San Antonio, TX, United States

Operational Technologies Corporation

www.otcorp.com
San Antonio, TX, United States

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Bruno J.G.,Operational Technologies Corporation
Pharmaceuticals | Year: 2013

The potential to emulate or enhance antibodies with nucleic acid aptamers while lowering costs has prompted development of new aptamer-protein, siRNA, drug, and nanoparticle conjugates. Specific focal points of this review discuss DNA aptamers covalently bound at their 3′ ends to various proteins for enhanced stability and greater pharmacokinetic lifetimes in vivo. The proteins can include Fc tails of IgG for opsonization, and the first component of complement (C1q) to trigger complement-mediated lysis of antibiotic-resistant Gram negative bacteria, cancer cells and possibly some parasites during vulnerable stages. In addition, the 3′ protein adduct may be a biotoxin, enzyme, or may simply be human serum albumin (HSA) or a drug known to bind HSA, thereby retarding kidney and other organ clearance and inhibiting serum exonucleases. In this review, the author summarizes existing therapeutic aptamer conjugate categories and describes his patented concept for PCR-based amplification of double-stranded aptamers followed by covalent attachment of proteins or other agents to the chemically vulnerable overhanging 3′ adenine added by Taq polymerase. PCR amplification of aptamers could dramatically lower the current $2,000/gram cost of parallel chemical oligonucleotide synthesis, thereby enabling mass production of aptamer-3′-protein or drug conjugates to better compete against expensive humanized monoclonal antibodies. © 2013 by the authors; licensee MDPI, Basel, Switzerland.


Grant
Agency: Department of Defense | Branch: Defense Health Program | Program: SBIR | Phase: Phase II | Award Amount: 709.98K | Year: 2011

Operational Technologies Corporation (OpTech) has completed a successful Phase I SBIR contract in which 72 new candidate DNA aptamer sequences against Leishmania major were developed and screened by ELISA-like plate assay. Several pilot assays were developed from the highest affinity candidate aptamers using a commercially available handheld and battery-operated fluorometer. In Phase II, OpTech plans to fully develop and deliver to the DoD two or more rapid (15 minute) ultrasensitive one-step plastic-adherent DNA aptamer-magnetic bead (MB) fluorescent sandwich assays (Bruno et al., J. Fluorescence 19:427-435, 2009) for L. major and L. infantum and a visceral leishmaniasis (39 kD kinesin) aptamer assay. In Phase I, this assay format demonstrated detection of Campylobacter and Listeria to<10 cfu/mL and between 10-100 Leishmania promastigotes. OpTech will optimize its lyophilized assay performance with respect to specificity, sensitivity and shelf life of>2 years at 35oC by systematic study of DNA aptamer, magnetic bead, and fluorescent nanoparticle reagent concentrations. OpTech will also evaluate trehalose or other excipient level effects along with innovative vacuum or air-tight packaging of the lyophilized assays. Finally, OpTech will test the assays against various species of live Leishmania and compare to gold standard ELISA and PCR assays.


Grant
Agency: Department of Defense | Branch: Defense Advanced Research Projects Agency | Program: SBIR | Phase: Phase I | Award Amount: 99.96K | Year: 2014

Operational Technologies Corporation proposes to develop non-immunogenic DNA aptamers against unfractionated snake, scorpion, spider, and insect venoms as well as snake phospholipase A2 (PLA2) and melittin (major components of venoms) in Phase 1. OpTech will then test each of the final round aptamers for the ability to inhibit PLA2 and prevent cell lysis due to various venoms in a human fibroblast culture. OpTech will also investigate the feasibility of developing aptamer-dendrimers to add weight and slow aptamer degradation and clearance in vivo during Phase 1. Various aptamers against each class of venom will be conjugated to the ends of existing dendrimers or built into the structure of self-assembling dendrimers to create a ?Universal? scavenger. In Phase 2 OpTech will test its aptamer-dendrimers in a murine challenge model and begin to seek FDA approval with the aid or the Texas BioAlliance. During Phase 2 OpTech will also develop aptamers against the more minor, albeit important, components of venoms (e.g., isolated ion channel blockers etc.) and experiment with DNAzyme development to breakdown non-enzymatic components of venoms (polyamines, histamine). Promising aptamers developed early in Phase 2 will also be added to dendrimers for further animal studies to determine if they enhance protective effects.


Grant
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase II | Award Amount: 999.47K | Year: 2015

DESCRIPTION provided by applicant Development of Anti OLAM Aptamers as Novel Analgesics Phase The management of pain remains a major health care problem due to an incomplete understanding of pain mechanisms TRPV a prominent member of the transient receptor potential TRP family of ligand gated ion channels detects noxious chemical and physical stimuli in peripheral tissues Both pharmacological and gene deletion studies have demonstrated a pivotal role for TRPV in inflammatory heat hyperalgesia and other pain conditions Oxidized linoleic acid metabolites OLAMs have been recently demonstrated to comprise a novel family of endogenous TRPV agonists that contribute to acute and inflammatory pain conditions Therefore compounds that block the OLAM system are likely to constitute a novel family of analgesics In direct support of this prediction Phase data provided herein demonstrate that injection of high affinity DNA aptamers developed against two of the major OLAMs HODE and HODE produced significant analgesia in patch clamp and rat behavioral models of heat pain Although these data provide evidence for proof of concept the aptamers must be developed into heavier conjugates in order to avoid rapid clearance by the kidneys and other major organs Accordingly Operational Technologies Corporation OpTech proposes to continue developing its successful high affinity anti OLAM aptamers from Phase into long lived aptamer conjugates in vivo which specifically bind to HODE and HODE and neutralize their pain producing activities for extended periods of time This would permit replacing anti HODE polyclonal antibodies with more specific less expensive and higher affinity DNA aptamers In Phase OpTech expects to complete several specific aims targeted toward enhancing pharmacokinetics PK by addition of ibuprofen to the andapos end ref thereby enabling association with serum albumin to add weight and retard renal and hepatic clearance Alternative but proven methods to slow in vivo clearance and protect aptamers in vivo including andapos polyethylene glycol PEG and covalent andapos rat albumin attachment will also be investigated Moreover dimensional molecular models of aptamer HODE interactions will be generated by the Southwest Research Institute SwRI to enable potential molecular engineering of improvements to aptamer affinity already exhibiting low nM KD values and specificity if possible using modified or unnatural deoxynucleotides In addition to in vitro patch clamp studies in the presence and absence of various doses of the aptamer conjugates rats will be studied for behavioral changes before and after injection and exposure to oC noxious heat with the various aptamer andapos conjugates If relatively long term on the order of hours PK is observed and heat burn analgesia can again be demonstrated OpTech will seek to file an IND or pre IND application with the FDA PUBLIC HEALTH RELEVANCE Development of Anti OLAM Aptamers as Novel Analgesics Phase Millions of patients suffer from pain and many available analgesic drugs andquot pain killersandquot suffer from either incomplete analgesia or unacceptable side effects This Phase SBIR proposal will further develop a novel class of long lived aptamer conjugate analgesics that work by blocking the endogenous capsaicin like substances that are released during tissue injury thereby specifically treating heat burn pain cancer pain and possibly counteracting some types of shock which are mediated by OLAMs oxidized linoleic acid metabolites including and hydroxyoctadecadienoic acid andquot HODEsandquot


Grant
Agency: Department of Defense | Branch: Army | Program: SBIR | Phase: Phase II | Award Amount: 729.99K | Year: 2011

Operational Technologies Corporation (OpTech) has completed a successful Phase I SBIR contract in which 124 new candidate DNA aptamer sequences against five different species of rickettsiae were developed and screened by ELISA-like plate assay. Several pi


Grant
Agency: Department of Agriculture | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 99.98K | Year: 2013

Most commercially available antibody-based test strips for the detection of foodborne pathogenic bacteria are rapid (work in minutes), but not very sensitive. Operational Technologies Corporation (OpTech) aims to improve the sensitivity of such test strips by using higher affinity and more specific DNA aptamers in place of the antibodies. In addition, OpTech will couple its aptamer-based test strips to fluorescent nanoparticles (FNPs) or quantum dots (QDs) because these have been shown to increase the visual sensitivity of test strips about ten-fold when an ultraviolet penlight is used as the fluorescence excitation source. OpTech will develop as many test strip prototypes as it can in Phase 1 using its existing patent-pending (U.S. patent application no. 13/136,820)DNA aptamer library for foodborne pathogens and toxins. OpTech will also develop new aptamers against the so called "Big 6" non-O157 E. coli which are known to cause disease and are now mandated for testing by the USDA. Finally, OpTech will investigate the use of its new Big 6 E. coli aptamers in fluorescent test strips. The end result of this research and development effort should be more sensitive foodborne pathogen and biotoxin detection in the popular and portable test strip format using fluorescence detection, leading to a safer food supply.


Grant
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 150.00K | Year: 2014

Development of novel antifungal drugs is spurring interest in the detection of pathogenic fungi in archival formalin-fixed tissues. While in situ hybridization and PCR-based methods for fungal detection in tissues exist. formaldehyde adducts on fungal DNA complicate or invalidate such detection methods. Operational Technologies Corporation (OpTech) has been developing DNA aptamers against a broad array of small molecule, protein and whole cell {bacterial and parasite) targets for the last decade. Much of OpTech's aptamer R & D work is published, patented, and the best (highest affinity and most specific) resultant DNA aptamers are sold on its website: www.OTCBiotech.com which has been profitable since inception in late 2012. In 2010-2011, OpTech developed and published DNA aptamers capable of specifically detecting and binding the N-acetylglucosamine (NAG) monomer of the chitin polymer in fungalcell walls for NASA under an SBIR contract to aid in protecting the recycled water supply aboard the InternationalSpace Station. OpTech now proposes to develop and characterize aptamers capable of specifically binding and detecting formalin-fixed Blastomyces, Histoplasma, Fusarium, and Scedosporium as well as the Mucormycetes (Mucor, Rhizopus, Absidia, and Cunninghamella) fungi at the genus level and species level in parffin-embedded tissue sections by fluorescence and/or peroxidase staining.


Grant
Agency: Department of Agriculture | Branch: | Program: SBIR | Phase: Phase II | Award Amount: 439.87K | Year: 2015

The lateral flow (LF) or immunochromatographic (IC) test strip is a staple test format in the food safety testing arsenal. However, the sensitivity of LF assays is limited partly due to antibody affinity and partly due to the visual detection limit of colloidal gold or colored latex particles. Operational Technologies Corporation (OpTech) has developed hundreds of high affinity candidate DNA aptamers against foodborne pathogenic bacterial surface markers such as E. coli intimins (eae gene products), O157 LPS, Campylobacter and Salmonella surface proteins, Listeria flagellin proteins and Listeriolysin O (LLO) many of which it has patented, published and now sells on its website (www.OTCBiotech.com). In Phase 1, OpTech developed and screened longer (200 base) "multivalent" DNA aptamers containing several of its highest affinity aptamer sequences by ELISA-like (ELASA) microplate assay. These lengthier aptamers appear to have enhanced avidity and specificity versus the older 72 base aptamers in preliminary ELASA screening. OpTech developed the new 200 base aptamers with USDA SBIR funding against the "Big 6" non-O157 Shiga-toxin producing E. coli (STEC) serogroups of lipopolysaccharides (LPS; O26, O45, O103, O111, O121, and O145) and has begun to incorporate these into LF strips having multiple detection lines per strip to help satisfy the new USDA requirement for "Big 6" non-O157 STEC detection.As a preliminary demonstration of how aptamer LF test strips could work, during Phase 1, OpTech screened a number of general botulinum toxin, Campylobacter, E. coli, Listeria, and Salmonella aptamer sandwich pairs in LF test strip formats using both colloidal gold and red quantum dots (Qdot 655). Promising aptamer pairs were identified for most of these analytes and the best studied E. coli system produced a limit of detection (LOD) of about 3,000 cells/test by colloidal gold and < 300 cells by Qdot 655-aptamer-based LF visual tests under a UV light. This work was published by Bruno in the open access journal Pathogens 3:341-355, 2014. Due to this clear enhancement of sensitivity via the use of red Qdots and aptamers, OpTech wishes to fully develop a whole line of fluorescent Qdot-aptamer foodborne pathogen LF test strips for commercialization via direct sales and/or product licensing to larger companies. OpTech is currently working with a food safety industry consultant (www.XgeneX.com) to develop partnering relationships and/or licensing deals.The broader impacts of this SBIR project will include enhanced food safety for the public as well as better on-site decision making ability for food processing and packing facilities to decide if foods can be sold or should be discarded. In addition, aptamer-based fluorescent LF test strips can be developed to detect a broad array of clinical, veterinary, environmental pollutant or other analytes. Initially, food processors will be OpTech's main customers. However, the customer base could expand dramatically as OpTech moves fluorescent aptamer-LF test strip technology into the homeland security and medical arenas to detect chemical and biological terrorist threat agents, infectious diseases in hospitals, patient wounds or contaminated raw materials in pharmaceutical production facilities, etc.


Grant
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 200.00K | Year: 2013

Operational Technologies Corp. (OpTech) proposes to develop DNA aptamers against ten or more phosphorylated peptides from NCI's list of proteomic targets. These aptamers will be biotinylated and coupled to streptavidin-coated magnetic microbeads (MBs) or covalently bound to MBs, if necessary. The aptamer-MB reagents will be used to bind and enrich for their specific peptides in chaotrope and trypsin-treated human plasma and serum. Aptamer-MB reagents should be directly transferred to systems such as SISCAPA(Stable Isotope Standards and Capture by Anti-Peptide Antibodies), but will have the advantage of: 1) grater lot-to-lot reproductivity because aptamer DNA sequences will be known and reproduced with very high fidelity, 2) lower cost and faster developmentthan comparable immunomagnetic reagents, 3) potentially greater affinity and specificity versus most antibodies. Aptamer-MBs can then release captured peptides by heating or acidification for mass spectral analyses in high throughput proteomic systems. OpTech will also attempt to precipitate target peptides from serum or plasma by chemically linking different aptamers at their ends to develop bi- and tridentate aptamers capable of linking peptides to form insoluble matrices. In Phase 2, OpTech will refineand expand the technology to include as may aptamers for peptide targets as possible for high-throughput proteomic use. PUBLIC HEALTH RELEVANCE


Grant
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 147.23K | Year: 2012

DESCRIPTION (provided by applicant): Development of Anti-OLAM Aptamers as Novel Analgesics The management of pain remains a major health care problem due to an incomplete understanding of pain mechanisms. TRPV1, a prominent member of the transient receptorpotential (TRP) family of ligand-gated ion channel, detects noxious chemical and physical stimuli in peripheral tissues. Both pharmacological and gene deletion studies have demonstrated a pivotal role for TRPV1 in inflammatory heat hyperalgesia and otherpain conditions. Oxidized linoleic acid metabolites (OLAMs) have been recently demonstrated to comprise a novel family of endogenous TRPV1 agonists that contributes to acute and inflammatory pain conditions. Therefore, compounds that block the OLAM systemare likely to constitute a novel family of analgesics. In direct support of this prediction, preliminary data provided herein demonstrate that injection of antibodies against two of the major OLAMs, 9-HODE and 13-HODE, produce significant analgesia in twopain models. Although these data provide evidence for proof-of-concept, the clinical development of rabbit polyclonal antibodies is not feasible due to adverse effects related to immunogenicity. Accordingly, Operational Technologies Corporation (OpTech) proposes to use combinatorial aptamer chemistry to discover DNA aptamers that specifically bind to these OLAMs and neutralize their pain-producing activities. This would permit replacing antibodies with more specific, less expensive and perhaps higher affinity DNA aptamers. In Phase 1, OpTech expects to complete two overall Specific Aims. Aim 1 will develop, clone, and sequence several highly specific DNA aptamers that bind 9-hydroxydecadienoic acid (HODE) and 13-HODE without binding to the precursor lipid, linoleic acid (Fig 1A). Aim 2 will evaluate the analgesic activity of the aptamers using several in vivo preclinical rat models of pain. In Phase 2, OpTech will refine, optimize, and begin commercialization of its anti-OLAM aptamer compounds. The Phase 2 optimization process will include 3-D modeling of putative aptamer binding pocket interactions with the OLAMs. Based on 3-D modeling findings, OpTech anticipates adding modified bases having various functional groups (e.g., primary amines, methyl, thiol groups, etc.) that are now commercially available for addition to oligonucleotides at the point of chemical synthesis. The effects of these additional functional groups on aptamer-OLAM binding affinity are expected to better emulate amino acid side chains andwill be studied by ELISA-like plate assays and surface plasmon resonance (SPR). The highest affinity and most specific unmodified or modified anti-OLAM aptamers will move into animal studies. The most effective aptamers in animal pain studies will be modified for longer in vivo lifetimes (3'-cholesterol addition and inclusion in liposomes, PEGylation, etc.) or time-released formulation and enter the FDA approval pipeline. PUBLIC HEALTH RELEVANCE: Development of Anti-OLAM Aptamers as Novel Analgesics Millions of patients suffer from pain and many available analgesic drugs ( pain killers ) suffer from either incomplete analgesia or unacceptable side-effects. The proposal will develop a novel class of analgesics that work by blocking the endogenouscapsaicin-like substances that are released during tissue injury.

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