CHEMICAL AND BIOLOGICAL DEFENSE PROGRAM
SBIR 10.1 Proposal Submission
In response to Congressional interest in the readiness and effectiveness of U.S. Nuclear, Biological and Chemical (NBC) warfare defenses, Title XVII of the National Defense Authorization Act for Fiscal Year 1994 (Public Law 103-160) required the Department of Defense (DoD) to consolidate management and oversight of the Chemical and Biological Defense (CBD) Program into a single office – Office of the Special Assistant, Chemical and Biological Defense and Chemical Demilitarization Programs. The Joint Science and Technology Office for Chemical and Biological Defense (JSTO-CBD), Defense Threat Reduction Agency (DTRA) provides the management for the Science and Technology component of the Chemical and Biological Defense Program. Technologies developed under the SBIR program have the potential to transition to the Joint Program Executive Office for Chemical and Biological Defense (JPEO-CBD) if the appropriate level of technology maturity has been demonstrated. The JSTO-CBD Science & Technology programs and initiatives are improving defensive capabilities against Chemical and Biological Weapons. The Small Business Innovation Research (SBIR) portion of the CBD Program is managed by the JSTO-CBD through the Army SBIR Program Management Office (PM, SBIR), Ft. Belvoir, VA.
The mission of the Chemical and Biological Defense Program is to ensure that the U.S. Military has the capability to operate effectively and decisively in the face of chemical or biological warfare threats at home or abroad. Numerously factors continually influence the program and its technology development priorities, including planning for war-fighting support to asymmetrical threats, the evolving geopolitical environment, development of new threat materials, the threat of global proliferation of chemical and biological weapons, and available DoD resources. Improved defensive capabilities are essential in order to minimize the impact of such weapons. The U.S. military requires the finest state-of-the-art equipment and instrumentation available that permits our warfighters to detect to warn and avoid contamination, if possible -- and to be able to sustain operations in a potentially contaminated environment through protection and decontamination. Further information regarding the DoD Joint Chemical and Biological Defense program is available at the DoD Counterproliferation and Chemical Biological Defense homepage at http://www.acq.osd.mil/cp
The overall objective of the CBD SBIR Program is to improve the transition or transfer of innovative Chem-Bio technologies to the end user – the warfighter – in addition to commercializing technologies within the private sector for mutual benefit. The CBD SBIR Program targets those technology efforts that maximize a strong defensive posture in a biological or chemical environment using passive and active means as deterrents. These technologies include chemical and biological detection for both point and stand-off capabilities; individual and collective protection; hazard mitigation (decontamination); information systems technology to include but not limited to modeling and simulation; medical pre-treatments (e.g., vaccine development and delivery); medical diagnostics; and medical therapeutics (chemical countermeasures and biological countermeasures).
Your entire proposal (consisting of Proposal Cover Sheets, the full Technical Proposal, Cost Proposal, and Company Commercialization Report) must be submitted electronically through the DoD SBIR/STTR Proposal Submission system located at www.dodsbir.net/submission. A hardcopy is NOT required for CBD. Hand or electronic signature on the proposal is also NOT required.
You must prepare a Company Commercialization Report through the Submission site and it will be included with your electronic submission; however, it does not count against the proposal page limit. Update your commercialization information if you have not done so in the past year. Please note that improper handling of the Commercialization Report may result in the proposal being substantially delayed and that information provided may have a direct impact on the review of the proposal. Refer to section 3.5d at the program solicitation for detailed instructions on the Company Commercialization Report.
Be reminded that section 3.5.a of this solicitation states: “If your proposal is selected for award, the technical abstract and discussion of anticipated benefits will be publicly released on the Internet; therefore, do not include proprietary or classified information in these sections”. Note also that the DoD web site contains timely information on firm, award, and abstract data for all DoD SBIR Phase I and II awards archived for several years. This information can be viewed on the DoD SBIR/STTR website at http://www.acq.osd.mil/sadbu/sbir/.
The CBD SBIR Program has enhanced its Phase I-Phase II transition process by implementing the use of a Phase I Option that may be exercised to fund interim Phase II activities while a Phase II contract is being negotiated. The maximum dollar amount for a Phase I feasibility study is $70,000. The Phase I Option, which must be proposed as part of the Phase I proposal, covers activities over a period of up to three months and at a cost not to exceed $30,000. All proposed Phase I Options must be fully costed and should describe appropriate initial Phase II activities, which would lead, in the event of a Phase II award, to the successful demonstration of a product or technology. The CBD SBIR program will not accept Phase I proposals which exceed $70,000 for the Phase I effort and $30,000 for the Phase I Option effort. Only those Phase I efforts selected for Phase II awards through the CBD SBIR Program’s competitive process will be eligible to exercise the Phase I Option. To maintain the total cost for SBIR Phase I and Phase II activities at a limit of $850,000, the total funding amount available for Phase II activities under a resulting Phase II contract will be $750,000.
Companies submitting a Phase I proposal under this Solicitation must complete the Cost Proposal using the on-line form within a total cost of $70,000 over a period of up to 6 months (plus up to $30,000 for the Phase I Option over a period of up to three (3) months). Phase I and Phase I Option costs must be shown separately.
Selection of Phase I proposals will be based upon the evaluation procedures and criteria discussed in section 4.2. The CBD SBIR Program reserves the right to limit awards under any topic, and only those proposals of superior scientific and technical quality in the judgment of the technical evaluation team will be funded.
Proposals not conforming to the terms of this solicitation, and unsolicited proposals, will not be considered. Awards are subject to the availability of funding and successful completion of contract negotiations.
CBD Program Phase II Proposal Guidelines
Phase II is the demonstration of the technology that was found feasible in Phase I. Only those Phase I awardees which achieved success in Phase I, as determined by the project technical monitor measuring the results achieved against the criteria contained in section 4.3, will be invited to submit a Phase II proposal. During or at the end of the Phase I effort, awardees will be invited to submit proposals for evaluation for a Phase II award based on the results of the Phase I effort. The invitation will be issued in writing by the organization responsible for awarding the Phase I effort. Invited proposers are required to develop and submit a commercialization plan describing feasible approaches for marketing the developed technology. Proposers are required to submit a budget for the entire 24 month Phase II period. During contract negotiation, the contracting officer may require a cost proposal for a base year and an option year, thus, proposers are advised to be mindful of this possibility. These costs must be submitted using the Cost Proposal format (accessible electronically on the DoD submission site), and may be presented side-by-side on a single Cost Proposal Sheet. The total proposed amount should be indicated on the Proposal Cover Sheet as the Proposed Cost. At the Contracting Officer’s discretion, Phase II projects may be evaluated after the base year prior to extending funding for the option year.
The CBD SBIR Program is committed to minimizing the funding gap between Phase I and Phase II activities. All CBD SBIR Phase II proposals will receive expedited reviews and be eligible for interim funding (refer to top for information on the Phase I Option). Accordingly, all Phase II proposals will be evaluated within a single multi-tiered evaluation process and schedule. Phase II proposals will thus typically be submitted within 5 months from the scheduled DoD Phase I award date (the scheduled DoD award date for Phase I, subject to the Congressional Budget process, is 4 months from close of the DoD Solicitation). The CBD Program typically funds a cost plus fixed fee Phase II award, but may award a firm fixed price contract at the discretion of the Contracting Officer.
Non-Proprietary Summary Reports
Not required for CBD SBIR Phase I or Phase II awards.
CBD SBIR Submission of Final Technical Reports
All Phase I and Phase II final technical reports will be submitted to the awarding organization in accordance with Contract Data Requirements List (CDRL). Companies should not submit final reports directly to the Defense Technical Information Center (DTIC).
Key Dates
10.1 Solicitation Pre-Release 12 November 2009 – 9 December 2010
10.1 Solicitation Open/Close 10 December 2009 – 13 January 2010
10.1 Solicitation Closes 13 January 2010; 6:00 a.m. ET
Phase I Selections March 2010
Phase I Awards May 2010*
Phase II Invitations October 2010
Phase II Proposals Due November 2010
*Subject to the Congressional Budget process.
CBD SBIR PROPOSAL CHECKLIST
This is a Checklist of Requirements for your proposal. Please review the checklist carefully to ensure that your proposal meets the CBD SBIR requirements. Failure to meet these requirements will result in your proposal not being evaluated or considered for award.
_____ 1. The Proposal Cover Sheets along with the Technical Proposal, Cost Proposal and Company Commercialization Report were submitted via the Internet using the DoD’s SBIR/STTR Proposal Submission website at http://www.dodsbir.net/submission.
_____ 2. The proposal cost adheres to the CBD Program criteria specified.
_____ 3. The proposal is limited to only ONE solicitation topic. All required documentation within the proposal references the same topic number.
_____ 4. The Project Abstract and other content provided on the Proposal Cover Sheet contains no proprietary or classified information and is limited to the space provided.
_____ 5. The Technical Content of the proposal, including the Option (if applicable), includes the items identified in Section 3.4 of the solicitation.
_____ 6. The Proposal Cover Sheets and technical proposal is 25 pages or less in length. The Cost Proposal and Company Commercialization Report do not count against the 25 page limit. Pages in excess of this length will not be considered for review or award.
_____ 7. The Company Commercialization Report is submitted online in accordance with Section 3.5.d. This report is required even if the company has not received any SBIR funding
_____ 8. The proposal contains no type smaller than 10-point font size (except as legend on reduced drawings, but not tables).
CBD SBIR 10.1 Topic Index
CBD10-101 Low/No Power Detector for Organophosphate Nerve Agents
CBD10-102 Improved Formulations to Enhance Bio-availability for Antisense Therapeutics
CBD10-103 In vitro Models Suitable for High-throughput Screening of Drug Toxicities in Human
Tissues
CBD10-104 MEMS Lamellar Based Interferometer for the Detection of Toxic Chemicals
CBD10-105 Focal Plane Array Technology for Passive Hyperspectral Standoff Detection
CBD10-106 Residual Life Indicator for Adsorptive and Reactive Single-Pass Filtration Systems
CBD10-107 High Throughput Screening to Identify Enzyme Variants with Increased Affinity for
Organophosphorus Compounds
CBD10-108 High-Throughput Screening of Multiple Enzymes in Human Blood Relevant to
Organophosphate Poisoning
CBD10-109 Blood Brain Barrier Drug Delivery of Therapeutics for Chemical Warfare Agents
CBD10-110 Nanofluidic Sensor Platforms for THz-Frequency Spectroscopic Fingerprinting of Bio-
Molecules
CBD SBIR 10.1 Topic Descriptions
CBD10-101 TITLE: Low/No Power Detector for Organophosphate Nerve Agents
TECHNOLOGY AREAS: Chemical/Bio Defense
OBJECTIVE: Develop a small, portable, no to low power, shelf-stable, rapid-detection field device that can differentiate between organophosphate nerve agents.
DESCRIPTION: Organophosphate (OP) nerve agents are the most dangerous chemical warfare agents and therefore pose a significant threat. The OP toxicity arises from the inhibition of acetylcholinesterase (AChE) by the agents. Existing treatments only offer limited protection and often variability of effectiveness exists due to the type of agent that is inhibiting AChE. Hazard mitigation response can also be more effectively tailored if the exact OP agent is known. Therefore a method to rapidly identify the specific OP agent involved in an incident could significantly facilitate the administration of the most effective therapeutic and allow for appropriate measures of protection and decontamination, thereby increasing survival of the warfighter.
Existing fielded technologies are capable of detecting the presence of an OP however, these technologies are often large in size and require power making the systems challenging for fieldability, or require specific equipment and technical expertise for data collection and analysis, which incurs a high cost. A simple, small, robust, sensitive, low to no- power device capable of rapidly distinguishing between OP agents that is shelf-stable and can operate in a variety of conditions, could provide a new capability to the warfighter.
In order to provide such capability, novel methods of detection of chemical agent may need to be applied. For example, protein or enzyme based recognition of agents that can distinguish between various OPs could be explored. The goal of this topic is to first develop the science (e.g. biological recognition) to support such a broad agent recognizing device and later in the course of the program, develop a detector that achieves the goals mentioned in this topic. The end product should ideally be hand-held or smaller, no/low power, have a long shelf-life (greater than or equal to 5 years), remain stable under storage and operation conditions of high and low temperatures [cold (-27 deg F/-33 deg C) to hot (160 deg F/71 deg C)] and detect OPs in the low mg/m-2 range from a variety of surfaces within 30 min. This device should also be capable of differentiating between target chemicals in the presence of interfering substances, and have a clear, simple readout.
PHASE I: Develop the science behind the recognition elements needed in the detector for capability to differentiate between four OPs (1 live agent and 3 simulants/pesticides) in the presence of interferents and from a variety of sources (surfaces, solutions, etc). Should demonstrate stability and recognition activity under the above mentioned temperature conditions and have data to support potential for long term stability. In addition, the limitations of detection under these conditions (both high and low concentrations of agent) should be determined.
PHASE II: Develop a device capable of distinguishing between four OP nerve agents (VX, GA, GB, and GD) and three simulants/pesticides. This phase must include live OP agent testing. This device should meet the requirements listed above for operation under a variety of thermal conditions and shelf-life conditions. In addition, this device must run on low to no power and be hand-held or smaller.
PHASE III DUAL USE APPLICATIONS: In addition to the Department of Defense, the technology would applicable to the Department of Homeland Security, state and local HAZMAT response/mitigation teams, and agricultural pesticide control.
REFERENCES:
1. Kuca, K., Jun, D., and Bajgar, J., (2007), Currently used cholinesterase reactivators against nerve agent intoxication: comparison of their effectivity in vitro. Drug Chem Toxicol 30, 31-40.
2. Hill, H. H., Martin S.J., (2002), Conventional analytical methods for chemical warfare agents. Pure Appl. Chem. 74, 2281-2291.
3. Seto Y, K.-K. M., Kouichiro T, Ohsawa I, Matsushita K, Sekiguchi H, Itoi T, Iura K, Sano Y, Yamashiro S. (2005) Sensing technology for chemical-warfare agents and its evaluation using authentic agents. Sensors and Actuators B 108, 193-197.
KEYWORDS: detection, organophosphate, chemical agent
CBD10-102 TITLE: Improved Formulations to Enhance Bio-availability for Antisense Therapeutics
TECHNOLOGY AREAS: Chemical/Bio Defense
OBJECTIVE: This topic seeks proposals to enhance the ease of use and bioavailability of antisense therapeutics.
DESCRIPTION: Antisense oligonucleotides offer enormous potential as a new class of anti-infective therapeutics. These are typically polar, charged hydrophilic compounds with poor oral bio-availability; routes of administration for systemic use are limited to intravenous or subcutaneous injection. This severely limits their utility for prophylaxis of mass casualties in a biological contingency. The Transformational Medical Technologies Initiative (TMTI) program is seeking improved formulations to support the development and licensure of anti-sense therapeutics. The preferred formulation would be for oral use; however, inhalation or transdermal routes of administration may also be acceptable. Formulations should be stable at ambient temperature and minimize the potential for introduction of immunogenic materials. The product should have a shelf life of at least two years, preferably up to five years; other considerations include minimized bulk and ease of dispensing. Approaches to enhance bioavailability may include, but are not limited to, chemical modifications of the oligonucleotide backbone, the addition of carrier molecules, novel formulation chemistries, the use of liposomes or other colloidal delivery systems, and the use of nanoparticles or other particulate technologies.
PHASE I: Identify a suitable candidate formulation that demonstrates product stability; design a proof of concept / feasibility study to establish enhanced bio-availability in an animal model.
PHASE II: Optimize candidate formulation for maximum bio-availability; prepare non-GMP batches of anti-sense oligonucleotide formulation; conduct initial pharmacokinetic / pharmacodynamic studies in an animal model.
PHASE III: Conduct an in vivo proof of concept of study; plan completion of IND-enabling preclinical studies.
PRIVATE SECTOR COMMERCIAL APPLICATIONS: Dual use commercial applications include enhancement of bio-availability of difficult to administer prescription drug formulations throughout the pharmaceutical industry.
REFERENCES:
1. Raoof A, Chiu P, Ramtoola Z. Oral bioavailability and multiple dose tolerability of an antisense oligonucleotide tablet formulated with sodium caprate. J Pharm Sci 2004;93: 1493-1499.
2. Geary RS, Khatsenko O, Bunker K, et al. Absolute bioavailability of 2’-O-(2-methoxyethyl)-modified antisense oligonucleotides following intraduodenal installation in rats. J Pharmacol and Exper Therap 2001;296: 898-904.
3. Wang H, Cai Q, Xiaofei Z, et al. Antitumor activity and pharmacokinetics of mixed-backbone antisense oligonucleotide targeted to Ria subunit of protein kinas A after oral administration. PNAS 1999;96:13989-13994.
4. Gelperina S, Kisich K, Iseman M, Heifets L. The potential advantages of nanoparticle drug delivery systems in chemotherapy of tuberculosis. Am J respire Crit Care Med 2005;172: 1487-1490.
5. Aouadi, M, Tesz, G, et. al. Orally delivered siRNA targeting macrophage Map4k4 suppresses systemic inflammation. Nature, 30 April 2009; 458: 1180-1185.
KEYWORDS: antisense therapy, drug formulation, oral bioavailability, nanoparticles, antisense therapeutics
CBD10-103 TITLE: In vitro Models Suitable for High-throughput Screening of Drug Toxicities in
Human Tissues
TECHNOLOGY AREAS: Chemical/Bio Defense
OBJECTIVE: This topic seeks in vitro platforms to assess the toxicity and drug-to-drug interactions of therapeutic candidates in preclinical development. These platforms may include, but are not limited to, cell culture and tissue culture based systems and cell imaging.
DESCRIPTION: The Transformational Medical Technologies Initiative (TMTI) program seeks enhanced in vitro screens to assess the toxicity and drug-to-drug interactions in human tissue models. Advantages include the ability to screen compounds early in the drug development process, reducing the need for animal studies and minimizing late-stage toxicity failures. The preferred platforms should be well characterized and representative of tissue types that are relevant to toxicological end points (liver, lung, kidney, cardiovascular). The ideal platform should be adaptable for high-throughput screening and data acquisition. Such data should be amenable to integration into systems biology architecture to provide for the identification of multiple toxicity targets.
PHASE I: Identify a candidate human tissue model system; design a proof of concept study to establish feasibility for use in drug toxicity screens.
PHASE II: Develop a prototype system for toxicity screening; conduct initial proof of concept studies utilizing drugs with known toxicities and interactions as validation. Identify appropriate datasets (e.g., gene expression, proteomic, etc.) that will be required for toxicological analysis.
PHASE III: This phase will include the conduct of additional validation studies and the adaptation of the prototype system for high-throughput screening.
PRIVATE SECTOR COMMERCIAL APPLICATIONS: Dual use commercial applications include drug discovery applications throughout the pharmaceutical industry.
REFERENCES:
1. Andersen ME, Krewski, D Toxicity Testing in the 21st Century: Bringing the Vision to Life, Toxicol Sci 2009: 107: 324-330.
2. Clarke E, Pereira C, Chaney R, et al. Toxicity testing using hematopoietic stem cell assays. Regenerative Medicine 2007;2:947-956.
3. O’Brien PJ, Irwin W, Diaz D. High concordance of drug-induced human hepatotoxicity with in vitro cytotoxicity measured in a novel cell-based model using high content screening. Arch Toxicol 2006;80:580-604.
4. Allen D, Caviedes R, Cardenas A M, et al. Cell lines as in vitro models for drug screening and toxicity studies. Drug Development and Industrial Pharmacy 2005;31:757-768.
KEYWORDS: in vitro toxicity, drug development
CBD10-104 TITLE: MEMS Lamellar Based Interferometer for the Detection of Toxic Chemicals
TECHNOLOGY AREAS: Chemical/Bio Defense
OBJECTIVE: Develop a miniature, MEMS enhanced tunable spectrometer sensor based on a lamellar interferometer for the detection and identification of chemical agents and toxic industrial compounds.
DESCRIPTION: The Joint Services have the need for miniature, highly sensitive and yet highly specific sensor for detection of toxic industrial chemicals. Infrared absorption spectroscopy has proven to be a very useful tool in the detection and precise identification of airborne chemicals. Pattern recognition is used to compare the infrared spectrum of library molecules against the infrared spectra of airborne contaminants. Infrared spectrometers have rapid response and clear-down times, which provide utility in cloud tracking or dynamic monitoring experiments. A miniature monolithic spectrometer would be extremely small and consume very little energy. MEMS based spectrometers are also extremely rugged. The components within the MEMS device have an extremely low mass, which results in a very low inertia. Thus a well-designed MEMS device can survive massive accelerations. Also, the structural vibrational modes of low mass devices are at very high frequencies, which are far away from vibrations found in most environments. Thus there is little or no vibrational coupling to the external environment.
Currently most infrared spectrometer in use by the Joint Service Chemical and Biological defense community are based on the Michelson interferometer design. However, Michelson interferometers exhibit certain disadvantages in MEMS devices. Michelson interferometers suffer from low efficiency and limited spectral range primarily due to the use of a beamsplitter. Typically a Michelson interferometer has an efficiency near 25%. In contrast Lamellar grating interferometers have efficiencies near 100% and can operate over a wide spectral range. Unlike the Michelson interferometer, in which the radiation is divided at the beamsplitter, the lamellar grating interferometer uses the principle of wavefront division. It consists of two sets of parallel interleaved mirrors, one set that can move in a direction perpendicular to the plane of the front facet with the other set fixed. When a beam of collimated radiation is incident onto the grating, half of the beam will be reflected back from the front facet and half of the beam will be reflected from the back facet. By moving one set of mirrors, the path difference between the two beams can be varied and an interferogram can be generated.
In the past lamellar grating have been used mainly in the very far infrared region. It is difficult to fabricate a lamellar grating that works well at higher frequencies. With recent advances in microelectromechanical system technology, it is now possible to extend the advantages of a lamellar interferometer based sensor to shorter wavelengths. The goal of this program is to develop a miniature chemical sensor system based on the lamellar interferometer and to determine the utility of a lamellar based sensor within the chemical and biological defense community.
PHASE I: Design a lamellar based infrared chemical sensor. Modeling of sensitivity expected and the limiting source of noise should be completed.
PHASE II: Fabricate the lamellar interferometer based chemical sensor. Measure the sensitivity to selected simulants and establish the limiting noise source. Using a recognition algorithm of choice, determine the variance possible in wavelength and amplitude vs. false alarm rate.
PHASE III DUAL USE APPLICATIONS: There are many environmental applications for a small chemical detector/identifier. A rugged, asensitive and flexible chemical detector will benefit the manufacturing community by providing very finely tuned monitoring of chemical processes. Also first responders such as Civilian Support Teams and Fire Departments have a critical need for a rugged, relatively inexpensive but versatile and rugged sensor that can be transported to the field to test for possible contamination by CW agents.
REFERENCES:
1. Richard T. Hall, Dale Vrabec, and Jerome M. Dowling, “A High-Resolution, Far Infrared Double-Beam Lamellar Grating Interferometer”, Applied Optics, Vol. 5, Issue 7, pp. 1147-1158 (1966).
2. Omar Manzardo, Roland Michaely, Felix Schädelin, Wilfried Noell, Thomas Overstolz, Nico De Rooij, and Hans Peter Herzig, “Miniature lamellar grating interferometer based on silicon technology”, Optics Letters, Vol. 29, Issue 13, pp. 1437-1439 (2004).
3. Fook Siong Chau, Yu Du and Guangya Zhou, “A micromachined stationary lamellar grating interferometer for Fourier transform spectroscopy”, Journal of Micromechanics and Microengineering, Volume 18, number 2, pp. 025023 (2008).
4. F. Montiel and M. Nevière “Differential theory of gratings: extension to deep gratings of arbitrary profile and permittivity through the R-matrix propagation algorithm”, Journal of the Optical Society of America A: Optics, Image Science, and Vision, Volume 11, Issue 12, pages 3241-3250 (1994).
5. C. Ataman, H. Ürey, and S. O. Isikman, “A MEMS based visible-NIR Fourier transform microspectrometer”, Proceedings of the SPIE, the International Society for Optical Engineering, Volume 6186, volume 6186, pages 61860C.1-61860C.9 (2006).
6. Feiwen Lee, Guangya Zhou, Hongbin Yu, and Fook Siong Chau, “A MEMS-based resonant-scanning lamellar grating Fourier transform micro-spectrometer with laser reference system”, Sensors and Actuators A: Physical, Volume 149, Issue 2, Pages 221-228 (2009).
KEYWORDS: MEMS, Infrared Absorption Spectroscopy, Interferometer, Chemical Agent Detection, Toxic Industrial Compounds
CBD10-105 TITLE: Focal Plane Array Technology for Passive Hyperspectral Standoff Detection
TECHNOLOGY AREAS: Chemical/Bio Defense
OBJECTIVE: Develop methods that enable the production of low cost LWIR (8 to 12 micron) focal plane array technology specialized for use with chemical imaging sensors.
DESCRIPTION: The Joint Services have a need for passive standoff systems that detect and classify areas contaminated with chemical and biological vapors, aerosols, liquids and solids. Military spectroscopy systems exist that have been applied to the passive interrogation of CB aerosols and chemical vapors. These systems include passive Fourier Transform IR embodied in the Joint Services Lightweight Standoff Chemical Agent Detector (JSLSCAD) for the detection of chemical vapor clouds. Recently, hyperspectral imaging systems have shown great promise for the detection, identification and real time display of chemical vapor and aerosol clouds. However, deployment of hyperspectral technology is limited by the cost, yield, and reliability of the infrared focal plane technology used in these sensors.
The infrared focal plane array technology required for passive standoff detection of chemical and biological signatures is significantly different from the technology optimized for thermal imaging. The subdivision of far field spectral radiance into discrete spectral bands requires array technology with higher sensitivity, shorter integration times, and lower noise figures than is typically achieved with un-cooled technology. Similarly, the need for spectral response cutoff ranges in the 10 to 12 micron wavelength range to access important CB and toxic industrial material signatures exceeds the requirements for cooled thermal imaging devices while placing an additional burden on the cryocoolers needed to achieve requisite noise levels.
Current military demand for broadband thermal imaging systems have limited investment in technology to support hyperspectral imaging. The present topic addresses the need to develop this technology and reduce its cost.
PHASE I: Examine methods for producing infrared focal plane arrays that are designed for using in standoff chemical and biological sensors. Conduct a study to define the spectral response range, pixel size and format, and noise requirements for focal plane array technology to be used across a range of standoff hyperspectral imaging system technologies. Examine focal plane array (FPA) design constraints and imaging systems requirements to achieve designs that improve yield and quality while meeting cost and performance targets. Develop a manufacturing improvement plan for the production of the technology that identifies further research and development needed for this effort.
PHASE II: Fabricate prototype focal planes, and assess FPA performance at the device level. Develop and implement the optical, cryocooler, and electronics designs required to effectively utilize the focal plane array technology. Package the prototype focal planes in an integrated detector dewar cryocooler assembly (IDDCA) for use with a hyperspectral imager and assess the performance of the system. Determine and document improvements in the system.
PHASE III: The primary barrier to the broader use of hyperspectral technology for remote sensing of hazardous materials has been the technology cost, which is primarily driven by the cost of the focal plane technology. First responders such as civilian support teams, fire departments, and military post-blast reconnaissance teams have a critical need for a rugged and versatile and low cost sensor that can be transported to the field to test for possible CB contamination. This effort will facilitate the transition of the technology to those applications.
REFERENCES:
1. D.A. Scribner, M.R. Kruer, and J.M. Killiany, “Infrared focal plane array technology”, Proceedings of the IEEE, Volume 79, Issue 1, pages 66-85 (1991).
2. T. J. de Lyon, J. E. Jensen, M. D. Gorwitz, C. A. Cockrum, S. M. Johnson, and G. M. Venzor, “MBE growth of HgCdTe on silicon substrates for large-area infrared focal plane arrays: A review of recent progress”, Journal of Electronic Materials, Volume 28, Number 6, Pages 705-711 (June 1999).
3. Werner Gross, Thomas Hierl, and Max Schulz, “Correctability and long-term stability of infrared focal plane arrays”, Optical Engineering, Volume 38, Issue 5, pages 862-869 (1999).
4. E. P. G. Smith, L. T. Pham, G. M. Venzor, E. M. Norton, M. D. Newton, P. M. Goetz, V. K. Randall, A. M. Gallagher, G. K. Pierce, E. A. Patten, R. A. Coussa, K. Kosai, W. A. Radford, L. M. Giegerich, J. M. Edwards, S. M. Johnson, S. T. Baur, J. A. Roth, B. Nosho, T. J. De Lyon, J. E. Jensen, and R. E. Longshore, “HgCdTe focal plane arrays for dual-color mid- and long-wavelength infrared detection”, Journal of Electronic Materials, Volume 33, Number 6, Pages 509-516 (June, 2004).
5. M. R. Kruer, D. A. Scribner, and J. M. Killiany, “Infrared focal plane array technology development for Navy applications”, Optical Engineering, Volume 26, Pages 182-190 (March 1987).
6. William E. Tennant, C. A. Cockrum, J. B. Gilpin, M. A. Kinch, M. B. Reine, and R. P. Ruth, “Key issues in HgCdTe-based focal plane arrays: An industry perspective”, Journal of vacuum science and technology. B, Volume 10, Issue 4, Pages1359-1369 (July 1992).
7. P. Ferret, J. P. Zanatta, R. Hamelin, S. Cremer, A. Million, M. Wolny, and G. Destefanis, “Status of the MBE technology at leti LIR for the manufacturing of HgCdTe focal plane arrays”, Journal of Electronic Materials, Volume 29, Number 6, Pages 641-647 (June, 2000).
8. Jose M. Arias, Majid Zandian, John G. Pasko, Jagmohan Bajaj, Lester J. Kozlowski, William E. Tennant, and Roger E. DeWames, “Molecular beam epitaxy (MBE) HgCdTe infrared focal plane array (IRFPA) flexible manufacturing”, Proceedings of SPIE, 1994, Volume 2274, pages 2-16 (1994).
KEYWORDS: Chemical detection, chemical identification, hyperspectral, focal plane, infrared spectrum, detector array
CBD10-106 TITLE: Residual Life Indicator for Adsorptive and Reactive Single-Pass Filtration
Systems
TECHNOLOGY AREAS: Chemical/Bio Defense
OBJECTIVE: Develop a generic residual life indicator that is able to probe the degradation in physical adsorption and reactive capacity arising from battlefield contaminants and exposure to the elements.
DESCRIPTION: Current air purification systems that provide breathable air typically employ highly adsorptive and reactive materials, such as impregnated activated carbon like ASZM-TEDA (Activated Carbon, Impregnated with Copper, Silver, Zinc, Molybdenum, and Triethlyenediamine), for the removal of toxic chemicals. Although such sorbent media initially provide extensive removal capabilities against a wide range of chemicals, the ability to filter toxic chemicals via chemical reaction decreases over time due to exposure to temperature, humidity, battlefield contaminants, and other common pollutants. The objective of this research is to develop, fabricate, and evaluate a novel system for assessing the residual life of sorbent-based air purification systems.
Although residual life indicators have been previously developed, they are either too bulky or detect specific gases/vapors eluting through the filter bed. This research seeks to develop a generic residual life indicator that is able to probe the degradation in physical adsorption and reactive capacity arising from battlefield contaminants and exposure to the elements. This research does not seek to develop technologies for detection of specific gases/vapors, such as toxic industrial chemicals (TICs) or chemical warfare agents (CWAs), but rather to assess the degradation in impregnants due to contaminants. At this time, proposals that focus on dosimeters to measure battlefield contaminants, dyes to induce colorimetric changes, and other methods that do not directly probe impregnants or adsorption capacity are not requested. Furthermore, methods using pulse gases and parallel elements are not requested in this topic. The residual life indicator should be capable of easily integrating into current Collective Protection (ColPro) systems such as the M98 and M48A1 filters. Furthermore, the technology should be able to report assess and report remaining filter life.
PHASE I: Research should focus on an initial feasibility analysis of the residual life indicator technology to demonstrate proof-of-concept. Specific objectives are to demonstrate the ability to quantitatively measure loss of physical adsorption capacity and impregnant activity of ASZM-TEDA due to temperature, humidity, and battlefield contaminants. At least 3 battlefield contaminants representing a variety of poisoning mechanisms must be used to evaluate the technology. The proposer must include plans for integration into full-scale, fielded systems.
PHASE II: Phase II should first focus on the development of breadboard concepts of the technology followed by field-ready prototypes. Breadboard concepts will be evaluated for synergistic effects of the residual life indicator technology integrated into packed beds of ASZM-TEDA. Field-ready prototypes will be constructed and evaluated in a relevant environment for an extended period of time.
PHASE III: Field-ready prototypes will be constructed and evaluated in a relevant environment for an extended period of time. Technologies will be evaluated in shipboard, fixed-site, and vehicle applications. Commercial filtration applications will also benefit from this technology.
REFERENCES:
1. Peterson, G. W.; Jones, P.; Keller, J.; Weller, E. “Contaminant-Induced Degradation of ASZM-TEDA Reactivity.” Limited Distribution. ECBC-TR-629. AD-B341 218. May 2008.
2. Peterson, G. W.; Jones, P.; Karwacki, C. J. “Cyanogen Chloride Filtration Performance of ASZM-TEDA from Fixed Site Filter Systems.” Limited Distribution. ECBC-TR-445. AD-B321 134. August 2006.
3. Deitz, V. R.; Puhala, R. J.; Stroup, D. B.; Dickey, G. F. “Influence of Atmospheric Weathering on the Performance of Whetlerite.” AD A118519. August 1982.
4. Rossin, J.; Petersen, E.; Tevault, D.; Lamontagne, R.; Isaacson, L. “Effects of Environmental Weathering on the Properties of ASC-Whetlerite.” Carbon Vol. 29, No. 2, pp. 197-205, 1991.
5. Rose-Pehrsson, S. L.; Williams, Monica L. “Integration of Sensor Technologies into Respirator Vapor Cartridges as End-of-Service-Life Indicators: Literature and Manufacturer’s Review and Research Roadmap.” ADA434905. 06 May 2005.
6. Friday, D.; Shrewsbury, M.; Deibert, S.; Peterson, G. W. “A Residual Life Indicator (RLI) for Physical Adsorption Capacity of Nuclear, Biological, and Chemical Filters. Part III. A Novel RLI Design for Collective Protection Demonstrated Using Breakthrough and Chemical Pulse Data.” ECBC-TR-658. AD-A491 483. November 2008.
KEYWORDS: Residual life indicator, activated carbon
CBD10-107 TITLE: High Throughput Screening to Identify Enzyme Variants with Increased Affinity
for Organophosphorus Compounds
TECHNOLOGY AREAS: Chemical/Bio Defense
OBJECTIVE: Design and develop a robust screening process for use with large panels of enzymes to identify variants with higher affinity (reduced KM values) for organophosphorus substrates.
DESCRIPTION: The Department of Defense (DoD) has a need for enzymes with enhanced catalytic activity against organophosphorus (OP) nerve agents to be used as in vivo pretreatments for nerve agent exposure. Two parameters define the catalytic efficiency of an enzyme: kcat, which quantifies the maximum rate at which an enzyme can promote catalysis of a given substrate, and KM, which can be used as a rough approximation of the binding affinity of an enzyme for a given substrate. To afford in vivo protection against poisoning by OP compounds, a catalytic scavenger must be capable of reducing the concentration of an OP in the blood to sublethal levels before the poison distributes to peripheral and central sites of toxic effect. While candidate scavenger enzymes with kcat values predicted to be sufficient for protection have been identified, to date none of the candidates have KM values that approximate the predicted lethal concentration of OPs in the blood. To be effective, a scavenger enzyme is predicted to require a KM value at least as low as the lethal concentration of OPs in the blood (less than or equal to 10 micromolar for most OPs). Molecular biology techniques allow for the generation of large libraries of candidate proteins, but currently there is no method for high throughput screening of the resulting variants to identify those with reduced KM (increased affinity) values.
Thus, development of a system or process for high throughput screening of enzymes to identify those with increased affinities is necessary. The technology/process must be sensitive enough to screen the activity of small amounts (nanogram quantities) of candidate enzymes, must operate under physiologically relevant conditions (aqueous, pH 7.4, 20-40°C), must be either a closed system (to contain both fluid and vapor) or amenable to use under the engineering controls necessary for safe use of dilute quantities of OP nerve agents, and must be sufficiently robust to withstand routine cleaning with highly basic and alcoholic solutions. The resulting process will provide the DoD with an improved capability to develop novel scavengers of OP nerve agents, and will provide improved warfighter performance in chemically contaminated environments while maintaining operational tempo and reducing logistical dependence on MOPP gear.
PHASE I: Develop a high throughput screening process/device for the identification of enzyme variants with reduced KM values for OP compounds. Demonstrate proof of concept and feasibility using candidate OP binding enzymes. Pesticides or nerve agent simulants are suggested at this stage to reduce risk while maintaining both the sensitivity and the applicability of the approach to nerve agent research. The process must be robust, capable of screening large numbers of variants, and able to be relocated or otherwise established at the site of use.
PHASE II: Process/device will be further developed, refined, and validated using OP pesticides and/or nerve agent simulants, followed by validation with bona fide OP nerve agents. Testing with OP nerve agents is required during Phase II development; as small business firms are unable to handle and utilize chemical weapon agents (CWAs), a testing facility approved to utilize these materials must be identified. Variants of candidate enzymes to be tested using the process/device may be designed and produced by the small business offeror or will be supplied by the United States Army Medical Research Institute of Chemical Defense (USAMRICD), or both. Candidates with sufficiently reduced KM values (less than or equal to 10 micromolar) will be expressed in large scale by the offeror for chemical agent testing both in vitro and in vivo for efficacy as nerve agent bioscavengers.
PHASE III: Advanced in vivo safety and efficacy testing in one or more FDA-approved animal models will be performed. Best candidate enzyme(s) for high yield commercial production and formulation as a candidate for submission to the FDA will be identified for assessment to be granted IND status as a pre-exposure treatment against OP nerve agent intoxication.
REFERENCES:
1. Masson, P, Nachon, F. et al. (2008). "A collaborative endeavor to design cholinesterase-based catalytic scavengers against toxic organophosphorus esters." Chem Biol Interact 175(1-3): 273-280.
2. Ashani Y, Pistinner S. (2004). "Estimation of the upper limit of human butyrylcholinesterase dose required for protection against organophosphates toxicity: a mathematically based toxicokinetic model." Toxicol Sci. 77(2): 358-367.
3. Saxena A, Sun W, Luo C, Myers TM, Koplovitz I, Lenz DE, Doctor BP. (2006). “Bioscavenger for protection from toxicity of organophosphorus compounds.” J. Mol. Neurosci. 30(1-2): 145-148.
4. Lenz DE, Yeung D, Smith JR, Sweeney RE, Lumley LA, Cerasoli DM. (2007), “Stoichiometric and catalytic scavengers as protection against nerve agent toxicity: a mini review.” Toxicology 233(1-3):31-39.
KEYWORDS: bioscavenger, organophosphorus nerve agent, enzyme, high throughput screening
CBD10-108 TITLE: High-Throughput Screening of Multiple Enzymes in Human Blood Relevant to
Organophosphate Poisoning
TECHNOLOGY AREAS: Chemical/Bio Defense
OBJECTIVE: The objective is to develop a high-throughput minimally invasive blood protocol to measure the human profile of enzymes that are affected by organophosphate chemical warfare agents (OP-CWA). The complete assay would, at least, measure in whole blood acetylcholineste-rase (AChE), butyrylcholinesterase (BChE), and paraoxonase (PON1). The primary purpose of pre-screening the warrior for these enzymes is to ensure that a full complement is on-board to provide natural protection to OP-CWAs. This diagnostic tool will allow the screening of individ-uals with reduced enzyme activities and preclude them from entering a conflict zone where OP exposure might be present (ref #1).
DESCRIPTION: Nerve agents (OPs) are extremely toxic chemicals that cause effects by inhi-biting the enzyme acetylcholinesterase (AChE), allowing excess acetylcholine to accumulate. This excess neurotransmitter then produces overstimulation and causes hyperactivity in muscles, glands, and nerves, and ultimately death at high doses of OPs. The nerve agents are GA (tabun), GB (sarin), GD (soman), GF, and VX. Pesticides and surrogates such as diisopropylfluorophos-phate (DFP) exhibit similar effects. Their routes of exposures are skin, respiratory system, and GI.
The current screening performed by many clinical and research laboratories use the colorimetric Ellman assay based on the hydrolysis of acetylthiocholine. CHPPM (US Army Center for Health Promotion and Preventive Medicine, ref #2) uses a slower and older delta pH method (Michel method) to screen more than 35,000 DOD personnel each year. Unlike this single enzyme con-ventional screening of red blood cell acetylcholinesterase, the full complement of a warrior’s or first responder’s OP sensitive enzymes should be quantified to provide a complete analysis of their suitability to potential OP exposure. Simultaneous AChE and BChE has been developed (ref #3). To permit high-throughput screening in the laboratory and ultimately field deployment, the multiple enzyme assay should not rely on the addition of selective inhibitors, use a single non-invasive blood collection technique (e.g. a small volume of blood and be minimally invasive), not be unusually labor intensive or otherwise complicated, and produce results rapidly. These systems will provide the army and general public health clinics with units capable of screening and confirming exposure of soldiers/agricultural workers/first responders to chemical warfare agents/pesticides/or other such toxic chemicals. In addition, these systems are critical in the discovery of new therapeutic agents for prophylaxis, neuroprotection, and treatment of chemical warfare agent poisoning.
Screening of several enzymes is proposed to provide a comprehensive description of the warrior’s capacity for low level OP exposure with minimal consequences. First, the enzymes AChE and BChE are the direct targets of OP-CWA. However currently, only AChE is measured for baseline levels. AChE is necessary for sustaining proper neuromuscular transmission. BChE (plasma/serum cholinesterase or pseudocholinesterase) has an unknown function, but is potent bioscavenger for OPs, likely protecting AChE from OP-induced inhibition. Absence of BChE, while rare, or low levels of BChE, would render the warrior more susceptible to OP poisoning, those individuals with reduced levels of the enzyme exhibit no deleterious effects (ref #4). The second class of enzymes, including CE and PON1, provides protection to organophosphate com-pounds such as OP nerve agents and pesticides by hydrolyzing them to forms that are then ex-creted. Notably, different types (isotypes) and amounts of these enzymes dramatically differ in their ability to neutralize OPs. Thus, the different responses of warriors and civilians to OPs, e.g., Gulf War illness or victims of the Tokyo subway incident in 1995, respectively, likely reflects the different types and activities of BChE, CE, and PON1 present in the blood and tissues.
PHASE I: Design, validate, and demonstrate the efficacy and robustness of a quantitative sys-tem for screening human unprocessed whole blood for, at a minimum, AChE, BChE, and PON1 levels (ref #5. Other enzymes under consideration may be the esterase activity of albumin, but not CE in humans in contrast to rodents (see ref #5). The detection assay should be able to be ported to clinical laboratory instrumentation. The effort includes evaluation of the combined as-say methodology and techniques for reliably obtaining minimal blood volumes and stor-ing/preserving such blood samples (to mimic time blood would be stored prior to testing).
PHASE II: Extend the technology developed in Phase I to fully automated equipment with for-ward thinking for an additional format that would be hand-held and field deployable. Demon-strate the reliability, detection limits, accuracy, precision, and linearity of the clinical laboratory methodology for at least AChE, BChE, PON1 enzymes. Test the assays with samples treated ex-vivo under a variety of conditions with OP stimulants such as DFP and pesticides, and eventually with CWAs. The method will also evaluate and sample blood from a varied human population to provide normal enzyme distributions.
PHASE III DUAL USE COMMERCIALIZATION: The public and agricultural workers face health risks associated with organophosphates during the use of commercially available pesti-cides, as do first responders to a terrorist attack. More than 23,000 emergency room visits per year in the United States can be accounted for by pesticide poisoning. The method would enable screening of the public for anesthesiology sensitivity (potentially fatal outcome), the result of a mutant BChE occurring in about 10% of the population. The technology developed for screening of organophosphate exposed soldiers would have direct applicability to the screen-ing/conformation of the public exposed to pesticides as described above and the public exposed to nerve agent as in the 1995 Tokyo subway incident. The possibility exists that terrorists may use this type of chemical agent at sporting events or other such gatherings with large number of civilian populations
REFERENCES:
1. Examining Possible Causes of Gulf War Illness RAND Policy Investigations and Reviews of the Scientific Literature, (and documents therein); HTTP://WWW.RAND.ORG/PUBS/RESEARCH_BRIEFS/RB7544/INDEX1.HTML (last accessed 19 June 2009).
2. TB MED 590; Red Blood Cell-Cholinesterase Testing and Quality Assurance, at http://chppm.com/ (last accessed 19 June 2009).
3. Haigh JR, Lefkowitz LJ, Capacio BR, Doctor BP, Gordon RK. Chem Biol Interact. 2008 Sep 25;175(1-3):417-20. Advantages of the WRAIR whole blood cholinesterase assay: comparative analysis to the micro-Ellman, Test-mate ChE, and Michel (DeltapH) assays.
4. Lockridge O, Masson P. Neurotoxicology. 2000 Feb-Apr;21(1-2):113-26. Pesticides and sus-ceptible populations: people with butyrylcholinesterase genetic variants may be at risk.
5. Li B, Sedlacek M, Manoharan I, Boopathy R, Duysen EG, Masson P, Lockridge O. Biochem Pharmacol. 2005 Nov 25;70(11):1673-84. Butyrylcholinesterase, paraoxonase, and albumin este-rase, but not carboxylesterase, are present in human plasma.
6. Examining Possible Causes of Gulf War Illness RAND Policy Investigations and Reviews of the Scientific Literature, (and documents therein); HTTP://WWW.RAND.ORG/PUBS/RESEARCH_BRIEFS/RB7544/INDEX1.HTML (last accessed 19 June 2009).
7. Makhaeva GF, Malygin VV, Strakhova NN, Sigolaeva LV, Sokolovskaya LG, Eremenko AV, Kurochkin IN, Richardson RJ. Hum Exp Toxicol. 2007 26(4):273-82. Biosensor assay of neuropathy target esterase in whole blood as a new approach to OPIDN risk assessment: review of progress.
8. Quistad GB, Klintenberg R, Casida JE. Toxicol Sci. 2005 86(2):291-9. Blood acylpeptide hydrolase activity is a sensitive marker for exposure to some organophosphate toxicants.
9. Casida JE, Nomura DK, Vose SC, Fujioka K. Chem Biol Interact. 2008 175(1-3):355-64.
Organophosphate-sensitive lipases modulate brain lysophospholipids, ether lipids and endocannabinoids.
KEYWORDS: Mass screening, automated laboratory analysis, organophosphate chemical warfare agents, human, acetylcholinesterase (AChE), butyrylcholinesterase, (BChE) carboxylesterase (CE), paraoxonase (PON1), albumin esterase.
CBD10-109 TITLE: Blood Brain Barrier Drug Delivery of Therapeutics for Chemical Warfare Agents
TECHNOLOGY AREAS: Chemical/Bio Defense
OBJECTIVE: The objective is to develop general carrier(s) for delivery of therapeutics to the central nervous system. Chemical warfare agents (CWAs), specifically organophosphate (OP) nerve agents, readily penetrate the CNS where they inhibit cholinesterases. The FDA approved reactivating therapeutic pralidoxime chloride (2-PAM), however, is excluded by the blood brain barrier (BBB). Therefore, therapeutics that can rapidly cross the BBB and penetrate to the CNS are needed to treat the OP-exposed warfighter.
DESCRIPTION: Nerve agents (OPs) are extremely toxic chemicals that cause effects by inhibiting the enzyme acetylcholinesterase (AChE), allowing excess acetylcholine to accumulate, yielding a cholinergic crisis, and ultimately death at high doses of OPs. Current treatment for acute organophosphate poisoning includes a combined administration of a cholinesterase reactivator (oxime, FDA approved 2-PAM), a muscarinic receptor antagonist (atropine) and an anticonvulsant (diazepam) (ref#1). However, oximes, AChE reactivating therapeutics such as 2-PAM, HI-6, MMB-4, or obidoxime, are all charged small molecules that do not penetrate the blood-brain barrier. Delivery of the oxime to the brain to achieve therapeutically beneficial concentrations should reactivate CNS AChE, limit the accumulation of excessive ACh and the consequent negative sequelae including status epilepticus (SE) (ref #2). SE is prolonged seizure leading to permanent brain damage, documented in animals and more recently in man in survivors from the 1995 Tokyo subway incident with the OP sarin (ref #3).
The greatest need for improvement in cholinesterase reactivation is the delivery of reactivator to the brain. Traditionally, drug companies have tried to produce uncharged lipophilic compounds as CNS penetrating drugs because they have the greatest probability of getting across the BBB. However, novel potentially more efficacious oximes being developed are generally charged molecules that are unlikely to penetrate the BBB (ref #4). Thus, various transport systems (ref #5) such as nanoparticles, liposomes, cyclodrexins, and oxime derivatives with lipophylic or active transport directed moieties need to be developed to carry a payload of any oxime, FDA approved today or tomorrow’s more efficacious oxime, across the BBB in a rapid manner to reactivate CNS AChE. The result of this effort will be the restoration of effective neural transmission in the CNS using oximes in a carrier that can penetrate the BBB and resolving a cholinergic crisis.
PHASE I: A) Utilize creativity and innovation to design, validate, and demonstrate an in vitro blood brain barrier model to be used to evaluate formulations of oxime carriers. B) Test at least two formulations of carriers for incorporation and stability of oxime and penetration of the BBB model at therapeutic doses and at relevant rates to prevent long term brain damage. Oxime to in-corporate is 2-PAM, the FDA approved therapeutic for OP/pesticide exposure.
PHASE II: 1. Extend the novel technology developed in Phase I to testing of additional BBB penetrating oxime carriers. 2. Evaluate penetration and therapeutic efficacy of the down-selected oxime carrier in an animal model of OP poisoning (e.g., rat, guinea pig). For example, therapeutic efficacy in the CNS could be quantified by a) reactivation of CNS AChE and b) protection afforded for SE using EEG radiotelemetry and brain histopathology (ref #6). While CWAs for testing are highly recommended, suitable surrogates for OPs such as diisopropylfluorophos-phate (DFP) at relevant doses to invoke equivalent OP poisoning can be considered.
PHASE III: The lead candidate oxime carrier will be evaluated in two animal species, in collaboration with a biotechnology company, for efficacy against chemical warfare agents. The final stage will require initiation of clinical trials. Due to the potential commercial as well as DoD use, a commercialization strategy will be required.
PHASE III DUAL USE COMMERCIALIZATION: The public and agricultural workers face health risks associated with OPs during the use of commercially available pesticides, as do first responders to a terrorist attack (ref #7). More than 23,000 emergency room visits per year in the United States can be accounted for by pesticide poisoning. The BBB penetrating oxime carrier would provide a new CNS therapeutic for OP poisoning and replacement for the oxime 2-PAM, which is excluded from the brain.
REFERENCES:
1. Newmark, J., 2004a: Therapy for nerve agent poisoning, Arch. Neurol., 61(5), 649-652.
2. Carpentier, P., Foquin, A., Rondouin, G., Lerner-Natoli, M., De Groot, D., and Lallement, G., 2000: Effects of atropine sulphate on seizure activity and brain damage produced by soman in guinea-pigs: EcoG correlates of neuropathology, Neurotoxicol., 21(4), 521-540.
3. Yamasue, H., Osamu, A., Kasai, K., Motomu, S., Iwan¬ami, A., Haruyasu, Y., Tochigi, M., Ohtani, T., Rog¬ers, M., Sasaki, T., Aoki, S., Kato, T., and Kato, N., 2007: Human brain structural change related to acute single exposure to sarin, Ann. Neurol., 61, 37-46.
4. Bajgar, J., Fusek, J., Kuca, K., Bartosova, L., and Jun, D., 2007: Treatment of organophosphate intoxication using cholinesterase reactivators: facts and fiction, Mini Rev. Med. Chem., 7(5), 461-466.
5. Denora N, Trapani A, Laquintana V, Lopedota A, Trapani G. Recent advances in medicinal chemistry and phar-maceutical technology--strategies for drug delivery to the brain. Curr Top Med Chem. 2009;9(2):182-96.
6. Tetz, L. M., Rezk, P. E., Ratcliffe, R. H., Gordon, R. K., Steele, K. E., and Nambiar, M. P., 2006: Development of a rat pilocarpine model of seizure/status epilepticus that mimics chemical warfare nerve agent exposure, Tox-icol. Ind. Health, 33(6), 255-266.
7. Marrs, T. C., Rice, P., and Vale J. A., 2006: The role of oximes in the treatment of nerve agent poisoning in civi-lian casualties, Toxicol. Rev., 25(4), 297-323.
KEYWORDS: Liposomes, cyclodextrins, nanoparticles, oximes, blood brain barrier, status epilepticus, cholinesterases, reactivators
CBD10-110 TITLE: Nanofluidic Sensor Platforms for THz-Frequency Spectroscopic Fingerprinting
of Bio-Molecules
TECHNOLOGY AREAS: Chemical/Bio Defense
OBJECTIVE: Design, fabrication, and demonstration of an electrophoretic capillary nanofluidic integrated sensor platform effective for the label-less "THz-frequency spectroscopic fingerprinting" of biological molecules in detection, identification, and classification applications.
DESCRIPTION: Terahertz (THz) frequency sensing and imaging has been an area of growing interest within the scientific community for many years and the U.S. Department of Defense (DoD) has long supported THz-related research topics that have relevance to defense and security applications [1]. In particular, the U.S. Army and the U.S. Defense Threat Reduction Agency (DTRA) have supported numerous investigations into the utility of THz spectroscopy for the detection and characterization of biological materials and agents. While much theoretical and experimental evidence has been generated to suggest that many biological macromolecules (e.g., DNA, proteins, and polysaccharides) display unique THz absorptive signatures that are useful for detection and identification, a number of challenging phenomenology factors (e.g., weak signatures, sample stability, etc.) have prevented the elevation of THz-based capabilities to levels that exist in the infrared region [2]. However, it has been demonstrated recently that nanofabrication of extremely small fluidic channels can be used for the direct manipulation and effective analysis of bio-molecules such as nucleic acids, proteins, enzymes, and other species. This work (see Ref. [3]) which was supported by a prior CB SBIR project clearly documents the importance of controlling the orientation of the molecular targets in achieving highly repeatable spectral characteristics, and produced very sharp THz spectral signatures (i.e., 10 GHz) that rival any ever reported for solids or liquids at room temperature. These results, which were obtained by a first generation THz-compatible sensing platform that provided for only partial injection and control of bio-molecules within a large number of nanofluidic channels, illustrate the significant benefits of designing and developing more elaborate integrated lab-on-a-chip THz sensors. More specifically, these results motivate the implementation and refinement of state-of-the-art microfluidic techniques [4] for sample injection, processing and control into a THz-compatible spectroscopic sensor platform that will provide for highly sensitive and repeatable fingerprinting of bio-molecules. Here nanofabrication will be applied to produce the required nanoscale portals, control structures and channels useful for the introduction, stretching and processing of bio-molecules (e.g., such as genomic DNA) and their accurate spectroscopic characterization. In order to uniformly stretch and/or process chain-like DNA molecules, the dimensions of the nanofluidic structures should be of the order or smaller than the persistence length of double stranded DNA (i.e., 50-nm to 100-nm). To sufficiently enhance the interaction of the THz radiation with the absorption signature it is expected that the bio-chip platforms must possess high-density arrays in excess of 10-million nanofluidic channels within the THz spectroscopic window.
The goal of this project is the development and demonstration of a functioning prototype device that will: (1) completely validate the effectiveness of nanofluidic sensor platforms for the label-less "THz-frequency spectroscopic fingerprinting" of biological molecules; and, (2) enable a detailed assessment of THz-based detection, identification, and classification of biological materials and agents. A successful technology demonstration will represent a major enabler to biological (and possibly chemical) sensing and monitoring in defense, security, biomedical, pharmaceutical, food quality, and environmental relevant scenarios for civilian, military, government, and commercial sectors around the world.
PHASE I: The Phase I effort of the program should focus on the specification of the required structural and electronic/photonic features along with the development of a design for a prototype nanofluidic sensor platform that will allow for the acquisition of THz-frequency spectroscopic features from bio-molecules (e.g., DNA, RNA, etc.). Requirements for the combined system include, but may not be limited to: capabilities for injecting and/or preprocessing of molecular targets; methodologies for transporting and/or positioning molecules (e.g., electro-kinetic motion mechanisms, such as, capillary electrophoretic diffusion control) within a properly defined spectroscopic window; technology components for executing transmission and/or reflection spectroscopic techniques to maximize the optical efficiency and THz radiation interaction with the molecular targets. This initial phase should also include preliminary experimental investigations for handling bio-targets, along with the electrical characterization of the relevant platform and bio-target materials.
PHASE II: The Phase II effort of the program should implement, test and refine (if necessary) a completely functioning nanofluidic sensor platform. The research and development work should include an extensive assessment of the prototype’s effectiveness in the THz-frequency spectroscopic fingerprinting of relevant bio-materials and/or bio-agent targets. The prototype nanofluidic sensor platform should be package in a form factor that readily interfaces with commercially available THz spectroscopy systems. The sensor system should afford stand-alone operation such as the sample injection and fluidic and electrophoretic control is self-sustained in one enclosure to facilitate a transparent operation by the user. It is strongly suggested that spectroscopic characterization studies be executed with U.S. Army or DoD research center or laboratory that possesses expertise in bio-materials/agents production, handling and general characterization capabilities.
PHASE III DUAL USE APPLICATIONS: Once an effective prototype sensor is achieved, the unit will be amenable to numerous applications areas related to biological sensing and characterization. Specifically such a nanofluidic THz sensor will have relevance to scientific studies on biological materials and structures, to the detection and identification of biological threats, to medical diagnostics of biological induced diseases, to the monitoring of commercial consumables for biological contamination, just to name a few possibilities.
REFERENCES:
1. “Terahertz Frequency Sensing and Imaging: A Time of Reckoning Future Applications?” Dwight Woolard, et. al., Proceedings of the IEEE, 93, pp. 1722-1743 (2005).
2. “Reliability Analysis of THz Characterization of Modified and Unmodified Vector Sequences,” Tatiana Globus, et. al., accepted IEEE Sensors Journal (2009).
3. “Narrow THz Spectral Signatures through an RNA Solution in Nanofluidic Channels,” Elliott Brown, et. al., accepted IEEE Sensors Journal (2009).
4. “Purification of Nucleic Acids in Microfluidic Devices,” Jian Wen, et.al., Analytical Chemistry, Volume 80 , Issue 17, pp 6472–6479 (2008).
KEYWORDS: nanofluidic sensor platform, terahertz frequency spectroscopy, spectral fingerprinting, biological molecules, biological agents