CHEMICAL AND BIOLOGICAL DEFENSE PROGRAMFY18.1: Small Business Innovation Research (SBIR)

Component Specific Proposal Submission Instructions

The approved FY18.1 topics included in the Chemical and Biological Defense (CBD) Small Business Innovation Research (SBIR) Program are listed below. Offerors responding to this Announcement must follow all general instructions provided in the Department of Defense (DoD) SBIR Program Broad Agency Announcement. Specific CBD SBIR requirements that add to or deviate from the DoD SBIR Program Announcement are provided below with references to the appropriate section of the DoD SBIR Announcement.

General Information

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 Assistant Secretary of Defense for Nuclear, Chemical and Biological Defense 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 portfolio of the Chemical and Biological Defense Program. Technologies developed under the CBD 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 (S&T) programs and initiatives are improving defensive capabilities against Chemical and Biological Weapons of Mass Destruction. The CBD SBIR portion of the CBD S&T Program is managed by the JSTO-CBD and is not part of the DTRA SBIR Program.

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.  Numerous factors continually influence the program and its technology development priorities.  Improved defensive capabilities are essential in order to mitigate the impact of Chemical and Biological Threats. The U.S. military requires the most up-to-date and state-of-the-art equipment and instrumentation to permit our Warfighters to detect to warn and avoid contamination, if possible, and to be able to sustain operations in a potentially compromised environment.  Further information regarding the DoD Joint Chemical and Biological Defense Program is available at the DoD Counter-proliferation and Chemical Biological Defense homepage at http://www.acq.osd.mil/cp.

The overall objective of the CBD SBIR Program is to identify, obtain and provide technical solutions from small businesses and 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 management, modeling, simulation, and analysis; medical pre-treatments (e.g., vaccine development and delivery); medical diagnostics and disease surveillance; and medical therapeutics (chemical countermeasures and biological countermeasures).

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Submitting Your Phase I CBD SBIR Proposal

The entire SBIR proposal submission (consisting of a Proposal Cover Sheet, the Technical Volume, Cost Volume, and Company Commercialization Report) must be submitted electronically through the DoD SBIR/STTR Proposal Submission system located at https://sbir.defensebusiness.org/.

The Proposal Technical Volume must be no more than 20 pages in length. The Cover Sheet, Cost Volume and Company Commercialization Report do not count against the 20-page Proposal Technical Volume page limit. Pages in excess of this length will not be evaluated and will not be considered for review. The proposal must not contain any type smaller than 10-point font size (except as legend on reduced drawings, but not tables).

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 information in these sections. Classified information is never permitted to be included in any SBIR proposal.

The CBD SBIR Program will fund a Phase I proposal up to $150,000 for a six-month period of performance (independent of Discretionary Technical Assistance funding, if requested). The CBD SBIR Program no longer uses a Phase I Option period and will not accept any Phase I proposals that include an option period. Additionally, the CBD SBIR Program will not accept Phase I proposals which exceed $150,000 (exclusive of Discretionary Technical Assistance; see below).

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. The offeror must be responsive to the topic requirements, as solicited.

Proposals not conforming to the terms of this Announcement, and any unsolicited proposals, will not be considered. Awards are subject to the availability of funding (subject to the Congressional Budget process) in addition to successful completion of contract negotiations. Note the Chemical and Biological Defense Program is not responsible for any funds expended by the offeror prior to contract award.

CBD Program Phase II Proposal Guidelines

Phase II is the demonstration of the technology that was found feasible in Phase I. Phase I awardees may submit a Phase II proposal without invitation; however, it is strongly encouraged that a Phase II proposal not be submitted until sufficient Phase I progress can be evaluated and assessed based on results of the Phase I proof-of-concept/feasibility study Work Plan and no earlier than a recommended five months from date of contract award. All Phase II proposal submissions must be submitted electronically through the DoD SBIR/STTR Proposal Submission system at https://sbir.defensebusiness.org/. At the DoD SBIR Proposal Submission Web site, Phase II proposals MUST be submitted to ‘CBD SBIR’ regardless of which DoD contracting office negotiated and awarded the Phase I contract. Additional instructions regarding Phase II proposal submission process including submission key dates will be provided to Phase I awardees after Phase I contract award and also can be found at https://www.cbdsbir.net.

CBD SBIR Phase II Cost Volumes must contain a budget for the entire 24-month Phase II period not to exceed the total maximum dollar amount of $1,000,000. These costs must be submitted using the Cost Volume format (accessible electronically on the DoD SBIR Proposal Submission Web site).  The total proposed amount should be indicated on the Proposal Cover Sheet as the proposed cost with approximately 50% of the total Phase II budget for Year 1 and the balance of the funding budgeted for Year 2.

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The CBD SBIR Program is committed to minimizing the funding gap between Phase I and Phase II activities. The CBD SBIR 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.

Discretionary Technical Assistance In accordance with the Small Business Act (15 U.S.C. 632), the CBD SBIR Program Office will authorize the recipient of a Phase I and/or a Phase II SBIR award to purchase technical assistance services (Discretionary Technical Assistance, DTA), such as access to a network of scientists and engineers engaged in a wide range of technologies, or access to technical and business literature available through on-line databases, for the purpose of assisting such concerns as:

making better technical decisions concerning such projects;

solving technical problems which arise during the conduct of such projects;

minimizing technical risks associated with such projects; and

developing and commercializing new commercial products and processes resulting from such projects.

If your small business firm is interested in proposing use of a vendor for Discretionary Technical Assistance, you must provide a cost breakdown in the Cost Volume under “Other Direct Costs (ODCs)” and provide a one-page description of the vendor you will use and the specific technical assistance services to be provided. The proposed amount may not exceed $5,000 for Phase I, and $5,000 for each year of a Phase II project. The DTA description should be included as the LAST page of the Technical Volume. This description will not count against the Phase I or Phase II proposal page limit and will NOT be assessed against SBIR proposal evaluation criteria. Approval of technical assistance is not guaranteed and is subject to review of the Contracting Officer.

CBD SBIR Projects Requiring Animal and Human Subjects

Companies should plan carefully for any research involving animal and/or human subjects in addition to the use of any chemical or biological warfare agents. The brief Phase I Period of Performance may preclude plans requiring the use of these materials as well as animal and/or human subjects prior to obtaining all necessary approvals.

The offeror is expressly forbidden to use or subcontract for the use of laboratory animals in any manner without the express written approval of the U.S. Army Medical Research and Material Command's (USAMRMC), Animal Care and Use Review Office (ACURO). Written authorization to begin research under the applicable protocol(s) proposed as part of the CBD SBIR program will be issued after contract award in the form of an approval letter from the USAMRMC ACURO to the recipient. Furthermore, modifications to already approved protocols require approval by ACURO prior to implementation.

Research under CBD SBIR awards involving the use of human subjects, to include the use of human anatomical substances or human data, shall not begin until the DTRA Research Oversight Board (ROB) provides authorization that the research protocol may proceed. Written approval to begin research protocol will be issued from the ROB, under separate notification to the recipient. Written approval from the ROB is also required for any sub-recipient that will use funds obtained from any CBD SBIR awards to conduct research involving human subjects.

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Changes in research involving human subjects shall be conducted in accordance with the protocol submitted to and approved by the ROB. Non-compliance with any provision may result in withholding of funds and or termination of the award.

Key Dates

18.1 Announcement Pre-Release 28 November 2017 – 7 January 201818.1 Announcement Open/Close 8 January 2018 – 7 February 2018 (submission deadline:

8:00 pm Eastern Time on closing date)Phase I Evaluations February – April 2018Phase I Selections No Later Than 6 May 2018Phase I Awards October 2018 (see Note 1)

Phase II Proposal Submission Recommend proposal submission no earlier than approximately fivemonths from date of Phase I contract award. Additional instructions regarding Phase II proposal submission process including key dates will be provided to Phase I awardees after Phase I contract award and also can be found at https://www.cbdsbir.net.

(Note 1) Subject to the Congressional Budget process.

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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 Sheet along with the Technical Volume, Cost Volume, and Company Commercialization Report were submitted via the Internet using the DoD SBIR Proposal Submission Web site at https://sbir.defensebusiness.org/. _____ 2. The proposal cost adheres to the CBD SBIR Program criteria specified.

_____ 3. The proposal is limited to only ONE SBIR topic. All required documentation within the proposal references the same topic number.

_____ 4. The proposal is responsive to the requirements addressed in the topic.

_____ 5. The Project Abstract and other content provided on the Proposal Cover Sheet does not contain any proprietary or classified information and is limited to the space provided.

_____ 6. The Technical Volume of the proposal includes the items identified in Section 5.3.c of the DoD SBIR Program Announcement.

_____ 7. The Proposal Technical Volume must be 20 pages or less in length. The Cover Sheet, Cost Volume and Company Commercialization Report do not count against the 20-page Proposal Technical Volume page limit. Technical Volume pages in excess of 20-pages will not be evaluated and will not be considered for review.

_____ 8. The Company Commercialization Report is submitted online in accordance with Section 5.4.e of the DoD SBIR Program Announcement. This report is required even if the company has not received any prior SBIR funding.

_____ 9. The proposal must not contain any type smaller than 10-point font size (except as legend on reduced drawings, but not tables).

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CBD SBIR 18.1 Topic Index

CBD181-001 3D Printed Manufacturing of Respiratory ProtectionCBD181-002 Innovative Respiratory Protection for Low Threat EnvironmentsCBD181-003 Durable Stretch Barrier MaterialsCBD181-004 Extended Release Formulations for Fielded Nerve Agent PretreatmentCBD181-005 Dual Formulation of Atropine/Scopolamine with Enhanced StabilityCBD181-006 Field Portable Mass Spectrometry for Small Molecule Drugs in Clinical SamplesCBD181-007 Bi-specific Antibodies Targeting Disease Caused by Encephalitic Alphaviruses

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CBD SBIR 18.1 Topic Descriptions

CBD181-001 TITLE: 3D Printed Manufacturing of Respiratory Protection

TECHNOLOGY AREA(S): Chemical/Biological Defense, Materials/Processes

OBJECTIVE: Develop additive manufacturing processes to produce a fully functional Air Purifying Respirator (APR). Demonstrate advantages of 3D printed manufacturing to provide innovations such as custom sizing and use of multifunctional elastomeric composites.

DESCRIPTION: Many respiratory protection systems are designed for a high threat environment. These threat levels and quality assurance requirements posed upon respiratory protection devices currently drive the utilization of expensive and timely production processes such as rubber and plastic injection molding. Limitations of injection molding include cost, time, and an inability to produce multifunctional composite materials. Also, small run respirator modifications or customizations are typically cost prohibitive. Additive manufacturing allows for improvement in manufacturing cost and time but does not currently account for the production of materials that offer the strength or permeation resistance required for respiratory protection systems. Also, layered 3D printing techniques do not result in smooth surfacing and some resin based options do not have the print bed or working time requisite for a full facepiece respirator.

This effort seeks to overcome these challenges by pursuing a technological manufacturing innovation that is critical to a strong manufacturing sector in the U.S. economy. The focus of this effort is to develop a respiratory protective device utilizing innovative additive manufacturing technologies and processes. Innovative technologies and multifunctional elastomeric composite materials are needed to allow custom sized manufactured respirators to be produced at a cost effective price and in a timely manner. Additive manufacturing processes of composite elastomeric materials offer many potential advantages. Potential advantages could include a respirator facepiece consisting of a soft internal comfort layer near the face and a rugged threat agent resistant layer on the exterior. Even further, a composite layer providing ballistic, fire, or other operationally tailored levels protection could be achieved. Materials developed shall have threat agent permeation resistance characteristics required by the NIOSH Statement of Standard for Chemical, Biological, Radiological, and Nuclear (CBRN) Full Facepiece Air Purifying Respirator (APR). Durability and tensile requirements shall compare favorably to existing military respirator materials and shall comply with requirements set forth in MIL-STD 810G.

PHASE I: Investigate innovative elastomeric and/or composite materials and additive manufacturing processes that would result in eight vertically printed coupons (4” diameter; .05” thickness) that are resistant to chemical warfare agent materials as required by the NIOSH Statement of Standard for CBRN Full Facepiece Air Purifying Respirator APR. Provide eight coupons to the Government for testing. Perform and demonstrate in house durability and tensile strength testing of additional coupons of simple and complex geometries. Referencing Government-owned and provided M40 APR technical data, develop designs that illustrate in detail how these elastomeric and/or composite materials would be used to develop a full facepiece system. Show how the necessary production quality attributes can be met, especially those related to a good respirator to face seal. Assess materials chosen for human safety.

PHASE II: Refine the material and production technology chosen to develop a full elastomeric facepiece of size equivalent to the M40 APR. Technology demonstrated will be capable of printing a minimum of 10” (L) x 8” (W) x 8” (H) print volume and will exhibit smooth sealing surfaces. Exact APR face seal size and shape produced will be based on head and facial anthropometric characteristics of five test subjects. A full facepiece APR will be constructed using these custom facepieces and government provided M40 component hardware, to include the filter. The ability to co-print multiple materials to reduce hand assembly will be demonstrated. A preliminary laboratory respiratory protection level (LRPL) test of these five constructed APRs on the chosen test subjects will be performed at this stage. The LRPL of the APR shall be 2000 or greater, for 95% of trials, sampled in the breathing and ocular zone of the respirator. The APRs developed should comply with the NIOSH Statement of Standard for CBRN Full

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Facepiece Air Purifying Respirator APR.

PHASE III: Improve the APR design by demonstrating the incorporation of customized hardware, allowance for improved human factors, and operational tailoring. Demonstrate the scalability of the technology to print a NIOSH compliant APR in under 12 hours.

PHASE III DUAL USE APPLICATIONS: Potential alternative applications include industrial, pharmaceutical, healthcare, international, and other commercial respiratory protection uses as well as protection systems beyond the respirator.

REFERENCES:1. Anonymous. (2017, June 29). “3D printers start to build factories of the future.” The Economist, retrieved from http://www.economist.com

2. Anonymous (2017, June 29). “3D printers will change manufacturing.” The Economist, retrieved from http://www.economist.com

3. Exec. Order No. 13329 (2004). Encouraging Innovation in Manufacturing.

4. Melnikova, R., Ehrmann A., and Finsterbusch, K., (2014). 3D printing of textile-based structures by Fused Deposition Modelling (FDM) with different polymer materials. Paper presented at 2014 Global Conference on Polymer and Composite Materials.

5. Military Standard, MIL-STD-810G, Environmental Engineering Considerations and Laboratory Tests, Revision G (DOD 31 Oct 2008).

6. National Institute for Occupational Safety and Health (2004). Statement of Standard for Chemical, Biological, Radiological, and Nuclear (CBRN) Full Facepiece Air Purifying Respirator (APR). Retrieved from https://www.cdc.gov/niosh/npptl/standardsdev/cbrn/apr/standard/aprstd-a.html

7. Rudykh, S., Ortiz, C. and Boyce, M. (2015). Flexibility and protection by design: imbricated hybrid microstructures of bio-inspired armor. Soft Matter, 11, 2547-2554.

8. Yakout, M. and Elbestawi, M. (2017) Additive Manufacturing of Composite Materials: An Overview. Paper presented at the International Conference on Virtual Machining Process Technology.

KEYWORDS: individual protection, respiratory protective mask, 3D printing, additive manufacturing

CBD181-002 TITLE: Innovative Respiratory Protection for Low Threat Environments

TECHNOLOGY AREA(S): Chemical/Biological Defense

OBJECTIVE: To develop a pressurized respiratory protection device for use in low threat environments without the use of rubber facial or neck seals.

DESCRIPTION: Current military respiratory protection systems such as the M50 are designed for a high threat environment. In part, the threat level requirements drive the utilization of rubber sealing surfaces onto the wearer’s face and/or neck, which can be cumbersome, uncomfortable, and may impede wearer task performance. This effort seeks to develop a respiratory protective device that is not completely reliant on elastomeric sealing surfaces for protection.

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Loose Fitting Powered Air Purifying Respirators (PAPR) utilize the positive pressure of high flow blowers to provide respiratory protection without rubber face seals. However, these loose fitting PAPR systems require significant power and blowers of significant size and weight.

Technologies and innovative respirator designs are needed that can remove the rubber from the wearer’s face or neck in low threat environment and provide pressurization without significant power needs, weight, or bulk. This technology would be used in low level chemical or biological hazardous threat environments, would not require protection from highly volatile organic compounds, and could allow for mission durations less than current military air purifying respirators.

The measured laboratory respiratory protection level (LRPL) shall be 2000 or greater, for 95% of trials, sampled in the breathing and ocular zone of the respirator. The system blower and battery combined, excluding filtration, shall weigh no more than one (1) pound and be no larger than 42 cubic inches. The battery and blower can be body mounted if needed. The system must be able to withstand a large range of temperature and environmental extremes and must be constructed of materials resistant to low level chemical and biological agents. Noise levels generated by the respirator, at each ear location and at the maximum airflow obtainable, shall not exceed 60 decibels (dBA).

PHASE I: Investigate alternative protection and pressurization options that would allow for the development of the innovative respiratory protection system, as described. Identify performance and human factors metrics for the system when worn. Perform a benchtop demonstration of the technology to illustrate adequate pressurization/protection in the breathing and ocular zone on an anthropomorphic mannequin exposed to an inert aerosol challenge and with simulated breathing conditions.

PHASE II: Refine the protection and pressurization technology as identified in Phase I. Develop a full scale prototype system. Demonstrate protection factor levels > 2000 using a human subject LRPL assessment.

PHASE III: Complete system refinement. Demonstrate the technology is durable and suitable for military combat applications.

PHASE III DUAL USE APPLICATIONS: Potential alternative applications include industrial, pharmaceutical, healthcare, international, and other commercial respiratory protection uses.

REFERENCES:1. http://www.avon-protection.com/military/m50.htm

2. https://www.cdc.gov/niosh/npptl/respstandards/pdfs/cbrn_aper.pdf

KEYWORDS: individual protection, respiratory protective mask, powered air purifying respirator

CBD181-003 TITLE: Durable Stretch Barrier Materials

TECHNOLOGY AREA(S): Chemical/Biological Defense, Materials/Processes

OBJECTIVE: Commercially available Chemical, Biological, Radioactive, and Nuclear (CBRN) protective ensembles are bulky and with a high thermal burden due to the lack of stretch of the materials used. The bulk of the garment prevents agile movements of arms and legs needed while performing critical tasks. Materials with stretch enable a more conformable garment to be designed, enabling ease of movement and better comfort for the wearer. This topic addresses the technical challenges and innovative solutions needed to create durable, stretch materials with barrier capabilities that can be integrated into novel CBRN protective ensembles.

DESCRIPTION: Chemical, Biological, Radioactive, and Nuclear (CBRN) protective ensembles provide the first line of defense for personnel exposed to victims and/or materials during assessment, extrication, rescue, triage,

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decontamination, treatment, site security, crowd management, and force protection operations at incidents involving CBRN terrorism agents. Current protective ensembles are designed to meet National Fire Protection Association (NFPA) 1994 Class 1 protection standards [1].

Commercially available CBRN protective ensembles are bulky and not sufficiently conformal or tactile to the wearer. It is well known that a simple body movement may extend the body skin by 50% or more and the fabric must be able to accompany the stretch and recover on relaxation.

Elastic fabrics are an important route to achieve comfort, tactility and freedom of movement of the body with a conformal fit [2]. Elastic garments used in athletics and sports are assumed to improve athlete’s performance in cycling, swimming, and so on. This type of fabric enables freedom of body movement by reducing the fabric resistance to body stretch. Elastic fibers and fabrics under trade names such as Lycra and Spandex are widely used in many areas where elasticity is needed such as tights, sportswear, swimwear, wet suits, SCUBA dive skins, etc. Stretch fabric materials are also of interest in wearable electronics [3] and tactile sensing applications [4]. It would be highly beneficial if performance can be extended by the CBRN protective material by adding stretch to provide agility and comfort to the wearer with a more conformable garment design. The challenge is the development of novel stretch materials that could also deliver the stringent protection function and durability. It is also recognized that repetitive biaxial stretching could affect the functional properties of the stretch materials and any material design should account for this challenge as well [5].

This topic solicits the following innovative technology requirements:

a) The stretch barrier material should satisfy the threshold barrier properties against vapor and liquid challenges with appropriate simulants selected for the study, with standard test protocols specified above.b) The basic mechanical properties to be evaluated include the stress-strain behavior under biaxial stretching and the extent of reversibility of stretching. Materials that can be stretched isotropically with 12% or more along each linear dimension are preferred, recognizing that 12% is the threshold and 20% is the preferred objective. Stretch properties should be tested per ASTM D2594 in both linear dimensions as well as on the bias.c) The material should possess durability in barrier function after being subjected to flexing, abrasion and cycles of biaxial stretching and relaxation. This should be demonstrated by testing for the barrier properties after subjecting the stretch material to the different stressors defined above.d) Since the stretch material forms a part of the protective ensemble and is not expected to provide all needed functionalities such as fire or flame resistance (FR), it will be combined with other materials eventually to create final product of interest. Keeping that in view, the stretch materials should be designed so that it would be possible to bond/integrate the material with FR fabrics like Nomex, for example. Similarly, the potential for the material to manufacture garments should be demonstrated by overcoming problem areas such as seams.

The barrier materials should offer protection against vapor and liquid chemical agent challenges tested using American Society for Testing and Materials (ASTM) F739. The threshold level of permeation resistance should be cumulative permeation mass of less than 6 micrograms/cm2 (for industrial chemicals, 1.25 micrograms/cm2 for Soman and 4.0 micrograms/cm2 for distilled mustard) when challenged with 20 grams per meter squared (g/m2) of liquid chemical agent or 1% agent in gas phase. The permeation is to be tested after subjecting the material to 100 cycles of flexing per ASTM F392 and 10 cycles of abrasion with 600 grit paper as per ASTM D4157. The objective level of permeation is the same cumulative permeation mass limits as above but when challenged with 100 g/m2 of liquid chemical agent or 100% agent in gas phase. Also, the permeation is to be tested after subjecting the material to 100 cycles of flexing per ASTM F392 and 25 cycles of abrasion with 80 grit paper per ASTM D4157.

PHASE I: Conduct research on novel concepts for stretch barrier materials to achieve both stretch and barrier functions. Upon completion of Phase I, samples of the stretch material should be made available for independent evaluation of stretch and barrier properties. The system should meet the threshold goal of 12% linear strain that is reversible and barrier properties at the threshold level against selected chemical simulants based on liquid and vapor challenges. The barrier testing for Phase I can be limited to the ‘as-produced’ stretchable material. Tests for durability in barrier properties after subjecting the stretch barrier material to stresses like flexing, abrasion or repetitive biaxial stretching are not required for Phase I. The detailed conditions for testing must be approved by the

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Government Technical POC.

PHASE II: Further improvements in stretch should be made to reach as high a value as possible near the objective of 20% reversible strain level. The barrier properties should meet the threshold level for both liquid and vapor challenges for selected chemical agent simulants, and improvements in barrier performance towards the objective should be demonstrated. The testing should be conducted after subjecting the stretch barrier material to flexing, abrasion and repetitive biaxial stretching. The detailed conditions for testing must be approved by the Government Technical POC. Methods must be developed to bond/integrate the stretch barrier material with other functional materials such as Nomex FR fabric, identified by the Government Technical POC. At the conclusion of Phase II, swatches of stretch barrier fabric samples obtained from continuous pilot scale production should be made available for independent evaluation.

PHASE III: The stretch barrier material successfully demonstrated in Phase II will be integrated into CBRN protective ensemble. Materials should be made in full width production and issues in garment manufacture that may arise, such as seams, will be addressed.

PHASE III DUAL USE APPLICATIONS: First responder and anti-terrorism personnel would also benefit from the use of improved protective garments that are more conformal and stretchable, allowing for improved agility and comfort. The stretch fabric can be used not only in protective clothing, but also in other applications such as respirator face seals, face masks in hospitals, schools, and other buildings when high levels of protection are desired.

REFERENCES:1. NFPA 1994 Standard on Protective Ensembles for Chemical/Biological Terrorism Incidents 2001 Edition, National Fire Protection Association (NFPA), Quincy, MA 02269, USA. http://www.disaster-info.net/lideres/english/jamaica/bibliography/ChemicalAccidents/NFPA_1994_StandardonProtectiveEnsemblesforChemicalBiologicalTerrorismIncidents.pdf

2. M. Senthilkumar, N. Anbumani and J. Hayavadana, Elastane fabrics – A tool for stretch applications in sports, Indian J. Fiber and Textiles Research 36 (2011) 300-307.

3. C. Wang, M. Zhang, K. Xia, X. Gong, H. Wang, Z. Yin, B. Guan and Y. Zhang, Intrinsically Stretchable and Conductive Textile by a Scalable Process for Elastic Wearable Electronics, ACS Appl. Mater. Interfaces 9 (2017) 13331−13338.

4. G. H. Büscher, R. Kõiva, C. Schürmann, R. Haschke, and H. J. Ritter, Flexible and stretchable fabric-based tactile sensor, Robotics and Autonomous Systems 63 (2015) 244–252.

5. H. S. Kim and C. H. Park, Effect of biaxial tensile extension on superhydrophobicity of rayon knitted fabrics, RSC Adv. 6 (2016) 48155.

KEYWORDS: stretch fabrics, barrier materials, chem-bio protection, biaxial stress-strain, durability, permeation resistance, elastic stretching; elastic relaxation

CBD181-004 TITLE: Extended Release Formulations for Fielded Nerve Agent Pretreatment

TECHNOLOGY AREA(S): Biomedical, Chemical/Biological Defense

OBJECTIVE: Develop formulations of pyridostigmine bromide (PB) that afford pretreatment to nerve agent exposure by reducing the number of daily administrations of the fielded product, while maintaining a consistent reduction in blood acetylcholinesterase activity.

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DESCRIPTION: Military operations occur under a wide-range of temperatures and austere conditions. The current and future joint service members are globally deployed and are executing missions across the full spectrum of military operations. A rebalancing of forces will focus on Asia-Pacific and Middle Eastern regions. The Joint Chemical and Biological Defense Program wants to make the brigade combat teams leaner and more nimble while retaining capability. Expeditionary missions require medical solutions that allow maximum flexibility and agility. These members must protect themselves against all nerve agent threats during deployments, sustainment operations, and re-deployments. Non-state actors and terrorists may develop or acquire nerve agents, and may use them abroad and in the homeland, and joint forces may be the first to respond. Prophylaxes used in conjunction with other treatments, would counter the threat of performance degradation and death caused by nerve agent threats.

Organophosphorus nerve agents exert their toxic effects by binding to and inactivating the enzyme acetylcholinesterase (AChE). These nerve agents bind very tightly to AChE, and after a certain amount of time, ranging from minutes to hours, this binding becomes irreversible. If sufficient amount of the enzyme is bound up, the effects can be lethal. The rationale behind the use of PB is that it temporarily binds to AChE, thereby preventing nerve agents from binding to, and permanently inactivating AChE. However, unlike the nerve agents, PB binding is reversible. Thus PB creates a pool of reversibly bound AChE that is essentially protected against the irreversible binding of nerve agents.

The only pretreatment medication currently FDA-approved for use against the lethal effects of nerve agent (e.g., soman) poisoning is pyridostigmine bromide (PB) tablets. PB tablets are procured by the DoD as Soman Nerve Agent Pretreatment Pyridostigmine (SNAPP). PB is an orally active cholinesterase inhibitor. The dose of pyridostigmine is one 30 mg tablet every 8 hours, to be started at least several hours prior to nerve agent exposure. In humans, this dosing of PB inhibits red blood cell cholinesterase (i.e., AChE) by 20-40%, a level that protects animals from the lethal effects of soman. The product must be stored in refrigeration between 2-8°C (36-46°F) and protected from light. A unit of issue is one blister pack of 21 tablets. NSN 6505-01-178-7903 represents a package of 10 blister packs. The tablets are hygroscopic. Once issued to a soldier, the blister pack must be discarded after three months as it is no longer maintained under refrigerated temperatures. Pyridostigmine also has been approved by the Medicines Control Agency, the United Kingdom equivalent of the U. S. Food and Drug Administration (FDA), for the Ministry of Defence as a pretreatment for organophosphorus inhibitors of AChE.

Based on current operation tempo, there is interest in obtaining a solution for administering a nerve agent pretreatment with reduced frequency than the SNAPP product, as well as a product/packaging that could maintain activity during prolonged exposure to both desert temperatures and high humidity.

Mestinon® TIMESPAN® tablets contain 180 mg PB in a specially coated tablet for sustained release. The product is stored at 25°C (77°F), with excursions permitted to 15-30°C (59-86°F). This form provides uniformly slow release, hence prolonged duration of drug action; it facilitates control of myasthenic symptoms with fewer individual doses daily. However, dosing of this product to achieve the desired 20-40% reduction in blood AChE activity for use as a nerve agent pretreatment has not been conducted.

The ultimate goal is to develop PB formulation data that could be used in a future supplement to the pyridostigmine bromide tablet New Drug Application 20-414.

PHASE I: Alterations to the PB tablet formulation will be studied that provide a prolonged blood level of PB, sufficient to generate a steady-state 20-40% reduction in red blood cell AChE activity. The initial goal is a single oral administration sufficient to maintain these levels minimally up to 24 hours. Proof-of-concept formulations should be tested in appropriate animal models (e.g., guinea pig, rat). The route of administration should be justified if other than oral in the animal model. Preclinical pharmacokinetic and pharmacodynamics data for proof-of-concept formulations will be assessed for suitability to meet steady-state 20-40% reduction in blood AChE activity for minimally 24 hrs.

Ideally, candidate formulations will be assessed for stability at elevated temperatures encountered by military personnel operating overseas.

PHASE II: Establish formulation strategy to accelerate entry into early stage clinical development. Design and perform formulation prototype development studies. Use FDA quality by design approach to formulation concept

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design based upon clinical targets. The desired goal is to use excipients that are clinical route of administrate appropriate, and generally recognized as safe. Produce prototype formulation(s) using state-of-the-art and scalable technologies in particle processing and drug physical form manipulation. Screen for and manufacture soluble intermediates. Perform risk assessment to identify potentially high risk formulation and process variables; propose/conduct studies to increase knowledge and reduce risk. Identify and mitigate any stability and processing issues during small scale-up manufacture of prototype formulation. Initiate real-time stability data to support 6-month stability of prototype formulations across a range of temperatures and humidity; define excursion limits.

PHASE III: Develop a tablet formulation and manufacturing process that ensures the quality, safety and efficacy of modified-release tablet. Define the quality target product profile (QTPP) of the modified-release PB tablet based on drug substance properties, consideration of the PB label and intended population. Continue risk assessment and propose a control strategy, which should include in-process and finished product specifications. Develop and optimize modified-release tablets compressed into scored tablets to achieve all of the attributes in the PB QTPP. Conduct necessary ICH stability studies. Optimize the extended-release final tablet process development using drug layering, ER polymer coating, blending and lubrication, and compression, as appropriate.

PHASE III DUAL USE APPLICATIONS: Given the unforeseen/unanticipated use of nerve agents on the homeland, and in contrast to military personnel, civilians are unlikely to be pretreated with pyridostigmine and protected by personal protective equipment. Extended pretreatment of a civilian population is not desirable nor feasible given the possibility of overdose and toxic effects in special populations. However, there may be instances where it would be desirable to pretreat both medical and emergency aid personnel (e.g., first responders) and first receivers (e.g., hospital personnel) following a known/suspected nerve agent attack. Healthcare-providers, as valuable assets, would have an additional layer of protection against the lethal effects of nerve agents during mass casualty decontamination (possibility for secondary contamination of medical personnel), evacuation and medical intervention activities.

The military counterparts to the DoD, notably the United Kingdom and NATO allies, along with members of the CBR MOU, use pyridostigmine bromide tablets. The armed forces of these countries would reap the benefits of a less frequent dosing regimen as well as a formulation that could withstand extended elevated temperatures and humidity.

Finally, additional PB extended release formulations for the treatment of myasthenia gravis could be of commercial interest to benefit this patient population.

REFERENCES:1. ATP 4-02.285 Multiservice Tactics, Techniques, and Procedures for Treatment of Chemical Agent Casualties and Conventional Military Chemical Injuries, 19 Aug 2015. Chapter 3: Nerve Agents.

2. Capstone Concept for Joint Operations: Joint Forces 2020. 10 Sep 2012.

3. Drug Approval Package, Pyridostigmine bromide tablet, NDA 20-414.

4. https://www.accessdata.fda.gov/drugsatfda_docs/nda/2003/20414_pyridostigmine.cfm

5. Herkert NM, Eckert S, Eyer P, Bumm R, Weber G, Thiermann H, et al. Identical kinetics of human erythrocyte and muscle acetylcholinesterase with respect to carbamate pre-treatment, residual activity upon soman challenge and spontaneous reactivation after withdrawal of the inhibitors. Toxicology 2008, 246:188–192.

6. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. Q1A-F Stability.

7. Scott, L. 2007 Pretreatment for Nerve Agent Poisoning. In: Marrs, T.C., Maynard, R.L., Sidell, F.R. (Eds.), Chemical Warfare Agents: Toxicology and Treatment. John Wiley & Sons Ltd., Chichester, pp. 343–353.

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8. Quality by Design for ANDAs: an Example for Modified-Release Dosage Forms. Available at FDA.gov: https://www.fda.gov/downloads/drugs/developmentapprovalprocess/howdrugsaredevelopedandapproved/approvalapplications/abbreviatednewdrugapplicationandagenerics/ucm286595.pdf

9. Thompson J, Rehn M, Lossius HM, Lockey D. Risks to emergency medical responders at terrorist incidents: a narrative review of the medical literature. Critical Care 2014, 18:521.

10. TRADOC Pamphlet 525-3-1 The U.S. Army Operating Concept:2020-2040, Win in a Complex World. 31 Oct 14.

KEYWORDS: chemical nerve agent, pretreatment, extended-release, drug delivery, medical countermeasure

CBD181-005 TITLE: Dual Formulation of Atropine/Scopolamine with Enhanced Stability

TECHNOLOGY AREA(S): Biomedical, Chemical/Biological Defense

OBJECTIVE: Develop stable, combined atropine/scopolamine formulations to exploit complementary pharmacological profiles for optimal muscarinic receptor blockade and anticholinergic activity within the peripheral and central nervous systems. The goal is to develop improved countermeasures against organophosphorus (OP) nerve agent (NA) intoxication.

DESCRIPTION: The current United States military standard treatment for NA intoxication is immediate administration of medications by autoinjector. Up to three Antidote Treatment for Nerve Agents Autoinjectors (ATNAA), each consisting of 2.0 mg atropine and 600 mg oxime (2-pralidoxime [2-PAM]), to reactivate OP-inhibited acetylcholinesterase [AChE]) may be administered to counter nerve agent symptoms. An autoinjector containing 10 mg of the anticonvulsant diazepam (Convulsant Antidote for Nerve Agents [CANA]) is administered following three ATNAA administrations. Medics in the field can administer an additional 20-30 mg of atropine and up to 30 mg of diazepam in the first hour in cases of severe intoxication. Animal studies demonstrated scopolamine could serve as a useful adjunct treatment for NA intoxication. Scopolamine enhanced the effectiveness of human equivalent doses of the standard medical countermeasures atropine, 2-PAM and diazepam. Unlike atropine, scopolamine rapidly penetrates the blood brain barrier and has greater potency (7.5 times atropine activity in the central nervous system). The addition of scopolamine to the NA treatment regimen appears to be advantageous for reducing seizures, enhancing survival, and recovery from OP NA exposure. The Joint Chemical and Biological Defense Program would like to make the brigade combat teams leaner and more agile while retaining capability. As such, minimizing to the extent feasible, the number of soldier-carried ATNAAs and medic-carried medical countermeasures is desirable. Support medical personnel carry Atropen® autoinjectors, containing the equivalent of 2.0 mg atropine sulfate (AS), to treat personnel with severe NA intoxication. A stable, combined formulation of atropine and scopolamine would increase the therapeutic efficacy of the current treatment regimen as well as reduce the requirement for additional atropine in cases of severe NA poisoning. The objective is to develop a formulation that has, minimally, stability for two years at room temperature. The desired final formulation is to be a stable, intramuscular (IM) injectable containing both atropine and scopolamine.

PHASE I: Establish preliminary specifications for a combined formulation under International Conference on Harmonization (ICH) Pharmaceutical Development Guidelines. Explore scopolamine hydrobromide dose ranges between 0.5 to 3.0 mg, combined with a fixed dose equivalent of 2.0 mg atropine sulfate. Generate initial specifications for a dual formulation designed to maximize compatibility and stability of the combined drug substances. The dual prototype formulation may be a single phase liquid mixture or a dual/multiphase system utilizing lyophilized drug powder(s) and vehicle/diluent components. The desired final volume is to be no more than 2.0 ml. Generate initial stability profile of the dual prototype formulation(s).

PHASE II: Continue assessment of the dual prototype formulation stability based on mass balance techniques under ICH Pharmaceutical Development Guidelines. Studies will employ forced degradation strategies based on specific

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chemical degradation. Stress conditions should include heat, acid, base, light, and the presence of oxidizing agents. Conduct necessary ICH stability studies, to include room temperature and accelerated conditions. Conduct a pharmacokinetic (PK) study of the optimized dual formulation prototype in rodents. Determine plasma levels of both drugs after a single IM injection and characterize standard pharmacokinetic parameters to include Cmax, AUCtotal, Tmax, and elimination half-life.

PHASE III: Conduct a 14 day toxicity and toxicokinetic (TK) study of a single dose IM dual formulation in non-human primates (Rhesus macaques). The TK of atropine/scopolamine will be evaluated using plasma drug concentrations, construction of concentration time profiles, and TK parameters. Toxicity will be evaluated using survival, clinical observations (including injection site reactions), body weights, food consumption measurements, functional observational battery assessments, clinical pathology (hematology, serum chemistry, coagulation, and urinalysis), and anatomic pathology including organ weights, gross observations at necropsy, and histopathology of selected tissues.

PHASE III DUAL USE APPLICATIONS The Department of Health and Human Services (HHS) has a similar need for next generation anticholinergics to potentially include an atropine/scopolamine dual formulation for human use. Successful completion of all three phases under this solicitation will support small business valuation by confirming technical merit that invites further investment. This award mechanism will bridge the gap between laboratory-scale innovation and entry into a recognized U. S. Food and Drug Administration (FDA) regulatory pathway leading to commercialization.

REFERENCES:1. Capacio, B.R., and Shih, T.-M. “Anticonvulsant actions of anticholinergic drugs in soman poisoning,” Epilepsia, 1991, 32, 604-615.

2. McDonough, J.H., Zoeffel, L.D., McMonagle, J., Copeland, T.L., Smith, C. D., and Shih, T.-M. “Anticonvulsant treatment of nerve agent seizures: Anticholinergics versus diazepam in soman-intoxicated guinea pigs,” Epilepsy Research, 2000, 38, 1-14.

3. Shih, T.-M., Duniho, S.M., and McDonough, J.H. “Control of nerve agent-induced seizures is critical for neuroprotection and survival,” Toxicol. Appl. Pharm., 2003, 188, 69-80.

4. Shih, T.-M., Rowland, T.C., and McDonough, J.H. “Anticonvulsants for nerve agent-induced seizures: The influence of the therapeutic dose of atropine,” J. Pharm. Exp. Ther., 2007, 320 (1), 154-161.

5. Ketchum, J., Sidell, F., Crowell Jr, E., Aghajanian, G., and Hayes Jr, A. “Atropine, scopolamine, and ditran: Comparative pharmacology and antagonists in man,” Psychopharmacologia, 1973, 28 (2), 121-145.

6. Koplovitz, I., and Schulz, S. “Perspectives on the use of scopolamine as an adjunct treatment to enhance survival following organophosphorus nerve agent poisoning,” Mil. Med., 2010, 175 (11), 878-882.

7. International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use. ICH Harmonized Tripartite Guideline. Pharmaceutical Development Q8 (R2). 2009.

KEYWORDS: atropine, scopolamine, chemical nerve agent, medical countermeasure, drug formulation

CBD181-006 TITLE: Field Portable Mass Spectrometry for Small Molecule Drugs in Clinical Samples

TECHNOLOGY AREA(S): Biomedical, Chemical/Biological Defense

OBJECTIVE: To provide a rapid, easy to use method for human clinical sample testing that is effective for the detection and identification of novel designer drug combinations in non-traditional sample types. Capabilities

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sought should encompass the end-to-end method to include low invasive clinical sample collection and be suitable for use in a field environment. Desired end-state is an U.S. Food and Drug Administration (FDA) cleared in vitro diagnostic device, but interim phases of capability development, to include environmental sample types and data sets sufficient for a Government supported Emergency Use Authorization submission, are acceptable.

DESCRIPTION: The U.S. Department of Defense (DoD) requires the capability to rapidly assess Warfighter exposures to a rapidly evolving range of drugs that affect the central nervous system (e.g., anesthetics, sedatives, analgesics, etc.) and, furthermore, to monitor the efficacy of medical countermeasures throughout the course of patient treatment. Current mass spectrometry instrumentation for drugs-of-abuse testing are fragile, bulky, heavy, and expensive, while methods often rely upon volunteered clinical urine samples. Furthermore, although there are many commercially available, portable, field-rugged mass spectrometers, none were developed or produced within a regulatory framework supporting future FDA-regulated applications and nor include end-to-end analytical methods suitable for the intended use in field environments. Immunoassay-based approaches (such as lateral flow immunoassays) may not have sufficient inclusivity, specificity, or sensitivity and involve a development pathway too long to keep pace with the rapidly emerging and evolving novel compounds and mixtures of concern.

This topic seeks to develop novel approaches to provide practical patient diagnostics and screening capabilities at field deployment locations from least-invasive clinical sample types while maintaining desirable operational suitability characteristics. Sample types that may be collected rapidly under any condition immediately after suspected exposure or from non-ambulatory personnel are essential (e.g., capillary blood, saliva, etc.) and appropriate pairing to the mass spectrometer sample inlet method (e.g., solid-phase microextraction, paper spray ionization) is a requirement. Incorporation of semi-automated capillary blood sample collection tools into the analytical method (i.e. arm stick collection devices) would further enhance operator safety and ease of use when wearing personal protective equipment. The topic may be inclusive of the development of a new mass spectrometer intended for the diagnostic market or the incorporation of an existing instrument into a new method and medical device.

PHASE I: Demonstrate efficacy, within a laboratory environment, of new methodology and/or instrumentation or instrument modifications/integration for mass spectrometry-based assays for the detection and identification of novel designer drug combinations that affect the central nervous system (e.g., anesthetics, sedatives, and analgesics) in non-traditional sample types simulating those which could be collected in a minimally invasive manner from exposed humans (e.g., capillary blood and saliva) and prepared in a field environment. Provide experimental evidence indicating the potential for this assay to be applied effectively using either existing, modified, or novel field-rugged mass spectrometry systems.

Use of human or animal subjects is not intended, or expected, in order to establish/achieve the necessary proof-of-concept in Phase I.

PHASE II: Develop initial small molecule compound libraries suitable to the analytical method. Demonstrate the end-to-end analytical method, relative to the device maturity, sufficient to indicate the operational suitability of the final device and method for the intended operational environment. Demonstrate efficacy using contrived surrogate samples or from an appropriate animal exposure model. Develop an in vitro diagnostic Target Product Profile and, relative to the maturity of the device, develop and submit a U.S. FDA pre-submission application describing the proposed regulatory approach for FDA clearance.

PHASE III: Complete device analytical method development and transition to full-rate manufacturing under current Good Manufacturing Practices (cGMP). Conduct reliability and ruggedness qualification testing appropriate to the intended operational environment. Conduct analytical studies and clinical trials to support a de novo or 510(k) submission. Expand the compound library and range of analytical methods to additional defense and dual use environmental, medical surveillance, and diagnostic applications.

PHASE III DUAL USE APPLICATIONS: Potential alternative applications include drug enforcement applications within the civilian sector and the protection of law enforcement and laboratory personnel.

REFERENCES:

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1. CLIA Categorizations (n.d.), retrieved December 2nd, 2014 from http://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/IVDRegulatoryAssistance/ucm393229.htm

2. Field Manual 4-02 Army Health System, August 26th 2013, retrieved Dec 17th from http://armypubs.army.mil/doctrine/DR_pubs/dr_a/pdf/fm4_02.pdf , pages 1-9, 7-7 and 7-8

KEYWORDS: mass spectrometry, Opioid, Fentanyl, Designer Drugs, in vitro diagnostic, point of care, small molecule

CBD181-007 TITLE: Bi-specific Antibodies Targeting Disease Caused by Encephalitic Alphaviruses

TECHNOLOGY AREA(S): Biomedical, Chemical/Biological Defense

OBJECTIVE: The objective of this effort is to identify and develop bi-specific antibodies that demonstrate the capability to cross the blood brain barrier and neutralize encephalitic strains of Alphaviruses.

DESCRIPTION: There are no approved medical countermeasures that are available for the treatment of encephalitic alphavirus infections of interest to the Department of Defense (DoD). Innovative technologies are sought to explore the bi-specific antibody platform as a potential therapeutic modality targeting Encephalitic alphavirus infections. Additional bi-specific platform technologies will also be considered.

This topic is intended to solicit responses from small businesses to develop Medical Countermeasures (MCMs) for these specific viruses. This topic, as solicited, includes acceptance criteria for the definition of project success and is intended to increase the number of potential MCMs in the anti-Alphavirus pipeline.

Currently, there is a capability gap for the effective treatment of viral induced encephalitis. It is widely acknowledged that viruses such as those represented in the Alphavirus family, e.g. Eastern, Western, and Venezuelan Equine Encephalitis viruses, pose significant risk to the warfighter. The Alphaviruses are recognized as potential biological warfare agents. There are no approved or licensed medical countermeasures against diseases caused by Alphaviruses. Currently, the only Medical Countermeasure is supportive care. Alphavirus infections can cause two distinct clinical presentations. In the mode of lesser concern, the infection can exhibit symptomology ranging from virtually asymptomatic to typical ‘flu-like’ symptoms. However, Alphavirus infections can also lead to encephalitis and it is this manifestation that is of grave concern, as viral encephalitis often leads to death, or at the very least, prolonged incapacitation.

The specific approach of the study and the pathogenic targets are to be determined by the individual investigators.

PHASE I: The Phase I proof-of-concept must demonstrate the offeror can produce a high binding (pM) bi-specific antibody. Successful Phase I proof-of-concept is defined as a demonstration that a bi-specific antibody directed at either Eastern, Western, or Venezuelan Equine Encephalitis virus has nanomolar binding capability. The offeror need not show efficacy against a challenge in vivo; just that the antibody can bind to alphavirus and an appropriate target for crossing the blood brain barrier (BBB).

PHASE II: Building upon a successful Phase I proof-of-concept technology demonstration, Phase II requires demonstration of permeability of the blood brain barrier as well as neutralization of the virus. Demonstration of the antibody crossing the BBB can be performed in an artificial system. Additionally the bi-specific antibody must demonstrate post exposure efficacy in vivo. The offeror will need to use a well-defined animal model that mimics human encephalitic disease; however, the development of the animal model is outside the scope of this topic. The successful demonstration could be for one or more of the encephalitic Alphaviruses. A candidate is considered successful if there is a statistically significant increase in survival of animals. Delayed time to treat will be further evaluated in Phase III.

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Phase II tasks should include demonstration of BBB permeability, efficacy proof-of-concept, chemistry, manufacturing, and controls (CMC) considerations, pilot lot production proof-of-concept experiments that successfully demonstrate the feasibility to manufacture the MCM, and a preliminary stability study. Other considerations for Phase II work includes formulation (route of administration), tissue cross reactivity (TCR), and pharmacokinetics (PK) evaluation.

PHASE III: With successful completion of Phase II, Phase III will focus on refinement and expansion of the concept. A successful Phase III effort will culminate in a product that is effective against one or more Alphaviruses, to include Eastern, Western, and Venezuelan Equine Encephalitis viruses. A “tri-valent” medical countermeasure would represent a distinct advantage, but is not an absolute requirement. A dosing regime in line with an FDA Phase I human safety trial will be determined, as will delayed time to treat. The technology will be evaluated in animal models using lethal doses of Eastern, Western, or Venezuelan Equine Encephalitis virus, separately. Time points for delayed time to treat should include 48, 72, 96 hrs post clinical signs of infection, with statistically significant target rescue rates of 80%, 50%, and 35%, respectively.

After the successful completion of the Phase III activities, additional tasks required to make the product marketable will include a pre-investigational new drug (IND) meeting with the FDA, and IND filing. This would require completion of all necessary IND enabling toxicology, PK studies required and cGMP manufacturing to support Phase I safety trials in healthy human volunteers.

PHASE III DUAL USE APPLICATIONS: Beyond DoD use, the product would most certainly have world-wide acceptance in the medical community for a treatment of encephalitis caused by Eastern, Western, or Venezuelan Equine Encephalitis virus infection. Moreover, the product would likely prove effective against treating infection of the aforementioned pathogens prior to the encephalitic stage.

REFERENCES:1. Julia C. Frei, Elisabeth K. Nyakatura, Samantha E. Zak, Russell R. Bakken, Kartik Chandran, John M. Dye & Jonathan R. Lai. Bispecific Antibody Affords Complete Post-Exposure Protection of Mice from Both Ebola (Zaire) and Sudan Viruses. Scientific Reports 6, Article number: 19193 (2016).

2. Pace, C. S. et al. Bispecific antibodies directed to CD4 domain 2 and HIV envelope exhibit exceptional breadth and picomolar potency against HIV-1. Proceedings of the National Academy of Sciences of the United States of America 110, 13540–13545, 10.1073/pnas.1304985110 (2013).

3. Elisabeth K. Nyakatura, Alexandra Y. Soare & Jonathan R. Lai. Bispecific antibodies for viral immunotherapy. Human Vaccines & Immunotherapeutics. 2017: 13 (4) 836-842.

4. Jitra Kriangkum, Biwen Xu, Les P. Nagata, R.Elaine Fulton, Mavanur R. Suresh, Bispecific and bifunctional single chain recombinant antibodies, Biomolecular Engineering, Volume 18, Issue 2, 2001, Pages 31-40, ISSN 1389-0344.

5. Tirado, Sol M. Cancel, and Kyoung-Jin Yoon. "Antibody-dependent enhancement of virus infection and disease." Viral immunology 16.1 (2003): 69-86.

6. Long, M. C., Nagata, L. P., Ludwig, G. V., Conley, J. D., Suresh, M. R., & Fulton, R. E. (2000). Construction and characterization of monoclonal antibodies against western equine encephalitis virus. Hybridoma, 19(2), 121-127.

7. Abulrob, A., Sprong, H., En Henegouwen, P. V. B. and Stanimirovic, D. The blood–brain barrier transmigrating single domain antibody: mechanisms of transport and antigenic epitopes in human brain endothelial cells. Journal of Neurochemistry (2005), 95: 1201–1214.

8. Banerjee, J., Shi, Y., and Azevedo, H.S. In vitro blood–brain barrier models for drug research: state-of-the-art and new perspectives on reconstituting these models on artificial basement membrane platforms, Drug Discovery

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Today, 2016 Jun 6.

9. Gabathuler, R. Approaches to transport therapeutic drugs across the blood-brain barrier to treat brain diseases. Neurology of Disease (2010) 37 (1), 48-57.

10. Steele, K.E., Twenhafel, N. A. REVIEW PAPER: Pathology of Animal Models of Alphavirus Encephalitis, Vet. Pathol., September 2010 47: 790-805.

11. Hamers-Casterman, C., Atarhouch, T., Muyldermans, S., Robinson, G., Hamers, C., Bajyana Songa, E., Bendahman, N., and Hamers, R. Naturally occuring antibodies devoid of light chains. Nature Vol 363, 3 June 1993.

12. Lajoie, J.M. and Shusta, E.V. Targeting receptor-mediated transport for delivery of biologics across the blood-brain barrier. Annual Review of Pharmacology and Toxicology, Vol 55: 613-631, January 2015.

13. Mäger, I., Meyer, A.H., Li, J., Lenter, M., Hildebrandt, T., Leparc, G., Wood, M.J.A. Targeting blood-brain-barrier transcytosis – perspectives for drug delivery, Neuropharmacology, Available online 22 August 2016.

14. Neves, V., Aires-da-Silva, F., Corte-Real, S., Castanho, M. Antibody approaches to treat brain diseases, Trends in Biotechnology, Volume 34, Issue 1, January 2016, Pages 36-48.

15. Stanimirovic, D.B., Bani-Yaghoub, M., Perkins M., and Haqqani, A.S. Blood-brain barrier models: in vitro to in vivo translation in preclinical development of CNS-targeting biotherapeutics. Expert Opin Drug Discov. 2015 Feb;10(2):141-55.

16. Watts, R. J., & Dennis, M. S. (2013). Bispecific antibodies for delivery into the brain. Current opinion in chemical biology, 17(3), 393-399.

17. Yu, Y. J., Atwal, J. K., Zhang, Y., Tong, R. K., Wildsmith, K. R., Tan, C., ... & Bumbaca, D. (2014). Therapeutic bispecific antibodies cross the blood-brain barrier in nonhuman primates. Science translational medicine, 6(261), 261ra154-261ra154.

KEYWORDS: Alphavirus, Blood Brain Barrier, BBB, Eastern Equine Encephalitis Virus, Encephalitis, Nanobody, Venezuelan Equine Encephalitis Virus, Western Equine Encephalitis Virus

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