17.6 Å. Also disclosed are small molecules fitting the pharmacophore model and compositions and methods of using thereof.

The present invention relates to in vitro and ex vivo methods of screening for modulators, homologues, and mimetics of lethal factor mitogen activated protein kinase kinase (MAPKK) protease activity, as well as methods of treating cancer by administering LF to transformed cells.

The present invention provides methods of specifically targeting compounds to cells overexpressing matrix metalloproteinases, plasminogen activators, or plasminogen activator receptors, by administering a compound and a mutant protective antigen protein comprising a matrix metalloproteinase or a plasminogen activator-recognized cleavage site in place of the native protective antigen furin-recognized cleavage site, wherein the mutant protective antigen is cleaved by a matrix metalloproteinase or a plasminogen activator overexpressed by the cell, thereby translocating into the cell a compound comprising a lethal factor polypeptide comprising a protective antigen binding site.

This invention relates to a method of inhibiting lethal factor (LF) or for treating anthrax and other conditions related to anthrax infection comprising co-administration of an effective amount of an LF inhibitor and a vaccine to a patient in need of such treatment. Such co-administration unexpectedly provides an effective immune response.

Plasmids encoding anti-HIV and anti-anthrax therapeutic agents are disclosed. Plasmid pWKK-500 encodes a fusion protein containing DP178 as a targeting moiety, the ricin A chain, an HIV protease cleavable linker, and a truncated ricin B chain. N-terminal extensions of the fusion protein include the maltose binding protein and a Factor Xa protease site. C-terminal extensions include a hydrophobic linker, an L domain motif peptide, a KDEL ER retention signal, another Factor Xa protease site, an out-of-frame buforin II coding sequence, the lacZα peptide, and a polyhistidine tag. More than twenty derivatives of plasmid pWKK-500 are described. Plasmids pWKK-700 and pWKK-800 are similar to pWKK-500 wherein the DP178-encoding sequence is substituted by RANTES- and SDF-1-encoding sequences, respectively. Plasmid pWKK-900 is similar to pWKK-500 wherein the HIV protease cleavable linker is substituted by a lethal factor (LF) peptide-cleavable linker.

Compositions and methods effective for eliciting an immune response for preventing or reducing infection or improving clinical outcomes caused by Bacillus anthracis are provided. The compositions include a naturally occurring or synthetic protein, peptide, or protein fragment containing all or an active portion of an antigenic epitope associated with anthrax toxin proteins optionally combined with a pharmaceutically acceptable carrier. The preferred antigenic epitopes correspond to immunogenic regions of protective antigen, lethal factor or edema factor, either individually or in combination. In addition, methods and compositions containing antibodies for reducing the effects of anthrax toxins are described. The methods involve administering to a human or animal the compositions described herein in a dosage sufficient to elicit an immune response or treat the anthrax infection.

The inventive subject matter relates to a competitive method for estimating the concentration in a sample of a Bacillus anthracis protein or antibody thereof selected from the group consisting of protective antigen (PA), lethal factor (LF) and edema factor (EF). The method may employ the use of Fluorescence Polarization, FLT or FRET. The competitive methods are capable of detecting a target protein within 5 minutes within the range of 0.1 to 10.0 nM. The methods provide for the rapid detection and quantitation of bacteria, bacterial antigen or antibody in culture media or broth of growing cultures of bacteria, including B. anthracis by fluorescent methods.

Plasmids encoding anti-HIV and anti-anthrax therapeutic agents are disclosed. Plasmid pWKK-500 encodes a fusion protein containing DP178 as a targeting moiety, the ricin A chain, an HIV protease cleavable linker, and a truncated ricin B chain. N-terminal extensions of the fusion protein include the maltose binding protein and a Factor Xa protease site. C-terminal extensions include a hydrophobic linker, an L domain motif peptide, a KDEL ER retention signal, another Factor Xa protease site, an out-of-frame buforin II coding sequence, the lacZa peptide, and a polyhistidine tag. More than twenty derivatives of plasmid pWKK-500 are described. Plasmids pWKK-700 and pWKK-800 are similar to pWKK-500 wherein the DP178-encoding sequence is substituted by RANTES- and SDF-1-encoding sequences, respectively. Plasmid pWKK-900 is similar to pWKK-500 wherein the HIV protease cleavable linker is substituted by a lethal factor (LF) peptide-cleavable linker. Plasmid pWKK-21 is similar to pWKK-500 wherein the highly truncated ricin B chain is substituted by a one-domain ricin B chain. Oligonucleotide cassettes encoding an HIV protease-cleavable peptide linker and a method of making modified plasmids encoding modified fusion proteins are also described.

The invention provides compounds that can efficiently and specifically inhibit bacterial toxins, such as inhibit the lethal factor (LF) protease activity of anthrax toxin and / or botulinum neurotoxin type A. The invention also provides methods for inhibiting proteases, such as lethal factor protease, as well as methods for treating bacterial infections, such as anthrax and botulinum.

The invention discloses an antibody of a humanized anti-anthrax protective antigen PA and an application thereof. According to the antibody, a heavy chain variable region has an amino acid sequence as shown in SEQ ID NO:4; a light chain variable region has an amino acid sequence as shown in SEQ ID NO:4; a heavy chain constant region has an amino acid sequence as shown in SEQ ID NO:6; and a light chain constant region has an amino acid sequence as shown in SEQ ID NO:8. The antibody can be specifically combined with the anthrax protective antigen (PA) and can be used for preventing a lethal factor LF so as to play a protective role.

This invention relates to compounds of formula (I), and a method for treating anthrax or inhibiting lethal factor by administrating a composition containing a compound of formula (I) and a pharmaceutically acceptable carrier. This invention further relates to the use of the compounds of formula (I) to treat other conditions related to an anthrax infection.

The present invention provides methods for inhibiting tumor associated angiogenesis by administering a mutant protective antigen protein comprising a matrix metalloproteinase-recognized cleavage site in place of the native protective antigen furin-recognized site in combination with a lethal factor polypeptide comprising a protective antigen binding site. Upon cleavage of the mutant protective antigen by a matrix metalloproteinase, the lethal factor polypeptide is translocated into cancer and endothelial cells and inhibits tumor associated angiogenesis.

Compositions and methods effective for eliciting an immune response for preventing or reducing infection or improving clinical outcomes caused by Bacillus anthracis are provided. The compositions include a naturally occurring or synthetic protein, peptide, or protein fragment containing all or an active portion of an antigenic epitope associated with anthrax toxin proteins optionally combined with a pharmaceutically acceptable carrier. The preferred antigenic epitopes correspond to immunogenic regions of protective antigen, lethal factor or edema factor, either individually or in combination. In addition, methods and compositions containing antibodies for reducing the effects of anthrax toxins are described. The methods involve administering to a human or animal the compositions described herein in a dosage sufficient to elicit an immune response or treat the anthrax infection.

The present invention is directed to a method for promoting or stimulating a cell-mediated immune response to an antigen, by administering a target antigen (such as a protein) with a transport factor that contains a fragment of a bipartite protein exotoxin, but not the corresponding protective antigen. Preferred transport factors include the protective antigen binding domain of lethal factor (LFn) from B. anthracis, consisting of amino acids 1-255, preferably a fragment of at least 80 amino acids that shows at least 80% homology to LFn, and a fragment of about 105 amino acids from the carboxy portion that does not bind PA. The target antigen can include any molecule for which it would be desirable to elicit a CMI response, including viral antigens and tumor antigens.

The invention belongs to the field of biology engineering technologies, and particularly relates to a method for improving bacillus subtilis strains. The method includes the steps of manually constructing a gene sequence not containing spore production lethal factor genes, electric shock conversion, sieving verifying and others. In the prior art, although part of basic research is already implemented on the spore production lethal factor genes (skf), whether the yield of endogenous enzymes can be increased through deleting the spore production lethal factor genes is still not determined, and meanwhile for different microorganism strains, after the spore production lethal factor genes are deleted, whether improvements on the fermentation capacity of the strains are the same lacks more research. With the detailed bacillus subtilis strains being examples, new strains lacking spore production lethal factor genes (skf) are successfully constructed, when the strains are concretely used for fermenting and producing amylase, determination results show that the enzyme activity of the constructed new strains is improved well, and good application prospects are achieved.

The present invention generally relates to HIV compositions and methods of use. One aspect of the present invention relates to a composition comprising a pharmaceutically acceptable carrier and an antigen preparation, the antigen preparation comprising an HIV polypeptide or fragment thereof and a Bacillus anthracis lethal factor (LF) polypeptide, such as a LFn polypeptide. In some embodiments, the LF polypeptide can be fused or otherwise associated with the HIV polypeptide. Other aspect of the present invention relate to use of vaccine comprising a HIV polypeptide and a Bacillus anthracis lethal factor (LF) polypeptide in methods to enhance efficacy traditional antiretroviral therapy.

One major problem in diagnosis methods presently available for anthrax is that these methods require several days to produce a result. The only existing treatment for anthrax requires administration soon after infection at a time when patients are exhibiting only mild flu-like symptoms. Thus, a patient may be days beyond the time when treatment would be effective by the time a diagnosis is made. The present invention reduces diagnosis time to as little as four hours providing same day identification of anthrax radically increasing the odds of delivering proper treatment and patient recovery. The rapid identification of anthrax lethal factor activity exhibited by the instant invention is also amenable to in vivo screening protocols for the discovery and development of anthrax vaccines and lethal factor inhibitors. The instant invention isolates and concentrates lethal factor and lethal toxin from nearly any biological sample. By capitalizing on the endopeptidase activity of lethal factor the present invention amplifies output signals producing reliable detection of picomolar concentrations of lethal factor. The instant invention involves novel purification and detection techniques and substrates for rapid, reproducible, and quantitative measurements of anthrax lethal factor in biological samples.

The present invention provides methods of specifically targeting compounds to cells overexpressing matrix metalloproteinases, plasminogen activators, or plasminogen activator receptors, by administering a compound and a mutant protective antigen protein comprising a matrix metalloproteinase or a plasminogen activator-recognized cleavage site in place of the native protective antigen furin-recognized cleavage site, wherein the mutant protective antigen is cleaved by a matrix metalloproteinase or a plasminogen activator overexpressed by the cell, thereby translocating into the cell a compound comprising a lethal factor polypeptide comprising a protective antigen binding site.

A novel group of aminoglycosides which share some structural features of currently available aminoglycosides with regard to the backbone, while also having significant structural differences, is disclosed. The similarity enables these aminoglycosides to be highly potent and effective antibiotics, while the significant differences enable these aminoglycosides to reduce or even block antibiotic resistance. The aminoglycosides of the present invention are suitable for inhibition of antrax lethal factor, hence are suitable for use as a cure for anthrax.