Contact PD/PI: Harty, Ronald N.

PI: HARTY, RONALD N Title: Development of Small Molecule Therapeutics Targeting Hemorrhagic Fever 

Viruses

Received: 09/08/2020 Opportunity: PA-20-265 Council: 01/2021

Competition ID: FORMS-F FOA Title: PHS 2020-2 Omnibus Solicitation of the NIH for Small Business 

Technology Transfer Grant Applications (Parent STTR [R41/R42] Clinical Trial Not 

Allowed)

2R42AI138630-03 Dual: Accession Number: 4489979

IPF: 10020652 Organization: FOX CHASE CHEMICAL DIVERSITY CENTER, INC

Former Number: 3R41AI138630-02 Department: Pathobiology and Microbiology

IRG/SRG: ZRG1 BCMB-G (10)B AIDS: N Expedited: N 

Subtotal Direct Costs Animals: Y New Investigator:

(excludes consortium F&A) Humans: Y Early Stage Investigator:

Year 3: Clinical Trial: N 

Year 4: Current HS Code: 30

Year 5: HESC: N 

HFT: N

Senior/Key Personnel: Organization: Role Category:

Ronald Harty Ph.D University of Pennsylvania PD/PI

Jay Wrobel Ph.D Fox Chase Chemical Diversity Center, 

Inc.

Co-Investigator

Olena Shtanko Texas Biomedical Research Institute Co-Investigator

John Kulp Ph.D Fox Chase Chemical Diversity Center, 

Inc. Department: Divisi

Co-Investigator

Allen Reitz Ph.D Fox Chase Chemical Diversity Center Co-Investigator

Always follow your funding opportunity's instructions for application format. Although this application 

demonstrates good grantsmanship, time has passed since the grantee applied. The samples may not reflect the 

latest format or rules. NIAID posts new samples periodically:

https://www.niaid.nih.gov/grants-contracts/sample-applications

The text of the application is copyrighted. The awardee provided express permission for NIAID to post this 

grant application and summary statement for educational purposes. The awardee allows you to use the

material (e.g., data, writing, graphics) they shared in the applications for nonprofit educational purposes only, 

provided the material remains unchanged and the principal investigators, awardee organizations, and NIH 

NIAID are credited.

Freedom of Information Act (FOIA). NIAID is strongly committed to protecting the integrity and confidentiality 

of the peer review process. When NIH responds to FOIA requests for grant applications and summary 

statements, the material will be subject to FOIA exemptions and include substantial redactions. NIH must

protect all confidential commercial or financial information, reviewer comments and deliberations, and personal 

privacy information.

Note on Section 508 Conformance and Accessibility. We have reformatted this sample to improve 

accessibility for people with disabilities and users of assistive technology. If you have trouble accessing the 

content, contact the NIAID Office of Knowledge and Educational Resources at deaweb@niaid.nih.gov.
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

OMB Number: 4040-0010 

Expiration Date: 12/31/2022 

APPLICATION FOR FEDERAL ASSISTANCE 

SF 424 (R&R) 

3. DATE RECEIVED BY STATE State Application Identifier 

1. TYPE OF SUBMISSION* 4.a. Federal Identifier 

R41AI138630 

❍Pre-application ❍Application ●Changed/Corrected 

Application 

b. Agency Routing Number 

2. DATE SUBMITTED Application Identifier c. Previous Grants.gov Tracking Number 

GRANT13199627 

5. APPLICANT INFORMATION Organizational DUNS*: 

Legal Name*: Fox Chase Chemical Diversity Center Inc 

Department: 

Division: 

Street1*: Pennsylvania Biotechnology Center 

Street2: 

City*: Doylestown 

County: Bucks 

State*: PA: Pennsylvania 

Province: 

Country*: USA: UNITED STATES 

ZIP / Postal Code*: 18902-8400 

Person to be contacted on matters involving this application 

Prefix: First Name*: Allen Middle Name: Last Name*: Reitz Suffix: Ph.D 

Position/Title: Chief Executive Officer 

Street1*: Pennsylvania Biotechnology Center 

Street2: 

City*: Doylestown 

County: Bucks 

State*: PA: Pennsylvania 

Province: 

Country*: USA: UNITED STATES 

ZIP / Postal Code*: 18902-8400 

Phone Number*: Fax Number: Email: 

6. EMPLOYER IDENTIFICATION NUMBER (EIN) or (TIN)* 

7. TYPE OF APPLICANT* R: Small Business 

Other (Specify): 

Small Business Organization Type ❍Women Owned ❍Socially and Economically Disadvantaged 

8. TYPE OF APPLICATION* 

❍New ❍Resubmission 

●Renewal ❍Continuation ❍Revision 

If Revision, mark appropriate box(es). 

❍A. Increase Award ❍B. Decrease Award ❍C. Increase Duration 

❍D. Decrease Duration ❍E. Other (specify) : 

Is this application being submitted to other agencies?* ❍Yes ●No What other Agencies? 

9. NAME OF FEDERAL AGENCY* 

National Institutes of Health 

10. CATALOG OF FEDERAL DOMESTIC ASSISTANCE NUMBER 

TITLE: 

11. DESCRIPTIVE TITLE OF APPLICANT'S PROJECT* 

Development of Small Molecule Therapeutics Targeting Hemorrhagic Fever Viruses 

12. PROPOSED PROJECT 

Start Date* Ending Date* 

04/01/2021 03/31/2024 

13. CONGRESSIONAL DISTRICTS OF APPLICANT 

PA-001 

Funding Opportunity Number: PA-20-265 . Received Date: 

2020-09-08T12:43:45.000-04:00 Tracking Number: GRANT13199760 

Page 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

SF 424 (R&R) APPLICATION FOR FEDERAL ASSISTANCE Page 2 

14. PROJECT DIRECTOR/PRINCIPAL INVESTIGATOR CONTACT INFORMATION 

Prefix: First Name*: Ronald Middle Name: N. Last Name*: Harty Suffix: Ph.D 

Position/Title: Professor 

Organization Name*: University of Pennsylvania 

Department: Pathobiology and Microbiology 

Division: School of Veterinary Medicine 

Street1*: 

Street2: 

City*: Philadelphia 

County: 

State*: PA: Pennsylvania 

Province: 

Country*: USA: UNITED STATES 

ZIP / Postal Code*: 19104-0000 

Phone Number*: Fax Number: Email*: 

15. ESTIMATED PROJECT FUNDING 

a. Total Federal Funds Requested* $ 

b. Total Non-Federal Funds* $0.00 

c. Total Federal & Non-Federal Funds* $ 

d. Estimated Program Income* $0.00 

16.IS APPLICATION SUBJECT TO REVIEW BY STATE 

EXECUTIVE ORDER 12372 PROCESS?* 

a. YES ❍ THIS PREAPPLICATION/APPLICATION WAS MADE 

AVAILABLE TO THE STATE EXECUTIVE ORDER 12372 

PROCESS FOR REVIEW ON: 

DATE: 

b. NO ● PROGRAM IS NOT COVERED BY E.O. 12372; OR 

❍ PROGRAM HAS NOT BEEN SELECTED BY STATE FOR 

REVIEW 

17. By signing this application, I certify (1) to the statements contained in the list of certifications* and (2) that the statements herein 

are true, complete and accurate to the best of my knowledge. I also provide the required assurances * and agree to comply with 

any resulting terms if I accept an award. I am aware that any false, fictitious, or fraudulent statements or claims may subject me to 

criminal, civil, or administrative penalties. (U.S. Code, Title 18, Section 1001) 

● I agree* 

* The list of certifications and assurances, or an Internet site where you may obtain this list, is contained in the announcement or agency specific instructions. 

18. SFLLL or OTHER EXPLANATORY DOCUMENTATION File Name: 

19. AUTHORIZED REPRESENTATIVE 

Prefix: First Name*: Kathleen Middle Name: Last Name*: Czupich Suffix: 

Position/Title*: Chief Financial Officer 

Organization Name*: Fox Chase Chemical Diversity Center, Inc. 

Department: 

Division: 

Street1*: 

Street2: 

City*: Doylestown 

County: Bucks 

State*: PA: Pennsylvania 

Province: 

Country*: USA: UNITED STATES 

ZIP / Postal Code*: 18902-8400 

Phone Number*: Fax Number: Email*: 

Signature of Authorized Representative* Date Signed* 

09/08/2020 

20. PRE-APPLICATION File Name: 

21. COVER LETTER ATTACHMENT File Name: 

Funding Opportunity Number: PA-20-265 . Received Date: 

2020-09-08T12:43:45.000-04:00 Tracking Number: GRANT13199760 

Page 2 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

424 R&R and PHS-398 Specific 

Table Of Contents 

SF 424 R&R Cover Page 1 

Table of Contents 3 

Performance Sites 4 

Research & Related Other Project Information____________________________________________________ 6 

Project Summary/Abstract(Description)________________________________________________________ 7 

Project Narrative 8 

Facilities & Other Resources 9 

Equipment 19 

Research & Related Senior/Key Person________________________________________________________ 23 

Research & Related Budget Year - 1 46 

Research & Related Budget Year - 2 49 

Research & Related Budget Year - 3 52 

Budget Justification 55 

Research & Related Cumulative Budget _______________________________________________________ 96 

Research & Related Budget - Consortium Budget (Subaward 1)_____________________________________ 97 

Research & Related Budget - Consortium Budget (Subaward 2)____________________________________ 108 

Total Direct Costs Less Consortium F&A ______________________________________________________ 120 

SBIR STTR Information ___________________________________________________________________ 121 

Commercialization Plan 123 

PHS398 Cover Page Supplement____________________________________________________________ 135 

PHS 398 Research Plan 137 

Specific Aims 138 

Research Strategy 139 

Progress Report Publication List___________________________________________________________ 151 

PHS Human Subjects and Clinical Trials Information___________________________________________ 152 

Study 1: Development of Host-Oriented Small Molecule Therapeutics for Filoviruses and Arena 

viruses 154 

Inclusion Enrollment Reports _________________________________________________________ 160 

Vertebrate Animals 167 

Select Agent Research 173 

Bibliography & References Cited 175 

Consortium/Contractual Arrangements______________________________________________________ 178 

Letters of Support 180 

Resource Sharing Plan(s) 189 

Authentication of Key Biological and/or Chemical Resources ____________________________________ 190 

Table of Contents Page 3 
 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. OMB Number: 4040-0010 

Expiration Date: 12/31/2022 

Project/Performance Site Location(s) 

Project/Performance Site Primary Location ❍I am submitting an application as an individual, and not on behalf of 

a company, state, local or tribal government, academia, or other type of 

organization. 

Organization Name: Fox Chase Chemical Diversity Center, Inc. 

Duns Number: 

Street1*: Pennsylvania Biotechnology Center 

Street2: 

City*: Doylestown 

County: Bucks 

State*: PA: Pennsylvania 

Province: 

Country*: USA: UNITED STATES 

Zip / Postal Code*: 

Project/Performance Site Congressional District*: PA-001 

Project/Performance Site Location 1 ❍I am submitting an application as an individual, and not on behalf of 

a company, state, local or tribal government, academia, or other type of 

organization. 

Organization Name: University of Pennsylvania 

DUNS Number: 

Street1*: 

Street2: 

City*: Philadelphia 

County: 

State*: PA: Pennsylvania 

Province: 

Country*: USA: UNITED STATES 

Zip / Postal Code*: 19104-6205 

Project/Performance Site Congressional District*: PA-002 

Page 4 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265. Received Date: 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Project/Performance Site Location 2 ❍I am submitting an application as an individual, and not on behalf of 

a company, state, local or tribal government, academia, or other type of 

organization. 

Organization Name: Texas Biomedical Research Institute 

DUNS Number: 

Street1*: 8715 W. Military Dr. 

Street2: 

San City* Antonio 

County: 

State*: TX: Texas 

Province: 

Country*: USA: UNITED STATES 

Zip / Postal Code*: 78227-5302 

Project/Performance Site Congressional District*: TX-020 

Additional Location(s) File Name: 

Page 5 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265. Received Date: 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

OMB Number: 4040-0010 

Expiration Date: 12/31/2022 

RESEARCH & RELATED Other Project Information 

1. Are Human Subjects Involved?* ● Yes ❍ No 

1.a. If YES to Human Subjects 

Is the Project Exempt from Federal regulations? ❍ Yes ● No 

If YES, check appropriate exemption number: 1 2 3 4 5 6 7 ___ 

If NO, is the IRB review Pending? ● Yes ❍ No 

IRB Approval Date: 

Human Subject Assurance Number none 

8 

2. Are Vertebrate Animals Used?* ● Yes ❍ No 

2.a. If YES to Vertebrate Animals 

Is the IACUC review Pending? ● Yes ❍ No 

IACUC Approval Date: 

Animal Welfare Assurance Number none 

3. Is proprietary/privileged information included in the application?* ● Yes ❍ No 

4.a. Does this project have an actual or potential impact - positive or negative - on the environment?* ❍ Yes 

4.b. If yes, please explain: 

4.c. If this project has an actual or potential impact on the environment, has an exemption been authorized or an ❍ Yes 

environmental assessment (EA) or environmental impact statement (EIS) been performed? 

4.d. If yes, please explain: 

● No 

❍ No 

5. Is the research performance site designated, or eligible to be designated, as a historic place?* ❍ Yes 

5.a. If yes, please explain: 

● No 

6. Does this project involve activities outside the United States or partnership with international ❍ Yes 

collaborators?* 

6.a. If yes, identify countries: 

6.b. Optional Explanation: 

● No 

Filename 

7. Project Summary/Abstract* Summary_Harty_Final.pdf 

8. Project Narrative* Narrative_Harty_Final.pdf 

9. Bibliography & References Cited References-Harty_Final_2.pdf 

10.Facilities & Other Resources Facilities_FCCDC.9_combined.pdf 

11.Equipment Equipment_FCCDC.2_combined.pdf 

Page 6 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265. Received Date: 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Summary: The ultimate goal of this Phase II application is to develop novel small molecule, broad-spectrum 

therapeutics against viral infections caused by filoviruses, arenaviruses, and other viruses that depend on the 

PPxY L-domain motif for egress and spread of infection. Some of these viruses, including Ebola (EBOV), 

Marburg (MARV), and Lassa fever (LAFV) viruses, are highly pathogenic and classified as Category A bioterror 

pathogens. We and others have determined that efficient budding of these emerging human pathogens depends 

on the subversion of host proteins, such as neural precursor cell expressed developmentally down-regulated 

protein 4 (Nedd4), by PPxY L-domains in the matrix proteins of these RNA viruses. The identification and 

development of small molecule inhibitors that interfere with these virus-host interactions should effectively block 

virus egress, disease progression, and transmission. In these efforts we have discovered several chemical series 

of small molecule inhibitors of the host Nedd4/virus PPxY complex important for viral egress which led to one 

analog possessing proof of concept in vivo activity in a Marburg virus challenged mouse model. As FDA 

approved therapeutic agents for the treatment of these most of these viral infections are not available, our 

identification of virus-host inhibitors that may prevent virus spread will fill a significant unmet need. Moreover, 

these inhibitors will be broad-spectrum, and therefore will likely be effective against newly emerging viruses as 

well as viral variants. As described below, we will use a rigorous multifaceted approach to identify, develop, and 

validate PPxY budding inhibitors identified in Phase I as potent, broad-spectrum antivirals. The goal of this Phase 

II STTR grant application is to optimize our lead inhibitors of VP40 PPxY-Nedd4 interactions to generate fullfledged predevelopment drug candidates ready for IND directed studies. This will be accomplished by combining 

the pharmaceutical and medicinal chemistry expertise of the scientists at the Fox Chase Chemical Diversity 

Center, Inc. (FCCDC) with the expertise and experience in the experimental aspects of antiviral therapy of the 

Harty Lab at the University of Pennsylvania. We will realize this goal by optimizing our existing series of inhibitors, 

exemplified by in vivo active FC-10696, for improved potency and oral drug properties (Aim 1), evaluating new 

compounds based on two potent series for their ability to specifically inhibit PPxY-Nedd4 interactions and 

subsequent VLP and surrogate virus egress (Aim 2), identifying compounds having suitable drug properties and 

selectivity using in vitro and in vivo ADMET evaluation (Aim 3), and evaluating compounds for their antiviral 

efficacy against authentic BSL-4 viruses in vitro and in vivo (Aim 4). 

Project Summary/Abstract Page 7 
 

 

Contact PD/PI: Harty, Ronald N. 

Narrative: 

There is an urgent need to develop antiviral therapy against emerging human RNA viruses that represent 

potential agents of bioterrorism (Ebola, Marburg, and Lassa fever). We have discovered small molecule 

compounds that disrupt budding and spread of live viruses; a process that is critical for virus dissemination and 

disease progression. Here, the Harty group at the University of Pennsylvania, experts in the antiviral technology 

of this proposal, have partnered with the small business Fox Chase Chemical Diversity Center, Inc. to further 

develop these broad spectrum antiviral budding inhibitors by using medicinal chemistry, live virus budding 

assays, and live cell imaging techniques. 

Project Narrative Page 8 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Resources – Laboratories of Fox Chase Chemical Diversity Center, Inc. 

Fox Chase Chemical Diversity Center, Inc. (FCCDC) is located in a 112,000 ft2 facility known as the 

Pennsylvania Biotechnology Center (PA Biotech Center) at Doylestown, PA. The Center 

is a recently renovated, $14 million, non-profit business incubator, funded in part by a grant from the 

Commonwealth of Pennsylvania, whose mission is to nurture entrepreneurship and biotechnology in the region. 

It is home to ~30 biotechnology companies, and the non-profit Blumberg Institute. The FCCDC has 1,600 ft2 of 

laboratory space within the facility including six chemistry fume hoods and one BSL2 incubator, and 2,400 ft2 of 

office space. FCCDC also conducts laboratory operations at a second nearby facility at 3700 Horizon Dr., King 

of Prussia, PA, the former Bachem site, where it leases 6,600 ft2 of combined lab and office space including 11 

chemistry fume hoods. 

Offices. The main FCCDC offices are located within the PA Biotech Center. They are on the same floor as the 

laboratories and with dedicated administrative assistance available within a reception area. The offices are 

equipped with a computer, conferencing-capable phone system to facilitate collaborative discussions, and 

locking file cabinets for storage of confidential study information. The office suite includes two conferences 

rooms, one of which is set-up for computer based presentations. FCCDC staff are fully supported by an 

associated administrative infrastructure (copy machines, fax, reception, video conferencing, etc.) A computer 

network connected by T1 lines for scientific writing/editing, data analysis, and graphics/illustration is available at 

the PA Biotech Center. 

FCCDC laboratories have the following equipment that will be specifically used for supporting the work 

described in this grant application: two 300-MHz 1H NMRs with multinuclear probe capability (13C, 31P, 18F), four 

LC/MSs including three Micromass ZQ mass spectrometers with Waters 2695 HPLC with diode array detectors, 

multiple ISCO and Biotage personal chromatography purification equipment, multiple Gilson 215 semipreparative HPLC systems, Biotage Initiator microwave, two Parr hydrogenators, 18 rotary evaporators, 

Genevac EZ2 Evaporation System, Mettler microbalances, JChem temperature controllers, Innova platform 

shaker, miscellaneous smaller glassware, PCs, printers, and related equipment, BSL2 incubators for cell culture, 

laminar flow biosafety cabinets, multiple pipettors, common refridgerators and freezers. 

Chemical compound information is stored within an MS Excel workbook and is accessible via Instant JChem 

which is also used to calculate biophysical properties such as LogP/D and tPSA (www.chemaxon.com). FCCDC 

has a companylicense for the structure and literature searching tool SciFinder-n, and free on-site access via the 

Pennsylvania Drug Discovery Institute to >30,000 reagents and starting materials (including >800 boronic acids) 

that are binned in functionality appropriate trays and can be searched electronically. 

Records Retention. All FCCDC final compound samples are stored in a locked repository. All laboratory 

notebooks are signed daily and witnessed on a monthly basis. We observe standard industry practices in the 

area of confidentiality. Laboratory notebooks are kept under lock and key in the evenings and weekends. NMR 

data is stored on a primary server which is backed up to a second server. We have a license to ACD software, 

allowing us to print out coupling constants, multiplicities and chemical shifts of NMR spectra quickly, which is 

very useful when writing patent applications or manuscripts for publication. 

Collaborative Scientific Environment. Fox Chase Chemical Diversity Center, Inc. holds regular and 

periodicgeneral research review and specific project review meetings. Project meetings would be held with the 

relevant research staff at our academic partner either in person, by go-to-meeting web review, or by phone. The 

PA Biotech Center and the non-profit Pennsylvania Drug Discovery Institute hosts scientific seminars on 

approximately a monthly basis, including eminent researchers in the area of drug discovery. Speakers have 

included Bill Greenlee, Magid Abou-Gharbia, Donna Huryn, and Dan Skronovsky. 

Facilities & Other Resources Page 9 
 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Facilities and Resources 

Harty, Ronald N.

Laboratory: 1,008 sq. ft. of BSL-2 laboratory space including a tissue culture area. The laboratory can 

accommodate up to 7 people and is equipped with one 6-ft and one 4-ft biohazard hoods for cell culture and 

virus experiments, three tissue culture incubators, two ultra low freezers, two 37° C floor shaker/incubators, 

two Beckman L90 ultracentrifuges, two Beckman Avanti J-E centrifuge, and several refrigerators/freezers. An 

additional shared equipment suite and darkroom are available on the floor. 

Animal: N/A 

Clinical: N/A 

Computer: I have a Mac Desktop, a MacBook Pro laptop, and an HP Laserjet M1212 laser printer/scanner/fax 

in my office. The laboratory is equipped with two Dell Optiplex GX260 systems and a Mac Desktop with all 

necessary software along with two laser printer/scanner/fax units. All computers have internet and email 

access. 

Office: I have a 92 sq. ft. office within my research lab. 

Other: Secretarial services are provided by the Department of Pathobiology. Building facilities, information 

technology (IT), and glassware services are located in the building, along with a meeting/conference room. 

Scientific Environment: The laboratory is within the School of Veterinary Medicine at the University of 

Pennsylvania. There are a plethora of active virology, immunology, molecular biology, biochemistry and 

parasitology laboratories within the School of Veterinary Medicine and University wide with an abundance of 

both pre- and postdoctoral trainees. 

Biohazards: The work in my laboratory involves studies approved for BSL-2 conditions. 

Penn Vet Imaging Facility: The Veterinary School Core Imaging Facility houses a two-photon/confocal (4 laser) 

spectral imaging Leica SP5 system on a DM6000 microscope with tandem scanner, five internal spectral 

detectors, and four external non-descanned detectors and motorized XY stage. This system is optimized for 

multicolor intravital imaging. The core also has a Yokagawa CSU X1 spinning disk confocal head on a Leica 

DMI4000 microscope and an LMM5 dual laser excitation source, a Hamamatsu 512 EM camera, and an ASI 

motorized and computer controlled XYZ stage. This instrument is housed within an environmental (temperature 

and CO2 controlled) chamber and is ideal for long term imaging of live cells. We just acquired a new Leica SP5 

spectral imaging microscope on an inverted stand (DMI6000) that is equipped with 3 continuous wave lasers, 3 

pulsed lasers and a Leica/Picoquant Fluorescence Lifetime Imaging (FLIM) system. This instrument is 

optimized for live cell imaging of cells and specifically for real time measurements of protein-protein 

interactions. This instrument is the only one of its kind at the University of Pennsylvania and is ideally 

configured for the FLIM measurement of virus-host interactions. The core also has several software packages 

for image analysis including Metamorph from Molecular Devices, Volocity from Improvision, and the LAS-AF 

suite from Leica. 

Facilities & Other Resources Page 10 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

FACILITIES AND OTHER RESOURCES 

Texas Biomedical Research Institute 

ENVIRONMENT: Texas Biomedical Research Institute (Texas Biomed) was founded as a private, not-forprofit research institution in 1941 by Thomas B. Slick Jr., with a goal to unravel the mysteries of disease 

through innovative thinking, creative problem solving, and cutting edge technologies. Located on a 200-acre 

campus on the northwest side of San Antonio, Texas, our scientists work each and every day to improve 

human health by being a world leader in infectious disease research. Texas Biomed partners with hundreds 

of researchers and institutions, targeting advances in the fight against infectious diseases impacting our 

communities around the world, including AIDS, TB, hepatitis, malaria, hemorrhagic fever viruses, parasitic 

infections and many other infectious diseases. We also study co-morbidities of infection and vulnerable 

populations, such as the very young and the elderly and those suffering from cardiovascular diseases and 

metabolic disorders. 

Texas Biomed is staffed by more than 350 employees including a multidisciplinary team of approximately 65+ 

doctoral-level scientists – physiologists, geneticists, virologists, immunologists, molecular biologists, cell 

biologists, physicians and veterinarians – who lead more than 200 major research projects. Texas Biomed 

also supports 150 technical staff and an administrative support staff of more than 100 professionals. 

Texas Biomed recently underwent a strategic planning process and defined its focus as an institute that studies 

infectious diseases and conditions that make an individual susceptible to infectious diseases (aging, metabolic 

syndrome, etc.). The Institute developed three new programmatic areas: Host-Pathogen Interactions is the 

study of the basic biology of infection in humans and animals – the immediate intersection of host and 

pathogen. Disease Intervention & Prevention is the development of diagnostics, treatments and vaccines to 

reduce the severity of/or cure infection. Population Health is the identification of correlates of disease 

susceptibility or resistance to infectious diseases on a population level. Each program has two faculty leads 

who focus on the academic development of the programs and their themes. Administrative responsibilities are 

supported through the Office of the Vice President for Research, unencumbering scientists to pursue the 

academic mission of Texas Biomed. 

Texas Biomed supports a culture of learning and development. The Institute holds a weekly seminar that hosts 

an external guest approximately monthly, and internal presentations from faculty and trainees at all other times. 

Attendance at seminar ranges between 50 and 80 faculty and research staff. Administrative staff are also 

frequently in attendance to learn about the latest research at Texas Biomed. Texas Biomed supports a monthly 

journal club that rotates across the three programmatic areas that typically has an attendance of 20-40 

researchers. Individual laboratories also hold their own journal clubs, frequently shared across multiple 

scientific groups. The Vice President for Research runs a monthly training course for Staff Scientists on how 

to manage a laboratory, and a monthly training course for a subset of Staff Scientists on how to write a grant. 

Staff Scientists at Texas Biomed are the equivalent of Research Faculty. Human Resources provide training 

through a Lunch & Learn lecture series and individualized training programs on management and leadership. 

Communication at Texas Biomed is facilitated by a monthly institute-wide newsletter and video blogs. 

Texas Biomed is part of a city wide, 4-institute collaborative (with University of Texas Health Science Center, 

University of Texas San Antonio, and Southwest Research Institute) that works synergistically to increase the 

biomedical reputation of the city of San Antonio. Initiatives include the San Antonio Vaccine Development 

Center and the Precision Therapeutics Program, which are supported by and foster collaboration across the 

four institutions. Administrative collaboration includes negotiated rates for use of each institutions cores, 

participation in graduate and training programs, and input on faculty search committees including 

consideration for dual hire appointments. 

RESEARCH FACILITIES: Our faculty and staff scientists conduct their research amid 550,000 square feet 

(sq. ft.) of space that includes laboratories and core facilities, offices, auditorium, animal hospital, library, and 

animal facilities, including a six-acre corral. Research laboratories are spaced across three main buildings for 

a total of 55,361 sq. ft. of space. 

Facilities & Other Resources Page 11 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Moorman (building 35; 31,954 sq. ft.; where Shtanko lab is located): The Moorman building has 12 

laboratories totaling 9,459 sq. ft. for in vitro BSL2 studies. Each lab has a dedicated technician office. Faculty 

offices (approx. 300 sq. ft.) are peripheral to the laboratories and directly adjacent to offices and cubicles for 

staff and students. The Moorman can support between 6-12 faculty, depending on space requirements. The 

Moorman also supports approximately 1,200 square feet of select agent BSL3 laboratory, and a 2,100 sq. ft. 

of ABSL4 suite laboratory (1,200 operational, remaining space is mechanical and annex space). 

Dr. Shtanko has 800 sq. ft. of lab space which is outfitted with common equipment necessary for studies in 

cell biology, molecular biology, and immunology. She also has private office space located in the same building 

within 80 feet of the lab. 

Earl Slick (building 71; 59,556 square feet): The Earl Slick building has 14 laboratories totaling 12,298 sq. ft. 

for in vitro BSL2 studies, 1,099 square feet for BSL2+ studies (for HIV), 1,840 sq. ft. of BSL3 prep space, and 

10,500 sq. ft. of office space. Faculty offices (approx. 250 sq. ft.) and offices for support staff and trainees are 

in close proximity to associated laboratory space. Earl Slick can support between 7-14 faculty, depending on 

space requirements. Laboratories for individual faculty are outfitted with common equipment necessary for 

studies in cell biology, molecular biology, and immunology. 

There is a dedicated blood draw room immediately next to the BSL2 lab space, to ensure privacy during 

phlebotomy procedures. This room is equipped with a refrigerator for juice boxes for donors and a phlebotomy 

chair with upholstered arms and foot rest, and can recline to the Trendelenburg position. Only trained individuals 

will serve as phlebotomists. 

Urschel-Slick (building 12; 57,875 sq. ft.): The Urschel-Slick building has 16 laboratories totaling 11,995 sq. 

ft. for BSL2 studies, 7,500 sq. ft. for BSL3 studies, including 1,840 sq. ft. of BSL3 prep space, and 10,500 sq. 

ft. of office space. Faculty offices (approx. 250 sq. ft.) and offices for support staff and trainees are in close 

proximity to associated laboratory space. Urschel-Slick can support between 8-16 faculty, depending on space 

requirements. Urschel-Slick is the location of a recently renovated 7,500 sq. ft. ABSL3 and rodent vivarium. 

Ledford (building 38; located 4,876 sq. ft.): The Ledford building is a free-standing building with 2,579 sq. 

ft. of laboratory space and 1,305 sq. ft. of adjacent office space. The Ledford building houses the institute 

Central Core Facility and is no more than 3 minutes walking distance from any research space at Texas 

Biomed. The building is keycard access restricted and monitored. Core staff are located within the building to 

provide support for any instrument. The Ledford building has a darkroom for individual light sensitive 

equipment. 

UT Health San Antonio (10 min drive) has the following cores available to the PI: Mass Spectrometry, 

Bioanalytics & Single cell, Biobanking & Genome Analysis, Biomolecular NMR, Flow Cytometry, 

Macromolecular interactions, Microt CT, Electron Microscopy, Optical Imaging, and X-ray Crystallography 

(see http://research.uthscsa.edu/RCL/cores.shtml). For detailed UT Health San Antonio Core Resources, see: 

http://research.uthscsa.edu/RCL/Resourcepages.shtml 

CLINICAL: Human subject studies are reviewed by UT Health San Antonio Institutional Review Board through 

a joinder agreement. UT Health is located approximately a 15-minute drive from Texas Biomed and is affiliated 

with University Hospital, Audie L. Murphy VA Hospital, Robert B. Green Campus, and the County Public Health 

Hospital. 

ANIMAL: Texas Biomed has an independent Institutional Animal Care and Use Program which is responsible 

for monitoring vertebrate animal research to be compliant with USDA and AAALAC standards, and adheres 

to the Animal Welfare Act, Public Health Service Policy on Humane Care and Use of Laboratory Animals, and 

the Guide for the Care and Use of Laboratory Animals. Animal Welfare Assurance #A308201. Texas Biomed 

has been accredited by AAALAC continuously since 1973 (AAALAC File #000246). 

A dedicated rodent vivarium is located in the Slick-Urschel building with current capacity to house rodents on 

ventilated racks or in static caging. Space is available for expansion and to accommodate additional species. 

The vivarium is supported by a dedicated supervisor and staff, and veterinary care. The vivarium staff also 

support rodent studies at ABSL3. 

Facilities & Other Resources Page 12 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

COMPUTER: In addition to personal computers, there are 57 desktop and 31 laptop computers in addition to 

printers and fax machines available within the Moorman and adjacent buildings, and the Library to be used 

for instrument operation, data management, word processing, graphics and manuscript preparation. These 

computers are connected to the central network and are protected by individual UPS devices. There is open 

access through the Internet to library resources at Texas Biomed. 

Scientists can use the institutional accounts of the Slack discussion platform and the Dropbox file share service 

to increase communication and data sharing internally and externally with collaborators. On servers, scientists 

have access to a wide variety of software and programming and scripting languages such as FORTRAN, C, 

C++, Java, Tcl, Perl, Python, Ruby, bash, R, etc. Statistical and mathematical packages available include 

GraphPad, Maple, S-PLUS, and R. Software available for genetic analysis includes PAP, S.A.G.E., MENDEL, 

FISHER, LINKAGE, CRI-MAP, PREST, Merlin, GeneHunter, SimWalk2, Loki, PLINK, GATK, vcftools, 

Ingenuity Pathway Analysis, kinship2, and SOLAR. Specific software or packages can be installed on demand. 

A dedicated staff of 9 persons led by the Vice President for Information Technology maintains and updates 

current systems and assists scientists in specific requests (installation of software, libraries and packages, 

help in debugging failed compilation). A ticket system is in place to efficiently address issues. The technical 

staff has expertise in database management in both SQL-based software and the PEDSYS database system, 

developed in-house for management and analysis of genetic, demographic, epidemiological and laboratory 

data. 

Data and System Security 

We have implemented extensive security measures to protect our computers and data files against 

unauthorized access. Our security measures are compliant with IRB requirements for human data. Our 

internet connection is a 100 Mb/s Ethernet connection, using a leased router managed by our internet provider, 

and protected by our Palo Alto Networks next generation security appliances, in a redundant configuration. 

Users connect to the system over the network using either SSH/SFTP or VPN connections. Access to network 

is removed on or before the day that an individual's employment is terminated, unless prior arrangements 

have been made. Access logs are checked periodically for repeated login failures which might suggest an 

attempt to break into our system. Countermeasures are taken if evidence of such an attempt is found. Access 

to central files containing research, clinical, support, or administrative data is restricted to a list of approved 

users. Separate lists are maintained for each research or administrative unit (laboratory, project, etc.). 

Disk-based daily incremental and monthly full backups of software and data are performed using the Veritas 

NetBackup system. Long-term data archiving is provided by a single six drives, 113-slot 800 GB tape library, 

and four 25-slot 400 GB tape libraries. 

REDCap (Research Electronic Data Capture) is available for sharing and managing data across computer 

platforms and between research institutes. REDCap is a secure, web-based application for building and 

managing online surveys and databases. REDCap provides automated export procedures for seamless data 

downloads to Excel and common statistical packages (SPSS, SAS, Stata, R), as well as a built-in project 

calendar, a scheduling module, ad hoc reporting tools, and advanced features, such as branching logic, file 

uploading, and calculated fields. 

BIOSAFETY: 

ABSL4 laboratory 

Texas Biomed is home to the only operational ABSL4 lab owned by a private institution. The ABSL4 lab supports 

basic research as well as vaccine and therapeutics testing of agents with no cure. This unique resource has 

supported the research of faculty in Texas Biomed in emerging diseases and bioterror agents including the 

hemorrhagic fever viruses Ebola virus and Marburg virus. 

This full-suit lab is CDC and USDA certified for work on human and animal pathogens. Access to the ABSL4 

area is secured via three successive magnetic card/keypad controlled airlock passages: The first is located at 

the entry foyer to the BSL3 suite, the second at the entry foyer to the outer changer room, and the third at the 

entry foyer to the ABSL4 area. The ABSL4 area is equipped with an entry and exit decontamination airlock 

and an emergency exit/decontamination chamber. The latter also serves as an equipment decontamination 

Facilities & Other Resources Page 13 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

chamber to allow for the repair, removal and/or replacement of faulty equipment without requiring a complete 

laboratory decontamination process. All supply and exhaust air for the ABSL4 passes through dual (and 

tandem) HEPA filters. 

Critical parameters (air supply and exhaust flow rates, pressure differentials, normal and emergency breathing 

air, decontamination systems, etc.) are continuously monitored. Personnel within the lab communicate, and 

are actively monitored by outside personnel, through a scrambled UHF communication system worn inside 

the laboratory suit. The 500 sq. ft. ABSL4 laboratory staging area includes an air locked suit change room, two 

inner change rooms with pass-through showers, and an outer “clean” change room equipped with sinks, 

lockers, toilets, etc. Critical mechanical support devices are located in an attached 3,000 sq. ft. mechanical 

support complex. All critical equipment and devices (valves, etc.) are redundant, including duplicate air-supply 

and exhaust systems, decontamination shower systems, as well as the compressors, pumps, valves, heater 

cores, etc. that comprise the liquid waste decontamination system. All critical mechanical devices operate on 

a system with automatic emergency electricity backup generators. The ABSL4 area (including mechanical 

support rooms) is equipped with an elaborate intrusion detection system that includes electronic door and roof 

hatch sensors, motion detectors, etc. 

The laboratory contains 3 x class IIB biological safety hoods, 4 x water-jacketed CO2 incubators, low-, highand ultra-speed centrifuges equipped with both analytical and preparative scale rotors, a microfuge, 2 x 4°C 

refrigerators, 2 x liquid nitrogen storage systems, and 3 x -80°C freezers. The ABSL4 also has an optical 

ELISA plate reader, MAGPIX multiplexing 96-well format unit, luminometer, Innova 4000 environmental 

shaker, microplate shaker, BioFlo III fermenters with chillers, Evos fluorescent microscope, pass-through 

autoclave, dunk tank, facsimile machine and networked computers. 

Texas Biomed maintains an experienced and trained staff of scientists, veterinarians, research technicians 

and veterinary technicians available to perform studies at high biocontainment. These individuals have 

demonstrated proficiency at conducting in vitro and animal studies with the agent identified in the proposal. 

The ABSL4 Operations and Safety Manuals specify policies, procedures, and standard operating procedures 

(SOPs) for the safe handling of biological materials in biosafety laboratories. The policies, procedures, and 

SOPs comply with applicable Federal, State, and municipal regulations and with the guidelines “Biosafety in 

Microbiological and Biomedical Laboratories” issued by the CDC and the National Institute of Health (NIH). 

Employees are trained from these manuals on each facilities mechanical systems, biosafety, biocontainment 

and security. Employees are also trained according to project specific and SOPs. These procedures apply to 

all that use, generate, store, or dispose of potentially infectious materials in Texas Biomed biosafety 

laboratories and to persons who must enter these laboratories to perform services. 

Select agent use, transfer or possession is forbidden without the permission of the Responsible Official (RO) 

and until the required forms filed and written approval received from the CDC Select Agent Program. Upon 

approval, ABSL4 investigators desiring to work on a BSL4 project must also submit an application to the 

Biohazard and Safety Committee for approval. The Committee is responsible for evaluating the facility, 

equipment, and staff capabilities to perform work in a safe manner. 

Infectious cultures and inventory stocks are stored inside the ABSL4 laboratory in locked refrigerators and 

freezers. An electronic Inventory (eInventory) system is used to document usage of infectious stocks. All 

infectious material stored in refrigerators or freezers is properly labeled and stored in containers capable of 

withstanding thermal shock of freezing and thawing. Each container is labeled with the identity of the infectious 

agent, the date of the preparation, and a barcode that links the material to the more inclusive information 

contained in the inventory database. 

When work is completed, all infectious cultures are removed from workbenches and cabinets and stored in 

a designated refrigerator or freezer. Materials to be discarded are placed in a sealable container filled with 

a suitable disinfectant. The container is placed in a discard pan containing the disinfectant. Discard pans 

are placed on a cart and transported to the autoclave. Labware containing infectious liquids are stored and 

transported in leak-proof containers large enough to contain the fluid in case of leakage. 

Facilities & Other Resources Page 14 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Biocontainment Program 

Texas Biomed has established an institution-wide comprehensive BSL3/BSL4 and CDC certified Select Agent 

BSL3 facilities program. The development of a centralized program for the campus ensures that 1) the growth 

and development of BSL4/BSL3/BSL2 facilities will remain well-coordinated both scientifically and 

administratively at all levels of the program; 2) spaces will be developed in the most effective manner based 

on an accurate assessment of need and cost; 3) technical support will be coordinated among the facilities; 4) 

training will be optimized, including sending key personnel to important national meetings for training and 

education on an ongoing basis, a critical need considering the rapidly changing landscape regarding security 

requirements and controls; and 5) assurance that broad expertise on campus is represented in BSL4/3/2 

discussions regarding a) work with bacteria, viruses, toxins, etc; b) entry criteria for users, including the 

CDC/FBI program; c) exposure protocols for users; d) proper use and maintenance of animals; e) user issues 

and concerns; and f) infrastructure issues and concerns. 

Institutional BSL3/4 compliance oversight is provided by Environmental Health and Safety (EHS). EHS 

provides the following services for the entire institute operation: radiation safety, environmental compliance, 

occupational health and safety, research safety and biosafety. Texas Biomed has a program director for the 

BSL3/BSL4/Biosecurity program, a Responsible Official and alternate for the Select Agent Program and a 

Biohazardous and Safety Committee (BSC). 

Complementing and interacting with the regulatory and compliance arm of the BSL3/BSL4/Biosecurity 

activities on campus is the centralized research oversight structure. The Biocontainment Program is managed 

by a BSL3 Director, BSL4 Director, and two biocontainment program coordinators. These staff work closely with 

the Director of Maximum Containment Contract research. Program operations include an institute-wide 

Biosafety-3/4 Advisory Committee (BAC) that consists of the EHS Director, BSL3 Director, BSL4 Director/IBC 

Chair, and coordinators. The VP for Research/RO attends the meeting monthly. The charge of the BAC is to 

develop and review institution BSL3/4 rules and regulations, develop and refine BSL3/4 user entry 

requirements for certification, evaluate the use and efficiency of campus BSL3/4 facilities, develop and refine 

the BSL3/4 medical surveillance program, handle user issues as they arise, and provide oversight for the 

operation of the facilities. A complete Texas Biomed BSL3/Select agent/BSL4 education and entry program 

for users is in place. Texas Biomed users meet as a group (BUG) with leadership on a monthly basis. 

Access to BSL3/4 facilities is granted only when personnel receive thorough biosafety and biosecurity training 

and proper on-site training. Training materials are reviewed by the Responsible Officer (RO) and/or Alternative 

Responsible Officer (ARO). Biosafety and biosecurity training emphasizes the facility design and system in 

place as well as the current rules and regulations users must follow. Refresher training is provided on an annual 

basis and time sensitive issues are discussed in the monthly BSL3/4 user group (BUG) meeting. On-site 

training is focused on demonstrating proper application of personal protective equipment (PPE), facility 

features, and proper usage of equipment. Junior scientists must be accompanied and supervised by senior 

scientists who are experienced in BSL3/4-related techniques to ensure proper handling of infectious materials. 

The BSL3/4 program at Texas Biomed ensures that all BSL3/4 users are fully trained and supported by 

operational staff, relieving some of the burden of training and oversight from the PI. 

Select Agent Research 

Texas Biomed is a select agent registered entity with Health and Human Services, Centers for Disease Control 

and Prevention and U.S. Department of Agriculture, Animal Plant Health Inspection Service, National Select 

Agent Program. The Institute has been inspected by the CDC National Select Agent Program for use of HHS 

Select Agents and Toxins, Overlap Select Agents and Toxins and USDA Select Agents and Toxins. The 

registration number is C20171120 1976. 

Facilities & Other Resources Page 15 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

OTHER INSTITUTIONAL RESOURCES: 

Biology Core instrumentation and services 

The Biology Core, housed in the Central Core facility, is operated by a Director and two PhD level Managers. 

Microscopy: The Biology Core operates a Zeiss 3-Channel Confocal System Microscope LLC LSM800 and a 

Nikon C2+ Confocal Module microscope. Microscopes are each located within 2 separate buildings (Earl Slick 

and Moorman) to be in close proximity to the most frequent users. 

Microscopy in Moorman building is performed using a Nikon Ti Eclipse microscope housed in a separate, 

dedicated microscopy room. The Nikon Ti Eclipse microscope is necessary for high resolution and high-speed 

image acquisition of cells and fluorescently labeled virus-like particles. It is fitted for both high speed widefield 

image capture as well as confocal capabilities using the Nikon C2+ scan head with high sensitivity 

photomultipliers. The microscope has automated image acquisition ability with electronic XYZ piezo stage and 

Nikon elements software suitable for high content screening of drug libraries, siRNA and expression plasmids. 

Objective lenses are of the highest quality including oil immersion 40x NA 1.30 lens, 100x oil immersion lambda 

NA 1.45 lens as well as conventional air 4x, 10x, 40x, and 60x lenses. The scope is fitted with a heated, CO2 

fed, incubator stage for live cell work. The camera for widefield imaging is an ANDOR iXon3 EMCCD camera, 

which provides rapid, highly sensitive acquisition of 14-bit and 16-bit images with high linear sensitivity. Software 

is a key part of quantitative image analysis. AutoQuant X3 software provides image deconvolution and 

interfaces with Imaris (Bitplane) to identify and measure image features, including measuring the distance 

between features in 3D. CellProfiler software is used to count and measure object intensity in multiple images 

and interfaces with FCSExpress to perform image cytometry, which allows gating of cell populations like 

FACS. The latter allows analysis of cell features after gating which is an advantage. ImageJ processing 

software is also available. 

BD FACSymphony: Available since September of 2019, the 5-laser BD FACSymphony cytometer is capable 

of 28 color acquisition, and it is equipped with a High Throughput Sampler. This advanced instrument features 

an ultra-quiet VPX electronics system that supports up to 50 high-performance photomultiplier tubes (PMTs) 

and improves detection sensitivity to enable the identification and analysis of rare cell types and events. The 

version at the Core is equipped with a 355 nm (UV), 405 nm (violet), 488 nm (blue), 561 nm (yellow-green) 

and 640 nm (red) lasers. When analyzing panels designed with BD Horizon Brilliant dyes, this platform helps 

to overcome research challenges, such as collecting maximal information from a precious sample, and 

increases lab throughput with broad phenotyping panels that combine multiple cell-line-specific panels. This 

highly customizable platform can be configured so that you can select from multiple laser wavelengths and 

power ratings, and choose the positions of decagon detection arrays to address the requirements of your 

specific research application. 

Beckman Coulter Cyan ADP: The CyAnTM LX Flow Cytometer provides three excitation lines, eleven 

parameters, complete compensation, and high-speed acquisition, in a bench-top configuration. The 405 nm, 

488 nm, and 633 nm lasers are fixed and alignment-free, with spatially separated beams for minimal optical 

crosstalk. Spatially separated optical apertures, high-performance solid-state detectors, and an obscuration 

bar minimize unwanted laser radiation. User-interchangeable filters provide experimental flexibility. Eleven 

acquisition channels (including forward scatter, side scatter, and 9 colors) provide parallel processing of pulse 

height, pulse length, area, and log for each parameter. 

BD Accuri C6 Plus: The 2-laser BD Accuri C6 plus cytometer is capable of 4-color acquisition. The BD Accuri 

is dedicated to BSL2+ HIV studies. The BD Accuri C6 Plus is equipped with a blue and red laser, two light 

scatter detectors, and four fluorescence detectors with optical filters optimized for the detection of many 

popular fluorochromes. 

BD FACSAria cell sorter: The 4-laser FACSAria II sorter is capable of 18-color, 4-way sorting and has an 

automated cell deposition unit for 6, 12, 24, 48 and 96 well sorting. The BD FACSAriaTM cell sorter is a fixedalignment cuvette flow cell that can detect up to 18 colors, for a total of 20 parameters. Cells passing through 

the cuvette flow cell are excited by four solid state lasers at 407-nm, 488-nm, 561-nm, and 633-nm. Advanced 

digital electronics improve sort performance and provide significant advantages in instrument ease-of-use. 

This cell sorter is contained within a Baxter High Containment Biosafety cabinet within one of the BSL3 

laboratories. 

Facilities & Other Resources Page 16 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

BDInflux cell sorter: The 5-laser BDInflux cell sorter is capable of 6-way sorting. The BDInflux is primarily 

dedicated to single cell sorting of malaria parasites and sorting of pluripotent stem cells. 

Luminex 100 and 200: The Luminex with xMAP (multi-analyte platform) technology systems measure analytes 

(cytokines, chemokines, etc.) on a multiplex platform. These instruments are operated with dedicated 

computers running Luminex Xponent software; analysis of reactivity is done with Masterplex QT software. 

Molecular Services Core instrumentation and services 

The Molecular Services Core (MSC) is located in the Central Core Facility and is operated by a Director, a 

full-time manager, and technical support staff. The MSC is dedicated to assist researchers in the development 

and implementation of state-of-the-art molecular approaches in the support of their research. The MSC has 

an active Users’ Group that periodically reviews available services, technology, and operations. The MSC is 

equipped with instrumentation to conduct most genomic and gene expression applications, and MSC 

personnel are trained in numerous technical and bioinformatic approaches. 

Equipment: The MSC provides access to an Illumina iSeq 100 and Illumina MiSeq sequencer for low- to midoutput NGS applications. Fully automated sequencing sample preparations of 8-96 samples are supported 

with a Sciclone G3 NGSx automated liquid handling system. A 10X Genomics Chromium controller and 

Fluidigm C1 are available for single cell NGS applications. Additional instrumentation includes a Sage 

PippinHT for automated size selection and PCR purification, an Agilent 2100 Bioanalyzer and 4200 

TapeStation for determining RNA, DNA, and protein integrity, and a Nanodrop spectrophotometer and 

Invitrogen Qubit Fluorometer for DNA, RNA and protein sample quantitation. Multiple additional thermal 

cyclers are available to support these instruments. 

The MSC also provides access to an Illumina iScan with autoloader to allow 24 h service without user intervention 

for high-throughput genotyping and gene expression analyses (allowing the processing of up to 96 multisample BeadChips per day). The iScan System supports rapid, sensitive, and accurate imaging of Illumina's 

array-based genetic analysis products. A Tecan Freedom Evo liquid handling platforms is also available for 

automation of all Illumina protocols. 

Multiple PCR machines (996- and 384-well blocks), two AB QuanStudio 5 and one QuantStudio 6 Real Time 

PCR Systems and an AB QuantStudio 3D Digital PCR System are available to support real-time PCR, TaqMan 

genotyping, gene expression, absolute quantification, and rare allele detection assays. The Core Facility also 

has a TECAN GENios multimode plate reader with fluorescence, absorbance and glow luminescence modes 

and a high-performance Tecan plate washer for both 96- and 384-well plates. 

Additionally, a BioRad Chemidoc gel documentation system with UV and chemiluminescence capabilities is 

available in an electrophoresis clean room. Two Beckman ultracentrifuges are available as well. 

A dedicated computer server for sequence analysis is available and consists of a Dell PowerEdge R830 server 

with two Xenon E5-4640 2.1GHz processors (48 cores), 32TB of useable space, and 256 GB RAM. Partek 

Flow Genomic Analysis Software is supports start-to-finish analysis for next generation sequencing data 

applications. 

Services: Services provided by the MSC facility include RNA and DNA isolation, quality determination, and 

quantitation from a variety of sample sources including blood, tissues, cells, and body fluids, sequencing library 

preparations supporting whole genome, transcriptome, exome, smRNA, ChIP, and targeted sequencing 

applications, onsite low and mid-output next generation sequencing services, and standard sequencing data 

analysis and bioinformatics for DNA, RNA, and smRNA sequencing. 

Pathology 

The pathology facility has the following rooms in Building 10: necropsy (288 sq. ft.), hematology (162 sq. ft.), 

chemistry (160 sq. ft.), tissue processing (162 sq. ft.), slide processing and administration (224 sq. ft.), 

refrigerator/freezer room (148 sq. ft.), storage room (132 sq. ft.), multi-use area (bacterial plating, automated 

immunohistochemistry, and additional clinical pathology microscopy station - 288 sq. ft.), four offices for the 

pathologists and technicians, and a room that houses the stereology microscope and visiting foreign and 

Facilities & Other Resources Page 17 
 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

American veterinarians and students. It also has a 332 sq. ft. climate controlled slide, block and wet tissue 

storage room in the quarantine building and a 216 sq. ft. walk in refrigerator located outside and adjacent to 

the necropsy room. 

The necropsy facility is organized to safely handle any necropsy (BSL2) or tissue share project. It has a downdraft 

stainless steel table, a mobile MOPEC HEPA filtered bone saw, and protective clothing for handling hazardous 

necropsies. Tissue samples can be collected in formalin, paraformaldehyde, or other fixatives as required by 

a given research project. Tissues can also be collected in various media, OCT embedded for frozen sectioning, 

snap frozen, or collected fresh. Perfusion fixation of tissues is also available. A Nikon D200 digital camera is 

used in the necropsy suite to record lesions. Digital gross and microscopic images are stored on the pathology 

server and are searchable by either pathology accession number or animal ID. The pathologists also provide 

on-site necropsy support for nonhuman primate studies in the BSL3 and BSL4 containment areas. 

The histology laboratory provides tissue processing, embedding, paraffin and frozen tissue sectioning, routine 

and special staining, and immunohistochemistry procedures. The laboratory is equipped with a Tissue-Tek 

processor and embedding center as well as automatic stainers for routine H&E and immunohistochemistry 

staining. The lab is also equipped with an automated coverslipper, centrifuge, cytospin, cryostat, oven, 

microscope, dissecting microscope, two microtomes with automatic features, refrigerators, scales, a safe for 

GLP and animal rule specimens, and other ancillary equipment. A Nikon DXM1200C camera on a Nikon 801 

microscope and an Olympus SC100 camera on an Olympus BX50 microscope are used to obtain images from 

stained slides. The histology laboratory has developed immunohistochemistry procedures for several 

antigens/pathogens, including filovirus, herpesvirus papio, baboon reovirus, alphavirus, flavivirus, amyloid 

beta- 42, herpes simplex virus, as well as antibody purification. Available antibodies include cluster of 

differentiation markers (CD) CD68 (KP1), CD68 (3F103), CD20, CD3, S-100, Ki-67, NeuN, Nurr1 (2 

antibodies), HSV1, progesterone receptor, estrogen receptor, Von Willebrand Factor, and TUNEL assay for 

apoptosis. Other IHC procedures are developed as needed. 

Facilities & Other Resources Page 18 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

EQUIPMENT (all onsite) – Fox Chase Chemical Diversity Center Inc 

NMR:Two Varian Mercury Plus 300-MHz Inova NMRs with multinuclear capability (1H, 13C, 31P, 18F). 

HPLC/MS: 

• Four Micromass ZQ Mass Specs. with Waters 2695 HPLC with 996 diode array detector. 

Chromatography: 

• Three Gilson 215 semi-prep HPLC systems, multi-wavelength, automated fraction collection. 

• Teledyne Isco CombiFlashRf automated chromatography system. 

• Three Isco Combiflash Sg 100c personal chromatography systems. 

Evaporation: 

• Genevac EZ2 Evaporation System. 

• FTS Systems Flexi-dry Lyophilyzer. 

• 18 Rotavapors with vacuum pumps (Buchi, Heidolph). 

• 2 VWR Sheldon 1400E Vacuum Ovens with Edwards oil pumps. 

Hydrogenation: Two Parr 3911 Shaker hydrogenators. 

Reaction: 

• Biotage Initiator microwave synthesizer with 60 position autosampler. 

• Innova platform shaker, 2000. 

• 4 J-Kem Gemini-2 dual temperature controllers with teflon-coated thermocouples. 

• 24Ika Magnetic Stirrer/Hotplates. 

BL2 Biological and Tissue Culture Labs: 

• Baker SG400 and Labconco A2 Class II biological safety cabinets. 

• ABI Prism 7000 Sequence Detection System. 

• New Brunswick Environmental incubator shaker and incubator. 

• Biotek Synergy 2 Multi-Detection Microplate Reader. 

• Molecular Devices SpectraMax 190 microplate reader. 

• TitertekMultidrop. 

• LabomediVU 1500 Microscope. 

• Nikon Eclipse (TE 2000) microscope interfaced with image analysis software 

• Fisher and VWR Carbon dioxide incubators. 

Computational Chemistry: 

• The PA Biotechnology Center has 600Mbs Internet access, more than sufficient. 

• Two AMD Threadripper 2950X 16 Core Workstations: Both workstations have 64GB of DDR4 RAM, 

1x Nvidia GeForce RTX 2070 GPU, and 1TB Samsung 970 Evo NVMe SSD. 

• One GPU server with 2x Intel Xeon Gold 5218 16 core/32 thread 2.3GHz, 96GB of DDR4 RAM, with 

5 GPUs (2x Nvidia Titan RTX, 2x Nvidia Titan V, 1x Nvidia 2080Ti), 1x 512GB Samsung 970 PRO 

NVMe SSD boot drive, and 2x 2TB Samsung 860 EVO SSDsTwo Dell Precision dual-processor Xeon 

5680 workstations with Nvidia 470GTX graphics cards; 

• Schrodinger’s Small Molecule Drug Discovery Suite (all modules); 

• GROMACS (molecular dynamics with customized scripts for automation); 

Storage, Miscellaneous: 

• Two (2) Justrite 45 Gallon Flammable Safety Cabinets. 

• Four (4) Refrigerator/Freezers, Frigidaire, GE, Revco. 

• SciFinder-n, latest, most powerful search engine for chemistry 

Reagents and starting materials collection available onsite: >30,000 members including >1,000 

boronic acids, bar-coded, searchable electronic database. 

Equipment Page 19 
 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Major Equipment 

Harty, Ronald N.

Major equipment in my laboratory and the adjacent equipment suite includes: 2 Beckman L90 ultracentrifuges, 

2 Beckman J21 centrifuges with appropriate rotors, DNA and protein electrophoresis systems, 4 heated water 

baths, 3 CO2 cell culture incubators, 2 cell culture bio-cabinets, 2 bacterial shaker/incubators, a fume hood, 3 

-85°C ultralow and 3 -20°C freezers, 2 refrigerators, a Savant speedvac concentrator, 2 PCR units, 2 rotator 

platforms, 3 microfuges, a gel dryer, 3 liquid nitrogen tanks, an inverted Olympus microscope, phosphoimager 

and related software, STORM scanner, Biotek Synergy microplate reader, X-ray film developer and darkroom. 

Equipment Page 20 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

MAJOR EQUIPMENT 

Texas Biomedical Research Institute 

Apart from the equipment located in Texas Biomed Core Facilities, Dr. Shtanko also has access to the following 

equipment: 

Shtanko Lab 

Class IIA 4 feet Biosafety cabinets (1) 

Tissue culture CO2 incubators (1) 

370 Incubators for bacterial work (1) 

Inverted microscopes (1) 

Refrigerated microcentrifuge (1) 

Microcentrifuge (1) 

Water baths (2) 

Heat blocks (2) 

Rockers (2) 

Neon transfection system (1) 

Geni syngene gel imaging system (1) 

Viaflo Assist Integra system (1) 

Impulse Sealer (1) 

Dryfast Welch 2014B-01 (1) 

Isotemp FisherScientific 205 (1) 

ABSL4 Facility 

Class IIB 6 feet Biosafety cabinets (3) 

Tissue culture CO2 incubators (4) 

Hydro Peroxide vapor decontamination systems (2) 

Beckman ultracentrifuge Optima (1) 

Refrigerated Benchtop fuge Eppendorf 5810R (1) 

Microcentrifuge (1) 

Multidrop Combi SMART dispenser (1) 

Milli-Q water purification system (1) 

Select agent tracking phase I and II (2) 

Select agent geneport system (1) 

Water baths (2) 

Heat blocks (7) 

Rockers (2) 

Optical ELISA plate reader (1) 

Luminometer microplate (1) 

Innova 4000 environmental shaker (1) Piccolo 

Evos fluorescent microscope (1) 

BioFlo III fermenters with chillers (2) 

Multidrop Combi SMART dispenser (1) 

Milli-Q water purification system (1) 

Agarose/PAGE gel apparatus and power packs (2) 

Countess Invitrogen C10281 (1) 

Vortexes (2) 

Thermoscientific legend XTR centrifuge (1) 

-20°C freezers (1) 

-80°C freezers (1) 

4°C refrigerators (1) 

Liquid nitrogen storage system (1) 

Biophotometer Eppendorf 6132 (1) 

Uninterrupted power supply (UPS) unit (2) 

iBlot 2 gel transfer device (1) 

Balances (2) 

Xpress Analyzer (2) 

Abaxis VetScan VS2 (3) 

Ventilated rabbit cage unit (2) 

One Cage 2100 ventilated racks and cages (2) 

bioBUBBLE containment enclosure (4) 

TissueLyser II and accessories Qiagen (2) 

Procyte hematology (1) 

DSI telemetry system large animal (2) 

NanoDrop One Microvolume UV-Vis 

Spectrophotometer (1) 

-80°C freezers (3) 

CytoSMART LUX system (2) 

4°C refrigerators (2) 

Liquid nitrogen storage system (2) 

MAGPIX 96-well unit (1) 

Moorman Research Building (where Dr. Shtanko lab is located) 

Laminar flow hoods (17) 

Tissue culture CO2 incubators (18) 

370 Incubators for bacterial work (3) 

Nikon Ti Eclipse confocal microscope system (1) 

Virus Counter 2100 (1) 

Cherry Biotech microscope system (1) 

NanoDrop One spectrophotometer (2) 

Inverted microscopes (7) 

Refrigerated microcentrifuges (5) 

Microcentrifuges (25) 

Benchtop medium speed centrifuges (14) 

Beckman ultracentrifuge Optima L100K (2) 

FACSAria flow cytometer (1) 

Milli-Q water purification systems (2) 

Countess Invitrogen C10281 (4) 

Touch thermal cycler C1000 (1) 

Equipment Page 21 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Cold room (1) 

Dark room (1) 

BSL2 labs (12) 

BSL3 labs (3) 

ABSL4 labs (1) 

4°C refrigerators (14) 

Liquid nitrogen storage units (7) 

-20°C freezers (12) 

-80°C freezers (10) 

pH meters (4) 

QStudio 6 Fix-Realtime PCR system (1) 

KrosFlo Research II TFF system (1) 

Image processing station for Nikon C2+ confocal 

module microscope (1) 

Odyssey SA IR imaging system (1) 

Illumina MiSeq System (1) 

Eppendorf epMotion M5073 automated DNA/RNA 

purification system (1) 

BenchPro plasmid processing station (1) 

HM Hematology analyzer (1) 

MagNa Lyser with rotors (1) 

Abaxis VetScan VS2 (5) 

Aero3G Biaera aerosol control platform (1) 

Aerodynamic particle sizer spectrometer 3321TSI 

(1) 

Aerosol Diluter 3302A TSI (1) 

Primate telemetry repeater (1) 

VS Blood Chemistry analyzer (1) 

iMark Absorbance microplate reader (2) 

Luminometer microplate readers (4) 

TECAN Genios multimode plate reader (1) 

Equipment Page 22 
 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. OMB Number: 4040-0010 

Expiration Date: 12/31/2022 

RESEARCH & RELATED Senior/Key Person Profile (Expanded) 

PROFILE - Project Director/Principal Investigator 

Prefix: First Name*: Ronald Middle Name N. Last Name*: Harty Suffix: Ph.D 

Position/Title*: Professor 

Organization Name*: University of Pennsylvania 

Department: Pathobiology and Microbiology 

Division: School of Veterinary Medicine 

Street1*: 

Street2: 

City*: Philadelphia 

County: 

State*: PA: Pennsylvania 

Province: 

Country*: USA: UNITED STATES 

Zip / Postal Code*: 19104-0000 

Phone Number*: Fax Number: 

E-Mail*: 

Credential, e.g., agency login: 

Project Role*: PD/PI Other Project Role Category: 

Degree Type: PhD Degree Year: 1991 

Attach Biographical Sketch*: File Name: Harty_Bio.pdf 

Attach Current & Pending Support: File Name: 

Page 23 

Funding Opportunity Number: PA-20-265 . Received Date: Tracking Number: GRANT13199760 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

PROFILE - Senior/Key Person 

Prefix: First Name*: Jay Middle Name Edward Last Name*: Wrobel Suffix: Ph.D 

Position/Title*: Vice President, Academic Relations 

Organization Name*: Fox Chase Chemical Diversity Center, Inc. 

Department: 

Division: 

Street1*: Pennsylvania Biotechnology Center 

Street2: 

City*: Doylestown 

County: 

State*: PA: Pennsylvania 

Province: 

Country*: USA: UNITED STATES 

Zip / Postal Code*: 18902-8400 

Phone Number*: Fax Number: 

E-Mail*: 

Credential, e.g., agency login: 

Project Role*: Co-Investigator Other Project Role Category: 

Degree Type: PhD Degree Year: 1983 

Attach Biographical Sketch*: File Name: Wrobel_biosketch_for_Harty.4.pdf 

Attach Current & Pending Support: File Name: 

PROFILE - Senior/Key Person 

Prefix: First Name*: Olena Middle Name Last Name*: Shtanko Suffix: 

Position/Title*: Staff Scientist I 

Organization Name*: Texas Biomedical Research Institute 

Department: Virology and Immunology 

Division: 

Street1*: 

Street2: 

City*: San Antonio 

County: 

State*: TX: Texas 

Province: 

Country*: USA: UNITED STATES 

Zip / Postal Code*: 782270000 

Phone Number*: Fax Number: 

E-Mail*: 

Credential, e.g., agency login: 

Project Role*: Co-Investigator Other Project Role Category: 

Degree Type: PHD Degree Year: 2010 

Attach Biographical Sketch*: File Name: 2._Bio_Shtanko_20-240_-_final.pdf 

Attach Current & Pending Support: File Name: 

Page 24 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265 . Received Date: 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

PROFILE - Senior/Key Person 

Prefix: First Name*: John Middle Name Last Name*: Kulp 

Position/Title*: Director of Computational Chemistry 

Organization Name*: Fox Chase Chemical Diversity Center, Inc. Department: Divisi 

Department: 

Division: 

Street1*: Pennsylvania Biotechnology Center 

Street2: 

City*: Doylestown 

County: 

State*: PA: Pennsylvania 

Province: 

Country*: USA: UNITED STATES 

Zip / Postal Code*: 18901-8400 

Suffix: Ph.D 

Phone Number*: Fax Number: 

E-Mail*: 

Credential, e.g., agency login: 

Project Role*: Co-Investigator Other Project Role Category: 

Degree Type: PhD Degree Year: 2006 

Attach Biographical Sketch*: File Name: Kulp_biosketch_for_Harty.pdf 

Attach Current & Pending Support: File Name: 

PROFILE - Senior/Key Person 

Prefix: First Name*: Allen Middle Name Bernard Last Name*: Reitz 

Position/Title*: CEO 

Organization Name*: Fox Chase Chemical Diversity Center 

Department: 

Division: 

Street1*: 

Street2: 

City*: Doylestown 

County: 

State*: PA: Pennsylvania 

Province: 

Country*: USA: UNITED STATES 

Zip / Postal Code*: 189020000 

Suffix: Ph.D 

Phone Number*: Fax Number: 

E-Mail*: 

Credential, e.g., agency login: 

Project Role*: Co-Investigator Other Project Role Category: 

Degree Type: PHD Degree Year: 1982 

Attach Biographical Sketch*: File Name: Reitz_Biosketch_for_Harty.2.pdf 

Attach Current & Pending Support: File Name: 

Page 25 

Funding Opportunity Number: PA-20-265 . Received Date: Tracking Number: GRANT13199760 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

OMB No. 0925-0001 and 0925-0002 (Rev. 03/2020 Approved Through 02/28/2023) 

BIOGRAPHICAL SKETCH 

Provide the following information for the Senior/key personnel and other significant contributors. 

Follow this format for each person. DO NOT EXCEED FIVE PAGES. 

NAME: Harty, Ronald N. 

eRA COMMONS USER NAME (credential, e.g., agency login): 

POSITION TITLE: Professor of Microbiology 

EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, 

include postdoctoral training and residency training if applicable. Add/delete rows as necessary.) 

INSTITUTION AND LOCATION 

DEGREE 

(if 

applicable) 

Completion 

Date 

MM/YYYY 

FIELD OF STUDY 

University of Lowell, Lowell, MA 

Louisiana State University Med. Ctr., Shreveport 

Louisiana State University Med. Ctr., Shreveport 

Mount Sinai School of Medicine, New York, NY 

B.S. 

Ph.D. 

Postdoctoral 

Postdoctoral 

06/1985 

08/1991 

01/1993 

08/1998 

Biology 

Virology 

Virology 

Virology 

A. Personal Statement 

The overall goal of my research on negative-sense RNA viruses is to understand how these viruses assemble 

and bud from cells, and how host protein recruitment and virus-host interactions contribute to these late stages 

of virus replication. My current research program is focused on understanding the molecular mechanisms of 

filovirus (Ebola and Marburg viruses) and arenavirus (Lassa fever) assembly/budding, so that we can identify 

broad-spectrum inhibitors of virus-host interactions and validate their efficacy in blocking virus budding. Indeed, 

through a highly collaborative and productive effort in partnership with Dr. Jay Wrobel’s team at Fox Chase 

Chemical Diversity Center, and Dr. Olena Shtanko at the BSL-4 laboratory at Texas Biomedical Research 

Institute, we have identified lead candidate budding inhibitors with potential broad-spectrum activity. In Phase 2, 

we will rapidly advance and develop these lead candidate therapeutics for NIAID Category A pathogens toward 

clinical trials. Overall, my expertise, experience, and demonstrated record of successful and productive research 

in the areas of RNA virus-host interactions and virus egress/transmission has prepared me to lead the proposed 

collaborative experiments described in this proposal. 

1. Harty, R. N., Brown, M. E., Wang, G., Huibregtse, J., and Hayes, F. P. 2000. A PPxY- motif within the 

VP40 protein of Ebola virus interacts physically and functionally with a ubiquitin ligase: Implications for 

filovirus budding. Proc. Natl. Acad. Sci. U.S.A. 97(25):13871-13876. PMCID: PMC17668. 

2. Han, Z., Madara, J.J., Herbert, A., Prugar, L.I., Ruthel, G., Lu, J., Liu, Y., Liu, W., Liu, X., Wrobel, J.E., 

Reitz, A.B., Dye, J.M., *Harty, R.N., and *Freedman, B.D. (*co-corresponding authors). 2015. Calcium 

regulation of hemorrhagic fever virus budding: Mechanistic implications for host-oriented therapeutic 

intervention. PLoS Pathog, 11(10):e1005220. PMCID: PMC4634230. 

3. Liang, J., Sagum, C.A., Bedford, M.T., Sidhu, S.S., Sudol, M., Han, Z., and Harty, R.N. 2017. Chaperonemediated autophagy protein BAG3 negatively regulates Ebola and Marburg VP40-mediated egress. PLoS 

Pathog, Jan. 11;13(1):e1006132. PMCID: PMC5226679. 

4. Han, Z., Dash, S., Sagum, C.A., Ruthel, G., Jaladanki, C.K., Berry, C.T., Schwoerer, M.P., Harty, N.M., 

Freedman, B.D., Bedford, M.T., Fan, H., Sidhu, S.S., Sudol, M., Shtanko, O., and Harty, R.N. 2020. Modular 

mimicry and engagement of the Hippo pathway by Marburg virus VP40: Implication for filovirus biology and 

budding. PLoS Pathog., Jan.6;16(1):e1008231. PMCID: PMC6977764. 

Biosketches Page 26 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

B. Positions and Honors 

1998-2004 Assistant Professor, Dept. of Pathobiology, School of Veterinary Medicine, University of 

Pennsylvania 

2004-2015 Associate Professor, Dept. of Pathobiology, School of Veterinary Medicine, University of 

Pennsylvania 

2014-present Head, Laboratory of Infectious Diseases & Immunology, Dept. of Pathobiology, School of 

Veterinary Medicine, University of Pennsylvania 

2015-present Professor, Dept. of Pathobiology, School of Veterinary Medicine, University of Pennsylvania 

Other Experience and Professional Memberships 

1987–present Member, American Society for Microbiology and American Society for Virology 

2001 Pfizer Award for Excellence in Research, University of Pennsylvania 

2002 Dean's Award for Leadership in Basic Science Education, University of Pennsylvania 

2003-2007 Member, Special Emphasis Panel, Biodefense and Emerging Pathogens, NIAID, NIH. 

2004-2005 Chair of the Filovirus Workshop Sessions at the American Society for Virology Meetings 

at McGill University, Montreal, Canada and at Penn State University, University Park, PA 

2004-present Member – Editorial Board, Journal of Virology; Ad Hoc Reviewer for Virology, Traffic, 

Molecular Cell, Vaccine, PNAS, PloS Pathogens, Cell. Micro., and J. of Gen. Virol. 

2005 Member - Drug Discovery and Mechanisms of Antimicrobial Resistance Special Emphasis 

Panel Study Section [ZRG1 DDR (01) Q], Infectious Diseases and Microbiology IRG, NIH. 

2005-2010 Member - Special Emphasis Panel, ZRG1 F13 NRSA Fellowships, NIH. 

2008-2009 Chair - Special Emphasis Panel, ZRG1 F13 NRSA Fellowships, NIH. 

2014 Ad Hoc Reviewer – NIH/NIAID Topics in Virology Study Section, ZRG1 IDM-W (applications 

having conflicts with standing VIR-A and VIR-B study sections) 

2014 Ad Hoc Reviewer – NIH/NIAID, Pathogenic Eukaryotes (PTHE) Study Section 

2015 Ad Hoc Reviewer – NIH/NIAID, Drug Discovery and Mechanisms of Antimicrobial Resistance 

(DDR) Study Section ZRG1 IDM-T (82) 

2015 Ad Hoc Reviewer – NIH, Special Emphasis Panel ZRG1 AARR-D (02) M 

2016 Chair – NIAID, Special Emphasis Panel to review an Ebola P01 application. 

2017 Ad Hoc Reviewer – NIH/NIAID VIR-B study section 

2018 Ad Hoc Reviewer – NIH R13 (Support for Scientific Conferences) Review Panel 

2019 Reviewer – FY19 Congressionally Directed Medical Research Programs (CDMRP), Peer 

Reviewed Medical Research Program (PRMRP) 

C. Contributions to Science 

1. Viral L-domain/host interactions regulate virus egress and spread. Viruses have developed unique and 

complex molecular mechanisms to ensure efficient egress of mature virions from infected cells. I focused my 

early studies on unraveling the intricate roles of both viral and host proteins in this process, and particularly the 

specific recruitment of host factors to promote efficient budding of infectious virus. A better understanding of 

these virus-host interactions and the mechanisms of virus budding not only will provide fundamental insights into 

the functions of both viral and host proteins, but also will lead to the emergence of novel strategies to inhibit 

virion egress and spread. The Late (L) budding domain motifs (PTAP and/or PPxY) are highly conserved in the 

matrix proteins of a wide array of RNA viruses (e.g. filoviruses, arenaviruses, rhabdoviruses, paramyxoviruses, 

henipaviruses, and retroviruses) and represent attractive and novel targets for the development of therapeutics 

having broad-spectrum antiviral activity. My lab was first to demonstrate that the Ebola virus VP40 protein plays 

a central and sufficient role in virion assembly and egress, due in part to the presence of overlapping PTAP and 

PPEY L-domains. We went on to show that independent expression of VP40 led to the production and egress 

of virus-like particles (VLPs) that accurately mimic budding of live infectious virus. We used VP40 VLPs and 

genetically engineered VSV recombinants that expressed WT and mutant L-domain motifs from Ebola virus 

VP40, to show that budding is dependent on viral L-domain mediated recruitment of host proteins associated 

with the ESCRT pathway (Tsg101 and Nedd4) for complete virus-cell fission or separation. Most recently, we 

have identified for the first time a third functional L-domain motif (YPx(n)L) within Ebola virus VP40 that serves as 

an alternative L-domain motif to enhance VP40 VLP release by recruiting and interacting with host ESCRTassociated protein Alix. 

a. Harty, R. N., Brown, M. E., Wang, G., Huibregtse, J., and Hayes, F. P. 2000. A PPxY- motif within the 

Biosketches Page 27 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

VP40 protein of Ebola virus interacts physically and functionally with a ubiquitin ligase: Implications for 

filovirus budding. Proc. Natl. Acad. Sci. U.S.A. 97(25):13871-13876. PMCID: PMC17668. 

b. Licata, J. M., Han, Z., Simpson-Holley, M., Wright, N. T., Paragas, J., and Harty, R. N. 2003. 

Overlapping motifs (PTAP and PPEY) within the Ebola virus VP40 protein function independently as 

late budding domains: Involvement of host proteins tsg101 and vps4. J. Virol. 77(3), 1812-1819. 

PMCID: PMC140960 

c. Han, Z., Schwoerer, M.P., Hicks, P., Liang, J., Ruthel, G., Berry, C.T., Freedman, B.D., Sagum, C.A., 

Bedford, M.T., Sidhu, S.S., Sudol, M., and Harty, R.N. 2018. Host protein BAG3 is a negative regulator 

of Lassa VLP egress. Diseases, Jul 13;6(3). pii: E64. 

d. Han, Z., Dash, S., Sagum, C.A., Ruthel, G., Jaladanki, C.K., Berry, C.T., Schwoerer, M.P., Harty, N.M., 

Freedman, B.D., Bedford, M.T., Fan, H., Sidhu, S.S., Sudol, M., Shtanko, O., and Harty, R.N. 2020. 

Modular mimicry and engagement of the Hippo pathway by Marburg virus VP40: Implication for filovirus 

biology and budding. PLoS Pathog., Jan.6;16(1):e1008231. PMCID: PMC6977764. 

2. Identification and development of host-oriented L-domain inhibitors of virus budding. Based in part on 

our steady progress in elucidating the molecular aspects and host involvement in virus budding from studies 

described above, and our long-term goal of developing antivirals, a current major effort in my lab is to identify, 

develop, and optimize small molecule compounds targeting viral L-domain/host interactions to inhibit virus 

egress and spread. We postulate that for viruses such as Ebolavirus, administration of such an antiviral 

therapeutic during an outbreak would inhibit virus dissemination and spread in infected individuals, thus slowing 

disease progression and allowing the individual’s immune system time to mount a robust response to effectively 

combat and clear the infection. Indeed, there is a vital need for the advancement and development of effective 

and safe therapeutics against emerging, high priority pathogens such as Ebola, Marburg, Lassa fever, and Junín 

viruses, and since these virus-host interactions represent a common mechanism in a range of emerging RNA 

viruses, we predict that they represent an Achilles’ heel in the life cycle of these RNA virus pathogens. Toward 

this end, we have ongoing, fruitful collaborations with pharmacologists and virologists at the BSL-4 labs at 

USAMRIID and with medicinal chemists at Fox Chase Chemical Diversity Center, which has led to the 

identification of two successful lead series of PTAP and PPxY budding inhibitors that exhibit on-target, broadspectrum antiviral activity against a wide array of RNA viruses. Specifically, we demonstrated that our current 

lead PTAP inhibitor, compound 0013, blocked egress of both Ebola and Junín VLPs at nanomolar 

concentrations, and our current lead PPxY inhibitors, compounds 4 and 5, blocked egress of Ebola, Marburg, 

and Lassa fever VLPs. Most exciting were our findings that these compounds could also block budding of live 

infectious viruses, including VSV and Junín (Harty lab), rabies (in collaboration with Dr. M. Schnell, TJU), Ebola 

(Kikwit) and Marburg (Ci67) (in collaboration with Drs. Andrew Herbert and J. Dye, USAMRIID, unpublished 

data). Currently, we are using Structure Activity Relationship (SAR) in collaboration with medicinal chemists Drs. 

J. Wrobel and A. Reitz (FCCDC) to identify analogs with enhanced potency and low cytotoxicity, as we seek to 

further transition one or more full-qualified L-domain inhibitors into more detailed IND-directed pharmacokinetic, 

pharmacodynamic and toxicity studies. 

a. Liu, Y., Lee, M. S., Olson, M. A., and Harty, R. N. 2011. Bimolecular complementation to visualize 

filovirus VP40-host complexes in live mammalian cells: toward the identification of budding inhibitors. 

Adv. Virol., pii: 341816. PMCID: PMC3217271. 

b. Lu, J., Han, Z., Liu, Y., Liu, W., Lee, M.S., Olson, M.A., Ruthel, G., Freedman, B.D., and Harty, R.N. 

2014. A host-oriented inhibitor of Junin Argentine hemorrhagic fever virus egress. J. Virol. 88(9):4736- 

4743. PMCID: PMC3993831. 

c. Han, Z., Lu, J., Liu, Y., Davis, B., Lee, M.S., Olson, M.A., Ruthel, G., Freedman, B.D., Schnell, M.J., 

Wrobel, J., Reitz, A., and Harty, R.N. 2014. Small molecule probes targeting the viral PPxY-host Nedd4 

interface block egress of a broad range of RNA viruses. J. Virol. 88(13):7294-7306. PMCID: 

PMC4054416. 

d. Loughran, H.M., Han, Z., Wrobel, J.E., Decker, S.E., Ruthel, G., Freedman, B.D., Harty, R.N., and 

Reitz, A.B. 2016. Quinoxaline-based inhibitors of Ebola and Marburg VP40 Egress. Bioorg. Med 

Chem Lett., 26:3429-3435. PMCID: PMC4955528. 

3. Calcium regulation of hemorrhagic fever virus budding. In non-excitable cells, Ca2+ signals are generated 

and maintained through a tightly regulated process termed “store-operated Ca2+ signaling”. As ER Ca2+ levels 

decrease, Ca2+ dissociates from C-terminal EF hands of the ER membrane-resident protein Stromal Interaction 

Molecule 1 (STIM1). Ca2+ dissociation from STIM1 initiates its oligomerization and localization to ER regions 

Biosketches Page 28 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

adjacent to the plasma membrane where it interacts with and activates Orai1 Ca2+ channels that control 

extracellular Ca2+ influx. While previous studies in my lab implicated Ca2+ broadly in control of Ebola VP40 VLP 

formation, we have now demonstrated for the first time that filovirus VP40 and arenavirus Z matrix proteins 

trigger host cell Ca2+ signals by activating the ER Ca2+ sensing protein STIM1, which then activates plasma 

membrane Orai1 (Ca2+) channels. Furthermore, we demonstrated that VP40 control of VLP production critically 

depends upon Orai1-mediated Ca2+ entry, as suppressing the expression or inhibiting the function of STIM1 and 

Orai1, or pharmacologically inhibiting Ca2+ permeation of Orai with selective channel blockers profoundly inhibits 

production of VLPs. Importantly, we also established that Orai1 mediated Ca2+ signals regulate transmission and 

spread of infectious Ebola, Marburg, Lassa fever, and Junín viruses in collaboration with John Dye’s group at 

USAMRIID. Together these results have broad and profound significance because of 1) the insight they provide 

into mechanisms that control virus egress and transmission, 2) their establishment of STIM1 and Orai1 as 

conserved, essential, and immutable host proteins that underlie Ca2+ signals critical for enveloped RNA virus 

egress, and 3) their validation of Orai blockers as the first of a family of broad-spectrum, host-oriented anti-viral 

therapeutics. Our ongoing research is focused on 1) defining the mechanism of virus induced calcium signaling, 

2) defining the role of calcium in both host and virus dependent steps of budding, and 3) pursuing the 

development of small molecule inhibitors of the Orai1 channel as a host-oriented therapeutic approach to block 

egress of a wide-array of RNA viruses. 

a. Han, Z and Harty, R. N. 2007. Influence of calcium/calmodulin on budding of Ebola VLPs: Implications 

for the involvement of the Ras/Raf/MEK/ERK pathway. Virus Genes, 35(3):511-520. PMID:17570046. 

b. Madara, J.J., Han, Z., Ruthel, G., Freedman, B.D., and Harty, R.N. 2015. The multifunctional Ebola virus 

VP40 matrix protein is a promising therapeutic target. Future Virol., 10(5):537-546. PMCID: 

PMC4480923. 

c. Han, Z., Madara, J.J., Herbert, A., Prugar, L.I., Ruthel, G., Lu, J., Liu, Y., Liu, W., Liu, X., Wrobel, J.E., 

Reitz, A.B., Dye, J.M., *Harty, R.N., and *Freedman, B.D. (*co-corresponding authors). 2015. Calcium 

regulation of hemorrhagic fever virus budding: Mechanistic implications for host-oriented therapeutic 

intervention. PLoS Pathogens, 11(10):e1005220. PMCID: PMC4634230. 

d. Freedman, B.D. and Harty, R.N. 2016. Calcium and filoviruses: A budding relationship. Future 

Microbiol., 11:713-715. 

4. Innate immune defenses of hemorrhagic fever virus infection. Innate immune responses to virus infection 

provide a critical first line of defense for the host against the invading pathogen. Understanding the complex 

interplay between the host innate immune defense mechanisms and counteraction by the filoviruses is crucial 

for developing novel antiviral strategies, vaccines, and therapeutics. We have been interested in the host innate 

immune response to filoviruses and have investigated several mechanisms of host innate immune mediated 

defenses involving host proteins TLR4, SOCS1, SOCS3, and ISG15. ISG15 is an interferon stimulated gene that 

has garnered much attention recently due to its broad-range of antiviral activity against a plethora of pathogens 

including DNA and RNA viruses. We demonstrated for the first time that ISG15 inhibited budding of EBOV VP40 

VLPs in a PPxY L-domain dependent manner, and that such inhibition involved impairment of host Nedd4 ligase 

activity. We continue to investigate whether ISGylation of additional VP40 host interactors (e.g. IQGAP1) can 

adversely affect VP40 function in budding. In addition, we are interrogating the effect of ISG15 expression on 

inhibition of Junín virus assembly and budding in vitro and in vivo using ISG15 -/- mice (in collaboration with Dr. 

A. Garcia-Sastre, MSSM). These studies will reveal new host innate immune defense mechanisms that may 

regulate the budding processes of several high priority NIAID Category A pathogens. Moreover, this information 

will be critical to identify strategies (therapies and/or vaccines) designed to tip the scale in favor of the host in 

the battle between viral pathogens and host innate immune defenses. 

a. Okumura, A., Pitha, P. M., and Harty, R. N. 2008. ISG15 Inhibits Ebola VP40 VLP Budding in an LDomain Dependent Manner by Blocking Nedd4 Ligase Activity. Proc.Natl. Acad. Sci. USA, 

105(10):3974-9. PMCID: PMC2268823. 

b. Okumura, A., Pitha, P. M., Yoshimura, A., and Harty, R. N. 2010. Interaction between Ebola virus 

glycoprotein and host TLR-4 leads to induction of pro-inflammatory cytokines and SOCS1. J. Virol., 

84(1):27-33. PMCID: PMC2798428. 

c. Lu, J., Qu, Y., Liu, Y., Jambusaria, R., Han, Z., Ruthel, G., Freedman, B., and Harty, R. N. 2013. Host 

IQGAP1 and Ebola virus VP40 interactions facilitate VLP egress. J. Virol., 87(13): 7777-80. PMCID: 

PMC3700276. 

d. Okumura, A., Rasmussen, A.L., Halfmann, P., Feldmann, F., Yoshimura, A., Feldmann, H., Kawaoka, 

Y., Harty, R.N., and Katze, M.G. 2015. Suppressor of cytokine signaling 3 is an inducible host factor 

Biosketches Page 29 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

during Ebola virus infection and 1 regulates virus egress. J. Virol., 89(20):10399-10406. PMCID: 

PMC4580175. 

Complete List of Published Work in MyBibliography: 

http://www.ncbi.nlm.nih.gov/sites/myncbi/ronald.harty.1/bibliography/44131509/public/?sort=date&dire 

ction=ascending. 

D. Additional Information: Research Support and/or Scholastic Performance 

Ongoing: 

R21 AI138052 Harty 01/15/18 – 12/31/20 

Modular Domains of Host Proteins Regulate Filovirus Maturation 

Major Goal – To elucidate the role of host protein BAG3 in filovirus egress. 

Role: PI 

R21 AI129890 Freedman 12/01/17 – 11/31/20 

The Membrane Repair Channel TRPML1 Regulates Ebola Virus Budding 

Major Goal – To elucidate the role of host channel TRPML1 in regulating Ebola virus egress 

Role: co-I 

R41 AI138630 Harty 06/01/18 – 05/31/21 

Small Molecule Therapeutics Targeting Hemorrhagic Fever Viruses 

Major Goal – To develop PPxY inhibitors of virus budding. 

Role: PI 

R21 AI139392 Harty 06/08/18 – 05/31/21 

Dueling PPxY Motifs of Filovirus VP40 and Host Angiomotin: Effects on Innate Immune Defenses and Tight 

Junction Integrity at Immune Privileged Sites. 

Major Goal – To examine the effect of VP40 on intestinal epithelium using a CACO2 model. 

Role: PI 

Major Goal – To test a unique series of budding inhibitors against Filovirus/Arenavirus infections. 

Role: PI 

R21 EY031465 Harty 04/01/20 – 03/31/22 

Predicted Role of Ebola VP40-Host Interactions in Ocular Pathology and Persistence 

Major Goal – To determine whether filovirus VP40 contributes to ocular pathology using an eye-on-a-chip 

model. 

Role: PI 

Completed:

R33 AI102104 Harty 09/01/12 – 06/30/19 

Host-Oriented Therapeutics Targeting Filovirus Budding. 

Major Goal – To elucidate the mechanisms and potency of PTAP L-domain inhibitors of RNA virus budding. 

Role: PI, No overlap with current proposal 

Biosketches Page 30 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

NAME: Jay Wrobel 

BIOGRAPHICAL SKETCH 

eRA COMMONS USER NAME (credential, e.g., agency login): 

POSITION TITLE: Vice President, Academic Relations, Fox Chase Chemical Diversity Center, Inc. 

EDUCATION/TRAINING 

INSTITUTION AND LOCATION DEGREE Completion 

Date FIELD OF STUDY 

University of Wisconsin-Milwaukee 

Cornell University 

B.S. 

Ph.D. 

1978 

1983 

Chemistry 

Organic Chemistry 

A. Personal Statement 

I am extremely well suited to for the medicinal chemistry leadership of this proposal. I have 34 years of 

experience in pharmaceutical industry focused on medicinal chemistry and drug design on a variety of drug 

discovery programs. In my current position at Fox Chase Chemical Diversity Center (FCCDC), I am involved in 

creation, leadership and management of medicinal chemistry and drug discovery partnerships with academic 

and biotech researchers to advance chemical hits to viable preclinical drug candidates up to and including the 

IND stage. 

I was formerly Senior Director of Medicinal Chemistry at Wyeth/Pfizer in Collegeville, PA where I 

mentored and guided the efforts of up to 31 medicinal chemists, and worked successfully with outside alliance 

partners in the design and synthesis of novel agents for multiple drug discovery programs (hit to Preclinical 

Development). I was directly involved in bringing forward nine development track candidates (Phase 0 and 

beyond). My hands-on expertise in research includes project and portfolio management, hit triage, hit-to-lead 

and lead optimization from hits found using enabling technologies such as high-throughput screening, fragment 

based drug discovery, and virtual screening. In these efforts I have employed ligand and structure based SAR 

methods, analytical chemistry, multi-step synthesis, parallel synthesis, data management. I obtain and use 

ADME and PK data in the design of our analogs. 

I have become involved with Prof. Harty on evaluation of egress inhibitors (namely mammalian Nedd4 / 

viral L domain inhibitors) of RNA viruses as an approach toward new antiviral drugs to combat Ebola, Marburg, 

Lassa and AIDS. In this capacity I have provided Prof. Harty with medicinal chemistry leadership on a Phase I 

STTR grant application and along with my colleagues at FCCDC have discovered new egress inhibitor leads 

that are more potent, stable and suitable for composition of matter patent status than what we (FCCDC and 

Harty lab) discovered prior to the STTR-funded work (see Journal of Virology, 2014, 88(13), 7294-7306). We 

have since published an SAR account of our further work (Bioorg. Med. Chem. Lett. 2016 26(15), 3429-35). We 

have obtained an issued U.S. patent on the initial series and have make great advances during the Phase I 

STTR period of study resulting in in vivo proof of concept efficacy in a mouse model of Marburg infection. 

I also have additional experience with infectious diseases including antiviral small molecules therapeutics 

(Nef inhibitors-HIV) and antibacterial agents against drug resistant pathogens. 

My role in this project would be to implement and oversee all aspects of the medicinal chemistry program 

including design of analogs (with participation of the bench chemists), day to day and long term management of 

medicinal chemists, directing all ADME, PK and drug safety studies with advice from our CRO partners. We 

will use a project flow scheme I devised to find the best compounds and I will also be involved in any major 

project decisions with PI Ron Harty. 

Loughran HM, Han Z, Wrobel JE, Decker SE, Ruthel G, Freedman BD, Harty RN, Reitz AB. Quinoxaline-based 

inhibitors of Ebola and Marburg VP40 egress. Bioorg Med Chem Lett. 2016 Aug 1;26(15):3429-35. doi: 

10.1016/j.bmcl.2016.06.053. Epub 2016 Jun 23. PubMed PMID: 27377328; PubMed Central PMCID: 

PMC4955528 

Han Z, Lu J, Liu Y, Davis B, Lee MS, Olson MA, Ruthel G, Freedman BD, Schnell MJ, Wrobel JE, Reitz AB, 

Harty RN. Small-molecule probes targeting the viral PPxY-host Nedd4 interface block egress of a broad range 

Biosketches Page 31 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

of RNA viruses. J Virol. 2014 Jul; 88(13): 7294-306. doi: 10.1128/JVI.00591-14. Epub 2014 Apr 16. PubMed 

PMID: 24741084; PubMed Central PMCID: PMC4054416. 

Emert-Sedlak LA, Loughran HM, Shi H, Kulp JL 3rd, Shu ST, Zhao J, Day BW, Wrobel JE, Reitz AB, Smithgall 

TE. Synthesis and evaluation of orally active small molecule HIV-1 Nef antagonists. Bioorg Med Chem Lett. 2016 

Mar 1;26(5):1480-4. doi: 10.1016/j.bmcl.2016.01.043. Epub 2016 Jan 21. PubMed PMID: 26852364; PubMed 

Central PMCID: PMC4756635. 

Rubin H, Selwood T, Yano T, Weaver DG, Loughran HM, Costanzo MJ, Scott RW, Wrobel JE, Freeman KB, 

Reitz AB. Acinetobacter baumannii OxPhos inhibitors as selective anti-infective agents. Bioorg Med Chem Lett. 

2015 Jan 15; 25(2): 378-83. doi: 10.1016/j.bmcl.2014.11.020. Epub 2014 Nov 22. PubMed PMID: 25496770. 

B. Positions and Honors 

1983-1987 Senior Scientist, Medicinal Chemistry, Ayerst, Inc., Princeton, NJ 

1987-1988 Research Associate, Medicinal Chemistry, Wyeth-Ayerst Research, Inc., Princeton, NJ 

1988-1996 Principal Scientist, Chemical Sciences, Wyeth-Ayerst Research, Inc., Princeton, NJ 

1996-2001 Associate Director, Chemical Sciences, Wyeth-Ayerst Research, Inc., Radnor, Pa 

2001-2007 Director, Chemical Sciences, Wyeth Research, Inc., Collegeville, Pa 

2007-2010 Senior Director, Chemical Sciences, Wyeth/Pfizer, Inc., Collegeville, Pa 

2011-2012- Senior Research Fellow, Fox Chase Chemical Diversity Center, Inc., Doylestown, Pa 

2012-present Vice President, Academic Relations, Fox Chase Chemical Diversity Center, Inc., Doylestown, Pa 

Member of the American Chemical Society 1978-present 

Alternate Councilor, American Chemical Society, Princeton, NJ Local Section, 2012-2014 

Member, DOD Study Section for review of applications, 2012, 2013. 

C. Contribution to Science 

In terms of discovering other novel drug candidates, I have worked extensively in cardiovascular areas 

on LXR modulators for atherosclerosis. As a LXR Modulator co-team leader and Medicinal Chemistry coordinator 

for the Wyeth/Karo-Bio LXR alliance, I delivered the first in-human LXR modulator (LXR-623) to phase I clinical 

studies for atherosclerosis indication in 2006. I also brought forth second and third generation LXR modulators 

devoid of mechanism and physiochemical-based side effects. 

Wrobel J, Steffan R, Bowen SM, Magolda R, Matelan E, Unwalla R, Basso M, Clerin V, Gardell SJ, Nambi P, 

Quinet E, Reminick JI, Vlasuk GP, Wang S, Feingold I, Huselton C, Bonn T, Farnegardh M, Hansson T, Nilsson 

AG, Wilhelmsson A, Zamaratski E, Evans MJ. Indazole-based liver X receptor (LXR) modulators with maintained 

atherosclerotic lesion reduction activity but diminished stimulation of hepatic triglyceride synthesis. J Med Chem. 

2008 Nov 27; 51(22): 7161-8. doi: 10.1021/jm800799q. PubMed PMID: 18973288. 

Quinet EM, Basso MD, Halpern AR, Yates DW, Steffan RJ, Clerin V, Resmini C, Keith JC, Berrodin TJ, Feingold 

I, Zhong W, Hartman HB, Evans MJ, Gardell SJ, DiBlasio-Smith E, Mounts WM, LaVallie ER, Wrobel J, Nambi 

P, Vlasuk GP. LXR ligand lowers LDL cholesterol in primates, is lipid neutral in hamster, and reduces 

atherosclerosis in mouse. J Lipid Res. 2009 Dec; 50(12): 2358-70. doi: 10.1194/jlr.M900037-JLR200. Epub 2009 

Mar 24. PubMed PMID: 19318684; PubMed Central PMCID: PMC2781308. 

Hu B, Unwalla RJ, Goljer I, Jetter JW, Quinet EM, Berrodin TJ, Basso MD, Feingold IB, Nilsson AG, Wilhelmsson 

A, Evans MJ, Wrobel JE. Identification of phenylsulfone-substituted quinoxaline (WYE-672) as a tissue selective 

liver X-receptor (LXR) agonist. J Med Chem. 2010 Apr 22; 53(8): 3296-304. doi: 10.1021/jm100034x. PubMed 

PMID: 20350005 

I was on a team that evaluated and brought in a FXR candidate from external alliance partner Exelixis. This 

candidate later became the first non-steroid based FXR agonist (FXR-450) taken to Phase I clinical studies for 

dyslipidemia in 2006. I later helped discover a FXR follow-on candidate with improved physiochemical properties 

(WYE-108635) that was taken to advanced preclinical animal toxicity evaluation in 2007. 

Biosketches Page 32 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Flatt B, Martin R, Wang TL, Mahaney P, Murphy B, Gu XH, Foster P, Li J, Pircher P, Petrowski M, Schulman I, 

Westin S, Wrobel J, Yan G, Bischoff E, Daige C, Mohan R. Discovery of XL335 (WAY-362450), a highly potent, 

selective, and orally active agonist of the farnesoid X receptor (FXR). J Med Chem. 2009 Feb 26; 52(4): 904-7. 

doi: 10.1021/jm8014124. PubMed PMID: 19159286. 

Lundquist JT, Harnish DC, Kim CY, Mehlmann JF, Unwalla RJ, Phipps KM, Crawley ML, Commons T, Green 

DM, Xu W, Hum WT, Eta JE, Feingold I, Patel V, Evans MJ, Lai K, Borges-Marcucci L, Mahaney PE, Wrobel JE. 

Improvement of physiochemical properties of the tetrahydroazepinoindole series of farnesoid X receptor (FXR) 

agonists: beneficial modulation of lipids in primates. J Med Chem. 2010 Feb 25; 53(4): 1774-87. doi: 

10.1021/jm901650u. PubMed PMID: 20095622. 

I co-invented the first non-steroidal PR antagonists that were delivered to development track (Phase 0 and 

beyond) for contraception or fibroid indications in 2003-2007. And I brought forth the first non-steroid based PR 

agonist (tanaproget) to human clinical trials for contraception indication in 2001. This compound successfully 

completed Phase II clinical trials and was licensed to Teva Pharmaceuticals. 

Fensome A, Adams WR, Adams AL, Berrodin TJ, Cohen J, Huselton C, Illenberger A, Kern JC, Hudak VA, 

Marella MA, Melenski EG, McComas CC, Mugford CA, Slayden OD, Yudt M, Zhang Z, Zhang P, Zhu Y, Winneker 

RC, Wrobel JE. Design, synthesis, and SAR of new pyrrole-oxindole progesterone receptor modulators leading 

to 5-(7-fluoro-3,3-dimethyl-2-oxo-2,3-dihydro-1H-indol-5-yl)-1-methyl-1H-pyrrole-2-carbonitrile (WAY-255348). J 

Med Chem. 2008 Mar 27;51(6): 1861-73. doi: 10.1021/jm701080t. Epub 2008 Mar 5. PubMed PMID: 18318463. 

Zhang Z, Lundeen SG, Slayden O, Zhu Y, Cohen J, Berrodin TJ, Bretz J, Chippari S, Wrobel J, Zhang P, 

Fensome A, Winneker RC, Yudt MR. In vitro and in vivo characterization of a novel nonsteroidal, species-specific 

progesterone receptor modulator, PRA-910. Ernst Schering Found Symp Proc. 2007; (1):171-97. PubMed PMID: 

18540573. 

Fensome A, Bender R, Chopra R, Cohen J, Collins MA, Hudak V, Malakian K, Lockhead S, Olland A, Svenson 

K, Terefenko EA, Unwalla RJ, Wilhelm JM, Wolfrom S, Zhu Y, Zhang Z, Zhang P, Winneker RC, Wrobel J. 

Synthesis and structure-activity relationship of novel 6-aryl-1,4-dihydrobenzo[d][1,3]oxazine-2-thiones as 

progesterone receptor modulators leading to the potent and selective nonsteroidal progesterone receptor agonist 

tanaproget. J Med Chem. 2005 Aug 11;48(16): 5092-5. PubMed PMID: 16078826. 

Zhang Z, Olland AM, Zhu Y, Cohen J, Berrodin T, Chippari S, Appavu C, Li S, Wilhem J, Chopra R, Fensome 

A, Zhang P, Wrobel J, Unwalla RJ, Lyttle CR, Winneker RC. Molecular and pharmacological properties of a 

potent and selective novel nonsteroidal progesterone receptor agonist tanaproget. J Biol Chem. 2005 Aug 5; 

280(31): 28468-75. Epub 2005 Jun 3. PubMed PMID: 15937332. 

I invented a mixed PPARα,γ,δ agonist that completed Phase II clinical trials for type II diabetes but was not 

selected to go further for business reasons. 

I have an excellent external scientific and technical record based on 75 papers and 81 presentations. I am an 

inventor on 79 issued U.S. Patents. My publications can be found online at 

http://www.ncbi.nlm.nih.gov/myncbi/collections/bibliography/47703212/?reload=deleteSuccess&dnum=1 

D. Research Support 

Current 

Development of Small Molecule Therapeutics Targeting Hemorrhagic Fever Viruses 

R41AI138630-01A1 (Role: Key Personnel) 06/01/18 – 05/31/21 (NCE) 

This is the Phase I STTR that provided most of the preliminary data that forms the basis of this Phase II STTR 

application. It was a two year Phase I which is currently in a third year no cost extension. 

Soft LXR Agonists for Idiopathic Pulmonary Fibrosis 

R43 HL154852-01 (Role: PI) 07/20/20 – 06/30/21 

Biosketches Page 33 
 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

This Phase I SBIR involves the discovery of soft drug LXR agonists for the treatment of Idiopathic Pulmonary 

Fibrosis (PDF). 

Overlap – none. 

Molecule Antagonists of PF4 for the Treatment and Prevention of HIT 

R42 HL12312602 (Role: PI) 09/15/2019 – 06/30/2022 

This Phase IIB SBIR is directed toward the optimization of novel small molecule PF4 tetramer antagonists for 

the treatment of Heparin-Induced Thrombocytopenia (HIT). 

Overlap – none. 

Completed 

Inhibitors of the PHD2 Zinc Finger to Treat Anemia 

R32HL137458 (Role: PI) 04/01/2017 – 3/31/2018 

This Phase I SBIR involves identification of SAR with intent to go to full-fledge drug discovery program in the 

Phase II portion for novel small molecule inhibitors the PHD2 Zinc Finger as a treatment for anemia 

Molecule Antagonists of PF4 for the Treatment and Prevention of HIT 

R42 HL12312602 (Role: Key Personnel) 10/01/2016 – 10/31/2018 

This Phase II STTR is a full-fledge drug discovery program toward the optimization of novel small molecule PF4 

tetramer antagonists for the treatment of heparin induced thrombocytopenia. 

Small Molecule Inhibitors of HIV1 Nef Virulence Factor for Treatment of HIV-Aids 

2 R42 GM112516-02A1 (Role: Key Personnel) 10/01/2016 – 10/31/2018 

This Phase II STTR is a full-fledge drug discovery program toward the optimization for novel small molecule 

inhibitors of the HIV-1 virulence factor, Nef. This viral protein is critical to HIV-1 replication in vivo, 

immune escape of HIV-infected cells, and AIDS progression 

Biosketches Page 34 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

OMB No. 0925-0001 and 0925-0002 (Rev. 03/2020 Approved Through 02/28/2023) 

BIOGRAPHICAL SKETCH 

Provide the following information for the Senior/key personnel and other significant contributors. 

Follow this format for each person. DO NOT EXCEED FIVE PAGES. 

NAME: Shtanko, Olena 

eRA COMMONS USER NAME (credential, e.g., agency login): 

POSITION TITLE: Staff Scientist I 

EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, 

include postdoctoral training and residency training if applicable. Add/delete rows as necessary.) 

INSTITUTION AND LOCATION 

DEGREE 

(if 

applicable) 

Completion 

Date 

MM/YYYY 

FIELD OF STUDY 

Harvard University, Cambridge, MA 

University of Wisconsin, Madison, WI 

Texas Biomedical Research Institute, San Antonio, 

TX 

A.L.B. 

Ph.D. 

Post-Doc 

06/2002 

09/2010 

12/2016 

Natural Sciences 

Microbiology 

Virology & Antivirals 

A. Personal Statement 

I am interested in understanding how filoviruses and other emerging viruses establish productive infection and 

trigger disease in affected individuals. My thesis research focused on the mechanisms of budding and 

incorporation of virus nucleocapsids into nascent virions of Mopeia virus, a close relative of Lassa Fever virus. 

As a Postdoctoral Scientist, I studied mechanisms of entry and replication of highly pathogenic viruses as well 

as developed and tested novel therapeutics to combat these pathogens. My work has identified cell factors 

critical for Ebola virus and Crimean-Congo hemorrhagic fever virus entry into cells. Since advancing to the Staff 

Scientist position, I have established my own laboratory to study mechanisms of pathogenesis and dissemination 

of filoviruses and other high-containment pathogens. I have also extensively collaborated with a number of 

investigators, including Dr. Harty, to coordinate and perform virus studies to identify host factors critical for 

replication and spread of filoviruses as well as test novel antiviral therapeutics using a mouse model of Ebola 

and Marburg virus disease. I provide sophisticated molecular biology and cell biology techniques for studies of 

highly pathogenic viruses. I am well-trained in BSL4 procedures and have been working in Texas Biomed’s 

ABSL4 laboratory since 2011. I will be responsible for all BSL4 studies as well as virus data analysis. 

B. Positions and Honors 

Positions and Employment 

2004-2010 Graduate Research Assistant, Department of Pathobiological Sciences, Advisor: Yoshihiro 

Kawaoka, University of Wisconsin, Madison, WI 

2011-2017 Postdoctoral Scientist, Department of Virology and Immunology, Advisor: Robert A. Davey, Texas 

Biomedical Research Institute, San Antonio, TX 

2017- Staff Scientist I, Department of Virology and Immunology, Texas Biomedical Research Institute, 

San Antonio, TX 

Professional Memberships 

2008- Member, American Society for Microbiology 

2009- Member, American Society for Virology 

2016- Member, American Society for Cell Biology 

Biosketches Page 35 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Honors 

2012 Douglass Foundation Post-doctoral Award 

2014 Travel Award recipient, American Society for Virology Annual Meeting 

2017 Texas Biomed Forum Research Support Grant 

2017 Texas Biomed Founder’s Council Grant 

C. Contributions to Science 

Characterization of mechanisms of assembly, egress, and spread of emerging viruses

Understanding how viruses invade and establish productive infection in host cells is essential to identify novel 

therapeutics and to reveal how the cell itself functions. My work on assembly of Mopeia virus, a close relative 

of Lassa Fever virus, has revealed that ALIX/AIP1, which is required for the biogenesis of multivesicular 

bodies (MVBs) of the cell, binds to and drives incorporation of virus nucleocapsids into nascent virions. 

Recently, in collaboration with Dr. Ronald Harty, I have helped to identify angiomotin (Amot), a key regulator 

of YAP nuclear/cytoplasmic localization and function, as essential for egress and spread of authentic Marburg 

and Ebola viruses. 

1. Shtanko O, Imai M, Goto H, Lukashevich IS, Neumann, G, Watanabe T and Kawaoka Y. A role for the 

C terminus of Mopeia virus nucleoprotein in its incorporation into Z protein-induced virus-like particles. J 

Virol. 2010; 84(10):5415-5422. PMID: 20200234. PMCID: PMC2863806. 

2. Shtanko O, Watanabe S, Jasenosky LD, Watanabe T and Kawaoka Y. ALIX/AIP1 is required for NP 

incorporation into Mopeia Z-induced virus-like particles. J Virol. 2011; 85(7):3631-3641. PMID: 

21248028. PMCID: PMC3067881. 

3. Han Z, Dash S, Sagum CA, Ruthel G, Berry CT, Freedman BD, Bedford M, Fan H, Sidhu S, Sudol M, 

Shtanko O and Harty RN. Modular mimicry and engagement of the Hippo pathway by Marburg virus 

VP40: implications for filovirus biology and budding. PLoS Path. 2020; 16(1):e1008231. PMID: 

31905227. PMCID: PMC6977764. 

4. Han Z, Ruthel G, Dash S, Berry CT, Freedman BD, Harty R and Shtanko O. Angiomotin regulates 

budding and spread of Ebola virus. J Biol Chem. 2020; 295(25):8596-8601. PMID: 32381509. PMCID: 

PMC7307192. 

Identification of host factors critical for entry and replication of high-containment viruses 

My work on Crimean-Congo Hemorrhagic Fever virus spearheaded the identification of the MVB as the site 

of release of viral genome into the cell cytoplasm. Since disruption of this compartment completely abolishes 

infection, I have been working on identifying compounds that interfere with MVB function. My research on 

EBOV has led to the finding that proteins that are known to initiate and coordinate autophagy, a catabolic 

process where cellular membranes serve as a source of vesicles whose role is to engulf cell contents for 

degradation, were also essential for macropinocytosis, a primary route of virus uptake. Interestingly, these 

cellular proteins appear to coordinate macropinosome formation close to the cell surface, an unexpected 

finding since these proteins were thought to function exclusively internally. I have been involved in several 

collaborative projects dedicated to mapping cellular networks critical for virus replication. For one such study, 

in collaboration with Dr. Schlesinger, I have developed assays to manipulate clinically relevant primary 

human cells to demonstrate the novel finding that E3 ubiquitin-protein ligase RBBP6 binds components of 

EBOV polymerase complex to modulate virus transcription and replication. 

1. Shtanko O, Nikitina RA, Altuntas CZ, Chepurnov AA and Davey RA. Crimean-Congo hemorrhagic 

fever virus entry into host cells occurs through the multivesicular body and requires ESCRT regulators. 

PLoS Pathog. 2014; 10(9):e1004390. PMID: 25233119. PMCID: PMC4169490. 

2. Shtanko O, Reyes AN, Jackson, WT and Davey RA. Autophagy-associated proteins control Ebola 

virus internalization into host cells. J Infect Dis. 2018; Jun 27. doi: 10.1093/infdis/jiy294. PMID: 

29947774. PMCID: PMC6249560. 

3. Batra J, Hultquist JF, Liu D, Shtanko O, Von Dollen J, Satkamp L, Jang GM, Luthra P, Schwarz TM, 

Small GI, Arnett E, Anantpadma M, Reyes AN, Leung DW, Kaake R, Haas P, Schmidt CB, Schlesinger 

LS, LaCount DJ, Davey RA, Amarasinghe GK, Basler CF and Krogan NJ. An Ebola virus-human 

protein-protein interaction map identifies the host ubiquitin ligase RBBP6 as a negative regulator of 

infection. Cell. 2018; 175(7):1917-1930. PMID: 30550789. PMCID: PMC6366944. 

Biosketches Page 36 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Antivirals discovery to combat filoviruses

As part of a multi-collaborative effort, I was involved in establishing methods to perform high-throughput 

screens to identify novel compounds with antiviral activity. I also initiated development of multiple assays to 

study the mechanism of virus entry into cells. These efforts have successfully identified novel potent inhibitors 

of macropinocytosis and consequently virus infection. 

1. Anantpadma M, Kouznetsova J, Wang H, Huang R, Kolokoltsov A, Guha R, Lindstrom AR, Shtanko O, 

Simeonov A, Maloney DJ, Maury W, LaCount DJ, Jadhav A and Davey RA. Large-scale screening and 

identification of novel Ebola virus and Marburg virus entry inhibitors. Antimicrob Agents Chemother. 

2016; 60(8):4471-4481. PMID: 27161622. PMCID: PMC4958205. 

2. Shtanko O, Sakurai Y, Noël R, Cintrat JC, Gillet D, Barbier J and Davey RA. Retro-2 and its 

dihydroquinazolinone derivatives inhibit filovirus infection. Antiviral Res. 2018; 149:154-163. PMID: 

29175127. 

3. Wu Y, Pons V, Noël R, Kali S, Shtanko O, Davey RA, Popoff MR, Tordo N, Gillet D, Cintrat JC and 

Barbier J. DABMA: A derivative of ABMA with improved broad-spectrum inhibitory activity of toxins and 

viruses. ACS Med Chem Lett. 2019; 10(8):1140-1147. PMID: 31413797. PMCID: PMC6691562. 

Macrophage signaling pathways to control acute infection

In collaboration with Dr. Wendy Maury, I investigated how acute infection with Plasmodium parasite, a 

causative agent of malaria, affected EBOV challenge in a BALB/c mouse model of virus disease. The work 

demonstrated that acute Plasmodium infection protected the animals from lethal viral challenge and that 

such protection was dependent on parasite-elicited IFN-γ and associated macrophage signaling events. In 

addition to elucidating a factor critical for the outcome of EBOV disease in malaria-virus coinfected 

populations, the study carefully analyzed what virus challenge dose was appropriate for research on EBOV 

pathogenesis. Inoculation of mice with either 1, 10, or 100 PFU resulted in uniform lethality and similar virus 

loads in blood and major tissues. Interestingly, the protective effect of acute parasitemia was apparent only 

in 1 PFU-inoculated animals, demonstrating the critical need for appropriate experimental set up. In a 

separate study, I co-investigated the mechanisms of tissue macrophage-mediated control of acute virus 

infections. The study identified a previously unappreciated role for macrophage-intrinsic CD40 signaling in 

controlling EBOV infection. 

1. Rogers K, Shtanko O, Vijay R, Mallinger L, Joyner C, Galinski M, Butler N and Maury W. Acute 

Plasmodium infection promotes interferon-gamma dependent resistance to Ebola virus infection. Cell 

Reports. 2020; 30(12):4041-4051. PMID: 32209467. PMCID: PMC7172281. 

2. Rogers K, Shtanko O, Stunz L, Mallinger L, White J, Schmidt M, Varga S, Butler N, Bishop G and Maury 

W. Frontline Science: CD40 signaling restricts RNA virus replication in MΦs, leading to rapid innate

immune control of acute virus infection. J Leukoc Biol. 2020; May 22. PMID: 32441445. 

For full publication record, see: https://www.ncbi.nlm.nih.gov/myncbi/1jeRfa5Kb3tko/bibliography/public/ 

D. Additional Information: Research Support and/or Scholastic Performance 

Ongoing Research Support

R41 AI138630 Harty (PI) 06/01/2018-05/31/2021 (2nd NCE) 

Development of small molecule therapeutics targeting hemorrhagic fever viruses 

NIH/NIAID 

The purpose of this study is to develop novel small molecule, broad-spectrum therapeutics against viral infections 

caused by filoviruses that depend on the PPxY L-domain motif for virus egress and spread of infection. 

Role: Subaward PI 

R21 AI144215 Maury (PI) 03/11/2020-03/10/2022 

CD40 regulation of acute virus infection 

NIH/NIAID 

The purpose of this study is to clarify the role of host macrophages and the CD40 signaling pathway in regulating 

early immune responses to filovirus infection. 

Role: Subaward PI 

Biosketches Page 37 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Completed Research Support

R21 AI139902 Maury (PI) 06/01/2018-05/31/2020 

Mechanisms and consequences of plasmodium/Ebola virus co-infections 

NIH/NIAID 

This project determined the impact of malaria on EBOV infection and defined the roles of specific pro- and antiinflammatory cytokines in regulating EBOV disease severity in a mouse co-infection model. 

Role: Subaward PI 

HDTRA118C0031 Westfall (PI) 02/01/2019-12/31/2019 

Grow, Expand, and Extract Total RNA 

Defense Threat Reduction Agency 

The purpose of this study was to develop a method to effectively extract and inactivate nucleic acids from select 

agents. 

Role: Subaward co-PI 

HHSN272201500015C McDonough (PI) 06/23/2015-5/31/2019 

Targeting therapeutics development to relieve bottlenecks: optimizing lead therapeutic compounds against 

infectious pathogens 

NIH/NIAID 

The purpose of this study was to characterize the efficacy of tetrandrine derivatives as inhibitors of filovirus 

disease using a mouse intraperitoneal Ebola virus exposure model. 

Role: Subaward PI 

Shtanko (PI) 12/01/2017-03/31/2019 

This grant explored how VPS34 complex regulates macropinocytosis and EBOV infection in primary human 

macrophages and iPSC-derived hepatocytes. 

Role: PI 

Biosketches Page 38 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

NAME: John L. Kulp III 

BIOGRAPHICAL SKETCH 

POSITION TITLE: Director of Computational Chemistry, FCCDC 

eRA COMMONS USER NAME (credential, e.g., agency login): 

EDUCATION/TRAINING 

INSTITUTION AND LOCATION DEGREE 

(if applicable) 

Completion Date 

MM/YYYY FIELD OF STUDY 

Drew University, Madison, NJ 

New York University, NY, NY 

Naval Research Laboratory, Washington, DC 

BA 

MS, 

PhD 

Postdoctoral 

05/2001 

05/2004, 

06/2006 

12/2009 

Chemistry, Mathematics 

Chemistry 

Chemistry 

A. PERSONAL STATEMENT 

I have 11 years of computational chemistry experience with 17 peer reviewed publications. My background 

includes extensive experience in computer-aided drug design with a specialty focus on computational 

fragment-based methods. In 2010, I joined BioLeap, a small business that specializes in protein-protein 

interactions and that has a proprietary fragment-based computational chemistry software platform. I 

participated in the development of novel inhibitors for DHFR, PCSK9, 11β-HSD, and recA. During that time, I 

integrated molecular dynamics and docking software into BioLeap’s proprietary fragment-based discovery 

platform. From 2013 to 2014, I was the project leader for BioLeap’s PCSK9 program, which consisted of 

modeling (BioLeap), assays (Broad Institute of MIT and Harvard), and chemical synthesis (WuXi and BioDuro). 

As Director of Computational Chemistry at FCCDC, I apply structural biology to SAR diversification efforts in 

areas of innovative pharmacology. For example, I work closely with Dr. Wrobel and his team on multiple 

projects currently. I will apply my knowledge and expertise in the area of pharmacophore development to 

achieves the potency, efficacy and ADME gains of this grant application targeting the host Nedd4:virus PPxY 

interaction for the treatment of Marburg, Ebola and related diseases. 

B. POSITIONS AND HONORS 

Positions and Employment

2009-11 Federal Staff Scientist, Naval Research Laboratory, Washington, DC 

2011-14 Distinguished Senior Scientist, BioLeap, Inc., Pennington, NJ 

2013- Assistant Professor, Baruch S. Blumberg Institute, Doylestown, PA 

2014- CSO, Small Molecule PPI Mimics LLC, College Station, TX 

2014- CEO, Conifer Point Pharmaceuticals LLC, Doylestown, PA 

2017- Director of Computational Chemistry, FCCDC 

Other Experience and Professional Memberships

2014 Consultant on NIH Phase I STTR grant entitled “Small Molecule Inhibitors of HIV1 Nef 

Virulence Factor for Treatment of HIV/AIDS” Fox Chase Chemical Diversity Center, R41 

GM112516-01. 

2015 Consultant on NIH Direct to Phase II SBIR entitled “Development of Small Molecule 

Therapeutics Against Smallpox and Other Poxviruses” Fox Chase Chemical Diversity 

Center, R44AI115759-01. 

Honors 

2002-6 MacCracken Fellowship, New York University 

2004 Kramer Fellowship, New York University 

2006-9 American Society for Engineering Educations Postdoctoral Fellowship 

2009 Young Investigator Award, American Peptide Society 

Biosketches Page 39 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

C. CONTRIBUTIONS TO SCIENCE 

1. My vision is to create better drugs by rigorous molecular design. Fragment-based lead discovery (FBLD) 

involves constructing drugs from small chemical building blocks (fragments) that bind to a protein target. 

Numerous companies, such as Abbott, Astex, and others, are pursuing this strategy by screening fragment 

libraries, followed by X-ray crystallography or NMR. In contrast, my research starts with a detailed hypothesis 

of what protein binding interactions are required of a drug for inhibition or activation. Commercial and 

proprietary design tools, some of which I created, are then used to search ligand-protein or fragment-protein 

simulation data for poses, ranked by predicted binding affinity, which can be assembled or optimized into 

custom compounds that satisfy the requirements. The designed compounds are synthesized and tested in 

assays. With success, this establishes a predictive model to guide subsequent lead optimization. This is a 

protein-centric strategy, complementary to ligand-centric SAR approaches. This methodology provides access 

to broad chemical diversity, which is crucial for solving difficult problems in lead identification and optimization. 

• "A Fragment-based Approach to the SAMPL3 Challenge." J.L. Kulp III, S.N. Blumenthal, Q. Wang, R. 

Bryan, F. Guarnieri, Journal of Computer-Aided Molecular Design 2012, 26(5), 583-594. 

• “Diverse Fragment Clustering and Water Exclusion Identify Protein Hot Spots,” J.L. Kulp III, J.L. Kulp Jr., 

D.L. Pompliano, F. Guarnieri, Journal of the American Chemical Society 2011, 133(28), 10740-1074. 

• “Simulations of Nanocylinders Self-Assembled from Cyclic β-Tripeptides,” N. Bernstein, J.L. Kulp III, M.A. 

Cato Jr., T.D. Clark, Journal of Physical Chemistry A, 2010, 114(44), 11948-11952. 

2. The field of foldamer design promises new routes to important compounds for use in sensors, smart 

materials, therapeutics and catalysts. The term “foldamer” refers to a molecule that folds into a structurally 

stable state in solution. Proteins and peptides are an important class of natural foldamers that carry out a host 

of essential functions in biology, including molecular recognition, information storage, catalysis, and controlled 

crystallization of inorganic materials. The desire to mimic such functions with synthetic molecules inspires the 

field of foldamer design. I developed cyclic β-peptides as stochastic sensing elements and as a model chiral 

system for studying surface interactions. Other work included a universal method for restraining short peptides 

in helical conformations by a main-chain hydrogen bond surrogate (HBS) approach. These HBS peptides 

inhibited a variety of pharmaceutically relevant protein-protein interactions and the resulting technology was 

licensed by Aileron Therapeutics for clinical development. 

• “Synthesis and characterization of cyclic peptides that are β-helical in trifluoroethanol.” K.P. Fears, S.J. 

Photiadis, J.L. Kulp III and T.D. Clark, Journal of Peptide Science 2014, 20(5), 366-374. 

• “Engineering a β helical D,L-peptide for folding in polar media.” J.L. Kulp III, T.D. Clark. Chemistry: A 

European Journal 2009, 15(44), 11867-11877. 

• “Trapping a Folding Intermediate of the α-Helix: Stabilization of the π-Helix.” R.N. Chapman, J.L. Kulp III, A. 

Patgiri, N.R. Kallenbach, C. Bracken, P.S. Arora, Biochemistry 2008, 47(14), 4189-4195. 

• “Evaluation of Biologically Relevant Short α-Helices Stabilized by a Main-Chain Hydrogen-Bond 

Surrogate.” D. Wang, K. Chen, J.L. Kulp III, P.S. Arora, Journal of the American Chemical Society 2006, 

128(28), 9248-9256. 

3. My postdoctoral fellowship and subsequent two year federal employment was in The Molecular Interfaces 

and Tribology Section of the Department of Chemistry, Naval Research Laboratory. This section studies 

interdisciplinary questions in basic and applied interface and surface science. Our group’s research stretched 

from advancing broad analytical methods, such as vibrational circular dichroism, to evaluating both prototypical 

and realistic interfaces to crafting and improving molecular surfaces and peptide architectures for applications 

such as controlling biological activity, anti-fouling, or lowering contact friction. One focus area was in the design 

and employment of bioinspired materials that conserve and utilize the exceptional characteristics of 

biomaterials while combining added functionality, such as fluorescence or conductivity, not originating from 

natural materials. 

• “Vibrational Circular-Dichroism Spectroscopy of Homologous Cyclic Peptides Designed to Fold into β

Helices of Opposite Chirality,” J.L. Kulp III, K.P. Fears, R. Lombardi, D.Y. Petrovykh, J.C. Owrutsky, L.A. 

Biosketches Page 40 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Nafie, T.D. Clark, Biointerphases, 2011, 6(1), 1-7. 

• “Characterization of the adhesive plaque of the barnacle Balanus Amphitrite: Anyloid-like nanofibrils are a 

major component,” D.E. Barlow, G.H. Dickinson, B. Orihuela, J.L. Kulp III, D. Rittschof, K.J. Wahl, 

Langmuir 2010, 26(9), 6549-6556. Highlighted in Chemical and Engineering News 2010, 88(9), 34. 

• “Molecular Recognition and Supramolecular Self-Assembly of a Genetically Engineered Gold Binding 

Peptide on Au {111}.” C. So, J.L. Kulp III, E.E. Oren, H. Zareie, H. Zareie, C. Tamerler, J.S. Evans, M. 

Sarikaya, ACS Nano 2009, 3(6), 1525-1531. 

• “Laser Ablation of Micropores for Formation of Artificial Planar Lipid Bilayers.” T.J. O’Shaughnessy, J.E. 

Hu, J.L. Kulp III, S.M. Daly, F.S. Ligler, Biomedical Microdevices 2007, 9(6), 863-868. 

4. The beginning of my computational chemistry career began when I was using nuclear magnetic resonance 

(NMR) spectroscopy to determine the structures of biological macromolecules like peptides, polypeptides, and 

proteins. To do this, I would rely on 1D, homo- and hetero-nuclear 2D, and triple resonance, 3D, spectra. 

Some other examples of this work are cited in scientific contribution 3, above. After extracting all available 

information about the atomic distances and torsion angles from the NMR spectra, computer-aided structure 

calculations would convert distance and torsion angle data (constraints) into the 3D atomic model. For these 

calculations, a molecular dynamics simulation was used with simulated annealing; increasing temperature to 

generate an ensemble of conformations then decreasing temperature with increasing constraints based on the 

NMR data. The result is a family of structures that span a relatively narrow conformational space (low RMSD), 

unless intrinsically flexible, and obeys the constraint criteria. 

• “Structural Properties of an Artificial Protein that Regulates the Nucleation of Inorganic and Organic 

Crystals.” J.L. Kulp III, T. Minamisawa, K. Shiba, M. Tejani, J.S. Evans, Langmuir 2007, 23(7), 3857-3863. 

• “Probing the Conformational Features of a Phage Display Polypeptide Sequence Directed Against Single 

Walled Carbon Nanohorn Surfaces.” J.L. Kulp III, K. Shiba, J.S. Evans. Langmuir 2005, 21(25), 11907-

11914. 

• “Affinity Selection of Peptide Phage Libraries Against Single-Wall Carbon Nanohorns Identifies a Peptide 

Aptamer with Conformational Variability” D. Kase, J.L. Kulp III, M. Yudasaka, J.S. Evans, S. Iijima, K. 

Shiba, Langmuir 2004, 20(20), 8939-8941. 

• “Molecular Characterization of a Prokaryotic Polypeptide Sequence that Catalyzes Au Crystal Formation.” 

J.L. Kulp III, M. Sarikaya, J.S. Evans, J. Mat. Chem.2004, 14(14), 2325-2332. 

Complete List of Published Work in MyBibliography: 

http://www.ncbi.nlm.nih.gov/sites/myncbi/john.kulp.2/bibliography/48391280/public/?sort=date&direction=ascen 

ding 

D. RESEARCH SUPPORT 

Current: 

Molecular Mechanism of the Dual Effects of Capsid Assembly Effectors on HBV Capsid Assembly and 

Disassembly (DoD/PRMRP). Role: Investigator 9/1/2017 – 8/31/2020 

The goal of this project is to obtain molecular insights on the mechanism of two distinct chemotypes of hepatitis 

B virus (HBV) core protein allosteric modulators (CpAMs) on the assembly and disassembly (or uncoating) of 

HBV nucleocapsids. 

Previous: 

Role: Investigator 7/1/2018 – 6/30/2019 

The goal of this project is to progress disubstituted aminothiazoles that demonstrate remarkable selective 

toxicity for HCC-derived cell lines versus non-HCC liver lines and most other cancer lines. 

Small Molecule Inhibitors of HIV1 Nef Virulence Factor for Treatment of HIV-Aids 

R42GM112516-01 (PI: Smithgall; Role: Key Personnel) 9/01/2016–8/31/2018 

This Phase II STTR involves the discovery of novel small molecule inhibitors of the HIV-1 virulence factor Nef, 

and has a substantial structural biology and computational chemistry component. 

Biosketches Page 41 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

BIOGRAPHICAL SKETCH 

Provide the following information for the Senior/key personnel and other significant contributors. 

Follow this format for each person. DO NOT EXCEED FIVE PAGES. 

NAME: Reitz, Allen Bernard 

eRA COMMONS USER NAME (credential, e.g., agency login): 

POSITION TITLE: Chief Executive Officer 

EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, 

include postdoctoral training and residency training if applicable. Add/delete rows as necessary.) 

INSTITUTION AND LOCATION DEGREE 

(if applicable) 

Completion 

Date 

MM/YYYY 

FIELD OF STUDY 

University of California, Santa Barbara 

University of California, San Diego 

University of Pennsylvania (Wharton) 

B.A. 

M.S. 

Ph.D. 

M.S. 

08/1977 

06/1979 

01/1982 

06/2003 

Biochemistry and 

Molecular Biology 

Chemistry 

Chemistry 

Management of 

Technology 

A. Personal Statement 

Dr. Reitz has had >38 years of experience in the pharmaceutical industry including ~26 years with Johnson & 

Johnson in various drug discovery research and management roles. He is an inventor of eight compounds that 

have entered clinical trials, including two from FCCDC which he founded in 2008 (www.fc-cdci.com). For 

example, he is a coinventor of troriluzole which emerged from an Phase I/II SBIR-funded project at FCCDC on 

which he was the Principal Investigator, and which is currently in Phase II/III clinical trial having recently passed 

an interim futility analysis for the treatment of Alzheimer’s disease. He is co-author of >160 scientific publications, 

co-inventor on 71 issued U.S. patents, Editor-in-Chief of the journal Current Topics in Medicinal Chemistry, on 

the Editorial Advisory Board of ACS Med. Chem. Lett. Dr. Reitz has extensive experience in project and portfolio 

management, target validation, hit triage, eADME profiling, and preclinical candidate selection. He is currently 

serving as an ad hoc reviewer on two of the rapid review COVID-19 study sections in NIAID. Dr. Reitz has 

worked with Drs. Wrobel and Prof. Harty since the inception of the FCCDC – U. Penn. collaboration of this novel 

host directed approach to treat Ebola and Marburg infection. He will continue to provide overall guidance as 

much as possible, working with Dr. Wrobel, and will seek to ensure a successful transition at the end of Phase 

II to further development and eventual commercialization including through taking advantage of various initiatives 

of the FDA such as the Neglected Disease Priority Review Voucher program. 

Loughran HM, Han Z, Wrobel JE, Decker SE, Ruthel G, Freedman BD, Harty RN, Reitz AB. Quinoxaline-based 

inhibitors of Ebola and Marburg VP40 egress. Bioorg Med Chem Lett. 2016 Aug 1;26(15):3429-35. doi: 

10.1016/j.bmcl.2016.06.053. PubMed PMID: 27377328; PubMed Central PMCID: PMC4955528 

Han Z, Lu J, Liu Y, Davis B, Lee MS, Olson MA, Ruthel G, Freedman BD, Schnell MJ, Wrobel JE, Reitz AB, 

Harty RN. Small-molecule probes targeting the viral PPxY-host Nedd4 interface block egress of a broad range 

of RNA viruses. J Virol. 2014 Jul; 88(13): 7294-306. doi: 10.1128/JVI.00591-14. PubMed PMID: 24741084; 

PubMed Central PMCID: PMC4054416. 

B. Positions and Honors 

Positions and Employment 

1982-1983 Postdoctoral Research Fellow, McNeil Pharmaceutical 

1983-1984 Research Scientist, McNeil Pharmaceutical 

1984-1987 Senior Scientist, Janssen Research Foundation 

Biosketches Page 42 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

1987-1992 Principal Scientist, R W Johnson Pharm. Res. Inst. 

1992-1998 Research Fellow, R W Johnson Pharm. Res. Inst. 

1999-2007 Senior Research Fellow, JNJPRD 

2008 Vice President, Research and Development, AlsGen, Inc. 

2008-present Founder and CEO, Fox Chase Chemical Diversity Center, Inc. 

2009-present Co-Founder and CEO, ALS Biopharma, LLC 

2010-present Founder and President, Pennsylvania Drug Discovery Institute (non-profit) 

Other Experience and Professional Memberships 

2015 National Co-Chair of the BIO Boot Camp, associated with the national BIO meeting 

2014-present Editorial Advisory Board member, ACS Med. Chem. Lett. 

2014-2016 Co-Editor, Technology Transfer and Entrepreneurship 

2008-present Adjunct Professor, Drexel University College of Medicine 

2008-2009 Consultant to the World Health Organization for the Tropical Diseases (TDR) 

2005-present Have served on >42 NIH Study Sections including Drug Discovery for the Nervous System (2012-

2014), and am currently an ad hoc member of COVID-19 rapid review study section study sections 

ZAI1 RG-W (S2) and ZAI1 RG-W (J2), July 14 and Oct. 15 respectively. 

2000-present Editor-in-Chief, Current Topics in Medicinal Chemistry 

1995-1996 Philadelphia Organic Chemists’ Club, Assistant Chairman, and then Chairman 

1985-1986 Philadelphia Organic Chemists’ Club, Assistant Secretary, and then Secretary 

Honors 

2016 Cornerstone Award CEO Honoree, Winner for Bucks County, PA 

2011 Gold Award, Best Early-Stage Company, Philadelphia Business Journal 

2006 Ronald J. Borne Distinguished Lectureship Award, in Recognition for Outstanding Contributions 

to Medicinal Chemistry, Department of Medicinal Chemistry, School of Pharmacy, University of 

Mississippi (April 18, 2006). Highlighted on pg. 6 of AACP News2006, 37(9). 

2003 Industrial Chemistry Award of the Philadelphia Organic Chemists’ Club.Poster Session, 

Reception, and Lecture: May 29, 2003 

1999 Johnson & Johnson Corporate Office of Science and Technology Excellence in Science Award 

1997 Johnson & Johnson Significant Achievement Award for Creating the POP Process to Rapidly 

Move New Compounds from the Laboratory into Clinical Trials 

1994 Philip B. Hofmann Research Scientist Award for Outstanding Achievement 

1990 Johnson & Johnson Achievement Award for Outstanding Contributions 

C. Contribution to Science 

1. Target Validation. Research in this area has been to understand the relation of structure to function among 

related compound libraries, using modern methods of synthetic chemistry and SAR development. Along 

those lines, FCCDC has submitted >12,000 new chemical entities in the context of individual collaborations 

with academic and biotech research partners. 

a. Punchi Hewage, A. N. D.; Yao, H.; Nammalwar, B. Gnanasekaran, K. K.; Lovell, S.; Bunce, R. A.; 

Eshelman, K.; Phaniraj, S. M.; Lee, M. M.; Peterson, B. R.; Battaile, K. P.; Reitz, A. B.; Rivera, M. Small 

Molecule Inhibitors of the BfrB-Bfd Interaction Decrease Pseudomonas aeruginona Fitness and 

Potentiate Fluoroquinoline Activity. J. Am. Chem. Soc. 2019, 141, 8171-8184. 

b. Karpova, Y.; Wu, C.; Divan, A.; McDonnell, M. E.; Hewlett, E.; Makhov, P.; Gordon, J.; Ye, M.; Reitz, 

A. B.; Childers, W. E.; Skorski, T.; Kolenko, V.; Tulin, A. V. Non-NAD-like PARP-1 inhibitors in prostate 

cancer treatment. Biochem. Pharmacol. 2019, 167, 149-162. 

c. Robinson, R. M.; Reyes, L.; Duncan, R. M.; Bian, H.; Reitz, A. B.; Manevich, Y.; McClure, J. J.; 

Champion, M. M.; Chou, C. J.; Sharik, M. E.; Chesi, M.; Bergsagel, P. L.; Dolloff, N. G. Inhibitors of the 

protein disulfide isomerase family for the treatment of multiple myeloma. Leukemia 2019, 33, 1011- 

1022. 

d. Martin, M. D.; Calcul, L.; Smith, C.; Jinwal, U. K.; Fontaine, S. N.; Darling, A.; Seeley, K.; Woitas, L.; 

Narayan, M.; Gestwicki, J. E.; Smith, G. R.; Reitz, A. B.; Baker, B. J.; Dickey, C. A. Synthesis, 

Stereochemical Analysis, and Derivatization of Myricanol Provide New Probes That Promote 

Autophagic Tau Clearance. ACS Chem. Biol. 2015, 10, 1099-1109. 

Biosketches Page 43 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

2. Hit to Lead Medicinal Chemistry. Much effort has focused on this area, in which libraries of related 

compounds are prepared in order to obtain maximum diversity based upon common themes dictated by the 

structure of the target, pharmacophore analysis, and the creation of new intellectual property. Multiple 

projects (e.g. BACE, EBNA1) have involved real-time X-ray co-crystallography collaboration and analysis. 

a. Baxter, E. W.; Conway, K. A.; Kennis, L.; Bischoff, F.; Mercken, M. H.; De Winter, H. L.; Reynolds, C. 

H.; Tounge, B. A.; Luo, C.; Scott, M. K.; Huang, Y.; Braeken, M.; Pieters, S. M. A.; Berthelot, D. J. C.; 

Masure, S.; Bruinzeel, W. D.; Jordan, A. D.; Parker, M. H.; Boyd, R. E.; Qu, J.; Alexander, R. S.; 

Brenneman, D. E.; Reitz, A. B. 2-Amino-3,4-dihydroquinazolines as Inhibitors of BACE-1 (β-Site APP 

Cleaving Enzyme): Use of Structure Based Design to Convert a Micromolar Hit into a Nanomolar Lead. 

J. Med. Chem. 2007, 50, 4261-4264. 

b. Huang, Y.; Strobel, E. D.; Ho, C. Y.; Reynolds, C. H.; Conway, K. A.; Piesvaux, J. A.; Brenneman, D. 

E.; Yohrling, G. J.; Arnold, H. M.; Rosenthal, D.; Alexander, R. S.; Tounge, B. A.; Mercken, M.; 

Vandermeeren, M.; Parker, M. H.; Reitz, A. B.; Baxter, E. W. Macrocyclic BACE Inhibitors: Optimization 

of a Micromolar Hit to Nanomolar Leads. Bioorg. Med. Chem. Lett. 2010, 20 ̧3158-3160. 

c. McDonnell, M. E.; Vera, M. D.; Blass, B. E.; Pelletier, J. C.; King, R. C.; Fernandez-Metzler, C.; Smith, 

G. R.; Wrobel, J.; Chen, S.; Reitz, A. B. Riluzole prodrugs for melanoma and ALS: design, synthesis, 

and in vitro metabolic profiling. 2012, Bioorg. Med. Chem., 20, 5642-5648. 

d. Rubin, H.; Selwood, T.; Yano, T.; Weaver, D. G.; Loughran, H. M.; Costanzo, M. J.; Scott, R. W.; 

Wrobel, J. E.; Freeman, K. B.; Reitz, A. B. Acinetobacter baumannii OxPhos inhibitors as selective 

anti-infective agents. Bioorg. Med. Chem. Lett. 2015, 25, 378-383. 

3. Lead Optimization (LO) Medicinal Chemistry Advancing to Preclinical and Clinical Development. 

Considerable emphasis has been placed on this phase of the drug discovery process, for which eight NMEs 

have entered human clinical trials. The 71 issued U.S. patents are largely devoted to LO SAR programs. 

There are three basic considerations for a medicinal chemist which apply to all indications: (1) route of 

administration (e.g. p.o, i.v., etc.), and (2) dosing regimen (acute, subchronic, chronic), and (3) standard of 

care (crowded marketplace or keen unmet need). 

a. Reitz, A. B.; Baxter, E. W.; Bennett, D. J.; Codd, E. E.; Jordan, A. D.; Maryanoff, B. E.; McDonnell, M. 

E. ; Ortegon, M. E.; Renzi, M. J.; Scott, M. K.; Shank, R. P.; Sherrill, R. G.; Vaught, J. L.; Wustrow, D. 

J. N-Aryl-N’-Benzylpiperazines as Potential Antipsychotic Agents. J. Med. Chem. 1995, 38, 4211-4222. 

[Description of mazapertine, which advanced to Phase II clinical trials.] 

b. Parker, M. H.; Smith-Swintosky, V. L.; McComsey, D. F.; Huang, Y.; Brenneman, D.; Klein, B.; 

Malatynska, E.; White, H. S.; Milewski, M. E.; Herb, M.; Finley, M. F. A.; Liu, Y.; Lubin, M. L.; Qin, N.; 

Iannucci, R.; Leclercq, L.; Cuyckens, F.; Reitz, A. B.; Maryanoff, B. E. Novel, Broad-Spectrum 

Anticonvulsants Containing a Sulfamide Group: Advancement of N-[(Benzo[b]thien-3-

yl)methyl]sulfamide (JNJ-26990990) into Human Clinical Studies. J. Med. Chem., 2009, 52, 7528-7536. 

[Anticonvulsant JNJ-26990990 which advanced into Phase I clinical trials.] 

c. Ross, T. M.; Battista, K.; Bignan, G. C.; Brenneman, D. E.; Connolly, P.. J.; Liu, J.; Middleton, S. A.; 

Orsini, M.; Reitz, A. B.; Rosenthal, D. I.; Scott, M. K.; Vaidya, A. H. A selective small molecule NOP 

(ORL-1 receptor) partial agonist for the treatment of anxiety. Bioorg. Med. Chem. Lett. 2015, 25, 602- 

606. [Anxiolytic ORL-1 receptor agonist that advanced into Phase I clinical trials.] 

d. Messick, T. E.; Smith, G. R.; Soldan, S. S.; McDonnell, M. E.; Deakyne, J. S.; Malecka, K. A.; Tolvinski, 

L.; van den Heuvel, A. P. J.; Gu, B. W.; Cassel, J. A.; Tran, D. H.; Wasserman, B. R.; Zhang, Y.; 

Velvadapu, V.; Zartler, E. R.; Busson, P.; Reitz, A. B.; Lieberman, P. M. Structure-based design of 

small-molecule inhibitors of EBNA1 DNA binding blocks Epstein-Barr virus latent infection and tumor 

growth. Sci. Tranl. Med. 2019, 11, pii: eaau5612. doi: 10.1126/scitranslmed.aau5612. [EBNA1 

inhibitor currently in Phase II clinical trials, discovered at FCCDC] 

4. Synthetic Organic Chemistry. Dr. Reitz was inspired by organic faculty as an undergraduate, which was 

reinforced by a positive experience as a graduate student with Murray Goodman at UCSD. Highly cited 

publications include a Chem. Rev. article on the Wittig reaction and Vol. 59 of Organic Reactions on reductive 

aminations using boron-containing reagents consisting of >700 pages and an entire volume. Research has 

been conducted in the areas of conformational analysis and new synthetic methods. 

Biosketches Page 44 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

a. Larsen, B. J.; Rosano, R. J.; Ford-Hutchinson, T. A.; Reitz, A. B.; Wrobel, J. A. A method for C2 

arylation of 1,3-indandiones. Tetrahedron 2018, 74, 2762-2768. 

b. Pelletier, J. C.; Velvadapu, V.; McDonnell, M. E.; Wrobel, J. E.; Reitz, A. B. Intramolecular 

rearrangement of α-amino acid amide derivatives of 2-aminobenzothiazoles.Tetrahedron Lett. 2014, 

55, 4193-4195. 

c. Maryanoff, B. E.; Reitz, A. B. The Wittig Olefination Reaction and Modifications Involving PhosphorylStabilized Carbanions. Stereochemistry, Mechanism, and Selected Synthetic Aspects. Chem. 

Rev.1989, 89, 863-927. 

d. Baxter, E. W; Reitz, A. B. "Reductive Aminations of Carbonyl Compounds with Borohydride and Borane 

Reducing Agents" In Organic Reactions: Wiley, New York, p. 1-714, Vol. 59, 2002. 

Complete List of Published Work in MyBibliography:

http://www.ncbi.nlm.nih.gov/sites/myncbi/1nyMq8FpqE0A5/bibliography/47313087/public/?sort=date&d

irection=ascending 

D. Research Support 

Ongoing Research Support 

Riluzole Prodrugs for Melanoma and ALS 

R44 CA156781-04A1 (PI: Reitz) 9/1/18-8/31/21 

This Phase II SBIR Bridge application involves the further pre-clinical development of troriluzole, an activelytransported (PepT1) prodrug of riluzole, . A Phase II clinical trial for the treatment of solid tumors is included in 

the grant, to be conducted at the Dana Farber Cancer Institute. Overlap – none. 

Troriluzole for the Treatment of Cocaine Addiction 

R41 DA047169-01 (PI: Rawls, Temple University; Role, Key Personnel) 07/01/18-7/30/21 (NCE) 

In this project, Prof. Rawls is testing whether troriluzole can counter the effects of cocaine in rats. Currently in 

the no cost extension phase. Overlap – none. 

Psychoactive bath salts and the glutamate system 

R01 DA039139-02 (PI: Rawls, Temple University; Role, Key Personnel) 8/1/16-7/31/21 

This R01 from Prof. Rawls at Temple University involves understanding the pharmacology of bath salts in CNS 

receptor and behavioral assays. Overlap – none. 

Targeting Telomerase for Pancreatic Cancer Therapeutics 

R01 CA208439-01 (PI: Skordalakes; Role, Key Personnel) 8/1/18-7/31/21 

This work is to discover new telomerase inhibitors for the treatment of pancreatic cancers. Overlap – none. 

DART2.0: comprehensive cell type-specific behavioral neuropharmacology 

RF1 MH117055-01 (PI: Tadross, Duke University; Role, Key Personnel) 9/14/18-9/13/21 

Prof. Tadross is the PI of a Brain R01 looking to identify new DART bifunctional probes to understand the 

localization and function of important receptor systems in the CNS. Our role is to perform synthetic chemistry to 

obtain suitable probes which are then studied in his laboratory. Overlap – none 

Biosketches Page 45 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

FCCDC Budget Justification 

Personnel 

Jay E. Wrobel, Ph.D. Vice President, Academic Affairs and Senior Research Fellow, Key Personnel, 

FCCDC Project Leader (Yrs 1-3: 3 cal. months). Dr. Wrobel has >35 years of experience in the pharmaceutical 

industry focused on drug design and discovery on a variety of drug discovery programs. He was formerly Senior 

Director of Medicinal Chemistry at Wyeth/Pfizer in Collegeville, PA where he mentored and guided the efforts of 

up to 31 medicinal chemists and worked successfully with outside alliance partners in the design and synthesis 

of novel agents for multiple drug discovery programs (hit to Preclinical Development). Dr. Wrobel was directly 

involved in bringing forward nine development track candidates (phase 0 and beyond). He has 78 peer reviewed 

articles and is an inventor on 83 U.S. Patents. Dr. Wrobel has expertise in project and portfolio management, 

hit triage, hit-to-lead and lead optimization from hits found using enabling technologies such as high-throughput 

screening, fragment based drug discovery, and virtual screening. In these efforts he has employed ligand and 

structure based SAR methods, analytical chemistry, multi-step synthesis, parallel synthesis, data management 

and ADME analysis. 

In his current position as VP Academic Relations at Fox Chase Chemical Diversity Center, Dr. Wrobel has 

collaborated and co-published with Professor Harty on in vitro and in vivo characterization of first-in-class novel 

inhibitors of filovirus PPxY interaction with host protein Nedd4 as an approach toward broad-spectrum antiviral 

therapy. Dr. Wrobel will be responsible for FCCDC project design, direction of the project and data interpretation. 

He will also organize regular project meetings among team members and consultants to maximize efficiency. 

Allen B. Reitz, Ph.D., CEO, Key Personnel (Yrs 1-3: 0.6 cal. months). Dr. Reitz has had >38 years of 

demonstrated accomplishment as a medicinal chemist in the pharmaceutical industry, including 26 years with 

Johnson & Johnson. He is a co-inventor as well as the project leader in most cases of nine compounds that 

have entered human clinical trials, two of which are currently in the clinic. He has >150 scientific publications 

and 71 issued U.S. patents, and is the Editor of the journal Current Topics in Medicinal Chemistry. He has 

extensive experience in project and portfolio management, target validation, hit triage, hit to lead and lead 

optimization medicinal chemistry, eADME profiling, and preclinical candidate selection. He is also Adj. Prof. at 

Drexel University, College of Medicine. Dr. Reitz founded FCCDC in 2008 to provide translational biomedical 

research in the area of early drug discovery research including medicinal chemistry hit to lead and lead 

optimization research. Dr. Reitz will assist Dr. Wrobel on project design and data interpretation. He will also pay 

particular attention to developing the required transition beyond Phase II, including a suitable project strategy, 

implementation of the Commercialization Plan, and interface with potential development partners and other 

interested parties. 

John Kulp III, Ph.D., Director of Computational Chemistry, Key Personnel (Yrs 1-3: 0.6 cal. months). Dr. 

Kulp has 9 years of computational chemistry experience with 17 peer reviewed publications. After his PhD and 

postdoctoral training he spent 4 years at BioLeap, a small business that specializes in protein-protein interactions 

and that has a proprietary fragment-based computational chemistry software platform. There he was the project 

leader for BioLeap’s PCSK9 program. He will perform computer-assisted drug design studies, and help to 

provide insight by the creation of computational pharmacophores during the course of the program. 

Katie B. Freeman, Ph.D., Director of Biology, FCCDC (Yrs 1-3: 0.6 cal. months/yr). Dr. Freeman has 

extensive experience with numerous in vitro assay formats, in microbiological methodologies, mammalian cell 

culture, bioanalytical methodologies. She will conduct cytotoxicity assays at FCCDC in liver HepG2 and 

endothelial HUVEC to compare the relative in vitro therapeutic index between series and individual compounds. 

In addition, she will perform formulations development as required to support the in vivo evaluation at TBRI. 

Hong Ye, Scientist (Yrs 1-3: 12 cal. months), Ms. Ye is an exceptionally talented bench medicinal chemist, 

who consistently ranks among the top staff at FCCDC in terms of productivity and creativity. She previously had 

20 years of experience at Johnson & Johnson in Spring House PA and other companies before joining FCCDC 

in June 2015. She has been the primary laboratory medicinal chemistry on the Harty Marburg/Ebola project and 

will continue to do so working under the direction of Dr. Wrobel with proper attention being paid to laboratory 

safety, confidentiality, productivity, notebook keeping and spectral data records retention. 

Budget Justification Attachment Page 55 
 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Leo J. Adalbert, M.B.A. Technical and Business Assistance (TABA). ($50,000 over three years 

requested) Mr. Adalbert is a Harvard MBA who will be our business development consultant for this project. 

He has more than 25 years of deep cross-functional commercial and clinical development experience across 

both small molecules and biologics. Prior to founding StraNexa LLC, Leo held various leadership roles across 

marketing, business development, and alliance leadership at leading firms such as AstraZeneca, Merck, Astra 

Merck, and Regeneron. Mr. Adalbert earned his MBA from Harvard Business School while on a fellowship, and 

holds executive leadership certifications from Northwestern (Kellogg) and the University of Pennsylvania 

(Wharton). He will interface with government agencies (FDA, BARDA) and investors as appropriate. 

Equipment

No new equipment is requested. 

Supplies

Materials will be purchased as needed for routine medicinal chemistry, including the following: 

Year 1 Year 2 Year 3 Total 

Solvent and reagents: 

Consumables lab supplies: 

Glassware: 

Total per year: 

Other Expenses

Contracted Research Organization Expenses: over three years 

Contract Research Organization (CRO) Expenses – ADMET and PK testing will be performed as in the following 

schedule. Contractor names are in parentheses, and have all worked effectively with FCCDC in the past. 

Budget Justification Attachment Page 56 
 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Indirect Cost and Fee 

Indirect costs of 40% are requested. A fee of 7% is requested. 

Justification for budget above statutory budget limitations

According to National Institutes of Health SBA-Approved SBIR/STTR Topics for Awards over Statutory Budget 

Limitations as listed in Appendix A of the omnibus 2020 solicitation 

(https://sbir.nih.gov/sites/default/files/NIH_Topics_for_Budget_Waivers.pdf), the topic “discovery and 

development of therapeutics for infectious diseases” (the prime concern of this proposal) is listed in NIAID, 

Division of Microbiology and Infectious Disease (DMD) as eligible for budgets over the statutory maximum 

amounts. 

Small molecule drug discovery is very expensive and labor intensive, involving the synthesis and evaluation of 

a large number of test compounds in an iterative fashion. In this STTR Phase II grant application, we embark on 

a full lead optimization program with the goal of obtaining preclinical drug candidates for the treatment of 

Marburg, Ebola and related viral infections. Our current lead series are novel and suitable for composition of 

matter patent protection, and a US patent has issued on the original series which will be supplemented by 

additional provisional patent applications over time. We expect the optimized compounds at the end of Phase II 

to have increased potency, and be safe and suitable for oral administration. 

Budget Justification Attachment Page 57 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

LE O J. AD A LBE RT 

G RE A T E R P HI L A D E L P HI A A RE A , PA 1890 1 

 www.stranexa.com 

BIO-PHARMA EXECUTIVE – MARKETING | STRATEGY | GLOBAL COMMERCIALIZATION | BD 

• Dynamic, creative, and strategic business leader with proven expertise in strategic commercialization, go-tomarket strategy, product launches, business development, and consulting across all phases of development, 

inclusive of biologics and small molecules. 

• Strong neuroscience, immunology, oncology, and rare disease/Orphan Drug commercialization and scientific 

expertise across multiple disease areas. 

• Outstanding track record of success in identifying and evaluating opportunities, developing a vision for growth, 

and building differentiated franchises. 

• Lead teams to exceed expectations utilizing high initiative, strong team-building skills, creative problem-solving, 

cross-functional leadership, and collaboration. Harvard MBA (Fellowship). 

CORE COMPETENCIES 

• Partnering & Collaboration • Leadership/Vision • New Product Launches & Promotion 

• Opportunity Identification • Customer, Competitor & Market Analysis • Product Development 

• Negotiations • Transactions • Capability Creation • Innovation/Medical Science 

• Strategic & Tactical Agility • Critical Thinking/Problem Solving • Talent Development 

• KOL and Advocacy Group Partnering • Metrics & Governance • Life-Cycle Management 

PROFESSIONAL EXPERIENCE AND KEY ACCOMPLISHMENTS 

STRANEXA, LLC (www.stranexa.com) (2014-Present) DOYLESTOWN, PA 

Founder and Principal 

Lead bio-pharmaceutical strategic commercialization consulting practice with focus on specialty and rare 

disease/Orphan Drug markets. Passion for building differentiated brands and capabilities transformative for 

patients and communities across for-profit and non-profit enterprises through a science-driven, market-led, patientfocused approach. 

• Led significant projects across multiple domains that consistently exceeded client expectations (portfolio and 

commercialization strategy, go-to-market strategy, product launch planning, BD/alliance leadership, competitor 

and market analyses, communications strategy and publications, non-profit market development). 

• Strong scientific foundation across diverse diseases and conditions. 

• Expert instructor with Biotech Primer www.biotechprimer.com (Strategic Commercialization Within BioPharma 

and Drug Development Immersion). 

CSL BEHRING, INC. (2012-2013) KING OF PRUSSIA, PA 

Global Marketing Director, Immunoglobulin (IgG) Marketing & Strategic Commercialization (’12-13) 

Consultant (’12) 

Led global marketing strategy, commercialization, launch planning and core tactics for global IgG portfolio with 

focus on HIZENTRA® and PRIVIGEN®. Led global commercial strategy for CSL362 (hematological tumors). 

• Chaired global business team and led major cross-functional initiatives guiding market leadership for key 

brands (globally PRIVIGEN® revenues up 17.9% to $1B+; HIZENTRA® revenues up 45.2% to $402MM). 

• Developed innovative strategies and major initiatives to build market leadership and differentiation. 

AXCAN PHARMA, INC. (2008-2011) BRIDGEWATER, NJ 

Director Business Development and Alliance Management (company restructured in 2011) 

• Successfully divested global PHOTOFRIN®/PHOTOBARR® oncology drug/device business, exceeding 

internal objectives (strong cash return, elimination of significant ongoing costs, and placement of commercial 

and R&D employees in acquiring firm). Led entire transaction process, including prospectus development, 

buyer identification, business case preparation, due diligence, negotiations and transition planning. 

• Created new Alliance Management capability (vision, strategies, metrics, tools and roll-out plan). Effectively 

managed ongoing and new alliance relationships. 

• Selected to lead LACTEOL® Global Product Team, and helped advance critical lifecycle initiatives and global 

presence in new markets. 

Page 1 of 4 

Budget Justification Attachment Page 58 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

LE O J. AD A LBE RT 

PROFESSIONAL EXPERIENCE AND KEY ACCOMPLISHMENTS (CONTINUED) 

 

ASTRAZENECA PHARMACEUTICALS (1999–2008) WILMINGTON, DE 

Advanced rapidly through a series of increasingly responsible roles, providing effective leadership in driving major 

commercialization and business development initiatives (multiple recipient of AstraZeneca Excellence Award). 

Alliance and Program Leader, Strategic Planning & Business Development 

• Spearheaded and managed collaboration to commercialize CRESTOR/fenofibrate fixed-dose combination. 

Collaboration progressed six months ahead of schedule and $3.5M under budget. 

• Instrumental in design and implementation of new Alliance Management capability within the firm. 

Senior Director, Licensing Commercial Evaluations 

Managed commercialization strategy in collaboration with Global Product Teams. Led evaluations of Phase II/III 

licensing/acquisition opportunities across multiple therapeutic areas. 

• Led product and company alliance and acquisition evaluations covering oncology and neuroscience, driving 

successful progression of target opportunities through formal diligence to deal completion. 

• Defined and implemented novel commercialization approaches for various CRESTOR® fixed dose 

combination opportunities. 

Senior Director, US Marketing & Commercial Operations – Oncology & Infection 

Directed commercialization strategy, market development, and licensing support for oncology/infection emerging 

brands portfolio (seven drug candidates from Discovery through Phase III clinical development covering lung, 

prostate, breast, and colorectal cancers). 

• Directed overall strategic direction, including creation of integrated target product profiles and patientcentered claims across entire brand portfolio encompassing multiple pathways (SRC kinase, VEGF/EGF, 

endothelin, vascular targeting agents, and prenylation inhibitors.) 

• Proactively identified new business opportunities and resolved strategic gaps with global product teams to 

consistently maintain momentum through key decision-points. 

Transaction Director, Licensing – North America 

Led transaction process through all stages involving analyzing, negotiating, closing, and managing transitions 

of select partnership opportunities (product divestments, co-promotions, and in-licenses.) 

• Directed all facets of buyer identification, business case prep, due diligence, negotiations, and transition 

planning. Successfully closed multiple divestment transactions under tight timelines, delivering significant 

operating income, tax benefits, and business efficiencies based on significant premiums to retention values. 

• Consistently earned Distinguished/Excellent performance ratings (top 2%) and achieved “Deal of the Year” 

recognition. 

EARLIER RELEVANT EXPERIENCE AND KEY ACCOMPLISHMENTS 

ELAN PHARMACEUTICALS, INC. SAN FRANCISCO, CA 

Director, Neuromuscular Marketing (business restructured via SEC obligations) 

Developed and led execution of strategic and tactical marketing plans to effectively support all stages of 

lifecycle/market entry. Developed comprehensive product pre-launch and launch deployment plans that 

expanded Élan’s neuromuscular franchise into attractive new market segments. 

ASTRA-MERCK PHARMACEUTICALS, INC. (JV dissolved) WAYNE, PA 

Director, Neuroscience Marketing & Business Strategy 

Marketing Manager, Neuroscience (’96-‘97) 

Directed franchise development of Astra-Merck’s neuroscience business as commercial lead in formulating 

strategic direction and integration activities with Astra Sweden corporate portfolio. 

• Established strategic alliances and launched market development activities with patient advocacy groups, 

medical associations, and payers. Recognized with Leadership Award by Huntington’s Disease Society. 

• Recipient of Astra-Merck Leadership Award for Excellence for development and implementation of an 

innovative, comprehensive business and licensing strategy encompassing multiple neurological and 

psychiatric diseases, indications and product opportunities globally. 

Page 2 of 4 

Budget Justification Attachment Page 59 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

LE O J. AD A LBE RT 

EARLIER RELEVANT EXPERIENCE AND KEY ACCOMPLISHMENTS (CONTINUED) 

 

RHONE-POULENC RORER PHARMACEUTICALS, INC. COLLEGEVILLE, PA 

Senior Product Manager – U.S. 

Led market development and successful introduction of RILUTEK®, the first and only medical therapy approved for 

the treatment of amyotrophic lateral sclerosis (Lou Gehrig’s Disease) prolonging survival. Achieved 95% formulary 

acceptance within six months and sales 121% to plan. 

REGENERON PHARMACEUTICALS, INC. TARRYTOWN, NY 

Manager, Market Development

Established commercial direction for the company’s first biologic product CNTF (ciliary neurotrophic factor) for the 

treatment of multiple neurodegenerative diseases. (Product failed in late-Phase III clinical trials.) 

MERCK HUMAN HEALTH U.S. – a Division of Merck & Company WEST POINT, PA 

Senior Promotion Manager – Promoted from Promotions Manager 

Directed $20M product promotion and market development pre-launch and launch program. Drove successful 

introduction of PROSCAR® in the U.S., the first medical therapy treatment of benign prostatic enlargement. 

• Physician and DTC Program acknowledged with several industry awards; achieved 119% to plan. 

Marketing Analyst – Promoted from Associate Marketing Analyst 

Designed, implemented, and analyzed strategic and product-specific marketing research. 

• Developed and presented recommendations to senior management based on research findings, resulting in 

>$20M in media efficiencies. 

• Recognized with Merck Leadership Award of Excellence for developing methodology to analyze the impact 

of changes in promotion mixes and budgets that decreased planning time and increased efficiencies by 33%. 

Professional Sales Representative, Western Pennsylvania Territory 

Managed $4 million office and hospital sales territory. Conducted educational forums, and presented product 

and clinical information to nurses, pharmacists and physicians in primary care, cardiology, neurology, 

orthopedics and ophthalmology. (Achieved 124% of goal) 

EDUCATION & PROFESSIONAL DEVELOPMENT 

HARVARD UNIVERSITY GRADUATE SCHOOL OF BUSINESS ADMINISTRATION BOSTON, MA 

Master of Business Administration (MBA) - Concentration: Marketing & Finance 

Awarded Harvard Business School Fellowship 

WEST VIRGINIA UNIVERSITY MORGANTOWN, WV 

Bachelor of Science in Business Administration – Highest Honors (Summa Cum Laude) / Rhodes 

Scholarship Finalist

Beta Gamma Sigma National Business Honor Society; Phi Kappa Phi & Mortar Board National Honor Societies; 

Phi Sigma Kappa Fraternity (Repeat Merit Scholar); Varsity Track Team (400 meters/relay). 

CONTINUING EDUCATION: 

• Harvard Medical School HMX Program (completed with Achievement Honors Pharmacology and Immunology; 

Genetics and Physiology in progress to complete 2019) 

• Villanova University/Bucks County Community College – Completion of full pre-med/health sciences program 

with labs (anatomy/chemistry/biology) - recognized with highest honors and selected for membership in Phi 

Theta Kappa 

• Harvard School of Public Health (TH Chan School) 

o Measurement, Design, and Analysis Methods for Health Economics Research 

o Navigating the US Bio-Pharmaceutical Sector – Policy, Politics, Pricing, and Payment 

o Leading in Health Systems: Integrating Effort, Improving Outcomes 

Page 3 of 4 

Budget Justification Attachment Page 60 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

LE O J. AD A LBE RT 

 

EDUCATION & PROFESSIONAL DEVELOPMENT (CONTINUED) 

• Northwestern University Graduate School of Management: Graduate of Kellogg Executive Scholar Program 

• Wharton School (University of Pennsylvania): Graduate of Professional Development Program 

• Center for Creative Leadership: Leadership Development Program 

• Dartmouth: Global Leaders Program 

• GAP International: Leadership for Breakthrough Outcomes 

• Harvard Law School: Negotiations Strategy Program 

• Merck & Company: Strategic Marketing Plan Development 

• MIT: Systems Thinking Program 

CERTIFICATIONS: Certified Licensing Professional & Certified Strategic Alliance Professional 

COMMUNITY / PERSONAL 

• Special Projects Leader – Castleman Disease Collaborative Network (CDCN) www.cdcn.org 

• Co-Creator & Executive Board member of Camden, NJ “Dream Center” (youth community outreach center) 

www.camdendreamcenter.org 

• Graduate of Chester County non-profit Leadership Development Program (partnership with United Way) 

• Competitive masters track and field athlete (400 and 800 meters) and road cyclist 

Page 4 of 4 

Budget Justification Attachment Page 61 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 1 of 20 

Quotation Date: 10 August 2020 Quotation Number: APQ-19-0548 

Sponsor 

Jay Wrobel, PhD 

Vice President, Academic Relations 

Fox Chase Chemical Diversity Center 

Doylestown, PA 18902 

Phone: 

Mobile: 

E-mail: 

Alliance Pharma, Inc. 

Ryan Klein, PhD 

Director, Business Development 

Malvern, PA 19355 

Mobile: 

Phone: 

Fax: 

Email: 

Web: 

Non-GLP ADME Studies of Compounds for Fox Chase Chemical Diversity Center 

(Three Year Plan) 

1 Plasma Protein Binding Study By Rapid Equilibrium Dialysis 

1.1 Scope of Service 

To assess the protein binding of compounds in mouse and human plasma using a rapid equilibrium 

dialysis (RED) device (Thermo Fisher Scientific). 

1.2 General Procedures 

The test compounds will be spiked into plasma samples to achieve a final concentration of 5 μM. 

Triplicate aliquots of the spiked plasma will be added to the red chambers of the RED device 

inserts, and phosphate buffered saline will be added into the white chambers of the inserts. The 

RED device plate will be sealed and incubated for 4 hours at 37°C with shaking. Warfarin, a known 

highly bound drug in plasma, will be tested as the positive control for this assay. 

After incubation, samples from both the red and white chambers will be analyzed by LC-MS/MS. 

The percentages of free and bound fractions will be determined using the peak area ratios of each 

analyte to internal standard. 

1.3 General Information and Sample Requirement 

• Molecular weights, including formula weights and exact mass units, of the compounds. 

• Sufficient amount of the compounds. 

• Stability information for the compounds such as light, temperature, and solvents, if available. 

• Safety information for handling the compounds. 

• Solubility of the compounds, if available. 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 65 
 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 2 of 20 

1.4 Deliverable 

A Microsoft Excel summary report including raw data, methodology, calculated percent bound, 

and percent recovery values will be provided. 

1.5 Turnaround Time 

Report will be delivered within 7 business days from receipt of the compounds and any additional 

items specific to the assay that need to be sourced externally. 

1.6 Documentation/Records 

Data from this non-GLP study will be preserved for 1 year after submission of the results to the 

Sponsor. 

1.7 Fee Schedule 

The cost of plasma protein binding analysis is per compound. 

Item Year 1 Year 2 Year 3 Cost 

Human Plasma 

Binding 

Number of compounds 2 2 2 

Cost 

Mouse Plasma 

Binding 

Number of compounds 1 1 1 

Cost 

Total: 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 66 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 3 of 20 

2 Metabolic Stability Studies (Intrinsic Clearance) in Liver Microsomes 

2.1 Scope of Service 

To assess the relative metabolic stability (intrinsic clearance) of compounds using mouse, rat, 

canine and human liver microsomes. 

2.2 General Procedures 

The test compounds at a concentration of 0.5 μM will be incubated with 0.5 mg/mL of liver 

microsomes and an NADPH-regenerating system (cofactor solution) in potassium phosphate 

buffer (pH 7.4). At 0, 5, 15, 30, and 45 minutes, an aliquot will be taken, and reactions will be 

quenched with an acetonitrile solution containing an internal standard. Additionally, controls will 

be measured that do not contain the cofactor solution. Following completion of the experiment, 

samples will be analyzed by LC-MS/MS. Results will be reported as peak area ratios of each 

analyte to internal standard. The intrinsic clearance (CLint) will be determined from the first-order 

elimination constant by nonlinear regression. 

2.3 General Information and Sample Requirement 

• Molecular weights, including formula weights and exact mass units, of the compounds. 

• Sufficient amounts of the compounds. 

• Stability information for the compounds such as light, temperature, and solvents, if available. 

• Safety information for handling the compounds. 

• Solubility of the compounds, if available. 

2.4 Deliverables 

A Microsoft Excel summary report of intrinsic clearance (CLint), half-life (t1/2), rate constant, 

peak area ratios of each analyte to internal standard, and graphic profiles will be provided. 

2.5 Turnaround Time 

Report will be delivered within 7 business days from Sponsor’s approval of the study and receipt 

of all necessary information and assay-specific materials/reagents by Alliance Pharma. 

2.6 Documentation/Records 

Data from this non-GLP study will be preserved for 1 year after submission of the results to 

the Sponsor. 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 67 
 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 4 of 20 

2.7 Fee Schedule 

The cost of metabolic stability analysis using liver microsomes is per compound. 

Item Year 1 Year 2 Year 3 Cost 

Microsomal 

Clearance (Mouse) 

Number of 

compounds 15 15 6 

Cost 

Microsomal 

Clearance (Rat) 

Number of 

compounds 2 2 2 

Cost 

Microsomal 

Clearance (Dog) 

Number of 

compounds 2 2 2 

Cost 

Microsomal 

Clearance 

(Human) 

Number of 

compounds 6 6 10 

Cost 

Total: 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 68 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 5 of 20 

3 Direct Cytochrome P450 (CYP 3A4) Inhibition Study 

3.1 Scope of Service 

To evaluate the direct inhibition potential of compounds on the major human hepatic CYP3A4 

isoform using pooled human liver microsomes and the isoform-specific probe substrate 

testosterone. 

3.2 General Procedures 

Pooled human liver microsomes will be used for this study. Incubation mixtures will contain 

a) potassium phosphate buffer, b) 0.1 mg/mL microsomal protein; c) CYP3A4 probe substrate 

testosterone (12.5 μM); d) specific concentrations of test compounds, blank solvent or positive 

controls; and (e) an NADPH-regenerating cofactor solution. The mixtures will be incubated at 

37°C and 5% CO2 with orbital shaking at 200 rpm for 5 minutes. Upon completion of the 

incubation, the reactions will be quenched with an acetonitrile solution containing an internal 

standard. The samples will be prepared and analyzed by LC-MS/MS to monitor for substrate 

metabolite formation. The enzyme activity in the presence of the compounds will be normalized 

with the enzyme activity in the absence of the compounds (controls) and expressed as a percentage 

of activity. The inhibitory potential (IC50) of the compounds will be determined using nonlinear 

regression. 

3.3 General Information and Sample Requirement 

• Molecular weights, including formula weights and exact mass units, of the compounds. 

• Sufficient amounts of the compounds. 

• Stability information for the compounds such as light, temperature, and solvents, if available. 

• Safety information for handling the compounds. 

• Solubility of the compounds, if available. 

3.4 Deliverables 

A Microsoft Excel summary report including IC50 summary tables and plots of test compound 

concentration versus peak area ratios will be provided. 

3.5 Turnaround Time 

Report will be delivered within 8 business days from Sponsor’s approval of the study and receipt 

of all necessary information and assay-specific materials/reagents by Alliance Pharma. 

3.6 Documentation/Records 

Data from this non-GLP study will be preserved for 1 year after submission of the results to 

the Sponsor. 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 69 
 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 6 of 20 

3.7 Fee Schedule 

The cost of CYP inhibition analysis is per compound, per probe substrate (one isoformspecific probe substrate used). 

Item Year 1 Year 2 Year 3 Cost 

Direct CYP IC50 

(1 substrate) 

Number of compounds 6 6 4 

Cost 

Total: 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 70 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 7 of 20 

4 Direct Cytochrome P450 (CYP) Inhibition Study – Full Panel 

4.1 Scope of Service 

To evaluate the direct inhibition potential of compounds on 7 major human hepatic 

cytochrome P450 (CYP) isoforms (1A2, 2B6, 2C8, 2C9, 2C19, 2D6 and 3A4) using pooled human 

liver microsomes and the isoform-specific probe substrates. 

4.2 General Procedures 

Pooled human liver microsomes will be used for this study. The incubation mixtures will contain 

the following: (a) potassium phosphate buffer; (b) 0.1 mg/mL microsomal protein; 

(c) isoform-specific probe substrates; (d) specific concentrations of the test compounds, 

blank solvent, or positive controls (isoform-specific inhibitors); and (e) an NADPH-regenerating 

cofactor solution. Table 1 below lists the inhibitors, substrates, and associated concentrations that 

will be used for each incubation. 

Table 1: Isoform-Specific Probe Substrates and Inhibitors 

CYP Isoform Inhibitor Final Concentrations 

(μM) Substrate Final Concentration (μM)

1A2 Fluvoxamine 0.0033, 0.001, 0.033, 

0.1, 0.33, 1, 3.3 Phenacetin 35 

2B6 Clopidogrel 0.033, 0.1, 0.33, 1, 3.3, 

10, 33 Bupropion 50 

2C8 Quercetin 0.033, 0.1, 0.33, 1, 3.3, 

10, 33 Amodiaquine 1.5 

2C9 Sulfaphenazole 0.0033, 0.01, 0.033, 0.1, 

0.33, 1, 3.3 Diclofenac 5 

2C19 Ticlopidine 0.033, 0.1, 0.33, 1, 3.3, 

10, 33 S-mephenytoin 25 

2D6 Quinidine 0.0033, 0.01, 0.033, 0.1, 

0.33, 1, 3.3 Dextromethorphan 5 

3A4 Ketoconazole 0.00033, 0.001, 0.0033, 

0.01, 0.033, 0.1, 0.33 Midazolam 1.25 

The mixtures will be incubated at 37°C and 5% CO2 with orbital shaking at 200 rpm for 5 minutes 

with reactions containing midazolam, for 10 minutes with reactions containing phenacetin, 

bupropion, diclofenac, or dextromethorphan, and for 20 minutes with reactions containing 

amodiaquine or S-mephenytoin. Upon completion of the incubation, the reactions will be quenched 

with an acetonitrile solution containing an internal standard. The samples will be prepared and 

analyzed by LC-MS/MS to monitor for substrate metabolite formation. The enzyme activity in the 

presence of the compounds will be normalized with the enzyme activity in the absence of the 

compounds (controls) and expressed as a percentage of activity. The inhibitory potential (IC50) of 

the compounds will be determined using nonlinear regression. 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 71 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 8 of 20 

4.3 General Information and Sample Requirement 

• Molecular weights, including formula weights and exact mass units, of the compounds. 

• Sufficient amounts of the compounds. 

• Stability information for the compounds such as light, temperature, and solvents, if available. 

• Safety information for handling the compounds. 

• Solubility of the compounds, if available. 

4.4 Deliverables 

A Microsoft Excel summary report including IC50 summary tables and plots of test compound 

concentration versus peak area ratios will be provided. 

4.5 Turnaround Time 

Report will be delivered within 8 business days from Sponsor’s approval of the study and receipt 

of all necessary information and assay-specific materials/reagents by Alliance Pharma. 

4.6 Documentation/Records 

Data from this non-GLP study will be preserved for 1 year after submission of the results to 

the Sponsor. 

4.7 Fee Schedule 

The cost of the full-panel CYP inhibition assay is per compound ( per compound, per 

probe substrate). 

Item Year 1 Year 2 Year 3 Cost 

Direct CYP IC50 

(7 substrates) 

Number of 

compounds 2 2 2 

Cost 

Total: 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 72 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 9 of 20 

5 Plasma Stability Studies 

5.1 Scope of Service 

To assess the relative stability of compounds in mouse and human plasma. 

5.2 General Procedures 

Test compounds will be incubated at a concentration of 1 μM in plasma samples. At 0, 15, 30, 60 

and 120 minutes, an aliquot will be removed and reactions will be quenched with an acetonitrile 

solution containing an internal standard. Following completion of the experiment, samples will be 

analyzed by LC-MS/MS. Results will be reported as peak area ratios of analyte to internal standard. 

5.3 General Information and Sample Requirement 

• Molecular weights, including formula weights and exact mass units, of the compounds. 

• Sufficient amounts of the compounds. 

• Stability information for the compounds such as light, temperature, and solvents, if available. 

• Safety information for handling the compounds. 

• Solubility of the compounds, if available. 

5.4 Deliverables 

A Microsoft Excel summary report including half-life (t1/2), elimination rate constant, peak area 

ratios of each analyte to internal standard, and graphic profiles will be provided. 

5.5 Turnaround Time 

Report will be delivered within 6 business days from Sponsor’s approval of the study and receipt 

of all necessary information and assay-specific materials/reagents by Alliance Pharma. 

5.6 Documentation/Records 

Data from this non-GLP study will be preserved for 1 year after submission of the results to 

the Sponsor. 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 73 
 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 10 of 20 

5.7 Fee Schedule 

The cost of plasma stability analysis is per compound. 

Item Year 1 Year 2 Year 3 Cost 

Mouse Plasma 

Stability 

Number of compounds 2 2 2 

Cost 

Human Plasma 

Stability 

Number of compounds 2 2 1 

Cost 

Total: 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 74 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 11 of 20 

6 Aqueous Solubility Study in Phosphate-Buffered Saline 

6.1 Scope of Service 

To assess the solubility of compounds in phosphate-buffered saline (PBS). 

6.2 General Procedures 

Approximately 1 mL of PBS buffer (pH 7.4) will be added to approximately 10 mg of each test 

compound. The solutions will be sonicated for 30 minutes and vortexed at a low speed for at least 

30 minutes. The solutions will then sit at room temperature for 16-28 hours. After such time, the 

solutions will be filtered through a 0.22-micron filter followed by diluting 1:10, 1:100, and 1:1000, 

and 1:10,000 in triplicate in 50% acetonitrile in water (1:1, v/v). The final sample solutions will 

be mixed with an internal standard solution (1:1) followed by LC MS/MS analysis. Solubility will 

be determined according to the calibration curve generated from 5 concentration standards. 

6.3 General Information and Sample Requirement 

• Molecular weights, including formula weights and exact mass units, of the compounds. 

• Approximately 10 mg of each compound (minimum requirement). 

• Stability information for the compounds such as light, temperature, and solvents, if available. 

• Safety information for handling the compounds. 

• Solubility of the compounds, if available. 

6.4 Deliverables 

A Microsoft Excel summary report including standard curve information and sample solubility 

data will be provided. 

6.5 Turnaround Time 

Report will be delivered within 7 business days from receipt of compounds by Alliance Pharma. 

6.6 Documentation/Records 

Data from this non-GLP study will be preserved for 1 year after submission of the report to 

the Sponsor. 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 75 
 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 12 of 20 

6.7 Fee Schedule 

The cost of aqueous solubility analysis is per compound. 

Item Year 1 Year 2 Year 3 Cost 

Aqueous Solubility 

Number of compounds 6 6 4 

Cost 

Total: 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 76 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 13 of 20 

7 Permeability in MDCKII-MDR1 cells 

7.1 Scope of Service 

To assess the permeability of compounds in MDCKII-MDR1 cells. 

7.2 General Procedures 

The permeability of the test compound in the MDCKII-MDR1 cell monolayer system will be 

assessed in duplicate in both apical to basolateral (A→B) and basolateral to apical (B→A) 

directions. 

MDCK cells will be seeded onto multiwell permeable supports in culture medium. The 

permeability study will be conducted approximately 4 days post-seeding. The Transepithelial 

Electrical Resistance (TEER) of the cell monolayer will serve as a quality control for monolayer 

integrity. On the day of the transport assay, TEER will be measured in all wells. 

Transport medium will be added to the basolateral and apical sides of the system for a 30-minute 

incubation to wash the monolayer. After removing transport medium, receiving solution and 

compound-specific dosing solution (containing Lucifer yellow) will be added to the appropriate 

wells of the system for A→B and B→A analysis. The multiwell system will be incubated at 37°C 

with 5% CO2. After incubation, samples will be collected from both the apical and basolateral 

sides of the system for LC-MS/MS analysis. Lucifer yellow will be fluorometrically determined 

in all receiving samples and will serve as an in-well quality control for monolayer integrity. 

7.3 General Information and Sample Requirements 

Sponsor shall provide the following: 

• Molecular weights, including formula weights and exact mass units, of the compounds. 

• Sufficient amount of each compound. 

• Stability information (e.g., light, temperature, and solvents) for the compounds, if available. 

• Safety information for handling the compounds. 

• Solubility of the compounds, if available. 

7.4 Deliverables 

A spreadsheet summary report (Microsoft Excel file) will be provided that includes the 

permeability coefficient of the compound from basolateral side to apical side (Papp, B to A) and from 

apical side to basolateral side (Papp, A to B). 

7.5 Turnaround Time 

The spreadsheet summary report will be delivered to the Sponsor within approximately 11 business 

days from Sponsor’s approval of the study and receipt of all necessary information and assayspecific materials/reagents by Alliance Pharma. 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 77 
 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 14 of 20 

7.6 Documentation/Records 

Data from this non-GLP study will be preserved for 1 year after submission of the results to the 

Sponsor. 

7.7 Fee Schedule 

The cost of MDCKII-MDR1 permeability analysis is per compound. 

Item Year 1 Year 2 Year 3 Cost 

Permeability 

Number of compounds 2 2 1 

Cost 

Total: 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 78 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 15 of 20 

8 Non-GLP in vivo Pharmacokinetic Studies in Balb/C Mice by PO Administration 

8.1 Scope of Service 

To provide the services of in-life animal dosing and sample bioanalysis for pharmacokinetic 

studies conducted in mice. 

8.2 General Procedures 

One group of 21 mice (n=3 mice per time point) will be administered an oral (PO) dose of the 

compound of interest. Dosing formulation will be prepared according to Sponsor-provided 

protocol. Terminal blood plasma samples will be collected from each animal at 7 established time 

points within a 24 hour period. After all study samples are collected, the samples will be analyzed 

by LC-MS/MS. Calculated concentrations will be reported to the Sponsor. 

8.3 General Information and Sample Requirement 

• Predicted concentration range and/or dosing, Cmax information, if available 

• Compound molecular weight and its purity information 

• Sufficient amount of standard compound 

• Sample stability information, such as light sensitivity, temperature and solvent, if available 

• Safety information for handling compounds/samples 

• Solubility of the compound, if available 

8.4 Deliverables 

A report of abnormal in-life observations will be provided to the Sponsor, if applicable. A 

Microsoft Excel spreadsheet summary of sample analysis results will be delivered to the Sponsor. 

8.5 Turnaround Time 

The pharmacokinetic study will be completed and results will be delivered within approximately 

10 business days from receipt of the test compound and all other necessary study materials. 

8.6 Documentation and Records 

Data from this non-GLP study will be preserved for 1 year after submission of the report to 

the Sponsor. 

8.7 Fee Schedule 

The cost of the in vivo pharmacokinetic study (dosing and bioanalysis) is per compound. 

Item Year 1 Year 2 Year 3 Cost 

In Vivo PO PK 

in Mice 

Number of 

compounds 1 1 1 

Cost 

Total: 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 79 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 16 of 20 

9 Non-GLP in vivo Pharmacokinetic Studies in Balb/C Mice by IV/PO Administration 

9.1 Scope of Service 

To provide the services of in-life animal dosing and sample bioanalysis for pharmacokinetic 

studies conducted in mice. 

9.2 General Procedures 

Two groups of 21 mice (n=3 mice per time point) will be administered an intravenous (IV) or oral 

(PO) dose of the compound of interest. Dosing formulation will be prepared according to Sponsorprovided protocol. Terminal blood plasma samples will be collected from each animal at 

7 established time points within a 24 hour period. After all study samples are collected, the samples 

will be analyzed by LC-MS/MS. Calculated concentrations will be reported to the Sponsor. 

9.3 General Information and Sample Requirement 

• Predicted concentration range and/or dosing, Cmax information, if available 

• Compound molecular weight and its purity information 

• Sufficient amount of standard compound 

• Sample stability information, such as light sensitivity, temperature and solvent, if available 

• Safety information for handling compounds/samples 

• Solubility of the compound, if available 

9.4 Deliverables 

A report of abnormal in-life observations will be provided to the Sponsor, if applicable. A 

Microsoft Excel spreadsheet summary of sample analysis results will be delivered to the Sponsor. 

9.5 Turnaround Time 

The pharmacokinetic study will be completed and results will be delivered within approximately 

10 business days from receipt of the test compound and all other necessary study materials. 

9.6 Documentation and Records 

Data from this non-GLP study will be preserved for 1 year after submission of the report to 

the Sponsor. 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 80 
 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 17 of 20 

9.7 Fee Schedule 

The cost of the in vivo pharmacokinetic study (dosing and bioanalysis) is per compound. 

Item Year 1 Year 2 Year 3 Cost 

In Vivo IV/PO PK 

in Mice 

Number of 

compounds 6 6 6 

Cost 

Total: Total: 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 81 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 18 of 20 

10 Total Fee Schedule 

Assay Year 1 Year 2 Year 3 

1. Plasma Protein Binding 

2. Microsomal Clearance 

3. Direct CYP IC50 (3A4 - Testosterone) 

4. Direct CYP IC50 (1A2, 2B6, 2C8, 2C9, 

2C19, 2D6 and 3A4-Midazolam) 

5. Plasma Stability 

6. Aqueous Solubility 

7. Permeability 

8. In Vivo PK in Mice (PO) 

9. In Vivo PK in Mice (IV/PO) 

Annual Total 

Grand Total 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 82 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 19 of 20 

TERMS AND CONDITIONS 

1. SERVICES 

1.1 Compliance — Alliance Pharma shall perform itself or, as the case may be, through its affiliate(s) for the benefit of 

Sponsor, the Services described herein in accordance with, as applicable: (i) this Proposal; (ii) applicable standard 

operating procedures; (iii) current standards of Good Laboratory Practice/Good Clinical Practice (GLP/GCP) as 

applicable in the country where the Services are performed; and (iv) any other applicable professional standards, codes, 

guidelines, laws, rules, and regulations, including, without limitation, those of any regulatory authority or agency having 

jurisdiction over the Services (collectively, the “Applicable Laws”). Performance of such Services shall only be 

initiated after the full execution of this Proposal. 

1.2 Mutual non-solicitation — During the term of this Proposal and for a two (2) year period thereafter, neither party nor its 

respective affiliates shall, directly or indirectly: (i) recruit, solicit or induce, or attempt to recruit, solicit or induce, any 

employee of the other party to terminate his or her employment or engagement with the other party, or (ii) hire any 

employee of the other party (including, without limitation, employees of Alliance Pharma that are providing services to 

Sponsor on behalf of Alliance Pharma at the time of hiring). 

NOTE.—This Proposal is valid for sixty (60) calendar days from quotation date. 

2. CHEMICAL AND BIOLOGICAL SAMPLES 

2.1 Delivery — Sponsor shall deliver to Alliance Pharma, at Sponsor’s risk and expense and at the location of Alliance 

Pharma identified in this Proposal or to such other location of Alliance Pharma as identified to Sponsor in writing, the 

“Chemical Samples” and “Biological Samples” in sufficient and necessary quantities for Alliance Pharma to perform the 

Services in compliance with the provisions of Section 1.1 above and as required under the Applicable Laws, along with 

instructions for the handling, storage, and safety requirements and conditions of such Chemical or Biological Samples. 

Alliance Pharma will receive shipment(s) of Chemical or Biological Samples only if the equipment required to safely 

process the samples exists at Alliance Pharma. 

2.2 Restricted Use — Alliance Pharma shall neither use nor analyze any Chemical or Biological Samples for any purposes 

other than to perform the Services without Sponsor’s prior written consent. 

2.3 Storage and Disposal of Chemical Samples— Alliance Pharma will store the Chemical Samples at the appropriate 

storage conditions as directed by the Sponsor for 3 months after submission of the final deliverable hereunder. Alliance 

Pharma will dispose of the Chemical Samples following the 3-month expiration date, unless requested otherwise by the 

Sponsor in writing prior to said date. 

2.4 Storage, Return or Destruction of Biological Samples— For regulatory (e.g., GLP/GCP) studies, Alliance Pharma will 

store the Biological Samples at the appropriate storage conditions as directed by the Sponsor for 6 months after submission 

of the final service deliverable herein. Alliance Pharma will continue to store the Biological Samples following the 6-

month expiration date at the rate of US $0.50 per Biological Sample per month, unless otherwise directed by the 

Sponsor in writing prior to said date. Sponsor will retain the option of having the Biological Samples destroyed, stored 

under Alliance Pharma’s responsibility, or returned to Sponsor at Sponsor’s expense. An authorization request form will 

be sent to Sponsor at sponsor’s request. For non-regulated studies, Alliance Pharma will destroy Biological Samples two 

(2) weeks after the Sponsor acknowledges receipt of final Service deliverables unless otherwise requested by the 

Sponsor in writing prior to such date. 

If Biological Samples are received by Alliance Pharma that are not intended for bioanalysis under the scope of this 

Proposal, such as back up samples, the Sponsor will be contacted and offered the option of having the Biological Samples 

destroyed, stored under Alliance Pharma’s responsibility, or returned to the Sponsor at Sponsor’s expense. If stored under 

Alliance Pharma’s responsibility, the samples, either regulatory or non-regulatory, will be stored and invoiced at the rate 

of US $0.50 per Biological Sample per month. 

All sample storage fees are based on standard 1.2 – 2 mL cryogenic storage vials with a maximum height of 49 mm and 

diameter of 13.5 mm. Larger sample containers may incur additional fees to be agreed upon between the parties to this 

Proposal. 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 83 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

STUDY PROPOSAL p. 20 of 20 

3. BUDGET AND PAYMENT TERMS 

3.1 Service Fees — In accordance with the following, Sponsor shall provide Alliance Pharma with a payment amount 

corresponding to the Service fees identified herein. The per-sample pricing provided in this Proposal is based on the 

number of samples requested by the Sponsor. Pricing may be renegotiated if the sample count decreases from the quoted 

count by more than 20%. Sample re-assays/repeats requested by Sponsor, dilutional rea-assays, and incurred sample 

reanalysis will be invoiced at the sample unit rate. Re-assays/repeats due to failure to meet the pre-defined criteria will be 

at no charge to Sponsor. However, any special requirements from the Sponsor not included in such budget will be invoiced 

to the Sponsor at additional cost. 

3.2 Pass-Through Costs — Pricing includes all standard laboratory consumables; however, the cost of non-standard items 

(e.g. chromatographic columns, reference standards, internal standards, SPE plates, commercial ELISA kits, enzymes, 

specific reagents etc.) purchased by Alliance Pharma will be passed-through to the study sponsor. The fees identified 

herein may include estimated costs that are expected at the start of the study; however, additional materials may be 

required. All pass-through costs are subject to a mark-up fee for purchasing, handling and accounting services. 

3.3 Installments - Alliance Pharma will invoice upon delivery of the first draft report for each study. (Any additional 

authorized dollars from contract modifications, if any, will be added to the next billing cycle.) 

3.4 Payment Terms — Sponsor shall pay each invoice within thirty (30) days of receipt thereof. Any amount owed by 

Sponsor under this Proposal that is not received by Alliance Pharma on or before the date due shall bear interest at a per 

annum rate of two percent (2%) above the prime rate in effect at Bank of America on the date due. Sponsor shall also 

pay all reasonable collection costs at any time incurred by Alliance Pharma in obtaining payment of amounts past due, 

including reasonable attorneys’ fees. If any portion of an invoice is disputed, the Sponsor shall pay the undisputed 

amounts as set forth therein, and the parties shall use good faith efforts to reconcile the disputed portion of the invoice 

within thirty (30) days of receipt by Sponsor of the applicable invoice. 

IN WITNESS WHEREOF, this Agreement has been accepted and executed by duly authorized 

representatives of both parties as follows: 

Alliance Pharma Inc. SPONSOR: Fox Chase Chemical Diversity Center 

By: By: 

Print Name: Ryan Klein, PhD Print Name: 

Title Director, Business Development Title: 

Date: Date: 

APQ-20-0548_DM_FoxChase_vFinal 

Budget Justification Attachment Page 84 
 

 

 

 Turnaround Time. 

 link) for more details. 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Quotation 

Customer Delivery Information Customer Invoice Information 

Fox Chase Chemical Diversity Center Inc Fox Chase Chemical Diversity Center Inc 

Doylestown, PA 18902, United States 

2, Doylestown, PA 18902, United States Name: Accounts Payable 

Name: Jay Wrobel Phone: 

Phone: Email: 

Email: 

Quotation Number Study Code Customer Account No Eurofins Contact 

Andrew Lord 

Date Issued Customer reference Payment Terms Eurofins Contact Phone 

04-Aug-2020 30 Days 

Quote Valid Until Promo Code Loyalty Category Eurofins Contact Email 

03-Sep-2020 Standard 

Summary 

Price Discount Charges Net Price Sales tax Grand Total Price Currency 

0.00 0.00 0.00 USD 

Details 

Item # Description Assay Cmpd Conc Rep Qty Option TAT* Unit price Disc unit Net Price 

Mode price 

PP241 SafetyScreen44 Panel, Panlabs Flexible 1 1 2 1.00 Standard 10 

Special Requirement: Default test concentrations 10 uM 

Cmpd = compounds; Conc = Concentrations; Rep = Replicates; Qty = Quantity; TAT =

*please see “Delivery Dates, Turnaround Time” section of Terms and Conditions (web

Additional Charges 0.00

Special Instructions: N/A 

Orders can be placed by purchase order or credit card. 

Purchase orders should detail the following company address: 

Eurofins Panlabs Discovery Services Taiwan Ltd 

New Taipei City, Taiwan 24891 

Please note that the address for sending check payment is as following: 

Birmingham, AL 35246-5588 

Please note that if arranging payment from a bank outside of Taiwan, any intermediary or sender’s charges should be borne by the sender. 

AUTHORIZATION AND ACCEPTANCE 

Submitting a purchase order with reference to this quotation number or sending a scanned copy of the signed quotation to the address stated below, is 

considered an acceptance of prices and all other special conditions listed on the quotation. 

Client Services 

Email: 

Fax: 

By authorization and acceptance of this quotation as indicated above, authorizes Eurofins to perform the work as described in this quotation, subject to the 

terms and conditions of the MSA by and between the Client and Eurofins or its affiliate, if any, or if none, or in the event that the MSA is not active or found to 

be invalid as of the date hereof, then the work described in this quotation shall be subject to the then current version of the Eurofins General Terms & 

Conditions of Sale of Products and Services, as amended from time to time, which can be viewed at the following link: 

https://www.eurofinsdiscoveryservices.com/cms/cms-content/misc/tc/order-terms-condition-products-and-services-dec19/. This quotation constitutes a separate 

and distinct contract between Client and Eurofins. In no event shall an affiliate of Eurofins be liable for its obligations under this quotation, and Client shall look 

exclusively to Eurofins in relation to any rights or remedies it may have with respect to this quotation. 

Eurofins Panlabs Discovery Services Taiwan, Ltd. 

Wugu District, New Taipei City, 24891, Taiwan 

Phone Fax 

Page 1 of Budget Justification Attachment 2 
 

Contact PD/PI: Harty, Ronald N. 

Quotation 

AGREED TO AND ACCCEPTED 

For: Fox Chase Chemical Diversity Center Inc 

By: 

Name: 

Title: 

Date: 

Eurofins Panlabs Discovery Services Taiwan, Ltd. 

Wugu District, New Taipei City, 24891, Taiwan 

Phone + Fax + 

Budget Justification Attachment www.eurofinsPdiascgoeve9ry3services.com Page 2 of 2 
 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Quotation 

Customer Delivery Information Customer Invoice Information 

Fox Chase Chemical Diversity Center Inc Fox Chase Chemical Diversity Center Inc 

3805 Old Easton Road Doylestown, PA 18902, United States 

PA 18902, United States Name: Accounts Payable 

Name: Jay Wrobel Phone: 

Phone: Email: 

Email: 

Quotation Number Study Code Customer Account No Eurofins Contact 

Andrew Lord 

Date Issued Customer reference Payment Terms Eurofins Contact Phone 

04-Aug-2020 30 Days 

Quote Valid Until Promo Code Loyalty Category Eurofins Contact Email 

03-Sep-2020 Standard 

Summary 

Price Discount 

0.00 

Charges 

0.00 

Net Price 

Details 

Sales tax 

0.00 

Grand Total Price 

Item # Description Assay 

Mode 

Cmpd Conc Rep Qty Option TAT* Unit price Disc unit 

price 

Currency 

USD 

Net Price 

PP223 SafetyScreen87 Panel, Panlabs Flexible 1 1 2 1.00 Standard 10 

Special Requirement: Default test concentrations 10 uM 

Cmpd = compounds; Conc = Concentrations; Rep = Replicates; Qty = Quantity; TAT = Turnaround Time. 

*please see “Delivery Dates, Turnaround Time” section of Terms and Conditions (web link) for more details. 

Additional Charges 0.00

Special Instructions: N/A 

Orders can be placed by purchase order or credit card. 

Purchase orders should detail the following company address: 

Eurofins Panlabs Discovery Services Taiwan Ltd 

New Taipei City, Taiwan 24891 

Please note that the address for sending check payment is as following: 

Birmingham, AL 35246-5588 

Please note that if arranging payment from a bank outside of Taiwan, any intermediary or sender’s charges should be borne by the sender. 

AUTHORIZATION AND ACCEPTANCE 

Submitting a purchase order with reference to this quotation number or sending a scanned copy of the signed quotation to the address stated below, is 

considered an acceptance of prices and all other special conditions listed on the quotation. 

Client Services 

Email: 

Fax: 

By authorization and acceptance of this quotation as indicated above, authorizes Eurofins to perform the work as described in this quotation, subject to the 

terms and conditions of the MSA by and between the Client and Eurofins or its affiliate, if any, or if none, or in the event that the MSA is not active or found to 

be invalid as of the date hereof, then the work described in this quotation shall be subject to the then current version of the Eurofins General Terms & 

Conditions of Sale of Products and Services, as amended from time to time, which can be viewed at the following link: 

https://www.eurofinsdiscoveryservices.com/cms/cms-content/misc/tc/order-terms-condition-products-and-services-dec19/. This quotation constitutes a separate 

and distinct contract between Client and Eurofins. In no event shall an affiliate of Eurofins be liable for its obligations under this quotation, and Client shall look 

exclusively to Eurofins in relation to any rights or remedies it may have with respect to this quotation. 

Eurofins Panlabs Discovery Services Taiwan, Ltd. 

New Taipei City, 24891, Taiwan 

Phone Fax 

www.eurofinsPdiascgoeve9ry4services.com Page 1 of 2 Budget Justification Attachment 
Contact PD/PI: Harty, Ronald N. 

Quotation 

AGREED TO AND ACCCEPTED 

For: Fox Chase Chemical Diversity Center Inc 

By: 

Name: 

Title: 

Date: 

Eurofins Panlabs Discovery Services Taiwan, Ltd. 

New Taipei City, 24891, Taiwan 

Phone Fax 

Budget Justification Attachment www.eurofinsPdiascgoeve9ry5services.com Page 2 of 2 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

SUBAWARD BUDGET JUSTIFICATION 

Senior/Key Personnel 

Ronald Harty, PhD, PI (25% effort – 3.0 ca mo). Dr. Harty is a Professor in the Department of Pathobiology 

at the University of Pennsylvania School of Veterinary Medicine and a member of the Cell and Molecular 

Biology Graduate Group. His laboratory is focused on the molecular events that lead to virus assembly and 

budding and has developed models of virus budding that recapitulate biological events occurring during 

infection with a number of Category A pathogens. Dr. Harty’s laboratory utilizes a wide-array of techniques for 

these studies such as, virus-like particle (VLP) budding assays, reverse-genetics to generate VSV 

recombinants, bimolecular complementation, ubiquitination and ISGylation assays, and fluorescence lifetime 

imaging microscopy (FLIM). Dr. Harty will direct studies to test lead compounds for their ability to block VLP 

egress from mammalian cells, block the specific virus-host interaction targeted by the lead compound using 

BiMC, and block budding of live virus using a surrogate model of filovirus and arenavirus infection. Dr. Harty 

will be responsible for the overall conduct of this study in the role of PI, dedicating 25% effort to the project and 

being responsible for communication/joint lab meetings with the Fox Chase Chemical Diversity Center and 

Texas Biomedical Research Institute to track the progress of the project. 

Other Personnel 

Ziying Han, MD, Ph.D. (100% effort – 12.0 ca mo). Budget is requested to support Research Specialist Dr. 

Han, who has provided all of the BSL2 preliminary virology and VLP budding data. Dr. Han will be responsible 

for maintaining the tissue culture models needed for the experiments and for all molecular biological 

techniques necessary for testing lead compounds for their ability to block VLP and live virus egress and 

specific virus-host interactions in live mammalian cells using the BiMC assay. 

Other direct costs: (Year 1), /year (Year 2-3) 

Materials and supplies: is requested in Year 1 to cover the cost of consumables and reagents 

necessary for the tissue culture and molecular virology studies associated with this project. These funds will 

also cover the costs of equipment or facility rental/user fees associated with using the University of 

Pennsylvania Microscopy Core facility. The imaging core operates on a fee for use cost recovery system that is 

heavily subsidized by departments and centers in the School of Veterinary Medicine. All users, regardless of 

their affiliation are charged the same fee for using the Leica SP5 confocal systems, which is /hr. This 

fee helps to defray maintenance and operation costs of the core. 

Fringe Benefits: 

Fringe benefits have been requested for the salaries in this budget at the DHHS negotiated rate, which is 

currently 29.5% 

Indirect Costs: (Year 1), /year (Year 2-3)

A total modified direct cost rate has been requested in this budget and is the DHHS negotiated rate. It is 

currently 62% through June 30, 2021 and then it changes to 62.5% starting July 1, 2020. 

Budget Justification Attachment Page 106 
 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

BUDGET JUSTIFICATION 

Texas Biomedical Research Institute 

PERSONNEL 

Olena Shtanko, Ph.D., Subaward Principal Investigator (2 calendar months/year): Dr. Olena Shtanko has 

extensive experience in researching and handling BSL4 agents, including Ebola, Marburg, and Lassa viruses, 

both in vitro and a mouse model of filovirus disease, as evidenced by her publications in the field. Dr. Shtanko 

is fully authorized to work with select agents and has the appropriate Homeland Security and CDC approvals for 

this work. Dr. Shtanko will be responsible for the overall experimental design and execution of the project at 

Texas Biomed. She will supervise the conduct of all studies at Texas Biomed, participate in review and 

interpretation of the data, and prepare all reports and communications with Dr. Harty. In addition, she will perform 

certain infection and all imaging assays. 

Gloria Rodriguez, B.S., Research Assistant (4 calendar months/year): Gloria Rodriguez, B.S., has training 

and experience in the cell culture and molecular biology experiments to be executed in the BSL2 setting. She is 

also fully authorized and experienced to work with replication-competent wild-type viruses in the Texas Biomed 

BSL4 facility and has been doing so for >5 years. Gloria Rodriguez will help with the extensive cell culture, qPCR, 

and compound treatments in the BSL2 facility, and will maintain supplies and reagents. She will also perform 

virus expansion, plaque assays, infection assays, virus inactivation and any other virus-associated work. 

SUPPLIES 

A total amount of is budgeted to cover costs of cell culture media, media supplements, culture vessels, 

and reagents for cell maintenance, donor recruitment to obtain peripheral blood cells, generation and 

characterization of virus stocks, testing compounds for antiviral activity and cytotoxicity, and plaque assays; 

molecular biology reagents; antibodies and materials associated with immunofluorescence assays. 

OTHER DIRECT COSTS 

In vivo toxicity studies

We request a total of for studies aimed to access toxicity of compound treatments in a mouse model. 

These costs include purchase and shipment of 540 4-5-week old BALB/c mice of either sex from the Jackson 

Laboratories, housing fees at Texas Biomed ABSL2 Vivarium, therapeutic dosing, necropsy, gross pathology, 

histopathology, required supplies and reagents, and veterinary personnel effort. 

Budget Justification Attachment Page 117 
 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

In vivo compound accumulation studies 

We request a total of for studies to determine the concentration and accumulation of compounds over 

time in blood and organs of treated mice. These costs include purchase and shipment of 360 BALB/c mice from 

the Jackson Laboratories, housing fees at Texas Biomed Vivarium, therapeutic dosing, blood and organ 

harvesting, isolation of plasma from blood, required supplies and reagents, and veterinary personnel effort. 

A total amount of is requested to ship animal samples to an external source for analysis of compound 

concentration. 

ABSL4 efficacy studies using a mouse model of Marburg virus disease

Animal procurement (a total of ) is budgeted to purchase and ship 180 4-5-week old BALB/c mice of 

either sex from the Jackson Laboratories. 

Efficacy studies (a total of ) cover animal housing fees, virus challenge, therapeutic dosing, necropsy, 

required supplies and reagents, and veterinary personnel effort. 

The indirect cost rate is the Texas Biomed Federal MTDC Rate of 98%. Fringe is calculated at 29.51% according 

to our federally negotiated rate. 

Budget Justification Attachment Page 118 
Contact PD/PI: Harty, Ronald N. 

Total Direct Costs less Consortium F&A 

NIH policy (NOT-OD-05-004) allows applicants to exclude consortium/contractual F&A costs when determining if an application falls at or beneath any applicable direct cost limit. When a direct 

cost limit is specified in an FOA, the following table can be used to determine if your application falls within that limit. 

Categories Budget Period 1 Budget Period 2 Budget Period 3 Budget Period 4 Budget Period 5 TOTALS 

Total Direct Costs less Consortium F&A 0 0 

 

 

 

Page 120 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265. Received Date: 2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

OMB Number: 4040-0010 

Expiration date: 12/31/2022 SBIR/STTR Information 

Agency to which you are applying (select only one)* 

❍ DOE ● HHS ❍ USDA ❍ Other: 

SBC Control ID:* 

Program Type (select only one)* 

❍ SBIR ● STTR 

❍ Both (See agency-specific instructions to determine whether a particular agency allows a single submission for both SBIR and STTR) 

Application Type (select only one)* 

❍ Phase I ● Phase II ❍ Fast-Track ❍ Direct Phase II ❍ Phase IIA ❍ Phase IIB ❍ Phase IIC 

❍ Commercialization Readiness Program (See agency-specific instructions to determine application type participation.) 

Phase I Letter of Intent Number: 

* Agency Topic/Subtopic: 

Questions 1-7 must be completed by all SBIR and STTR Applicants: 

1a. Do you certify that at the time of award your organization will meet the eligibility criteria for a small business as ● Yes ❍ No 

defined in the funding opportunity announcement?* 

1b. Anticipated Number of personnel to be employed at your organization at the time of award.* 28 

1c. Is your small business majority owned by venture capital operating companies, hedge funds, or private equity ❍ Yes ● No 

firms?* 

1d. Is your small business a Faculty or Student-Owned entity?* ❍ Yes ● No 

2. Does this application include subcontracts with Federal laboratories or any other Federal Government agencies?* ❍ Yes ● No 

If yes, insert the names of the Federal laboratories/agencies:* 

3. Are you located in a HUBZone? To find out if your business is in a HUBZone, use the mapping utility provided by ❍ Yes ● No 

the Small Business Administration at its web site: http://www.sba.gov * 

4. Will all research and development on the project be performed in its entirety in the United States?* ● Yes ❍ No 

If no, provide an explanation in an attached file. Explanation:* 

5. Has the applicant and/or Program Director/Principal Investigator submitted proposals for essentially equivalent ❍ Yes ● No 

work under other Federal program solicitations or received other Federal awards for essentially equivalent work?* 

If yes, insert the names of the other Federal agencies:* 

6. Disclosure Permission Statement: If this application does not result in an award, is the Government permitted to ❍ Yes ● No 

disclose the title of your proposed project, and the name, address, telephone number and email address of the official 

signing for the applicant organization to state-level economic development organizations that may be interested in 

contacting you for further information (e.g., possible collaborations, investment)?* 

7.Commercialization Plan: The following applications require a Commercialization Plan: Phase I (DOE only), Phase II (all agencies), 

Phase I/II Fast-Track (all agencies). Include a Commercialization Plan in accordance with the agency announcement and/or agencyspecific instructions.* 

Attach File:* Commercialization_Plan_Harty_Final.pdf 

Page 121 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265. Received Date: 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

OMB Number: 4040-0001 

Expiration date: 10/31/2019 SBIR/STTR Information 

SBIR-Specific Questions: 

Questions 8 and 9 apply only to SBIR applications. If you are submitting ONLY an STTR application, leave questions 8 and 9 blank and 

proceed to question 10. 

8. Have you received SBIR Phase II awards from the Federal Government? If yes, provide a company ❍ Yes ❍ No 

commercialization history in accordance with agency-specific instructions using this attachment.* 

Attach File:* 

9. Will the Project Director/Principal Investigator have his/her primary employment with the small business at the time ❍ Yes ❍ No 

of award?* 

STTR-Specific Questions: 

Questions 10 - 12 apply only to STTR applications. If you are submitting ONLY an SBIR application, leave questions 10 - 12 blank. 

10. Please indicate whether the answer to BOTH of the following questions is TRUE:* ● Yes ❍ No 

(1) Does the Project Director/Principal Investigator have a formal appointment or commitment either with the small business directly 

(as an employee or a contractor) OR as an employee of the Research Institution, which in turn has made a commitment to the small 

business through the STTR application process; AND 

(2) Will the Project Director/Principal Investigator devote at least 10% effort to the proposed project? 

11. In the joint research and development proposed in this project, does the small business perform at least 40% of ● Yes ❍ No 

the work and the research institution named in the application perform at least 30% of the work?* 

12. Provide DUNS Number of non-profit research partner for STTR.* 

Page 122 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265. Received Date: 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Development of Small Molecule Therapeutics against RNA Viruses 

Fox Chase Chemical Diversity Center, Inc. 

Commercialization Plan 

We have developed small molecule leads that impede the progression and transmission of RNA viruses including 

Marburg and Ebola viruses, both potential bioterrorism agents and an area of focus for National Institute of 

Allergy and Infectious Diseases (NIAID) small business funding. In addition, other viruses that may be 

therapeutic targets of our agents include Lassa and Human T Cell Leukemia virus. A therapeutic agent arising 

from our efforts will be taken orally alone or in combination with other agents by persons infected with these RNA 

viruses, or taken as prophylactic agents for individuals deemed to be in high risk situations such as military or 

healthcare workers. Our current lead candidate FC-10696 was shown to be a potent inhibitor of virus budding 

(egress) and transmission in vitro and is active in vivo (mouse). Its mechanism is novel (inhibition of the viral 

matrix protein interaction with mammalian target cell proteins). In this Phase II STTR application we plan to fully 

characterize preclinical candidates suitable for IND enabling toxicology studies. We present here our 

Commercialization Plan from our joint program of study with researchers at University of Pennsylvania and the 

Texas Biomedical Research Institute, covering the topics listed in the guidelines for STTR submissions. 

A significant portion of this report was taken from a proprietary Technology Niche Analysis supported 

by the NIH and prepared for FCCDC by Bruce Inwood of Foresight Science and Technology. 

1. Value of the STTR Project, Expected Outcomes and Impact. 

We review here our key technology objectives for this STTR, commercial need for our technology, our 

competitive advantage relative to standard of care, and the impact of this technology upon drug discovery and 

society as a whole. 

1.1 Key technology objectives, therapeutic benefit, commercial need and competitive advantage relative to 

current standard of care of the STTR project. 

Marburg and Ebola viruses cause hemorrhagic diseases that afflict humans and nonhuman primates. Our 

proposed drug candidate will be a novel small molecule oral agent used alone or in conjunction with other antiviral 

agents for individuals infected with filoviruses Marburg and Ebola or other RNA viruses, including arenaviruses 

(Lassa), which can cause severe hemorrhagic fever syndromes in humans with high mortality rates, or 

rhabdoviruses (Rabies). “Viral hemorrhagic fevers (Ebola, Lassa fever, etc.)” are a focus for NIAID SBIR/STTR 

research and funding (Omnibus Solicitation for SBIRs and STTRs, PHS 2020-, p. 34). These viruses depend 

on an interaction between their matrix protein PPxY motif and select host proteins for efficient virus egress and 

spread of infection. Thus, drugs that block or disrupt viral-host interactions required for virus budding are 

expected to effectively block virus transmission and disease progression. 

The 2014-2015 outbreak of Ebola in western Africa resulted in over 28,000 infected individuals and over 11,000 

deaths (WHO: Ebola Situation Report 2015). More recently the Democratic Republic of the Congo has second 

largest Ebola epidemic on record, which is ongoing, and as of June, 2020, with more than 2,200 lives lost, 3,400 

confirmed infections and 1,160 survivors since the outbreak was declared on 1 August 2018. These 

unprecedented epidemics have spurred a call to action to discover new, cost effect therapies that combat this 

deadly pathogen. Among these efforts are several vaccines and antiviral drug candidates. However, the current 

vaccines are not a complete defense since they must be given pre-exposure. Oral antiviral agents used alone 

or in combination may be of value for individuals who respond adversely to the vaccine, and could be of value 

as prophylactic agents for individuals deemed to be in high risk 

situations such as military or healthcare workers. Therefore, effective 

therapeutics are needed to safeguard the largely immunologically naive 

human population by providing immediate protection. 

We have discovered two novel series of small molecule early leads that 

have nanomolar potency at inhibiting RNA virus budding and are 

Commercialization Plan Page 123 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

effective at reducing spread of live viruses including Marburg and Ebola virus. We are currently pursuing both 

series in order to increase our chance of success. Our most advanced lead is compound, FC-10696 and close 

structural analogs have the following attributes, tested to the extent afforded by the Phase I level of funding: 

• Potent activity (IC50 < 100 nM) in the Marburg and Ebola VLP cellular budding (egress) assays, 

• Evidence that analogs of FC-10696 directly target the host cell and virus molecular machinery that 

effect viral cellular egress, 

• Live virus activity against RNA viruses (Marburg, Ebola), 

• Metabolic stability in mouse and human liver microsomes, 

• No cytochrome P450 3A4 activity which translates to a lower risk of drug-drug interactions, 

• Good drug levels upon IP administration in mice, and 

• Proof of concept in vivo reductions of Marburg virus (IP administration) in mouse models. 

However, it is important to note that improved analogs and additional novel composition of matter are expected 

to be discovered during the course of the Phase II STTR period of study, with fully characterized ADME 

properties suitable as antiviral oral drug candidates for preclinical development. 

1.2 Expected outcomes: path to commercialization including timelines. 

The goal of this STTR Phase II application is to produce one or more preclinical drug candidates by medicinal 

chemistry synthesis optimization strategies that would be ready for GMP scale up and IND enabling studies. At 

the end of this award period we would have one or more drug candidates that are: 

• Potent, orally available inhibitors of RNA virus egress, due to inhibition of the viral matrix protein PPxY 

interaction with cellular Nedd4, 

• Broad spectrum activity against RNA viruses (Ebola, Marburg, Ebola, Lassa, RSV, etc). 

• Are not cytotoxic at concentrations 20-fold or higher than the live virus assay IC50, 

• Have suitable ADME and PK properties in mice for once a day oral administration, 

• Have oral activity in a mouse antiviral assay, 

• Have little or no cytochrome P450 inhibitory (Cyp) activity against the seven Cyp isoforms most likely to 

cause exposure variability and drug-drug interactions (DDIs), 

• Have little or no activity in a manual patch clamp assay to assess hERG (human Ether-à-go-go-Related 

Gene product), suggesting a greatly lower risk of the compound causing cardiac arrhythmias, 

• Have little or no activity in an Ames testing with and without metabolic activation, suggesting a greatly lower 

risk of the compound being mutagenic, 

• Have chemical syntheses that are efficient and scalable, and 

• Provide the basis for one or more composition of matter U.S. patent applications. 

We anticipate being in an ideal position to pursue IND enabling studies with our candidate compound and will 

require additional funding to support IND filing and early clinical development. Many of the future IND-enabling 

activities would be funded from sources listed later in this application. Such post-Phase II SBIR activities include: 

• Preparation of 300 g of non-GMP active pharmaceutical agent, 

• Selection of a suitable salt form (if necessary), vehicle and formulation, and evaluation of crystallinity and 

the propensity for polymorph formation, 

• Analytical methods development, 

• Drug substance stability testing, including standard pharmaceutics development work-up involving shelflife and stress testing, 

• File additional provisional and PCT patent applications as necessary, and 

• Prepare 5 kg of GMP quality active pharmaceutical ingredient. 

Commercialization Plan Page 124 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

With the completion of GMP and non-GMP batches we plan to initiate the necessary studies to support an IND 

filing, specifically investigation of metabolites, in vitro toxicity and acute in vivo safety pharmacology and toxicity 

assessment. Activities include: 

• Metabolite ID (human, rat and monkey), characterization for toxicity and biological testing, 

• Full PK parameters in three relevant safety pharmacology species, mice, rabbit, and non-human primate 

(NHP), chosen since they are used in relevant in vivo pharmacology studies, 

• Investigation of Cyp450 inhibition and induction and transporter modulation in detail, for the potential for 

unexpected drug-drug interactions, 

• Enhanced survival in two or more GLP double blind placebo-controlled animal models of Ebola according 

to the Animal Rule of Efficacy (some of these studies might be done during the IND phase). Viral load and 

survival will also be assessed, 

• Completion of in vitro GLP toxicology (hERG, Ames, genotoxicity), 

• Conducting dose ranging and 28-day safety profiles in mice and NHPs, 

• Safety Pharmacology: Cardiovascular safety study in dogs, CNS safety in rats, respiratory safety in rats (if 

needed), 

• Reproductive toxicology in mice and rabbits (possibly do after/during Phase I studies). 

• Conducting a pre-IND meeting with the FDA to establish and understand expectations, and 

• Initiating preparation of an IND to facilitate the transition into clinical evaluation. 

We have established a world-class PPxY Project Leadership Team (PPLT) to interpret data and provide 

strategic direction on a real time basis. The PPLT will regularly provide advice and guidance (at least monthly) 

in person, by Zoom, or by email. The PPLT will interpret data most appropriately and coordinate experimental 

design in close coordination with the Harty group and FCCDC management. The PPLT will conduct more formal 

quarterly meetings that Prof. Harty will chair, and any member of the PPLT may add items to the agenda 

prepared in advance of the meeting. The PPLT shall consist of the following members: 

Ronald N. Harty, Ph.D., Professor, University of Pennsylvania. Dr. Harty is Professor of Pathobiology and 

Microbiology, at the School of Veterinary Medicine. His research interests are focused on understanding the 

molecular mechanisms of filovirus (Ebola and Marburg viruses) and arenavirus (Lassa fever and Junin) 

assembly/budding, in order to identify broad-spectrum inhibitors of virus-host interactions. His research has 

directly led to the discovery of the antiviral agents described in this application. He is also interested in 

understanding the innate immune responses to virus infection that provide a critical first line of defense for the 

host against the invading pathogen. Prof. Harty has co-authored >80 scientific publications. 

Jay E. Wrobel, Ph.D., Vice President, Business Development, FCCDC. Dr. Wrobel has spent 37 years as a 

medicinal chemist in the pharmaceutical industry, 26 years at Wyeth (later Pfizer), with his last position as Senior 

Director of Medicinal Chemistry at the Collegeville, PA facility. At Wyeth, he mentored and guided the efforts of 

31 Ph.D./MS medicinal chemists and worked successfully with outside alliance partners. He was directly involved 

in bringing forward nine development track candidates (Phase 0 and beyond) in a variety of therapeutic areas. 

He has co-authored 78 scientific publications and is an inventor on 83 issued US patents. 

Bruce Freedman, VMD, Ph.D., Associate Professor, University of Pennsylvania. Dr. Freedman is an 

Associate Professor of Pathobiology and Microbiology, at the School of Veterinary Medicine. Dr. Freedman is 

Director of the Microscopy Core facility at PennVet which houses state-of-the-art confocal and live cell imaging 

instruments that will be critical for assessing the efficacy of our budding inhibitors. Dr. Freedman is a longstanding collaborator with Dr. Ronald Harty, the PI on this proposal. 

Olena Shtanko, Ph.D., Staff Scientist, Texas Biomedical Research Institute. Dr. Shtanko researches 

fundamental molecular processes used by Ebola and Marburg viruses to disseminate within the host to 

understand the extraordinary capacity of these emerging viruses to trigger disease in affected individuals, with 

the aim to design and test antiviral therapies. Dr. Shtanko has over 16 years of conducting research on viruses, 

Commercialization Plan Page 125 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

with 9 years spent at the maximum containment laboratory (BSL-4) at Texas Biomedical Research Institute. Dr. 

Shtanko's work has identified factors critical for Ebola virus and Crimean-Congo Hemorrhagic Fever virus entry 

into cells. Dr. Shtanko also extensively collaborates with a number of investigators to coordinate and perform 

virus studies to identify host factors critical for replication and spread of high-containment viruses as well as test 

novel antiviral therapeutics using a small animal model of Ebola and Marburg virus disease. 

Allen B. Reitz, Ph.D. CEO, FCCDC. Dr. Reitz has spent 38 years as a medicinal chemist in the pharmaceutical 

industry, 26 years with Johnson and Johnson (Spring House, PA). At J&J he mentored and guided a staff of 16 

Ph.D./MS medicinal chemists and was co-inventor on 7 compounds that have entered human clinical trials. He 

has co-authored >150 scientific publications and is an inventor on 71 issued US patents. He is Editor-in-Chief 

and founding editor of Current Topics in Medicinal Chemistry. He has established two biotechnology companies: 

FCCDC and ALS Biopharma. FCCDC has already successfully partnered a separate technology platform for 

the treatment of Alzheimer’s disease, which has passed an interim futility analysis in Phase II/III. 

Leo J. Adalbert, M.B.A. Mr. Adalbert will be a business development consultant for this project. An insightful, 

collaborative, and creative leader, Leo carries more than 25 years of deep cross-functional commercial and 

clinical development experience across both small molecules and biologics. Core therapeutic areas of expertise 

include cardiovascular, neuroscience, oncology, immunology, and GI, as well as biomarkers. Prior to founding 

StraNexa LLC, Leo held various leadership roles across marketing, business development, and alliance 

leadership at leading firms such as AstraZeneca, Merck, Astra Merck, and Regeneron. Leo earned an MBA from 

Harvard Business School (fellowship), and executive leadership certifications from Northwestern University 

(Kellogg) and University of Pennsylvania (Wharton). He also completed extensive coursework in biological 

sciences (Villanova University/Bucks CC), earning high honors (Phi Theta Kappa distinction), and is a graduate 

of the HMX Program at Harvard Medical School (pharmacology and immunology). 

FCCDC has established a world-class Scientific Advisory Board, as posted on the company website, who are 

on call for questions regarding drug development strategy, flowchart design, chemistry and pharmacology. 

Nicholas A. Meanwell, Ph.D., Bristol-Myers Squibb Research & Development, Princeton, NJ. Nicholas A. 

Meanwell is currently Vice President, Discovery Chemistry Platforms, Small Molecule Drug Discovery where he 

has led drug discovery programs in the cardiovascular, neurosciences and virology therapeutic areas, work that 

has resulted in the advancement of over 30 clinical candidates for the prevention of thrombosis, the treatment of 

stroke and therapy for viral infections, including human immunodeficiency virus-1 (HIV-1), hepatitis C virus (HCV) 

and respiratory syncytial virus (RSV). Significant compounds in the HCV arena include daclatasvir (DaklinzaTM), 

asunaprevir (SunvepraTM), DaklinzaTM and SunvepraTM. Dr. Meanwell has authored/co-authored more than 250 

publications, review articles, book chapters and editorials and is named as an inventor/co-inventor on 138 issued 

U.S. Patents. Dr. Meanwell is the co-recipient of a PhRMA Research and Hope Award for Biopharmaceutical 

Industry Research, 2014 and the recipient of the 2015 Philip S. Portoghese Medicinal Chemistry Lectureship 

Award. He was inducted into the ACS Division of Medicinal Chemistry Hall of Fame on August 18th, 2015 and 

he is a member of the team that was recognized by a 2017 “Heroes of Chemistry” Award sponsored by 

the American Chemical Society. 

Eileen Jaffe, Ph.D., Fox Chase Cancer Center, Temple University School of Medicine & Drexel University 

School of Medicine. Eileen Jaffe received her PhD in Biochemistry from the University of Pennsylvania, working 

with Mildred Cohn. She was then a Postdoctoral Fellow at Harvard University with Jeremy Knowles. Since 2003 

she has been at the Fox Chase Cancer Center in Philadelphia, PA, and is currently also an Adjunct Professor of 

Biochemistry at Temple University School of Medicine and Drexel University College of Medicine. Her research 

interests lie in understanding protein structure-function relationships using both biochemical and biophysical 

approaches. Prof. Jaffe is currently focused on the roles of protein quaternary structure and quaternary structure 

rearrangements in the control of protein function as this relates to phenylalanine hydroxylase and 

phenylketonuria. She has been associated with FCCDC since 2008. 

Patrick Lam, Ph.D., Baruch S. Blumberg Institute, Drexel University College of Medicine & Lam Drug Discovery 

Consulting. Patrick Lam is currently a Distinguished Professor of Chemistry at Baruch S. Blumberg Institute and 

Adjunct Professor at Drexel University College of Medicine. He is responsible for the discovery of a total of eight 

Commercialization Plan Page 126 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

clinical candidates. At Bristol Myers Squibb, Dr. Lam was the group leader/co-inventor responsible for the 

discovery of Eliquis®/Apixaban, a novel Factor Xa anticoagulant with annual sales of >$2 B. Eliquis® was chosen 

as the “Best New Medicine of 2012” by Med Ad News. He and his team were awarded 2015 American Chemical 

Society Heroes of Chemistry Award for the discovery of Eliquis®. Patrick is also internationally known as the codiscoverer of the powerful Chan-Lam Coupling Reaction, which is complementary to the Nobel-prize winning 

Suzuki-Miyaura Coupling Reaction. Patrick has authored 98 papers/reviews/book chapters and is an inventor 

on 36 patents/patent applications. He has presented 110 invited seminars worldwide. 

Assuming that our candidate has progressed to IND status, our next objective will be clinical evaluation. 

Collectively, the management of FCCDC has been responsible for more than a dozen compounds that have 

entered human clinical trials, with one currently in Phase III and two in Phase II. Based on this experience, we 

propose the following path to commercialization with the associated timelines. 

• A pre-IND meeting will be held with the FDA prior to initiation of IND enabling studies, to obtain guidance as 

to proper preclinical and clinical study design and requirements. We expect to file an Investigational New 

Drug (IND) in ~2Q/2024, with First in Human (FIH) dosing to begin ~1Q/2025. If we have partnered with an 

appropriate mid-level or major pharmaceutical company, then our development partner will monitor and lead 

the drug development process. 

• Since our primary indication is as a Marburg antiviral medication, effectiveness of our candidate may not 

easily be determined in human clinical trials for ethical reasons, particularly if there is not an outbreak of the 

virus. Thus, we will resort to the “Animal Rule" (21CFR Part 314 Subpart 1) as the regulatory pathway for 

approval of this drug. 

• Initial Phase I clinical evaluation will take place in normal individuals looking for safety and tolerability in a 

single ascending and multiple ascending dose format and drug-drug interactions. Clinical read-outs will 

include standard hemodynamic parameters involved in cardiovascular safety testing. Adverse events would 

be monitored. The study would most likely take 12 months, enroll 40 participants and cost $3 million. 

• Phase II trial beginning ~1Q/2027 will be an intervention trial for healthy patients with human T cell leukemia 

virus, (a virus with a similar mode of egress to Marburg). A dose of candidate compound would be 

administered orally with a placebo. The endpoints would be change in viral load. The study would most likely 

take 10 months, enroll 140 participants and cost $10 million. 

• A New Drug Application (NDA) will be filed with the U.S. FDA containing all of the CMC and other ICH 

regulatory and safety pharmacology documentation required for commercialization. This would be schedule 

to occur at ~2Q/2029. 

• Initial FDA approval would be for use as an antiviral therapeutic agent for Marburg infection. This would occur 

on or about 1Q/2030. The Target Product Profile for our candidate is shown in Table 1. 

Table 1. Desired Product Profile 

Preferred Minimally acceptable 

Efficacy (% survive) >95% >85% 

Efficacy (virus) Undetectable levels of virus Undetectable levels of virus 

Adverse events Comparable to placebo Minor 

Administration Oral* Oral 

Dosing schedule Once daily Twice a day 

Drug resistance None None 

Stability No special packaging required Modest precautions 

acceptable 

* An IV formulation may be preferable for the extremely ill, and a liquid formulation useful for children 

Commercialization Plan Page 127 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

1.3. Impact on drug discovery and society. 

In the outbreak of a terrorist exposure to Marburg, Ebola or related virus, thousands if not greater numbers of 

people could die before vaccines become available to treat the affected population. Our technology, an oral 

antiviral medicine for Ebola, Marburg, Lassa and other deadly viruses, could be used alone or with other antiviral 

medications to protect unvaccinated individuals for the time required to mount an immune response to the 

vaccine. These agents may also be of value for individuals who respond adversely to the vaccine, and could be 

of value as prophylactic agents for individuals deemed to be in high risk situations such as military or healthcare 

workers. In addition, our agent may be effective against the related human T cell leukemia virus that can lead to 

a rare but highly aggressive cancer. This agent also has the potential to be used as an HIV medication used in 

conjunction with other HIV antiviral agents. Since current HIV medications can have reduced efficacy over time 

due to resistance mechanisms, other agents used in combination can reduce resistance and side effects, new 

drugs are of great need. Finally, our agents that block Nedd4 activity, such as FC-10696, have the potential to 

show clinical effectiveness against other areas of medical need including various cancers, and obesity (Huang, 

2019, Li, 2015; Wan, 2019, Ye, 2014; Zou, 2015). 

Continued successful prosecution of this project will allow Fox Chase Chemical Diversity Center to benefit 

financially and be able to expand and hire additional staff, thereby creating jobs. In addition, the development, 

clinical validation and commercial exploitation of this platform technology will serve as a great testament to the 

value of the SBIR/STTR program. In summary, patients will be treated with an effective and safe therapy that 

had not been available before, Fox Chase Chemical Diversity Center and partners will realize potentially 

substantial financial return, and the STTR program will have served its mission to society. 

2. Company 

Fox Chase Chemical Diversity Center, Inc., (FCCDC, www.fc-cdci.com) founded in 2008, is an emerging 

biotechnology company located in Doylestown, PA whose mission is to advance basic scientific discoveries by 

providing value-added early drug discovery and medicinal chemistry research support for the translation of viable 

preclinical drug and diagnostic candidates prior to eventual entry into human clinical trials. Our goal is to partner 

with suitable biomedical research collaborators on selected projects or act as their company’s proprietary 

medicinal chemistry research service. We bring a unique and highly experienced team of internal scientists and 

collaborative external partners in order to leverage cutting edge scientific advances. The staff at FCCDC has 

vast drug discovery experience, drawing from a diverse background of pharmaceutical and biotech companies 

including Johnson & Johnson, Wyeth and Merck. FCCDC has two fully functional, multisite medicinal chemistry 

laboratories with two 300-MHz NMRs, multiple LC/MS machines, microwave reactor, and multiple Gilson semiprep HPLCs for compound and library purification. We have prepared >12,000 new chemical entities (NCEs) so 

far, with two compounds advancing into human clinical trials, both funded initially by SBIR grant support. FCCDC 

has a >30,000 member onsite chemical inventory of reagents and starting materials for ready use, which are 

bar-coded, computer searchable, and segregated according to chemical class. 

As the basis for our internal technology platform, we have the ability to empower research programs at multiple 

stages of their development: provide insight into probe molecule identification and synthesis, in silico 

identification of custom and commercial high throughput screening libraries, perform hit triage, hit to lead 

medicinal chemistry, and lead optimization. Importantly, we also prepare or assist in preparation of intellectual 

property documents. We seek to improve efficiencies in early drug discovery by quickly derisking the overall 

process using modern methods and wisdom regarding drug suitability and compound library evaluation. 

3.Market, Customers and Competition 

3.1 Market. In response to credible scenarios of bioterrorist attacks using a filovirus or other infectious agents, 

the US Congress enacted Project BioShield legislation that was signed in 2004 which provided $5.6 billion over 

10 years to encourage the private sector to develop medical countermeasures against chemical, biological and 

other terrorism agents (Grotton, 2011). Following this legislation, the Biomedical Advanced Research and 

Development Authority (BARDA) was created two years later to oversee BioShield’s advanced development and 

procurement efforts. In 2013 Congress renewed laws to extend funding by passing the Pandemic All Hazards 

Commercialization Plan Page 128 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Preparedness Reauthorization Act that provided an additional $2.8 billion and extended BARDA until 2018 

(Genome Web Daily News, 2013). As an example, the US has granted Fast Track designation to the Arbutus 

Ebola drug and has funded $140 million for R&D on EVD (PR Newswire 2014). BioCryst was awarded $12 

million for development of BCX4430 (Barda 2015). In addition to the Barda program, the commercial market for 

future natural outbreaks is growing at a CAGR of 95.2% and the market is expected to reach $56 million by 2020 

(Allied Market Research 2015). 

Lassa fever is another viral disease treatable with our egress inhibitors. This malady is caused by the Lassa 

virus and results in 300,000 to 500,000 cases annually, mainly in western Africa and causes about 5,000 deaths 

each year (Ogbua 2007). Transmission of Lassa virus to humans occurs most commonly through ingestion or 

inhalation due to rodent infestation. 

A drug in this category could also be effective against human T cell leukemia virus (HTLV), which can lead to 

adult T-cell leukemia/lymphoma (ATLL). ATLL is a rare and fast-growing T-cell lymphoma that can be found in 

the blood (leukemia), lymph nodes (lymphoma), skin, or multiple areas of the body (Lymphoma Research 

Foundation, 2015). Around 20 million people worldwide are infected with HTLV. It is common in Japan, South 

and Central America, the Caribbean and Western Africa, and a large percentage (>30%) of African Americans 

are infected in certain areas of the US (e.g. New Jersey and New Orleans). It is transmitted through sexual 

contact or exposure to infected blood and blood products. Only a small percentage of carriers (<5%) actually 

come down with ATLL however, but there is no way to predict those that do. Survival depends on the ATLL 

subtype and varies from 4-6 months to more than five years (Gonçalves, 2010). Treatment options include 

standard anti-cancer protocols but at present no therapy targets the virus. Other diseases and disorders from 

HTLV include the incapacitating neurological disease (HTLV-associated myelopathy/tropical spastic paraparesis 

[HAM/TSP]), uveitis, rheumatic syndromes, and predisposition to helminth and bacterial infections (Gonçalves, 

2010). 

As mentioned earlier, the compounds in this application may block other biological functions of Nedd4. For 

example, Nedd4 is overexpressed in a variety of cancers including bladder, breast, prostate, gastric carcinomas, 

colorectal, breast and non-small cell lung carcinomas and inhibits several tumor suppressor systems (e.g. PTEN, 

Beclin). Suppression of Nedd4 has been shown to reduce tumor cell growth (Boase, 2015; Wan, 2019, Ye, 2014; 

Zou 2015). Thus, these compounds could find applications in the treatment of such cancers. 

Nedd4 also negatively regulates interferon (IFN)-induced transmembrane protein 3 (IFITM3) that restricts cellular 

infection by influenza virus. Therefore, blocking Nedd4 could lead to therapeutic treatment of influenza 

(Chesarino, 2015) 

3.2 Customers. As mentioned previously, BARDA is the most obvious customer for our RNA virus antiviral agent; 

however several potential governmental partners could provide development funds and/or resources toward 

development including the National Institute of Allergy and Infectious Diseases (NIAID), Office of Biodefense 

Research (OBRA), Department of Defense (DOD), The Defense Threat Reduction Agency (DTRA), The United 

Sates Army Medical Research Institute for Infectious Diseases (USAMRIID), Defense Science and Technology 

Laboratories (DSTL), and the Centers for Disease Control and Prevention (CDC). In addition to pursuing the 

government funding options outlined above, we may seek to partner with small biotechnology companies such 

as Siga, Tekmira, Chimerix, MappBio Pharm, Athene, Biocryst and Tetraphase, and large companies such as 

Merck, Johnson & Johnson, and GSK who currently are developing one or more therapeutics targeting a 

bioterrorism threat as most of these companies have a stated company goal of identifying and developing 

additional therapeutics in this area. 

3.3 Competition. Since there are currently no other agents that inhibit virus egress that are being developed as 

broad spectrum RNA antiviral agents, an oral drug product based on FC-10696 is potentially a first-in-class 

treatment either as a mono-therapeutic prophylactic agent, or in combination with other agents currently in 

development for treatment of RNA viral infections. With respect to Ebola efforts, presently there are several 

therapeutics agents being evaluated that are summarized here. 

Commercialization Plan Page 129 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

• Recombinant vesicular stomatitis virus–Zaire Ebola virus (rVSV-ZEBOV), sold under the brand name Ervebo 

(Merck) is a vaccine that prevents Zaire-strain Ebola. When used in ring vaccination, Ervebo has shown a 

high level of protection. Around half the people given the vaccine have mild to moderate adverse effects that 

include headache, fatigue, and muscle pain. Ervebo does not protect against other species of Ebolavirus or 

Marburg virus. Also, rural distribution has been a challenge and there is no data on longevity of the vaccine’s 

protection and how often an additional dose boost is required to extend immunity. 

• Johnson and Johnson is developing a vaccine regiment that consists of two doses leveraging different 

vaccines. One is the Ad26.ZEBOV monovalent vaccine based on adenovirus type 26 (Ad26) vector 

expressing the glycoprotein (GP) of the Ebola virus Mayinga variant (Ad26.ZEBOV). The other is multivalent 

Modified Vaccinia Virus Ankara (MVA) strain containing ZEBOV, SEBOV, Marburg Virus GP and Tai Forest 

nucleoprotein inserts (MVA-BN®-Filo). More than 6,500 individuals have now participated in clinical studies 

for the Ebola vaccine across the U.S., Europe, and Africa. So far, the data from these studies suggest that 

the vaccine stimulates a robust immune response and has a favorable safety profile. Janssen has also 

assembled a stockpile of 1.5 million vaccine regimens for potential use in public health emergencies. 

• ZMapp (MappBio USA) is a cocktail of three monoclonal antibodies with neutralizing activity against Ebola 

virus in animal models. Phase II trial is underway. A similar produce MIL-77 (MabWorks, China) is undergoing 

Phase I study. A trial conducted in 71 patients, 36 of whom received ZMapp in addition to the standard of 

care, did not show a significant effect on mortality. At 28 days, death had occurred in 84 of 169 (49.7%) of 

patients in the ZMapp group. 

• REGN-EB3 (Regeneron) is an experimental biopharmaceutical antiviral drug that is a cocktail of three 

monoclonal antibodies (REGN3470, 3471 and 3479) for treatment of Ebola virus disease. It is being 

developed, tested and manufactured as part of an agreement established in 2015 with BARDA, part of the 

Office of the Assistant Secretary for Preparedness and Response at the U.S. Department of Health and 

Human Services. REGN-EB3 is currently under clinical development and its safety and efficacy have not 

been fully evaluated by any regulatory authority. It was developed against the Zaire species of Ebola virus, 

but the Sudan and Bundibugyo strains have also caused outbreaks and it is unlikely that EB3 would be 

effective against these strains. It was tested in the PALM trial that had enrolled nearly 700 participants across 

four Ebola Treatment Centers in the DRC. At 28 days, death occurred in 52 of 155 (33.5%) patients in the 

REGN-EB3 group. 

• mAb114 is a monoclonal antibody (Ridgeback Biotherapeutics LP) that is being evaluated as a treatment for 

Ebola virus disease. A license for mAb114 was obtained by in 2018 from the National Institutes of HealthNational Institute of Allergy and Infectious Diseases. In the PALM trial that had enrolled nearly 700 

participants across four Ebola Treatment Centers in the DRC. At 28 days, death occurred in 61 of 174 

(35.1%) patients in the MAb114 group. 

• Galidesivir (BCX4430, BioCryst) is a broad-spectrum small molecule antiviral in advanced development 

under the Animal Rule for the treatment of viruses including Ebola, Marburg, Yellow Fever and Zika viruses. 

A Phase 1 clinical safety and pharmacokinetics study in healthy subjects has been completed, and in animal 

studies, galidesivir has demonstrated survival benefits. Galidesivir is a viral RNA-dependent RNA 

polymerase inhibitor that has also demonstrated broad-spectrum activity in vitro against more than 20 

ribonucleic acid (RNA) viruses in nine different families, including filoviruses, togaviruses, bunyaviruses, 

arenaviruses, paramyxoviruses, coronaviruses and flaviviruses. Galidesivir may be suitable for 

administration by intravenous, intramuscular, and oral routes. 

There remains a critical need for additional agents, particularly with alternative modes of action. First many of 

the therapeutic agents above, particularly among the vaccine, antibody, siRNA and/or antisense 

oligonucleotides are specific for individual viruses and sometimes only one or more strains within a virus. 

Biological agents require iv or infusion administration, and also need to be kept in the cold during storage, which 

are not ideal conditions in many remote areas of virus prevalence. Our product would be broad spectrum for 

RNA viruses that have PPxY motifs. Our product differentiates further from vaccines in that it would be an oral 

agent with a great probability of more economic manufacturing and ease of distribution. Our agent could be 

Commercialization Plan Page 130 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

used in conjunction with other oral agents, like the cocktail approach to HIV drug administration, as synergistic 

therapeutic effects and a decrease in the risk of drug resistance are possible. 

4. Intellectual Property Protection 

FCCDC and the Trustees of the University of Pennsylvania obtained U.S. Patent 10,160,756, December 25, 

2018. This application contains composition of matter and use claims specific to earlier analogs of FC-10696. 

FC-10696 and its present suite of analogs are outside the scope of this patent and will be subject to new 

composition of matter applications. We will seek to create additional value in this project by the continued 

prosecution of the other U.S. provisional patent applications and those that would be expected to emerge in the 

future. We will file PCT applications and pursue patent protection in major foreign markets, including in Europe, 

Asia and the BRIC countries (Brazil, Russia, India, China). No special restrictions or licenses have been or are 

imposed on the rights of the key personnel, FCCDC or the University of Pennsylvania that fall outside of the 

project description in this grant application. 

Additional new compositions of matter will be produced during our execution of this proposal. Therefore, new 

patent applications will be filed during the course of this grant award time period. The preparation of the 

applications will commence shortly after wrap-up of medicinal chemistry new analog synthesis. Our Project 

Leadership Team includes those with extensive pharmaceutical industry experience, who are well versed in the 

mechanics and opportunities afforded by patent filing and prosecution, with >100 issued U.S. patents between 

them. 

5. Finance Plan 

5.1. Initial funding strategy and current status. Our initial funding has been provided by1R41AI113952-01A1 (Ron 

Harty, PI) 03/01/2015 – 02/29/2016 for a total of . We obtained another Phase I STTR R41 AI138630-

01A1 (Ron Harty, PI) 06/01/2018 – 05/31/2020 for a total of . 

5.2. Models of commercialization. A likely outcome for these studies is licensure of the Ebola/Marburg 

therapeutic to the US government and Department of Defense. While there are challenges to this type of 

business model (Bolken 2008), several companies have demonstrated success, including SIGA, PharmAthene 

and more recently Regeneron. Furthermore, there appears to be substantial funds from the US government, 

primarily the Biomedical Advanced Research and Development Authority (BARDA), that would help support late 

stage development costs for IND- and NDA-enabling studies which would be needed once funding under this 

grant has stopped. Of direct relevance to this direct application, BARDA awarded SIGA a $433M 5-year base 

contract for development and licensure of the smallpox antiviral agent, tecovirimat, which could rise to $2.3B 

with additional treatment courses. Other potential sources for support are the Defense Threat Reduction Agency 

(DTRA), the Centers for Disease Control (CDC), the NIH and the Office of Biodefense Research Resources and 

Translational Research (OBRRTR).The types of support available from these agencies include direct research 

and development funding and/or work-in-kind where development studies are conducted within or under the 

direction of the respective agencies. We will also seek, when appropriate, to establish partnerships with the 

private sector to help complete the required development studies in a timely and cost-effective manner. There 

are a number of companies that perform highly specialized tasks important for regulatory approval, such as those 

addressing preclinical toxicity, safety pharmacology and genotoxicity, while other companies have substantial 

CMC expertise for drug substance and drug product manufacturing. As described previously, there is substantial 

pharmaceutical development expertise at FCCDC so we have hand-on experiences with many of these 

companies and can make informed decisions on the most appropriate partner for each task. During the course 

of the development phase, there are three primary models of commercialization. 

• Commercialization model #1. This involves licensing for all indications to a mid-level or large pharmaceutical 

company, for development within the partner company. This is routinely conducted, and these types of 

agreements typically involve an upfront cash payment followed by a combination of royalties and milestone 

payments as the asset progresses further. We believe that we need additional validation data such as 

provided in this Phase II SBIR program of study to sufficiently interest in such a partner. This model is a 

probable outcome of this Phase II SBIR grant research effort. 

Commercialization Plan Page 131 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

• Commercialization model #2. In this model, FCCDC continues the development program through the Animal 

Rule efficacy and Phase 1 safety studies. This type of derisking adds tremendous value to the asset and 

often multiple groups will show interest at this time. The value of a transaction at this point intime can be a 

substantial upfront payment with a combination of royalties and milestones later. The funding to get to this 

decision point could come from a corporate partner who has first rights of refusal, angel or venture capital 

support, government (e.g. NIH or DOD) or foundation (e.g. Wellcome Trust) support. NIAID offers a Phase 

IIB grant opportunity to assist small businesses such as FCCDC to continue their project further if they are 

not sufficiently advanced to be able to attract an investment partner. 

• Commercialization model #3. This is a hybrid of plans 1 and 2, in which transition or licensing to a 

development partner occurs prior to the conclusion of the Phase 1 safety studies, perhaps during the course 

of the Animal Rule efficacy studies or at IND filing. Model #3 is a model of opportunity, in that as we 

aggressively pursue alternate funding to empower our research program to advance a candidate into clinical 

evaluation, an unforeseen opportunity for joint development, acquisition or licensing may prove too good to 

pass up. 

Several general concepts permeate throughout all of our activities, irrespective of which Commercialization 

Model we adopt. These are: 

• Most important: Ebola and Marburg are disease targets that are eligible for the U.S. Neglected Disease 

Voucher program. In this program, if we can obtain an approved NDA for Ebola from the U.S. FDA for the 

treatment of Ebola-virus infection, we will obtain a saleable voucher for priority review of a subsequent drug 

candidate in any therapeutic category, even if in reality the treatment for Ebola is never used therapeutically 

for any reason. Such vouchers have recently been receiving a high premium on the secondary market, often 

in excess of $100 million. NDA approval from the U.S. FDA for Marburg or Ebola would involve the "animal 

rule" in which efficacy would be inferred from in vivo efficacy studies, and of course no human clinical trials 

would be conducted in human patients with the disease. 

• We will file for Orphan Drug designation for Marburg or Ebola infection since infection affects less than 

200,000 people/year in the US. This designation results in seven-year market exclusivity for companies that 

develop the orphan drug, tax credits equal to half of the development costs, grants for drug development, 

and fast-track approvals of drugs indicated for rare diseases. SIGA was successful in getting this designation 

for tecovirimat (Love, 2011) 

• FCCDC will actively solicit support from academic investigators and clinicians who manage anti-infective 

clinical trials. 

• FCCDC will seek to obtain financial support, consultation and coordination from various non-profits and 

charitable organizations working diligently to find new medicines for infectious disease. 

• The FDA frequently waives application fees for small businesses (<500 employees), especially when this 

would be a significant barrier to innovation. We will apply for these application waivers and expect no difficulty 

in obtaining them given the compelling need for new treatments for RNA virus infections as a result of 

bioterrorism. 

• We will seek to expand the therapeutic indications that can be treated by our RNA virus antiviral drug lead in 

order to address the unmet medical need of a greater patient population and to attract further investment 

and/or interest from suitable development partners. 

• We will actively cultivate relationships with the external licensing and evaluation offices of major 

pharmaceutical companies with established product pipelines and portfolios in the area of bioterrorism 

infectious diseases, such as Siga, Tekmira, Chimerix, MappBio Pharm, Athene Biocryst, and Tetraphase, 

Merck, Johnson & Johnson. 

6. Production and Marketing Plan. 

Our current laboratory capabilities enable us to prepare 100 g batches of any candidate, to a high level of purity 

(>99%). As we require GMP-grade drug substance, we will engage an appropriate contract research organization 

who is properly certified for this type of chemistry project. Once a facility is selected, we will perform a site visit 

to ensure that our projects are performed under standard good laboratory practices. We will increase awareness 

of our candidate and related platform compounds in the scientific community by publications in appropriate peerreviewed journals and presentations at national scientific meetings, such as the Infectious Disease Society of 

Commercialization Plan Page 132 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

America annual meeting. We will engage key opinion leaders in the area as we move forward, as part of the 

FCCDC Scientific Advisory Board, in order to select the appropriate clinical safety study design. 

We have established an internal business development function within FCCDC, to be able to effectively market 

and seek out partners or funding for transition into full development and commercialization. Our CFO (Kathleen 

Czupich, MBA) has also participated in multiple discussions with potential partners and investors, marketing our 

technology. If we are awarded the Phase II SBIR funding and successfully carry out the research described, we 

will have more derisking validation data with which to market the technology and attract substantial investment. 

7. Revenue Stream 

FCCDC and the University of Pennsylvania have negotiated and signed an Intellectual Property Management 

Agreement (IPMA) to codify the terms of distributions of milestones and royalties from this project, and to 

establish the mechanisms for project governance and patent protection. 

A primary source of revenue for this program will most likely be from licensure to the US government and the 

Department of Defense. As described in Section 5, there is relevant precedence for the potential of significant 

income with the smallpox antiviral agent, tecovirimat where SIGA was awarded from BARDA a $433M 5-year 

base contract for development and licensure that could rise to $2.3B with additional treatment courses. 

Additional examples of successful Biodefense licensures include $1.2B for the SparVax® anthrax vaccine, 

>$500M for the nerve agent bioscavenger, rBChE and >$500M for the anthrax anti-toxin, Valortim®. FCCDC 

intends to generate a revenue stream for our product platform, depending on our ultimate production and 

marketing strategy. The revenue stream we envision will be as follows: 

In the scenario where we license rights to our product to a development partner, we expect that additional trained 

and qualified business development staff will be hired in order to manage these and other relationships. This 

dedicated business development function will manage the commercial exploitation of our intellectual property 

platform. A licensing deal would generate income for FCCDC through a combination of upfront fees, milestone 

payments and royalties on production of additional treatment courses. 

If we subcontract components of manufacturing and develop our candidate and related drug candidates 

ourselves, we will hire additional staff as required to manage this greater revenue stream as appropriate. FCCDC 

will evaluate pursuing additional grant funding and pre-seed investments to continue development up to and 

through the Animal Rule efficacy studies and Phase 1 clinical safety studies. This path that will increase the 

value of the company and technology prior to negotiating a commercial agreement or private investment as 

advancements through the development stages continue. 

Pre-seed and seed-stage investment are not revenue, but are sources of working capital. This funding can be 

obtained through convertible debt or equity investments from early-stage angel and venture investment groups, 

a number of which are located in the Pennsylvania/New York/New Jersey corridor (Ben Franklin Technology 

Partners, BioAdvance, Jump Start New Jersey, Mid Atlantic Angel Group, Delaware Crossing, etc.), where many 

of the major pharmaceutical companies are also located. FCCDC would anticipate generating significant 

revenues from out-licensing or sale of the technology based upon the well-recognized need for new anti-infective 

drugs for biodefense applications. 

The major source of revenue anticipated for this program is the Neglected Disease Priority Review Voucher 

offered by the U.S. FDA for new agents to treat the neglected, tropical diseases such as Marburg, Ebola and 

related viral infections. This voucher could then be sold to a major pharmaceutical company for cash 

considerations, and in return the acquiring company can use the voucher to reduce the drug approval time for a 

different therapeutic in their clinical development pipeline. 

References for Commercial Plan 

Lymphoma Research Foundation: Adult T Cell Leukemia / Lymphoma, 

http://www.lymphoma.org/site/pp.asp?c=bkLTKaOQLmK8E&b=6300141 

Allied Market Research 2015: https://www.alliedmarketresearch.com/ebola-therapeutics-vaccines-market 

Commercialization Plan Page 133 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Barda 2015: http://www.hhs.gov/news/press/2015pres/03/20150331a.html 

Boase, N. A.; Kumar, S., NEDD4: The founding member of a family of ubiquitin-protein ligases, Gene 2015, 557, 113-122. 

CDC 2015: http://www.cdc.gov/vhf/lassa/transmission/index.html 

Chesarino, N. M.; McMichael, T. M.; Yount, J. S., E3 Ubiquitin Ligase NEDD4 Promotes Influenza Virus Infection by 

Decreasing Levels of the Antiviral Protein IFITM3, PLoS Pathog 2015, 11, e1005095. 

GenomeWeb Daily News, “Congress passes Pandemic Act, extending funding for Project Bioshield and BARDA”, March 6, 

2013, http://www.genomeweb.com/congress-passes-pandemic-act-extending-funding-project-bioshield-and-barda. 

Gilead 2016. Warren, T. K.; Jordan, R.; Lo, M. K.; Ray, A. S.; Mackman, R. L.; Soloveva, V.; Siegel, D.; Perron, M.; 

Bannister, R.; Hui, H. C.; Larson, N.; Strickley, R.; Wells, J.; Stuthman, K. S.; Van Tongeren, S. A.; Garza, N. L.; Donnelly, 

G.; Shurtleff, A. C.; Retterer, C. J.; Gharaibeh, D.; Zamani, R.; Kenny, T.; Eaton, B. P.; Grimes, E.; Welch, L. S.; Gomba, 

L.; Wilhelmsen, C. L.; Nichols, D. K.; Nuss, J. E.; Nagle, E. R.; Kugelman, J. R.; Palacios, G.; Doerffler, E.; Neville, S.; 

Carra, E.; Clarke, M. O.; Zhang, L.; Lew, W.; Ross, B.; Wang, Q.; Chun, K.; Wolfe, L.; Babusis, D.; Park, Y.; Stray, K. M.; 

Trancheva, I.; Feng, J. Y.; Barauskas, O.; Xu, Y.; Wong, P.; Braun, M. R.; Flint, M.; McMullan, L. K.; Chen, S.-S.; Fearns, 

R.; Swaminathan, S.; Mayers, D. L.; Spiropoulou, C. F.; Lee, W. A.; Nichol, S. T.; Cihlar, T.; Bavari, S., Therapeutic efficacy 

of the small molecule GS-5734 against Ebola virus in rhesus monkeys, Nature 2016, 531, 381-385. 

Gonçalves, D. U.; Proietti, F. A.; Ribas, J. G. R.; Araújo, M. G.; Pinheiro, S. R.; Guedes, A. C.; Carneiro-Proietti, A. B. F., 

Epidemiology, Treatment, and Prevention of Human T-Cell Leukemia Virus Type 1-Associated Diseases, Clinical 

Microbiology Reviews 2010, 23, 577-589. 

Gottron F, “Project BioShield: Authorities, Appropriations, Acquisitions, and Issues for Congress”, Congressional Research 

Service, May 27, 2011 https://www.fas.org/sgp/crs/terror/R41033.pdf 

Heald AE, et al., Safety and Pharmacokinetic Profiles of Phosphorodiamidate Morpholino Oligomers with Activity against 

Ebola Virus and Marburg Virus: Results of Two Single-Ascending-Dose Studies. Antimicrobial Agents and Chemotherapy, 

2014, 58(11): 6639-6647. 

Huang, X. et al. The many substrates and functions of NEDD4-1. Cell Death & Disease 10, 904, doi:10.1038/s41419-019-

2142-8 (2019). 

Li, J. J.; Jr, R. J. F.; Diao, S.; Xue, B.; Bahouth, S. W.; Liao, F.-F., Nedd4 Haploinsufficient Mice Display Moderate Insulin 

Resistance, Enhanced Lipolysis, and Protection Against High-Fat Diet-Induced Obesity, Endocrinology 2015, 156, 1283-

1291 

Love, J, “How the US government subsidized Ron Perelman’s smallpox drug: ST-246 (Tecovirimat)” Knowledge Ecology 

International blog, Nov. 13, 2011, http://keionline.org/node/1314 

Ogbua O, Ajuluchukwub E, & Uneke CJ, Lassa fever in West African sub-region: an overview. J Vector Borne Dis, 2007, 

44(1): 1–11. 

PRNewswire 2014: http://www.prnewswire.com/news-releases/global-potential-analysis-of-ebola-drug-and-vaccinesmarket-pipeline-analysis-drugs-vaccines-and-geography-through-2020-279953592.html 

Wan, L. et al. NEDD4 expression is associated with breast cancer progression and is predictive of a poor prognosis. Breast 

Cancer Research 21, 148, doi:10.1186/s13058-019-1236-7 (2019). 

WHO: Ebola situation report 2015: http://apps.who.int/ebola/ebola-situation-reports 

Ye, X.; Wang, L.; Shang, B.; Wang, Z.; Wei, W., NEDD4: A Promising Target for Cancer Therapy, Current cancer drug 

targets 2014, 14, 549-556. 

Zou, X.; Levy-Cohen, G.; Blank, M., Molecular functions of NEDD4 E3 ubiquitin ligases in cancer, Biochimica et 

Biophysica Acta (BBA) - Reviews on Cancer 2015, 1856, 91-106. 

Commercialization Plan Page 134 
 

 

If "No" to AVMA guidelines, describe method and provide scientific justification 

 

Contact PD/PI: Harty, Ronald N. 

PHS 398 Cover Page Supplement OMB Number: 0925-0001 

Expiration Date: 02/28/2023 

1. Vertebrate Animals Section 

Are vertebrate animals euthanized? 

If "Yes" to euthanasia 

● Yes ❍ No

● Yes ❍ No

Is the method consistent with American Veterinary Medical Association (AVMA) guidelines? 

2. *Program Income Section 

*Is program income anticipated during the periods for which the grant support is requested? 

❍ Yes ● No

If you checked "yes" above (indicating that program income is anticipated), then use the format below to reflect the amount and 

source(s). Otherwise, leave this section blank. 

*Budget Period *Anticipated Amount ($) *Source(s) 

Page 135 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265 . Received Date: 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

PHS 398 Cover Page Supplement 

3. Human Embryonic Stem Cells Section 

*Does the proposed project involve human embryonic stem cells? ❍ Yes ● No 

If the proposed project involves human embryonic stem cells, list below the registration number of the specific cell line(s) from the 

following list: http://grants.nih.gov/stem_cells/registry/current.htm. Or, if a specific stem cell line cannot be referenced at this time, 

check the box indicating that one from the registry will be used: 

❏ Specific stem cell line cannot be referenced at this time. One from the registry will be used. 

Cell Line(s) (Example: 0004): 

4. Human Fetal Tissue Section 

*Does the proposed project involve human fetal tissue obtained from elective abortions? ❍ Yes ● No 

If "yes" then provide the HFT Compliance Assurance 

If "yes" then provide the HFT Sample IRB Consent Form 

5. Inventions and Patents Section (Renewal applications) 

*Inventions and Patents: ❍ Yes ● No 

If the answer is "Yes" then please answer the following: 

*Previously Reported: ❍ Yes ❍ No 

6. Change of Investigator/Change of Institution Section 

❏ Change of Project Director/Principal Investigator 

Name of former Project Director/Principal Investigator 

Prefix: 

*First Name: 

Middle Name: 

*Last Name: 

Suffix: 

❏ Change of Grantee Institution 

*Name of former institution: 

Page 136 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265 . Received Date: 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

PHS 398 Research Plan OMB Number: 0925-0001 

Expiration Date: 02/28/2023 

Introduction 

1. Introduction to Application 

(for Resubmission and Revision applications) 

Research Plan Section 

2. Specific Aims 

3. Research Strategy* 

4. Progress Report Publication List 

Specific_Aims_Harty_Final.pdf 

Proposal_Harty_Final_2.pdf 

Progress_Report_Publication_List_none.pdf 

Other Research Plan Section 

5. Vertebrate Animals 

6. Select Agent Research 

7. Multiple PD/PI Leadership Plan 

8. Consortium/Contractual Arrangements 

9. Letters of Support 

10. Resource Sharing Plan(s) 

11. Authentication of Key Biological and/or 

Chemical Resources 

VAS_4_combined.pdf 

10._Select_agent_use2_20-240_-_final.pdf 

Combined_LOI.pdf 

Combined_LOS.pdf 

Resource_Sharing_Plan_FCCDC.3.pdf 

Authentication_for_Harty_combined.pdf 

Appendix 

12. Appendix 

Page 137 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265. Received Date: 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Specific Aims. The ultimate goal of this Phase II application is to discover and develop novel small molecule, 

broad-spectrum therapeutics against viral infections caused by filoviruses, arenaviruses, and others that depend 

on the PPxY L-domain motif for virus egress and spread of infection such as Marburg (MARV), Ebola (EBOV), 

and Lassa fever (LAFV) viruses which are highly pathogenic and classified as Category A bioterror pathogens. 

In our ongoing studies to identify broad-spectrum small molecule inhibitors of viral egress that can slow the 

dissemination of both existing and newly arising variants of these potentially lethal viruses in unvaccinated 

populations, we have discovered several chemical series that block the host Nedd4/virus PPxY complex, a 

critical regulator of viral transmission. Importantly, we have identified an analog capable of blocking in vivo activity 

in a Marburg virus challenged mouse model, providing essential proof of concept for this novel class of anti-viral 

therapeutics. As there are no commercially available small molecule therapeutic agents for the treatment of these 

viral infections, our identification of virus-host inhibitors that prevent virus egress and spread will fill a significant 

unmet need. The goal of this Phase II STTR grant application is to optimize our lead inhibitor series, exemplified 

by FC-10696 to generate full-fledged predevelopment drug candidates ready for IND directed studies. This will 

be accomplished by combining the pharmaceutical and medicinal chemistry expertise of the scientists at the Fox 

Chase Chemical Diversity Center, Inc. (FCCDC) with the expertise and experience of the Harty Lab at the 

University of Pennsylvania in the experimental aspects of antiviral therapy, and the lab of Olena Shtanko at 

Texas Biomedical Research Institute for evaluating compounds against live viruses in vitro and in vivo using 

established small animal models of infection under BSL-4 conditions. 

Specific Aim 1: Lead optimization medicinal chemistry. Our current generation of PPxY/Nedd4 inhibitors 

exhibits excellent potency in blocking egress and spread of VLPs and live viruses, and is more metabolically 

stable than earlier series. We will continue to develop FC-10696 by performing SAR lead diversification using 

cutting edge methods in medicinal chemistry and drug discovery. This series will serve as the starting point for 

improved analogs to increase solubility and improve potency, as well as ADME, PK and safety properties suitable 

for once-daily oral dosing in patients. These data will inform further modifications to advance SAR development. 

Specific Aim 2. Evaluate compounds for their ability to specifically inhibit PPxY-mediated virus egress and host 

interactions. We will determine whether new compounds (Aim 1) block PPxY-mediated budding of VLPs 

produced by EBOV and MARV VP40, and by LAFV Z proteins using our established budding assays and a 

bimolecular Nedd4:PPxY complementation (BiMC) approach. We will also evaluate lead candidate inhibitors for 

potential effects on Nedd4-mediated ubiquitination and ESCRT-associated functions. Lastly, we will use an RNA 

seq. approach to globally assess the effects of our inhibitors on the host transcriptome, including specific E3 

ubiquitin ligases to identify potential off-target effects. 

Specific Aim 3: In vitro and in vivo ADMET evaluation to ensure suitable drug properties and selectivity. We 

will evaluate up to 15 compounds per year that meet the criteria in Aims 1 and 2 using in vitro ADMET assays, 

including microsomal stability, inhibition of CYP 3A4, solubility, and plasma protein binding. We will evaluate 

three to six compounds in mouse PK studies via IV and PO administration. The 2-3 most advanced compounds 

meeting in vitro potency and ADME/PK criteria will be evaluated in in vitro non-GLP safety assays such as: a) 

inhibition of cytochrome P450 isozymes (1A2, 2C9, 2D6, 2B6, 2C8, and 2B19, and 3A4-midazolam) most likely 

to cause variable exposure and drug-drug interactions; b) manual patch clamp assays to assess the hERG 

channel potential for cardiac arrhythmias; c) Ames testing with/without metabolic activation; and d) PK evaluation 

in rat, dog and non-human primate. These data will identify the best compounds for in vivo testing (Aim 4). 

Specific Aim 4: In vitro and in vivo analysis of lead inhibitors against authentic hemorrhagic fever viruses. We 

will evaluate the antiviral activity of select lead compounds against live BSL-4 pathogens. In each year, we will 

evaluate up to 4 compounds developed through studies in Aim 2 for the ability to selectively inhibit EBOV, MARV, 

and LASV spread and egress in human cell cultures. We will use a mouse model to assess compounds with the 

highest potency for toxicity, body distribution and accumulation over time. With this invaluable information in 

hand, we will evaluate two lead compounds per year for antiviral efficacy against filoviruses in a mouse model 

of MARV or EBOV disease. 

Our team is a leader in the development of host-oriented, broad-spectrum inhibitors of RNA virus egress, and 

we have made tremendous progress toward this goal as described below in our Phase I findings, published 

manuscripts, and U.S. Patent 10,160,756. Completion of these aims will allow us to choose a predevelopment 

candidate for IND enabling studies, pursue multiple funding options including additional NIH support; and 

ultimately leading to clinical evaluation for Marburg and/or Ebola indications, both of which are eligible for an 

FDA Priority Review voucher currently worth ~$80-100 M. 

Specific Aims Page 138 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

A. Significance and Impact.

Premise. The ultimate goal of this Phase II application is to develop 

novel small molecule, broad-spectrum therapeutics against viral 

infections caused by filoviruses, arenaviruses, rhabdoviruses, and 

others that depend on the PPxY L-domain motif for virus egress and 

spread of infection. Some of these viruses, including Ebola (EBOV), 

Marburg (MARV), and Lassa fever (LAFV) viruses, are highly 

pathogenic and classified as Category A, bioterror pathogens. The 

symptoms of these pathogens are often severe and, although these 

outbreaks largely originate in Africa, they have the ability to spread 

globally as observed in 2014-2015 (see graphic to the right). In our 

ongoing studies to identify small molecule inhibitors of viral egress that 

can slow the dissemination of both existing and newly arising variants 

of these potentially lethal viruses in unvaccinated populations, we have 

discovered several chemical series that block the host Nedd4/virus 

PPxY complex, a critical regulator of viral transmission. Importantly, 

we have identified an analog capable of blocking in vivo activity in a 

Marburg virus challenged mouse model, providing essential proof of 

concept for this novel class of anti-viral therapeutics. 

We predict that the novel class of anti-viral products targeting 

EBOV, MARV, and LAFV will be used for treatment of infected 

individuals as well as in prophylactic treatment of soldiers, healthcare 

workers, or others at high risk. Emergency administration of such an 

antiviral therapeutic during an outbreak could potentially inhibit virus 

dissemination in infected individuals and reduce the efficacy of 

infection in newly exposed individuals, thus slowing disease 

progression, allowing for more effective viral clearance by the immune 

system, and preventing further viral transmission. Moreover, as EBOV 

and MARV can cross the blood-brain barrier and re-emerge months 

later in the CNS, semen, and other immunologically privileged sites 

that are inaccessible to antibody therapy, rapidly diffusible small 

molecular inhibitors will fill a unique niche in filling the significant unmet 

need for anti-viral therapeutics against these classes of virus. Finally, 

as these host-oriented inhibitors are broad-spectrum, they are likely to 

be effective against newly emerging viruses as well as viral variants. 

Indeed, we predict that targeting a virus-host interaction necessary for 

efficient virus egress and dissemination will greatly diminish or 

eliminate the occurrence of drug resistant viral mutations and may lead 

to a paradigm shift in the search for better antiviral drugs. 

Hfv-info@lanl.gov 

RNA Virus Egress as a Target for Host-Oriented Therapeutics. 

Late budding domains (PPxY and PTAP) are highly conserved in the matrix proteins of a wide array of RNA 

viruses (e.g. filoviruses, arenaviruses, rhabdoviruses, paramyxoviruses, henipaviruses, and 

retroviruses) and thus represent broad-spectrum targets for the development of novel antiviral therapeutics [1-

11]. Notably, independent expression of filovirus VP40 or arenavirus Z matrix proteins leads to the production 

and egress of virus-like particles (VLPs) that accurately mimic budding of live virus, and PPxY L-domains within 

these proteins play a central role in this process [8, 12-18]. Moreover, efficient egress of VLPs depends on viral 

L-domain mediated recruitment of host proteins required for complete virus-cell separation or pinching-off of 

virus particles [1-8, 19]. Here we focus on the interaction between Nedd4, a WW-domain containing cellular E3 

ubiquitin ligase associated with the host ESCRT1 complex and the viral PPxY L-domain motif. Based on the 

critical role of this viral-host interaction in efficient budding of filoviruses, arenaviruses, and rhabdoviruses [3, 4, 

7, 8, 13, 17, 19-29], we have used a multifaceted approach to identify, develop, and validate several independent 

series of inhibitors of Nedd4:PPxY complex as potent, broad-spectrum antivirals. For this grant, our team, that 

has worked together for a half dozen years and composed of experts in the fields of virology and medicinal 

chemistry, proposes to find a predevelopment candidate for future IND enabling studies by 1) drug property 

optimization of current of analogs via iterative design and synthesis in Aim 1; biological testing for potency and 

Research Strategy Page 139 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

selectivity in Aim 2; drug property evaluation for oral activity using state of the art ADME and PK assays in Aim 

3 and finally evaluation of antiviral efficacy under oral administration in a mouse challenged with Marburg and/or 

Ebola virus in Aim 4. Our efforts will result in preclinical antiviral drug candidates for IND enabling studies. We 

will ensure that all assay experiments will be completed in a rigorous, unbiased manner, and quantitative 

readouts from virological assays proposed below will be statistically validated as indicated. Our team continues 

to lead in the area of developing host-oriented, broad-spectrum inhibitors of RNA virus egress, and we have 

made tremendous progress toward this goal as described below in our Phase I findings, published manuscripts 

[5, 7, 30-37], and U.S. Patent 10,160,756. 

B. Innovation. Development of small molecule therapeutics with potent, broad-spectrum activity against 

emerging RNA viruses and NIAID Category A pathogens remains a high priority, but also represents a major 

challenge due to the inherent difficulty of working with a subset of these pathogens under BSL-4 conditions. 

Several major innovations from our group help to alleviate this obstacle. 

1) Our team is a leader in the innovative development of host-oriented, broad-spectrum inhibitors of RNA 

virus egress, and we have made tremendous progress toward this goal as described below in our Phase I 

findings, published manuscripts, and U.S. Patent 10,160,756. For example, the Harty lab is a world-leader in 

functional characterization of VP40-interacting host proteins that facilitate virus egress [31-34, 36, 38, 39] and 

have developed innovative VLP budding assays that accurately recapitulate authentic live filovirus and 

arenavirus budding under BSL-2 conditions and state-of-the-art bimolecular complementation (BiMC) 

approaches to detect and quantitate virus-host protein interactions important for budding in live mammalian cells 

[37, 40] and to assess the impact of inhibitors on these processes. The innovative use of VSV and VSV 

recombinants (e.g. VSV-M40) as surrogate viruses for BSL-4 pathogens like EBOV, MARV and LASV allows us 

to test our inhibitors for antiviral activity against live viruses under BSL-2 conditions prior to undertaking these 

experiments with more dangerous BSL-4 pathogens [19, 41]. 

2) Our medicinal chemistry experts at FCCDC have successfully used SAR to identify several 

independent series of novel Nedd4:PPxY inhibitors that target this virus-host interaction, and Dr. Shtanko’s team 

of BSL-4 virology experts demonstrated that our lead inhibitors blocked egress and spread of live authentic 

viruses in cell culture and protected mice in a MARV challenge model (see below). 

3) Overall, our approach is conceptually innovative in that it pursues a conserved host:virus complex 

for the development of broad-spectrum antiviral agents for the treatment of Marburg, Ebola and related virus 

infections, for which there are no small molecule inhibitors on the market today. The success of our project will 

validate a new strategy for to the development of antiviral therapy. 

Research Strategy Page 140 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

C. Progress to Date: Preliminary and Published Findings.

C1. SAR Plan and Design. The primary goal of our Phase I STTR grant was to improve metabolic stability in 

mouse microsomes and aqueous solubility of our acylurea series of compounds[30] (Fig, 1) for demonstration 

of proof of concept efficacy in a mouse model of either Marburg or Ebola infection. Our novel agents potently 

interrupt the host Nedd4: viral PPxY interaction, thereby inhibiting virus budding and spread. Our initial lead 

series had a quinoxaline scaffold with an acylurea side chain. This subseries was unstable to mouse liver 

microsomes and also had poor aqueous solubility, resulting in very poor blood levels in mice when administered 

IV or IP. Here we describe the SAR that led to FC-10696, a compound with good antiviral potency, vastly 

improved overall ADME and PK properties, and demonstrated proof of concept efficacy in a Marburg 

challenged mouse model.

Design Plan. We focused our efforts to improve the metabolic stability via both modification of the acyl 

urea side chain and quinoxaline scaffold (Fig. 1.). Crystallographic studies [42, 43] of compounds that contain 

the acylurea moiety show that the CH2CONHCONHAr group exists in a pseudo six membered ring due to internal 

H-bonding (see Fig. 1). We reasoned that this pseudo six-membered ring may also exist in the bioactive 

conformation of our acylurea subseries, thus we prepared compounds with covalent six membered rings, as 

shown in the “Biphenyl subseries” in Fig. 1. 

SAR Map. In support of our prediction that elimination of one or more H bonds in this design would 

improve stability, solubility and overall bioavailability, the biphenyl subseries provided many highly potent and 

stable analogs (Fig. 1). Over 100 compounds have been prepared in this subseries. We later found that a thiazole 

amine or thiazole methyl 

moiety could replace the 

acylurea in this subseries to 

afford the “Thiazole benzyl” 

subseries (Fig. 1), of which 

over 30 compounds have been 

prepared. 

Table 1 shows in vitro, 

in silico ADME and PK data for 

selected compounds and Fig. 

2 shows the structures of the 

selected compounds in Table 

1. The quinoxaline compounds 

represented by FC-10696 to 

FC-12004 generally show 

strong inhibition of Marburg 

VLP egress activity at 0.1 uM, 

the lowest concentration 

tested. These compounds are 

also potent inhibitors of Ebola 

VLP egress as well, although 

less potent than for Marburg 

egress. 

New Scaffolds. The 

scaffold is 

exemplified by FC-11659. All 

members with this scaffold 

showed high potency; 

however, they were plagued 

by poor metabolic stability in 

mouse microsomes. Very 

recently we discovered that we 

could replace the quinoxaline 

with a 

(FC-11716 and FC12048) and retain good egress 

inhibition potency. We found 

Table 1. In vitro, in silico, ADME and PK data for selected compounds 

FC# Marburga Ebolab MW 

Log D 

(pH 

7.4) 

TPSAb 

(Å2) 

MLMc 

stability 

HLMd 

stability 

P450 3A4 

inhibition Mouse PK, IP administrationf 

1 

uM 

0.3 

uM 

0.1 

uM 

0.3 

uM 

0.1 

uM 

t1/2 

(min) 

t1/2 

(min) (uM) Cmax 

(ng/ml) 

AUC 

(ng x 

ml/hr) 

t1/2 

(hrs) 

IP Bioavailbility (%) 

FC-10696 91 82 57 357.5 5.9 52 76.9 1670 >33 253 1378 1.5 42 

FC-10905 92 75 412.4 5.5 52 65.5 188 

519 

513 

549 

597 

710 

2.6 

1.8 

1.2 

35 

22 

41 

FC-10906 

FC-11017 

99 90 

95 

67 

85 

358.5 

403.6 

5.1 

6.0 

52 

39 

8.8 

7.7 

73.9 

70.7 

>33 

>33 

FC-11078 91 87 440.5 5.9 65 87.2 95 269 2.6 16 

FC-12004 89 348.5 4.9 26 

FC-11659 100 95 89 350.5 3.8 52 1.0 

FC-11716 99 28 358.5 5.6 33 25.9 

FC-12048 93 372.5 6.0 33 >180 

a% Reduction of Marburg VLP at listed compound conc, VLP = virus like particle, in HEK293T cell line. b% Reduction of Ebola VLP at listed 

compound conc.cTPSA = total polar surface area. dMLM mouse liver microsome. eHLM human liver microsome. fBalbC mice administered one 

time 10 mg/kg of drug with plasma collection time points of 0.25, 0.5, 1, 2, and 6 hr. IP Bioavailbility is the ratio of IP AUC to IV AUC x 100. IV 

PK parameters are not shown. 

Research Strategy Page 141 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

that the SAR of the sulfur containing side chain and the 3-methyl moiety of this new scaffold is the same as for 

the quinoxaline scaffold SAR, but the stability to mouse microsomes is superior. Thus, these series will be further 

pursued. 

ADME and PK in Mouse. Five compounds, FC-10696 to FC-11078, were evaluated in a single dose 

mouse PK experiment under IP administration (Table 1). Compounds containing 6, 7 di-F atoms on the 

quinoxaline scaffold (FCs 10905 and 11078) showed poor blood levels, despite higher metabolic stability, 

presumably due to lower plasma solubility, when compared to the des F analogs, possibly resulting in 

precipitation at the injection site. Three of the five compounds (FC-10696, FC-10906 and FC-11017) that were 

examined in mouse PK were deemed suitable for live virus and mouse efficacy experiments described in later 

sections as they showed good stability to human liver microsomes and did not inhibit cytochrome P450 3A4 at 

concentrations up to 33 uM, thus showing low risk for drug/drug interactions. Of the three analogs, FC-10696 

had superior PK properties. 

Thus, from our SAR efforts, we have found several series and many potent and stable analogs resulting 

in good live virus potency and proof of concept in vivo activity in a mouse model as shown below. Our knowledge 

of the SAR will allow us now to pursue our goals of analogs that are potent and orally available. 

C2. In Aim 2 of Phase I, we evaluated 

the compounds described above using 

VLP, BiMC, and live-virus budding 

assays to determine their specific 

inhibition of the PPxY-Nedd4 interaction 

and PPxY-mediated egress. Below, we 

highlight representative data to 

demonstrate that the PPxY-Nedd4 

WW/domain interaction is a viable and 

promising antiviral target. 

VLP Budding Assay. The wellestablished and validated filovirus VLP 

budding assay that recapitulates live Fig. 3. Representative Marburg and Ebola VP40 VLP budding assay 

virus budding under BSL-2 conditions [4 data. HEK293T cells were transfected with either mVP40 (A) or eVP40 - (B) plasmids in the presence of DMSO alone, or the indicated budding 7, 12] is our primary assay used to inhibitors at the stated concentrations. Two of our lead compounds are evaluate the antiviral potency of all new highlighted in red. mVP40 and eVP40 were detected by Western blot in compounds (see Table 1 and Fig. 2). cell extracts and VLPs at 24 hours post-transfection. mVP40 and eVP40 Representative results are shown in Fig. levels in VLPs were quantified using NIH Image-J software. “%” is 

3. Briefly, we transfected HEK293T cells percent of VLP after compound treatment relative to DMSO control set with either mVP40 or eVP40 expression at 100%. 

plasmids in the absence (DMSO 

alone) or presence of the indicated 

compounds at the stated 

concentrations (Fig. 3). Expression 

levels of mVP40 and eVP40 were 

detected in cell extracts and VLPs by 

Western blotting and quantified by 

Image-J software as described 

previously (Fig. 3) [30, 36]. Here we 

used FC-4005, one of our early 

“acylurea inhibitors [30], at 1uM as 

our positive control. Notably, we Fig. 4. Bimolecular Complementation (BiMC) Assay data. HEK293T cells observed robust inhibition of mVP40 were co-transfected with equal amounts of NYFP-Nedd4 + CYFP-mVP40 in and eVP40 VLPs in the presence of the presence of DMSO alone, or 0.5uM concentration of FC-11017 or FC- two of our leading candidate 10906. Representative cell images (out of ~100 each) are shown for each 

inhibitors (FC-10696 and FC-11017 condition and were obtained at 24 hrs post-transfection using fluorescence 

highlighted in red) at 0.3uM microscopy. NucBlue = stained nuclei. Green = a Nedd4/mVP40 interaction. 

compared to DMSO alone (set at 

100%) (Fig. 4). These results were consistent in multiple independent budding assays (data not shown), and 

highlight the enhanced antiviral potency and broad-spectrum nature of these newly identified compounds. 

Research Strategy Page 142 
Contact PD/PI: Harty, Ronald N.

BiMC Assay. We used our well-established 

and validated BiMC approach [30, 37] to 

detect and visualize a MARV VP40-Nedd4 

interaction in live mammalian cells in the 

absence (DMSO alone) or presence of FC11017 or FC-10906 (Fig. 4). Briefly, 

HEK293T cells were co-transfected with 

plasmids expressing NYFP-Nedd4 and 

CYFP-mVP40 in the presence of vehicle 

(DMSO) alone, or PPxY inhibitors FC11017 and FC-10906 at a concentration of

0.5 μM (Fig. 4). Total cell counts based on 

NucBlue staining confirmed equal numbers 

of cells in all assay samples. YFP 

fluorescent complementation (i.e. green 

cells) is indicative of a PPxY-mediated 

interaction between mVP40 and host 

Nedd4, and fluorescent cells were 

visualized and quantified using an inverted 

Leica Sp5-II confocal microscope and 

MetaMorph software, as described 

previously [30, 36] (Fig. 4). In multiple

independent experiments, we observed a 

significant decrease in the relative number 

of YFP-positive cells in samples treated 

with either FC-11017 and FC-10906 

compared to that in vehicle-alone control concentrations of the compounds, equal concentrations of DMSO, or no treatment, 

samples (Fig. 4). As we published 

previously [36], a PPxY L-domain mutant of 

mVP40 that does not interact with Nedd4 

served as a valid negative control (data not 

shown). These results demonstrate that our 

lead candidate inhibitors specifically block 

the PPxY-mediated interaction between 

mVP40 and Nedd4 in mammalian cells 

and, together with data provided below, 

provide compelling evidence for their 

further optimization in Phase II.

In Vitro Live Filovirus Assays. To 

assess safety of the three lead 

compounds, we first assessed their

cytotoxicity in a HeLa cell line, routinely 

used in anti-filoviral screens, and 

primary human macrophages, the initial 

targets of filovirus infection in the host 

[44, 45]. The compounds appeared to 

be more cytotoxic to HeLa cells than to 

macrophages, with FC-10696 being the Fig. 5. FC-10696 inhibits MARV egress from primary human 

least toxic among the three. macrophages. Supernatants of primary human macrophages challenged 

Assessment of efficacy revealed that all with MARV and treated with indicated concentrations of FC-10696 or 

compounds inhibited both MARV and equal amount of DMSO were titrated on Vero cells for 24 hrs. Samples 

EBOV replication; albeit inhibition of were treated with anti-MARV VLP antibody to detect infected cells (green) 

and Hoechst dye to detect nuclei (blue) and subsequently photographed MARV was more efficient. We using a Nikon imaging system. The bar in each image is 100 um. determined CC50 and IC50 values for all 

time points for each virus using nonlinear regression analysis using GraphPad 8 software and listed select values in Table 2 and images of MARV

Research Strategy Page 143
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

in Fig. 5. Notably, the selectivity 

index (SI50, calculated as 

CC50/IC50) for each compound 

was ≥10 in MARV virion egress

assay, signifying antiviral 

potency and selectivity of these 

compounds against this virus. 

In Vivo Toxicity Studies. Since 

BALB/c mice will be used in 

efficacy studies with live virus, 

we used this mouse strain to 

assess toxicity of our lead 

compounds at two different 

concentrations: 1) the highest 

resuspendable concentration of 

each compound (20 mg/kg for 

FC-10696, 10 mg/kg for both FC11017 and FC-10906) and 2) a 

concentration which was 5-fold 

lower. Dosing with FC-10696 

resulted in the smallest overall 

weight loss and only transient 

clinical signs of toxicity (Fig. 6). 

Based on these toxicity data and 

in vitro data (Table 2), we chose 

to test FC-10696 for protection 

against MARV disease in a 

suitable mouse model. 

Efficacy studies using a 

mouse model of MARV 

disease. BALB/c mice are highly 

susceptible to infection with 

mouse-adapted MARV, 

developing disease symptoms 

and high viremia approximately 

three days post-infection, and 

succumbing to the disease by 

day 6. Treatment with 20 mg/kg 

twice daily for 10 days delayed 

the onset of mortality (p=0.0182; 

Fig. 7A), weight loss (Fig. 7B), 

and virus load in serum 

(p=0.0255; Fig. 7D), providing 

strong support for this class of 

compounds for further 

development into potent

antivirals against filoviruses and 

possibly other viruses that 

require the PPxY/Nedd4 

interaction for productive 

infection. These groundbreaking findings represent the 

first proof-of-concept in vivo 

Fig. 6. In vivo toxicity studies. Compounds were resuspended in 30%PEG400/ 

2%DMSO/14%kleptose HPB parenteral grade (Roquette) a formulation at two 

different concentrations: 20 and 4 mg/kg (FC-10696); 10 and 2 mg/kg (FC11017, FC-10906), and administered to groups of five 4-week old female 

BALB/c mice twice daily (BID) via the intraperitoneal route (IP) for a period of 

10 days. Animals were monitored daily for signs of treatment-associated 

toxicity: weight loss (A); rough hair coat, discharge from eyes and nose, 

diarrhea, decreased food intake and activity (B); and mortality. Clinical scores 

for each group were recorded as a sum of all observations in the group. 

Fig. 7. Efficacy studies of FC-10696 treatment in a mouse model of MARV 

disease. Three groups of 10 4-week old female BALB/c mice were challenged 

with 1,000 plaque-forming units (PFUs, as determined on Vero cells) of mouseadapted MARV. IP dosing by vehicle or FC-10696 at either 20 or 10 mg/kg started 

6 h post-challenge and continued BID for 10 consecutive days. Animals were 

observed daily for mortality (A), weight loss (B), and clinical signs of disease (C) 

for 21 days post-infection. Clinical scores for each group were recorded as a sum 

of all observations in the group, and if a score of ≥12 was recorded for an 

individual animal, it was considered terminally ill and euthanized. On day 3 postchallenge, 3 animals/group were euthanized to collect serum for virus load 

assessment by the neutral red plaque assay (D). The remaining 7 mice/group 

were used to determine animal survival. 

activity for our lead host-oriented PPxY inhibitor. 

Research Strategy Page 144 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Highlights of what we have achieved with our analogs are listed below. 

• Low nanomolar functional activity (<100 nM in VLP Marburg and Ebola cell assay); 

• Low nM target engagement (<500 nM, inhibition of Nedd4/VP40 PPxY binding in cellular BiMC assay); 

• Inhibition of live virus cellular egress (<100 nM, Marburg); 

• Little to no cytotoxicity at effective antiviral concentrations; 

• Stable in human liver microsomes (low clearance and long in vivo half-life); 

• No inhibition of CYP 3A4 (lower probability of drug-drug interactions); 

• Suitable ADME and PK properties for IP administration in mice; 

• Tolerability in mice at doses up to 20 mg/kg; 

• Proof of concept in vivo activity in a mouse model of Marburg virus disease; and 

• Novelty in structure, resulting in composition of matter patent protection. 

Based on these favorable properties, additional analogs will be prepared and tested, with the goals of maintaining 

and improving favorable ADMET and PK properties using pharmaceutical industry standard in vitro and in vivo 

metrics, maintaining and improving VLP and live virus potency, and showing efficacy in a mouse model of Ebola 

or Marburg virus disease under oral administration. Medicinal chemists have routinely improved drug 

properties that overcome many issues including designing and identifying analogs with oral activity so we fully 

expect to accomplish the goals below. 

Criteria (Go/No Go) for analogs to move forward to preclinical candidate status include: 

• Low nM (<100 nM) potency for VLP and BiMC in vitro comparable to our current leads, FC-10696; 

• Inhibition of live virus proliferation (IC50 <100 nM); 

• Minimal compound effects on other cellular Nedd4 activities; 

• Little or no cytotoxicity in mammalian cell lines at 50-fold higher than minimum effective concentrations; 

• Suitable ADME and PK properties for once 

a day oral administration; 

• In vivo activity in a mouse model of Ebola 

or Marburg virus disease under oral 

administration (PO); 

• Composition of matter patent protection 

worldwide. 

D. Experimental Approach:

Specific Aim 1: Lead optimization medicinal 

chemistry. Our goal is to design and synthesize 

novel analogs based on FC-10696 and to identify 2-

3 that meet the above criteria. We will pursue 

acceptable ADME and PK properties suitable for 

PO administration and identify one or more of these 

compounds to move forward to IND-enabling 

studies. We will accomplish these objectives via an 

iterative SAR approach using medicinal chemistry 

with parallel synthesis methods according to the 

work flow diagram and timelines in Figure 8. We 

will use in silico property calculations and 

pharmacophore analysis to monitor ADME drug 

properties, such as Log D, solubility and 

permeability using Chem Axon and QikProp from 

the Schrodinger Small-Molecule Drug Discovery 

Suite. Orally administered drugs must maintain 

good aqueous solubility by maintaining lipophilicity 

(cLog D, cLogP) and topological polar surface area 

(tPSA) values <5 and < 120 Å2, respectively. These 

values predict optimal clearance properties, good 

cell-based activity and avoidance of off-target 

Research Strategy Page 145 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

effects associated both with high molecular weight and lipophilicity. All compounds will be synthesized, purified 

and characterized at FCCDC labs. FCCDC has the capacity to prepare up to 250 g of any chemical analog. We 

will scale up, in multigram amounts, analogs that meet the criteria on the flow scheme for in vivo or other nonGLP assays. In addition, we will synthesize possible metabolites found during the metabolite identification 

processes. 

Our mandate is to improve aqueous solubility of our analogs while continuing to improve their potency, 

therapeutic index, ADME, PK and safety properties. We will focus on the following four areas: 

1) Understand stereospecificity of FC-10696. Of our three lead compounds, FC-10696 has the best 

microsomal stability and solubility. Thus, we will synthesize 

both R and S enantiomers from the readily available starting 

materials S and R 1-[1,1'-biphenyl]-4-ylethanol, 

respectively. These will be evaluated for potency, stability 

and solubility to determine the most promising enantiomer 

for further studies. We will also determine IV and PO PK in 

mice of the best enantiomer to determine if it could be an 

early front runner in our pursuit of a preclinical candidate. 

2) Quarternary methylene analogs of FC-10696. 

Analogs C and or D in Fig. 9 avoid the chiral center of FC10696 altogether and we expect good potency in our 

primary assays based on previous SAR, as well as good 

stability in mouse microsomes. Our previous method of 

preparing compounds of this type utilized 4-(1-bromo-1-

methyl-ethyl)-biphenyl to alkylate a quinoxaline thiol and led 

to FC-12004 in Table 1 among others, which gave poor yields of desired products (< 5%). However, using a 

different synthetic approach outlined in Figure 9, much better yields are expected. The starting materials A [46] 

and B [47] should be easily synthesized from readily available commercial chemicals. 

Fig. 9. Quaternary methylene analogs of FC-10696 

3) Water soluble prodrugs of FC-10696. 

Despite our SAR studies to date, we have been 

unable to add appropriate basic or acidic 

heteroatoms or sp3 carbons that might increase 

water solubility, and also retain good potency. 

However, FC-10696 has a 2-NH2 moiety, which 

we could link to a water-soluble prodrug group, to 

give FC-10696 (or the active enantiomer of FC- Fig. 10. Potential water-soluble prodrugs of FC-10696. 10696) greater water solubility to aid intestinal 

absorption following oral administration. The 

prodrug moiety of the absorbed drug would be removed by plasma or liver esterases, amidases or phosphatases 

(depending on prodrug type) to provide good plasma levels of parent compound (FC-10696) [48-50]. FCCDC 

has considerable experience with prodrugs of riluzole, which 

contains a similar NH2 moiety from which the prodrug group 

emanates [51, 52]. One of our riluzole prodrug derivatives is 

currently in Phase II clinical trials. Some potential water-soluble 

prodrugs are shown in Fig. 10. 

4) New quinoxaline analogs. In a further effort to increase 

water solubility of quinoxaline analogs of FC-10696, we plan to 

find an appropriate area on our molecules to append a watersoluble group (WSG) as outlined in Fig. 11. We know from our 

wide-ranging SAR that the quinoxaline ring or 2-methyl moieties 

cannot tolerate extensive modifications. However, the two phenyl 

rings of the biphenyl side chain moiety can tolerate substitutions. 

Therefore, our plan is to prepare analogs such as A and B that 

walk the WSG around the two rings to identify the best position to 

accept these groups. In addition, we know the middle ring of the 

“biphenyl like” side chain of our analogs can be a pyridine ring. 

Thus, we will prepare the two pyridyl analogs C and D in Fig. 11, and this should provide additional solubility to 

common formulations when compared with FC-10696. 

Fig. 11. Quinoxaline analogs with water 

soluble groups and pyridyl moieties. 

Research Strategy Page 146 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

5) New scaffolds. As a backup compound 

strategy, we will pursue alternative scaffold 

compounds, such as the 

scaffold, that yielded the 

promising analog FC-12048 (Table 1, Figure 2). 

In addition, we will prepare the close analog 

A shown 

in Fig. 12, which will increase the basicity and Fig. 12. Potential New Scaffolds 

ability to form salts with simple acids to increase 

aqueous solubility. This compound should be readily synthesized from and 

. Other scaffolds we will pursue that fit the current SAR profile and have been vetted for synthetic 

feasibility include: respectively which can be prepared from 

readily available appropriately substituted (for B) or 

(for C) and . 

All new analogs will be evaluated in our filovirus VLP MARV budding assay (Aim 2) [4-6, 12] to determine 

the antiviral potency, and those meeting potency criteria will be further assessed as described the screening tree 

in Fig. 8 and in Aim 3 and 4. 

Specific Aim 2. Evaluate compounds for their ability to specifically inhibit the PPxY-Nedd4 interaction and 

subsequent virus egress. All of the assays, reagents, and expertise are in place and readily available for these 

studies [30, 36]. We will use a three-pronged approach under BSL-2 conditions to identify the best compounds 

to be advanced to the more challenging and hazardous studies in the BSL-4 laboratory. First, we will use our 

well-established filovirus and arenavirus VLP budding assays, which recapitulate live-virus budding [4-7, 12] (see 

Fig. 4), to determine the antiviral potency of the best compounds identified in Aim 1. For EBOV and MARV, we 

will also include co-expression of NP and GP (along with VP40) in our VLP budding assays to more closely 

mimic budding of authentic EBOV and MARV. We will quantify egress of VLPs, focusing initially on mVP40-WT 

or PPxY mutants of mVP40 as controls (e.g. mVP40-AAAA or mVP40-∆PY), in the presence of each compound, 

and identify those that robustly block PPxY-dependent VLP egress in a dose-dependent manner. Once we 

identify inhibitors having activity against PPxY-mediated mVP40 VLP budding, we will then test these 

compounds for broad-spectrum activity using both EBOV VP40 and LAFV Z VLP budding assays [30, 36]. 

Second, we have implemented a BiMC approach [37] to detect and visualize mVP40-Nedd4 interactions 

in live mammalian cells [30] . We will use BiMC to evaluate the relative ability of analogs identified in Aim 1 to 

disrupt mVP40-Nedd4 interactions (see Fig. 3). NYFP-Nedd4 and appropriate CYFP-WT or PPxY mutants of 

mVP40 will be co-expressed in HEK293T cells, and we will use an inverted Leica Sp5-II confocal microscope to 

evaluate the ability and potency of inhibitors to block fluorescent complementation of mVP40-Nedd4 complexes. 

Quantitative analysis of complementation will be achieved by scoring individual cells and the rate, localization, 

intensity, and kinetics of fluorescence accumulation to determine drug potency. A PPxY L-domain mutant of 

mVP40 that does not interact with Nedd4 will be utilized as a negative control [36], and MTT cell viability assays 

will be performed in triplicate for all potent analogs tested, as described previously [34, 36]. 

Third, we will test lead analogs for their ability to block PPxY-mediated budding of live BSL-2 viruses 

VSV-WT, VSV-M40, and PPxY mutants of each (21). While results of the VLP budding and BiMC assays will 

identify novel potent budding inhibitors, the ultimate test of efficacy will be their ability to inhibit live-virus budding. 

We will test lead compounds at various concentrations for their ability to block live virus budding and spread in 

cell culture, using confocal microscopy and virus titration assays to quantify our findings. Lead compounds with 

robust activity in VLP, BiMC, and VSV budding assays will be prioritized by their anti-budding potency and used 

to test for antiviral activity against authentic BSL-4 pathogens at TBRI as described in Aim 4. We do not anticipate 

any insurmountable pitfalls with these well established and routinely used experimental approaches. 

Evaluate lead candidate inhibitors for potential effects on Nedd4-mediated ubiquitination and ESCRT-associated 

functions. Nedd4 is a member of the HECT family of E3 ubiquitin ligases that mono-ubiquitinates ESCRT proteins 

as well as viral matrix proteins to regulate their functions [3, 4, 7, 8, 13, 17, 19-29]. Since our budding inhibitors 

target Nedd4 interactions, it is important to determine whether these inhibitors adversely affect the normal 

function(s) of Nedd4, which includes trafficking and degradation of cell surface receptors, as well as cytokinesis 

[54-58]. Cells proliferate normally in the presence of these inhibitors and the presence of a number of potentially 

compensatory HECT E3 ubiquitin ligases suggests that global cellular ubiquitination will be maintained. However, 

to verify that there is no inhibitor-induced defect in global cellular ubiquitination, we will assess total cellular 

Research Strategy Page 147 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

ubiquitination profile by Western analysis using HA-tagged ubiquitin in HEK293T cells treated with either DMSO 

alone or with working concentrations of our lead inhibitors. As Nedd4 regulates the ESCRT protein Tsg101- 

dependent turnover and degradation of transferrin receptor 1 (TfR1) and epidermal growth factor receptor 

(EGFR) [54, 55, 59-61], we will determine whether our lead PPxY inhibitors alter Nedd4-mediated protein 

trafficking events at concentrations that block virus budding by measuring total cellular and surface expression 

levels of TfR1 and EGFR using immunofluorescence and confocal microscopy. DMSO treated cells will be used 

as a negative control. 

Second, we will assess potential global effects on total cellular RNA expression in HEK293T cells treated 

with either DMSO alone or with working concentrations of our lead inhibitors using RNAseq in collaboration with 

Dr. Dan Beiting (letter attached). mRNA-seq libraries will be prepared using the TruSeq Stranded mRNA LT Kit 

(Illumina), and all sequencing will be carried out in-house on our Illumina NextSeq 500 (base calling accuracy of 

>99.9%). All data processing and analyses will be carried out using the R programming language (Version 3.2.2) 

and the RStudio interface (Version 0.99.489). We do not anticipate that our inhibitors will impact gene expression 

and our preliminary findings using RNA seq. revealed no significant difference in expression levels of the Nedd4 

gene in the presence of DMSO alone vs. 1μm of FC-11017 (data not shown), suggesting that the effects of our 

inhibitors are not due to decreased Nedd4 expression and instead work by blocking Nedd4 interactions with viral 

proteins. 

Specific Aim 3. In vitro and in vivo ADMET evaluation to ensure suitable drug properties and selectivity. FCCDC 

will evaluate potential drug suitability issues as another filter to choose the best compounds for in vivo study and 

preclinical candidate selection. An estimated10-15 compounds per year that meet activity criteria in the screening 

tree (high potency and suitable in silico property criteria; Fig. 8) will be evaluated in in vitro ADMET assays 

(performed by the CRO Alliance Pharma, Inc.). Specifically, we will use microsomal stability and plasma protein 

binding assessment as reliable indicators of clearance and half-life in vivo. In addition, measurement of CYP 

3A4 inhibition will give us an early indication as to whether our compounds may have variable exposure in vivo 

or a risk of drug-drug interactions. Aqueous solubility and permeability will also be assessed, because low 

solubility/stability in assay medium is detrimental to development of a sound SAR and will also hamper our ability 

to formulate analogs for in vivo studies. We will move compounds forward that have acceptable values according 

to standard drug property criteria (see Fig. 13). We will also evaluate the 5-8 best compounds for PK properties 

in male and female mice following IV and PO administration (performed by the CRO Alliance Pharma, Inc.) to 

assess PK properties for eventual study in our mouse model (Aim 4). We will evaluate each sex separately to 

determine whether sex is a biological variable. If there is no difference, we will use both sexes in later studies. 

We expect to see low to moderate clearance, moderate volume of distribution, moderate to long half-life and 

good oral bioavailability. This last point is important, because we envision that the ultimate clinical use of our 

agents will involve oral administration alone or in combination with other antiviral drugs. The 2-4 most advanced 

compounds that have met potency and selectivity criteria and have acceptable in vitro ADME and PK properties 

will be evaluated further in the following non-GLP safety assays: 

• IC50 values will be determined for the seven 

most 

likely to cause exposure variability and drug-drug interactions 

(Alliance Pharma). 

• Manual patch clamp assays will be performed to assess hERG 

(human Ether-à-go-go-Related Gene product) potential for 

cardiac arrhythmias (Cyprotex). 

• Ames testing will be performed with and without metabolic 

activation to assess mutagenic potential (Cyprotex). 

• Safety panel of 44 primary molecular targets that detects major 

potential adverse activity (Eurofins). Fig. 13. Standard drug property criteria 

Compounds that have successfully completed the in vivo live virus 

mouse model (Aim 4), will be evaluated for PK properties in male and female rats and dogs following IV and PO 

administration (performed by the CRO Absorption Systems, Inc.) in preparation for evaluation in IND enabling 

studies. The best overall compounds will also be incubated in mouse and human microsomes under phase I and 

II conditions to generate and identify metabolites. 

Research Strategy Page 148 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

These industry-standard preclinical ADMET/PK studies will ensure that the selected analogs have the 

best chance to complete the drug development process. We are confident that at least one preclinical candidate 

will ultimately proceed through IND-enabling, preclinical development studies prior to human clinical evaluation. 

For a detailed description of the activities and timeline surrounding our push to an IND application, please refer 

to the Commercialization Plan. 

Specific Aim 4: In vitro and in vivo analyses of lead inhibitors against authentic hemorrhagic fever viruses. All 

experiments with replication-competent EBOV, MARV, and LASV will be performed in the BSL4 laboratory of 

Texas Biomedical Research Institute (TBRI) by Dr. Olena Shtanko (letter of collaboration is attached). We will 

first determine the cytotoxic properties of lead compounds identified in Aim 2. HeLa cells will be left untreated or 

treated with the compounds at ≥12 2-fold serially diluted concentrations or DMSO for 24, 48, or 72 hrs. The 

number of metabolically active cells will be determined using a CellTiter-Glo kit as in Table 2 and described [62, 

63]. We will determine the CC50 value for each compound at each time point using non-linear regression analysis 

to select a non-toxic concentration range for antiviral tests. 

To test whether the compounds affect egress and spread of EBOV, MARV, and LASV (virus stocks 

available at TBRI repository), we will incubate HeLa cells with virus at MOI=0.01 for 1 hr to allow binding, then 

wash, and overlay with new medium containing compounds at ≥7 2-fold serially diluted concentrations, DMSO, 

or no treatment, for 24, 48, or 72 hrs to cover several cycles of virus replication. We will detect infected cells by 

treatment with EBOV GP antibody, MARV VLP antibody, or LASV GP antibody (clone 83.6; Dr. MartinezSobrido’s letter is attached) [64], and nuclei by staining with Hoechst dye. We will photograph samples using a 

Nikon automated system and use CellProfiler software to quantify virus spread. To assess virus egress, we will 

titrate cell supernatants at each time point on Vero cells. Virus infection and inactivation studies will be performed 

in BSL4 using approved protocols. All post-infection analysis will be conducted in BSL2. The IC50 (virus spread, 

egress) values for each compound will be determined by non-linear regression analysis. We will use the 

CC50/IC50 (selectivity index, SI50) to assess antiviral potential of the compounds; those with SI50≥10 in egress

assays will be tested for antiviral activity in primary human macrophages, the initial and clinically relevant target 

cells of filoviruses and arenaviruses [44, 45, 65]. 

Human monocytes will be obtained from peripheral blood, with a typical yield of ≥15-20 million cells per 

a pint of blood from each donor and monocyte-derived macrophages (MDMs) will be obtained by differentiation 

in autologous serum [66, 67] to more closely mimic the physiological environment. We will assess compound 

cytotoxicity and antiviral properties at 24, 48, and 72 hrs time points as described above and, for those with 

SI50≥10 in egress assays, determine the IC95 value to be used in the design of a treatment regimen in animals. 

For each experiment, ≥3 independent tests in triplicate will be performed. Experiments with human MDMs 

will be performed from ≥6 different healthy adult donors (50:50 female:male ratio). We expect the same outcome 

trend among donors, although there may be a difference in the magnitude of the response. All data will be 

analyzed for statistical significance using either a Student t-test to compare averages of 2 groups or one-way 

ANOVA with Tukey’s test to compare averages of >2 groups, with p≤0.05 being considered significant. We will 

inspect data for normality. Where deviations from normality are seen, we will use appropriate transformations, 

or non-parametric tests. 

In vivo analysis of toxicity and accumulation of lead inhibitors. All ABSL2 experiments in mice will be performed 

at the TBRI rodent vivarium, which is staffed with full-time veterinarians and a large number of support personnel. 

Up to 4 compound candidates will be assessed for toxicity in BALB/c mice each year. Due to their susceptibility 

to filoviruses infection and pathology [44, 68], these mice have been used extensively in efficacy studies. Males 

and females randomly assigned to groups of 5 animals will receive dose formulation containing compounds at 2 

different doses or vehicle PO, up to 2 times a day (BID) for 10 consecutive days. The starting dose level and 

dosing frequency for each compound will be selected based on the findings of the PK study (Aim 3) and IC95 

data, above, to continuously maintain blood concentrations sufficient to block virus replication. A lower 

concentration will be needed to assess whether any observed toxicity is dose-dependent. The following 

parameters and end points will be evaluated in each group: (i) mortality; (ii) clinical signs, including weight 

change, food consumption, decreased motor activity, eye and nasal discharge, dyspnea, and unresponsiveness; 

(iii) serum biochemistry, including aspartate aminotransferase, alanine transarninase, alkaline phosphatase, 

total and direct bilirubin, urea, and creatinine levels, at the end of the study; (iv) post-mortem gross examination 

of the external features of the carcass as well as cranial, thoracic, and abdominal cavities, at the terminal 

necropsy; and (v) post-mortem histopathological examination of major organs, including liver, kidney, spleen, 

brain, heart, lungs, and adrenal glands (to be performed by TBRI Pathology Core personnel). We will use 5 

Research Strategy Page 149 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

mice/experimental condition as determined by power calculation. A Student t-test (with proper data 

transformation if needed) will be used to compare the difference between two groups. A sample size of 5 

mice/experimental condition will provide at least 80% power to detect a 3 standard deviation difference at each 

condition between two groups at a significance level of 0.01 (up to 5 multiple comparison adjustments). Multiple 

group comparisons will be analyzed by one-way ANOVA with Tukey's test. A p≤0.05 will be considered

significant. The experiments will be performed at least 2 times to ensure experimental reproducibility. To evaluate 

any sex-specific differences in response to treatment, groups of both male and female animals will be used. We 

will require 540 mice total [5 mice per group x 9 treatments (vehicle, 4 compounds at 2 different concentrations) 

x 2 sexes x 2 (repetitions) x 3 (years)] to complete these studies. 

Based on the results of the toxicity studies, we will select up to 2 compounds per year to assess body 

distribution and accumulation over time. Treated mice will be used to collect blood and organs critical for filovirus 

replication (liver, kidney, spleen) on days 1, 3, 5, 7, and 11. Blood samples will be centrifuged to separate the 

plasma fraction and, together with tissues, shipped to Alliance Pharma, Inc. to quantify compound levels. We 

will determine whether the compounds (i) circulate systemically and (ii) are present at concentrations necessary 

to inhibit ≥95% of virus replication. In these tests, 3 animals are required to show uniform outcome by a Student 

t-test or one-way ANOVA. We will repeat each study at least once. These experiments will require 360 mice [3 

mice per time point x 5 time points x 2 compounds x 2 sexes x 2 (repetitions) x 3 (years)]. 

In vivo treatment efficacy studies. Based on the data obtained from the compound toxicity/accumulation studies 

above, we will test up to 2 different treatments each year for protection against filovirus challenge using a mouse 

model of MARV disease. TBRI maintains an experienced and trained staff of veterinarians and veterinary 

technicians available to perform rodent studies in the ABSL4 laboratory. Groups of 10 BALB/c mice will be 

challenged with a lethal dose of 1,000 PFU of mouse-adapted MARV by IP. PO dosing will start 6±2 hrs postchallenge and will continue for 10 days at a frequency determined above. Animals will be closely monitored at 

least twice daily for signs of viral disease (ruffled hair coat, hunch back, inappetence, weight loss, and decreased 

movement) and mortality for 21 days post-challenge. Group clinical scores will be recorded as the sum of all 

clinical observations for the group. If a clinical score of ≥12 is recorded for an animal, it will be considered 

terminally ill and euthanized. Three animals from each group will be euthanized on day 3 post-challenge to collect 

blood and organs (liver, spleen, and kidneys) to determine virus titer by a plaque assay and load by qPCR as 

we described [69] and in Fig. 7D. Based on our data, BALB/c mice challenged with 1,000 PFU of mouse-adapted 

MARV become ill at 3 days and succumb to disease 5-6 days post-challenge (Fig. 7A). Assuming that we aim 

to protect 70% of animals from the virus disease, a power calculation comparing two independent proportions 

(significance=0.05 and power=0.8) indicates that 6-7 animals would be required for the survival study. Since we 

require 3 animals to be euthanized on day 3 for evaluation of virus burden, the group size is set to 10. To evaluate 

any gender-specific differences in response to treatment, groups of both male and female animals will be used. 

Viral burden will be analyzed using a Student t-test or one-way ANOVA Tukey’s test, and survival analysis will 

be performed using a Log-rank (Mantel-Cox) test, with p≤0.05 considered significant in all analyses. The efficacy 

studies will require 180 mice [10 mice per group x 3 treatments (vehicle, 2 dosings) x 2 sexes x 3 (years)]. 

Summary. Our success in Phase I of identifying potent, in vivo active, filovirus egress and spread inhibitors has 

positioned us to execute an expanded novel drug discovery program in Phase II. Notably, the success of inhibitor 

FC-10696 in protecting mice against MARV disease upon IP administration highlights the tremendous potential 

for this class of host-oriented compounds to be further developed into potent broad-spectrum antivirals. In Phase 

II, we seek to further transition one or more full-qualified PPxY: Nedd4 complex inhibitors into more detailed INDdirected pharmacokinetic, pharmacodynamic and toxicity studies suitable for once-daily PO administration in the 

clinic. Successful completion of these goals will provide a comprehensive package to support advanced 

development, efficacy testing in nonhuman primates and an IND submission for Phase I clinical safety testing in 

normal volunteers. Our ultimate goal is to provide and oral small molecule pharmacotherapy to patients who may 

be infected with Marburg, Ebola or related viruses in the future. Such viral hemorrhagic fever diseases are listed 

as being a focus of interest for SBIR/STTR funded research in the 2020 omnibus SBIR/STTR solicitation. Clinical 

development would be simplified by the use of the animal rule, in which Phase II/III efficacy studies are not 

required in human patients as FDA approval would be based upon efficacy in relevant animal models followed 

by acceptable safety and tolerability in Phase I. Drug approvals for Marburg and Ebola would both qualify for a 

saleable Neglected Disease Priority Review Voucher, currently worth 

Research Strategy Page 150 
 

Contact PD/PI: Harty, Ronald N. 

Progress Report Publication List 

None 

Progress Report Publication List Page 151 
 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

PHS Human Subjects and Clinical Trials Information OMB Number: 0925-0001 

Expiration Date: 02/28/2023 

Use of Human Specimens and/or Data 

Does any of the proposed research in the 

application involve human specimens and/or data * 

Provide an explanation for any use of human 

specimens and/or data not considered to be 

human subjects research. 

● Yes ❍ No 

Are Human Subjects Involved ● Yes ❍ No 

Is the Project Exempt from Federal regulations? ❍ Yes ● No 

Exemption Number ❏1 ❏2 ❏3 ❏4 ❏5 ❏6 ❏7 ❏8 

Other Requested Information 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265 Received Date: 

2020-09-08T12:43:45.000-04:00 Page 152 
 

Contact PD/PI: Harty, Ronald N. 

Human Subject Studies 

Study# Study Title Clinical Trial? 

1 Development of Host-Oriented Small Molecule Therapeutics for Filoviruses and Arenaviruses No 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265 Received Date:

2020-09-08T12:43:45.000-04:00 Page 153 
 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Section 1 - Basic Information (Study 1) OMB Number: 0925-0001 

Expiration Date: 02/28/2023 

1.1. Study Title * 

Development of Host-Oriented Small Molecule Therapeutics for Filoviruses and Arenaviruses 

1.2. Is this study exempt from Federal ❍ Yes ● No Regulations * 

1.3. Exemption Number ❏1 ❏2 ❏3 ❏4 ❏5 ❏6 ❏7 ❏8 

1.4. Clinical Trial Questionnaire * 

1.4.a. Does the study involve human participants? ● Yes ❍ No 

1.4.b. Are the participants prospectively assigned to an intervention? ❍ Yes ● No 

1.4.c. Is the study designed to evaluate the effect of the intervention on the ❍ Yes ● No participants? 

1.4.d. Is the effect that will be evaluated a health-related biomedical or ❍ Yes ● No behavioral outcome? 

1.5. Provide the ClinicalTrials.gov Identifier (e.g. 

NCT87654321) for this trial, if applicable 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265 Received Date: 

2020-09-08T12:43:45.000-04:00 Page 154 
 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Section 2 - Study Population Characteristics (Study 1) 

2.1. Conditions or Focus of Study 

❍ Develop host-oriented therapeutics targeting filoviruses and arenaviruses 

2.2. Eligibility Criteria 

Must be 18-50 years old- Must weigh at least 110 pounds- Cannot be pregnant- Cannot be on a medication- No illness within 

10 days 

2.3. Age Limits Min Age: 18 Years Max Age: 50 Years 

2.3.a. Inclusion of Individuals Across the Lifespan 20-240_Inclusion_Across_Lifespan.pdf 

2.4. Inclusion of Women and Minorities 20-240_Inclusion_Women_Minorities.pdf 

2.5. Recruitment and Retention Plan 20-240_Recruitment_and_Retention_Plan.pdf 

2.6. Recruitment Status Not yet recruiting 

2.7. Study Timeline 20-240_Study_Timeline.pdf 

2.8. Enrollment of First Participant 04/01/2021 Anticipated 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265 Received Date: 

2020-09-08T12:43:45.000-04:00 Page 155 
 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Inclusion of Individuals Across the Lifespan 

Women and men between the ages of 18-50 years old will be enrolled onto the study. 

The venipuncture of healthy children (less than 18 years old) adds concerns and risks beyond those seen with 

adults. At the present time, there is no reason to believe that children would represent a unique population 

compared to adults in this study. The amount of blood required for the requisite number of cells essential for an 

experiment would be proportionally higher in children due to their decreased size. Anatomically, children have 

significantly smaller veins presenting increased challenges of drawing blood in a laboratory setting. The 

discomfort and initial pain of the blood draw procedure while minimal in adults would be more likely to incite 

increased psychological trauma and fear in children. Thus, the inclusion of children at the present time is not 

justified given the additional risks for the procedure. With additional data obtained from adults, in the future a 

new study directly comparing peripheral blood mononuclear cells from adults and children may be warranted. 

Venipuncture of aged adults (over 50 years old) adds concerns and risks, particularly of infection, beyond 

those seen with adults. Thus, the inclusion of aged adults at the present time is not justified given the 

additional risks for the procedures. It is well established that cells from aged and adult individuals exhibit 

different inflammatory phenotypes. It is beyond the scope of the current proposal to compare responses to 

aged and adult cells, these complex studies are better suited for dedicated studies once responses in adults 

have been clearly defined. 

The study team is well experienced in working with adults (18-50 years old) and facilities for working with these 

individuals are in place and being utilized for other studies. The developmental nature of this study does not 

require assessing the impact of aging on Marburg, Ebola, and Lassa virus infection. It is critical to develop in 

vitro models with adult cells prior to conducting studies directly comparing cells from adults to children 

and aged adults. 

Inclusion of Individuals Across the Lifespan Page 156 
 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Inclusion of Women and Minorities 

There will be no discrimination based on gender, race, or ethnicity. To ensure that the gender distribution is 

equal, recruitment will be directed at acquiring approximately 50% females and males. For venipuncture, 

9 females and 9 males will be recruited. Women who are currently pregnant will be excluded from the study, 

due to health concerns associated with donating blood. Recruitment will be monitored to assure that 

minority populations are represented in this cohort of subjects. The target percentages of racial minorities are 

based on the racial distribution in San Antonio, TX. The population of San Antonio, TX is 62.7% Hispanic/ 

Latino, 26.6% Caucasian, 6.8% African-American, 2.4% Asian, and less than 0.4% American Indian/Alaska 

Native, and, thus the study population may reflect a slight bias toward Hispanic/Latino to reflect the 

population demographics. These demographics are acceptable for studying macrophage responses during 

EBOV infection. If the usual recruitment methods yield only Caucasian volunteers, notices will be placed on 

bulletin boards where larger numbers of minority students are likely to see them, such as the AfroAmerican Culture Center. The notices could also be expanded to include a statement that “women and 

minority students are especially encouraged to apply.” 

Inclusion of Women and Minorities Page 157 
 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Recruitment and Retention Plan 

Recruitment 

Subjects are recruited from among healthy laboratory and office personnel, other Texas Biomed employees; 

students at the Colleges and Universities in San Antonio, including UT Health San Antonio and University of 

Texas San Antonio (UTSA); hospitals, including the San Antonio Metropolitan Health District; and the community. 

Subjects are largely recruited through flyers that are posted throughout the community, schools, hospitals, and 

Texas Biomed, as well as word of mouth. Recruitment activities also involve community events. Recruitment will 

be monitored to assure that minority populations are represented in this cohort of subjects. The target 

percentages of racial minorities are based on the racial distribution in San Antonio, TX. The population of San 

Antonio, TX is 62.7% Hispanic/Latino, 26.6% Caucasian, 6.8% African-American, 2.4% Asian, and less than 

0.4% American Indian/Alaska Native, and, thus the study population may reflect a slight bias toward 

Hispanic/Latino to reflect the population demographics. If the usual recruitment methods yield only Caucasian 

volunteers, notices will be placed on bulletin boards where larger numbers of minority students are likely to see 

them, such as the Afro-American Culture Center. The notices could also be expanded to include a statement 

that “women and minorities are especially encouraged to apply.” 

Retention 

Subjects are allowed to donate no more than 480 ml of blood in an 8-week time span. They will be allowed, but 

not expected, to participate multiple times, as long as they donate no more than 480 ml of blood in an 8-week 

time span, or only once. 

Recruitment and Retention Plan Page 158 
 

 

Contact PD/PI: Harty, Ronald N. 

Study Timeline 

Recruiting for the study will start on 04/01/21, following the approved UT Health San Antonio IRB 

protocol #HSC20180013H. Recruitment is expected to occur throughout the entire three year duration of 

the proposal. 

Study Timeline Page 159 
 

 

 

Contact PD/PI: Harty, Ronald N. 

2.9. Inclusion Enrollment Reports 

IER ID# Enrollment Location Type Enrollment Location 

Study 1, IER 1 Domestic Texas Biomedical Research Institute 

Funding Opportunity Number: PA-20-265 Received Date: Tracking Number: GRANT13199760 

2020-09-08T12:43:45.000-04:00 Page 160 
 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Inclusion Enrollment Report 1 

1. Inclusion Enrollment Report Title* : Development of Host-Oriented Small Molecule Therapeutics for Filoviruses and 

Arenaviruses 

2. Using an Existing Dataset or Resource* : ❍ Yes ● No 

3. Enrollment Location Type* : ● Domestic ❍ Foreign 

4. Enrollment Country(ies): USA: UNITED STATES 

5. Enrollment Location(s): Texas Biomedical Research Institute 

6. Comments: 

Planned 

Racial Categories 

Ethnic Categories 

Total Not Hispanic or Latino Hispanic or Latino 

Female Male Female Male 

American Indian/ 

Alaska Native 0 0 0 0 0 

Asian 0 0 0 0 0 

Native Hawaiian or 

Other Pacific Islander 0 0 0 0 0 

Black or African 

American 1 1 0 0 2 

White 2 2 6 6 16 

More than One Race 0 0 0 0 0 

Total 3 3 6 6 18 

Cumulative (Actual) 

Racial Categories 

Ethnic Categories 

Total Not Hispanic or Latino Hispanic or Latino 

Unknown/Not 

Reported Ethnicity 

Female Male 

Unknown/ 

Not 

Reported 

Female Male 

Unknown/ 

Not 

Reported 

Female Male 

Unknown/ 

Not 

Reported 

American Indian/ 

Alaska Native 0 0 0 0 0 0 0 0 0 0 

Asian 0 0 0 0 0 0 0 0 0 0 

Native Hawaiian or 

Other Pacific Islander 0 0 0 0 0 0 0 0 0 0 

Black or African 

American 0 0 0 0 0 0 0 0 0 0 

White 0 0 0 0 0 0 0 0 0 0 

More than One Race 0 0 0 0 0 0 0 0 0 0 

Unknown or 

Not Reported 0 0 0 0 0 0 0 0 0 0 

Total 0 0 0 0 0 0 0 0 0 0 

Page 161 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265 Received Date: 

2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Section 3 - Protection and Monitoring Plans (Study 1) 

3.1. Protection of Human Subjects 20-240_Human_Subjects_-_final.pdf 

3.2. Is this a multi-site study that will use the same protocol to ❍ Yes ● No ❍ N/A 

conduct non-exempt human subjects research at more than one 

domestic site? 

If yes, describe the single IRB plan 

3.3. Data and Safety Monitoring Plan 

3.4. Will a Data and Safety Monitoring Board be appointed for ❍ Yes ● No 

this study? 

3.5. Overall structure of the study team 

Funding Opportunity Number: PA-20-265 Received Date: Tracking Number: GRANT13199760 

2020-09-08T12:43:45.000-04:00 Page 162 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

PROTECTION OF HUMAN SUBJECTS 

1. Risks to Human Subjects 

Human Subjects Involvement, Characteristics, and Design 

The proposed research involves use of human subjects as a source of blood obtained by venipuncture for 

the isolation of peripheral blood mononuclear cells. Studies in primary human macrophages, initial targets of 

MARV, EBOV, and LASV, are important to understand whether antiviral treatment affects virus replication and 

spread in clinically relevant cells. Subjects will be limited to normal healthy adult (18-50 years old) volunteers 

without symptoms of recent infections. Both genders will be studied and there will be no discrimination 

of race or ethnicity. To ensure that the gender distribution is equal, recruitment will be directed at acquiring a 

50% female:male ratio. Participant recruitment will also be monitored to assure that minority populations 

are represented in this cohort of subjects. Individuals under the PI’s supervision, including students or staff, 

will be excluded. We will obtain venous blood samples from 6 volunteers/year for a total pool of 18 

individuals (see Targeted/Planned Enrollment Tables). 

All of the risks described below will be explained to the subject by the principal investigator or an 

appropriate physician representative. The subject will be given the opportunity to ask questions about the 

procedures and the risks. They will then be asked to sign a written consent form which summarizes the 

risks and also states that the subject is free to withdraw at any time. Special groups (pregnant females, 

fetuses, the elderly, children, prisoners and the cognitively impaired, etc.) will be excluded from this protocol. 

All studies will be conducted at Texas Biomedical Research Institute. 

Study Procedures, Materials, and Potential Risks 

Peripheral blood cells will be obtained from venipuncture, following standard procedures from healthy subjects. 

The date on which blood was given and the amount of blood withdrawn will be recorded in a notebook to 

be kept locked in the medical personnel’s research office. This information and the peripheral blood are the 

only things collected from the human subjects. Specimens and data will be labeled with a numerical code. 

The subjects’ name will not appear on any specimens and participation will remain confidential. The identity 

of the subjects will not be revealed in any publications, slides, presentations or other related materials. The 

subject’s name, phone number, and subject numerical designation will be recorded in a notebook to be 

kept in the medical personnel’s research office. This office is kept locked and only laboratory personnel 

have access to the human subject notebook. 

The possible risks associated with participating in this research project are as follows: most people experience 

transient pain on insertion of the needle. Occasionally, a small bruise will develop at the site of the blood drawing. 

Transient lightheadedness can develop and on rare occasions, fainting may occur. 

2. Adequacy of Protection against Risks 

Informed Consent and Assent 

Consent will be obtained at Texas Biomedical Research Institute, room L117 by lab personnel. All risks 

described above will be explained to the subject. The subject will be given the opportunity to ask 

questions about the procedures and the risks. They will then be asked to sign a written consent form which 

summarizes the risks and also states that the subject is free to withdraw at any time. 

Protections Against Risk 

Several procedures will be employed in order to minimize the risks. Venipuncture will be performed by 

a physician or a qualified technician. The antecubital vein will be used and ordinary sterile technique will 

be observed. The subject will be in a sitting or supine position. Following withdrawal of blood, firm pressure 

will be applied to the site for several minutes. The usual amount of blood will vary from 30-200 ml, and 

any single adult subject will be limited to a maximum of 480 ml in any two-month period. The identity of 

subjects will not be revealed in any publications, slides, presentations or other related materials. The date on 

which blood was given and the amount of blood withdrawn will be recorded in a notebook to be kept 

locked in the research office. The subjects' name will not appear on any specimens and participation will 

Protection of Human Subjects Page 163 
 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

remain confidential. Specimens and data will be labeled with a numerical code. 

Due to the nature of the study, no incidental findings are expected to occur. 

Vulnerable subjects 

Special groups (pregnant females, fetuses, the elderly, children, prisoners and the cognitively impaired, etc.) 

will be excluded from this protocol. 

3. Potential Benefits of the Proposed Research to Research Participants and Others 

For study participants, both the anticipated risks and benefits are very low. Society is expected to gain a 

long-term benefit from the results of this study, related to discovery of novel countermeasures against 

filoviruses and arenaviruses. 

4. Importance of the Knowledge to be Gained 

There will be no direct benefits to the individual subject, but expected long-term benefits to society include 

improved treatment of patients with filovirus and arenavirus infection. Thus, the potential benefits to society 

are high. 

Protection of Human Subjects Page 164 
 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Section 4 - Protocol Synopsis (Study 1) 

4.1. Study Design 

4.1.a. Detailed Description 

4.1.b. Primary Purpose 

4.1.c. Interventions 

Type Name Description 

4.1.d. Study Phase 

Is this an NIH-defined Phase III Clinical Trial? ❍ Yes ❍ No 

4.1.e. Intervention Model 

4.1.f. Masking ❍ Yes ❍ No 

❏Participant ❏Care Provider ❏Investigator ❏Outcomes Assessor 

4.1.g. Allocation 

4.2. Outcome Measures 

Type Name Time Frame Brief Description 

4.3. Statistical Design and Power 

4.4. Subject Participation Duration 

4.5. Will the study use an FDA-regulated intervention? ❍ Yes ❍ No 

4.5.a. If yes, describe the availability of Investigational 

Product (IP) and Investigational New Drug (IND)/ 

Investigational Device Exemption (IDE) status 

4.6. Is this an applicable clinical trial under FDAAA? ❍ Yes ❍ No 

4.7. Dissemination Plan 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265 Received Date:

2020-09-08T12:43:45.000-04:00 Page 165 
 

 

 

Contact PD/PI: Harty, Ronald N. 

Delayed Onset Studies 

Delayed 

Onset Study# 

Study Title Anticipated Clinical 

Trial? 

Justification 

The form does not have any delayed onset studies 

Page 166 

Tracking Number: GRANT13199760 Funding Opportunity Number: PA-20-265. Received Date: 2020-09-08T12:43:45.000-04:00 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

VERTEBRATE ANIMALS: MOUSE 

Texas Biomedical Research Institute 

1. Description of animals and how they will be used 

Suppliers 

Wild-type BALB/c mice for in vivo treatment toxicity, compound concentration and accumulation studies, and 

infections will be obtained from The Jackson Laboratory. 

Strain choice 

BALB/c mice are being generated and widely supplied by The Jackson Laboratory for research purposes. Wildtype BALB/c mice have been shown to be susceptible to infection with mouse-adapted Marburg virus and 

develop symptoms that are consistent with human infections. 

Age choice 

We will use 4-5-week old mice. 

Sex differences 

We will use a 50:50 female:male ratio distribution of sex to ensure our results are not confounded by sex 

differences. 

Experimental needs 

Experiments in this proposal require: 

1. Oral administration (PO) of compounds twice daily (BID) over a period of 10 days, followed by post-mortem 

gross examination and tissue isolation for ex vivo analysis for any pathobiological changes, to determine if the 

treatments have adverse effect in the host (270 males + 270 females = 540 animals). 

2. BID administration of compounds by PO and subsequent plasma and organ isolation (after 1 treatment; 3, 5, 

7, or 10 days of treatment) to determine compound distribution and concentration over time (180 males + 180 

females = 360 animals). 

3. Challenge with mouse-adapted MARV by intraperitoneal route and subsequent BID treatment by PO for 10 

consequent days to follow the course of infection and isolation of tissues for ex vivo analysis (90 males + 90 

females = 180 animals). 

The safety operations procedure (SOP) for filovirus infection and other procedures specified in this application 

have been filed and approved by Texas Biomed Biohazard & Safety Committee. The IACUC application 

#1706MU detailing procedures in this application has been approved by Texas Biomed Institutional Animal Care 

and Use Committee. The Texas Biomedical Research Institute animal assurance welfare number is D16-00048 

(A3082-01) under file with the NIH. 

A total of 1,080 mice (540 males + 540 females) is requested. 

2. Justification for the use of animals 

The use of animals is justified by the fact that the course of MARV infection and affect of antiviral treatment on 

the disease outcome cannot yet be reliably predicted by in vitro assays or computer simulations. BALB/c mice 

have been shown to be susceptible to infection with mouse-adapted Marburg virus and develop symptoms that 

are consistent with human infections. Because of this sensitivity, the high degree of similarity between the murine 

and human immune systems, and the large number of reagents available to study mice, the species used in the 

study would provide the most complete information possible. The small size of the animal requires smaller 

amounts of treatment reagents injected, thus maximizing reagents that may be in short supply. Because of the 

limitation of ABSL4 space, the smaller model facilitates proof-of-concept testing at a fraction of the cost of 

experiments in larger animals. The inbred nature of the mice also reduces variation in response to treatments 

and virus, allowing for limiting the size of test groups. 

Vertebrate Animals Page 167 
 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

3. Provisions to minimize discomfort, distress, pain, and injury 

To help minimize pain and distress, as well as minimize risk for care staff, all mice will be sedated with isofluorane 

for virus challenge or ketamine+xylazine for terminal blood collections. After challenge, animals will be closely 

monitored for signs of: (i) ruffled hair coat, (ii) hunch back, (iii) inappetence, (iv) weight loss, and (v) decreased 

movement. If the animal fulfills 2 of the above criteria, where animals are in distress as result of disease or 

treatment, it will be euthanized. If at any time the animal experiences labored breathing, the animal will be 

euthanized immediately. If at any time in the responsible veterinarian’s judgment, unnecessary suffering is 

occurring, the animal will be euthanized. Based on our preliminary data, BALB/c mice challenged with mouseadapted MARV become ill at 3 days and die 5-6 days post-challenge. 

Vertebrate Animals Page 168 
 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Prepared for: Fox Chase Chemical Vertebrate Animal Section Diversity Center 

(VAS) 

Date: July 18, 2018 

Point 1: Description of animals and how they will be used 

A total of 3 Sprague Dawley rat, male and female, aged approximately 6-8 weeks and weighing 200-400 grams will 

be used for each test article for the studies. Rats for pharmacokinetic evaluation will be serially bled over a 24 hour 

period. Route of administration for the test article will be intravenous (IV) at a dose TBD. Dosing of the test article 

will occur once via the corresponding dose route. Animals will be fasted prior to dosing with water provided ad 

libitum. All blood samples will be collected via tail vein and/or mandibular vein and then placed in tubes with 

anticoagulant. All samples will be transferred to the Absorption Systems Analytical Department for analysis. The 

blanket IACUC protocol approval number for this study is ASLP16-01 and was approved February 12, 2016. 

Point 2: Justifications for use of animals 

There are no known acceptable alternatives to the use of live animals to accomplish the purpose of this study. The test 

article is a novel drug and the study does not unnecessarily duplicate any previous work. The rat is a standard model 

for non-clinical studies, for which there is a large historical database, and has been found to be a suitable model in 

previous investigations with similar test articles. Examination of pharmacokinetics and distribution studies is critical 

for drug development. This study was designed to use the fewest number of animals possible consistent with the 

scientific objectives of the study, the goals of the sponsor, contemporary scientific standards, and in consideration of 

applicable regulatory requirements. The number of animals used is considered the minimum to make meaningful 

biological comparisons, and for statistical calculations. 

Point 3: Provisions to minimize discomfort, distress, pain and injury 

All animals will be observed by professional technicians trained in lab animal care and use. An attending veterinarian 

is available at all time for emergencies. In the event that any aspect of this study causes more than normally expected 

brief pain or distress to the animals, the study director shall determine, in consultation with the attending veterinarian, 

if administration of appropriate sedatives, analgesics, or anesthetics, or other therapies would be contraindicated by 

the objectives of the study and document the resultant course of action. Animals that experience severe or chronic 

pain or distress that cannot be relieved will be euthanized. 

Point 4: Euthanasia 

Animals will typically be sacrificed after the last clinical observation noted or blood sample has been collected. All 

rats will be euthanized by cervical dislocation under isoflurane anesthesia. Isoflurane ensures that the rats are 

unconscious, while dislocation ensures quick death. This minimizes animal distress, is effective, and efficient; it is 

consistent with the recommendations of the AVMA Guidelines on Euthanasia. 

Vertebrate Animals Page 171 
 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

VERTEBRATE ANIMALS SECTION (VAS) p. 1 of 1 

1 

2 

3 

4 

Description of procedures 

In vivo pharmacokinetics (PK) assessment will be performed to determine the dose and dosing 

schedule for additional in vivo studies. PK work will be performed at Alliance Pharma (Malvern, 

PA), a CRO specializing in drug metabolism and PK (DMPK). C57Bl/6 mice will be used. 

Justifications 

The proposed animal experiments are critical to determine PK parameters after intravenous (IV) 

and intratracheal (IT) drug administration, including half-life, peak plasma and lung tissue 

concentration (Cmax), and the integral of the concentration-time curve in both plasma and lung 

tissue (AUC, a measure of drug exposure) to determine the dosing schedules to be used in future 

studies, and to enable an understanding of how drug exposure relates to efficacy in models of lung 

disease. 

To assess the PK profile of each compound, mice will be dosed IV and/or IT with each compound, 

and plasma concentrations will be determined at up to eight time-points over 24 hours (n=3 mice 

per time-point) using LC-MS/MS. Total number of mice to be used for PK studies is 192 over a 

two year period. 

Minimization of Pain and Distress 

All procedures are designed to minimize discomfort and pain. Mice will be fed and watered 

appropriately throughout the entire protocol. Each cage also contains a nestlet for enrichment 

purposes. Levels of pain/distress and endpoints are discussed by the IACUC as an integral part of 

the protocol review process. The IACUC ensures that the procedures described in the protocol 

are categorized appropriately. As an IACUC member, the Attending Veterinarian reviews all 

animal care and use protocols with an emphasis on those that require any anesthetics or 

analgesics. The AV and investigators discuss the appropriate selection and regimen for 

anesthetics and analgesics for the animal procedures in the protocol. The recommendations are 

then transferred into the protocol for IACUC review and approval. 

Euthanasia 

Any mice exhibiting pain or discomfort lasting longer than a few seconds that cannot be instantly 

relieved will be immediately euthanized. No animals are euthanized in the presence of their 

conspecifics (same room). Animals are placed in a clear, plexiglass chamber of known size and 

CO2 is introduced at a flow rate of 10-30% (based on chamber size) via a calibrated flow meter. 

When animals are euthanized, death is assured by direct observation to ensure an absence of 

respiration. In addition, the chest is palpated to ensure an absence of a heartbeat. Methods of 

euthanasia used at Alliance are in compliance with the AVMA Guidelines for the Euthanasia of 

Animals: 2013 Edition. 

Vertebr

A

at

l

e nimal

V Page 172 liAance_ s AS VAS Alliance 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

SELECT AGENTS 

Texas Biomedical Research Institute (Texas Biomed) 

BSL4 agents licensed for use in this proposal. Low passage human isolates of Ebola virus variant Mayinga, 

Marburg virus strain Musoke, Lassa virus strain Josiah as well as mouse-adapted Marburg virus strain Angola 

will be used in this study and are available in the repository at Texas Biomed. Work with replication competent 

viruses will be performed in the Texas Biomed ABSL4 facility by Dr. Olena Shtanko, Ms. Gloria Rodriguez 

(Research Assistant), and ABSL4 veterinary staff. Dr. Shtanko, Ms. Rodriguez, and veterinary staff are 

authorized for work with these agents and have appropriate Homeland Security and CDC approval for this 

work. Dr. Shtanko is a CDC-registered PI and has been working with high-containment viruses for >9 years. 

Ms. Rodriguez has been working with filoviruses for >5 years. Animal work (mouse, guinea pigs and nonhuman primate) with Ebola, Marburg, and Lassa viruses has been performed at Texas Biomed for >15 years. 

The Biohazard & Safety Committee, Animal Care & Use Committee, and the Institutional Review Board to 

protect the rights and welfare of human research subjects have approved the protocols detailing the work 

described herein. 

Registration status of all entities where Select Agent(s) will be used. Select agent work will only be 

conducted at Texas Biomed. Texas Biomed is a select agent registered entity with Health and Human Services 

(HHS), Centers for Disease Control and Prevention (CDC) and U.S. Department of Agriculture, Animal Plant 

Health Inspection Service, National Select Agent Program. Texas Biomed has been inspected by the CDC 

National Select Agent Program for use of HHS Select Agents and Toxins, Overlap Select Agents and Toxins 

and USDA Select Agents and Toxins. Per the requirements of 42 CFR 73, the most recent certificate of 

registration was issued on November 20, 2017 for use of select agents at BSL2, 3, and 4 and ABSL3 and 4. 

Texas Biomed registration number is 20171120-1976. Texas Biomed has a Responsible Official (RO) and an 

Alternate Responsible Official. 

Facilities where the Select Agents will be used. We have one ABSL4 full-suit lab which is registered for 

CDC and USDA select agent work on human and animal pathogens. Access to the ABSL4 area is secured 

via three successive magnetic card/keypad controlled airlock passages: The first is located at the entry foyer 

to the BSL3 suite, the second at the entry foyer to the outer changer room, and the third at the entry foyer to the 

ABSL4 area. The ABSL4 area is equipped with an entry and exit decontamination airlock and an emergency 

exit/decontamination chamber. The latter also serves as an equipment decontamination chamber to allow for 

the repair, removal and/or replacement of faulty equipment without requiring a complete laboratory 

decontamination process. All supply and exhaust air for the ABSL4 passes through dual (and tandem) HEPA 

filters. The laboratory contains 3 x class IIB biological safety hoods, 4 x water-jacketed CO2 incubators, low-, 

high- and ultra-speed centrifuges equipped with both analytical and preparative scale rotors, a microfuge, 2 x 

4C refrigerators, 2 x liquid nitrogen storage systems, and 3 x -80C freezers. The ABSL4 also has 2 

CytoSMART Lux systems, an optical ELISA plate reader, MAGPIX multiplexing 96-well format unit, 

luminometer, Innova 4000 environmental shaker, microplate shaker, BioFlo III fermenters with chillers, Evos 

fluorescent microscope, pass- through autoclave, dunk tank, facsimile machine and networked computers. 

Housing for animals in the ABSL4 space includes 2 rodent racks able to accommodate 36 cages for guinea 

pigs or up to 480 mice. Non-human primate (NHP) housing consists of squeeze back cages to house up to 12 

marmosets and 24 macaques which are placed within a microisolated enclosure. Vetscan and Procyte 

hematology and blood chemistry Abaxis analyzers are available in the space for analysis of blood collected 

from infected animals. Coagulation parameters can be measured in the lab with Idexx analyzers. Specialized 

equipment for monitoring biopotentials of NHPs in the ABSL4 includes the DSI (Data Science International) 

Telemetry System with analysis software. A dedicated procedure area is located within the lab to perform routine 

animal sampling and full necropsies. 

Critical parameters (air supply and exhaust flow rates, pressure differentials, normal and emergency breathing 

air, decontamination systems, etc.) are continuously monitored. Personnel within the lab communicate, and 

are actively monitored by outside personnel, through a scrambled UHF communication system worn inside 

the laboratory suit. The 500 sq. ft. ABSL4 laboratory staging area includes an air locked suit change room, two 

inner change rooms with pass-through showers, and an outer “clean” change room equipped with sinks, 

Select Agent Research Page 173 
 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

lockers, toilets, etc. Critical mechanical support devices are located in an attached 3,000 sq. ft. mechanical 

support complex. All critical equipment and devices (valves, etc.) are redundant, including duplicate air-supply 

and exhaust systems, decontamination shower systems, as well as the compressors, pumps, valves, heater 

cores, etc. that comprise the liquid waste decontamination system. All critical mechanical devices operate on 

a system with automatic emergency electricity backup generators. The ABSL4 area (including mechanical 

support rooms) is equipped with an elaborate intrusion detection system that includes electronic door and roof 

hatch sensors, motion detectors, etc. 

Texas Biomed maintains an experienced and trained staff of scientists, veterinarians, research technicians 

and veterinary technicians available to perform studies at high biocontainment. These individuals have 

demonstrated proficiency at conducting in vitro and animal studies with the agent identified in the proposal. 

The ABSL4 Operations and Safety Manuals specify policies, procedures, and standard operating procedures 

(SOPs) for the safe handling of biological materials in biosafety laboratories. The policies, procedures, and 

SOPs comply with applicable Federal, State, and municipal regulations and with the guidelines “Biosafety in 

Microbiological and Biomedical Laboratories” issued by the CDC and the National Institute of Health (NIH). 

Employees are trained from these manuals on each facility mechanical systems, biosafety, biocontainment 

and security. Employees are also trained according to project specific and Departmental SOPs. These 

procedures apply to all that use, generate, store, or dispose of potentially infectious materials in Texas Biomed 

biosafety laboratories and to persons who must enter these laboratories to perform services. 

Select agent use, transfer or possession is forbidden without the permission of the Responsible Official (RO) 

and until the required forms filed and written approval received from the CDC Select Agent Program. Upon 

approval, ABSL4 investigators desiring to work on a BSL4 project must also submit an application to the 

Biohazard and Safety Committee for approval. The Committee is responsible for evaluating the facility, 

equipment, and staff capabilities to perform work in a safe manner. 

Infectious cultures and inventory stocks are stored inside the ABSL4 laboratory in locked refrigerators and 

freezers. An electronic Inventory (eInventory) system is used to document usage of infectious stocks. All 

infectious material stored in refrigerators or freezers is properly labeled and stored in containers capable of 

withstanding thermal shock of freezing and thawing. Each container is labeled with the identity of the infectious 

agent, the date of the preparation, and a barcode that links the material to the more inclusive information 

contained in the inventory database. 

When work is completed, all infectious cultures are removed from workbenches and cabinets and stored in a 

designated refrigerator or freezer. Materials to be discarded are placed in a sealable container filled with a 

suitable disinfectant. The container is placed in a discard pan containing the disinfectant. Discard pans are 

placed on a cart and transported to the autoclave. Labware containing infectious liquids are stored and 

transported in leak-proof containers large enough to contain the fluid in case of leakage. 

Select Agent Research Page 174 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

References: 

1. Bieniasz, P.D., Late budding domains and host proteins in enveloped virus release. Virology, 2006. 

344(1): p. 55-63. 

2. Calistri, A., et al., Role of multivesicular bodies and their components in the egress of enveloped RNA 

viruses. Rev Med Virol, 2009. 19(1): p. 31-45. 

3. Chen, B.J. and R.A. Lamb, Mechanisms for enveloped virus budding: can some viruses do without an 

ESCRT? Virology, 2008. 372(2): p. 221-32. 

4. Hartlieb, B. and W. Weissenhorn, Filovirus assembly and budding. Virology, 2006. 344(1): p. 64-70. 

5. Harty, R.N., No exit: targeting the budding process to inhibit filovirus replication. Antiviral Res, 2009. 

81(3): p. 189-97. 

6. Jasenosky, L.D. and Y. Kawaoka, Filovirus budding. Virus Res, 2004. 106(2): p. 181-8. 

7. Liu, Y. and R.N. Harty, Viral and host proteins that modulate filovirus budding. Future Virol, 2010. 5(4): 

p. 481-491. 

8. Urata, S. and J.C. de la Torre, Arenavirus budding. Adv Virol, 2011. 2011: p. 180326. 

9. Chen, H., et al., TSG101: a novel anti-HIV-1 drug target. Curr Med Chem, 2010. 17(8): p. 750-8. 

10. Jiang, Y., X. Liu, and E. De Clercq, New therapeutic approaches targeted at the late stages of the HIV-1 

replication cycle. Curr Med Chem, 2011. 18(1): p. 16-28. 

11. Adamson, C.S. and E.O. Freed, Novel approaches to inhibiting HIV-1 replication. Antiviral Res, 2010. 

85(1): p. 119-41. 

12. Harty, R.N., et al., A PPxY motif within the VP40 protein of Ebola virus interacts physically and functionally 

with a ubiquitin ligase: implications for filovirus budding. Proc Natl Acad Sci U S A, 2000. 97(25): p. 

13871-6. 

13. Harty, R.N., et al., A proline-rich motif within the matrix protein of vesicular stomatitis virus and rabies 

virus interacts with WW domains of cellular proteins: implications for viral budding. J Virol, 1999. 73(4): 

p. 2921-9. 

14. Irie, T., et al., Budding of PPxY-containing rhabdoviruses is not dependent on host proteins TGS101 and 

VPS4A. J Virol, 2004. 78(6): p. 2657-65. 

15. Licata, J.M., et al., Overlapping motifs (PTAP and PPEY) within the Ebola virus VP40 protein function 

independently as late budding domains: involvement of host proteins TSG101 and VPS-4. J Virol, 2003. 

77(3): p. 1812-9. 

16. Urata, S. and J. Yasuda, Regulation of Marburg virus (MARV) budding by Nedd4.1: a different WW 

domain of Nedd4.1 is critical for binding to MARV and Ebola virus VP40. J Gen Virol, 2010. 91(Pt 1): p. 

228-34. 

17. Wirblich, C., et al., PPEY motif within the rabies virus (RV) matrix protein is essential for efficient virion 

release and RV pathogenicity. J Virol, 2008. 82(19): p. 9730-8. 

18. Zhadina, M. and P.D. Bieniasz, Functional interchangeability of late domains, late domain cofactors and 

ubiquitin in viral budding. PLoS Pathog, 2010. 6(10): p. e1001153. 

19. Irie, T., J.M. Licata, and R.N. Harty, Functional characterization of Ebola virus L-domains using VSV 

recombinants. Virology, 2005. 336(2): p. 291-8. 

20. Dolnik, O., et al., Tsg101 is recruited by a late domain of the nucleocapsid protein to support budding of 

Marburg virus-like particles. J Virol, 2010. 84(15): p. 7847-56. 

21. Martin-Serrano, J., T. Zang, and P.D. Bieniasz, HIV-1 and Ebola virus encode small peptide motifs that 

recruit Tsg101 to sites of particle assembly to facilitate egress. Nat Med, 2001. 7(12): p. 1313-9. 

22. May, E.R., et al., The flexible C-terminal arm of the Lassa arenavirus Z-protein mediates interactions with 

multiple binding partners. Proteins, 2010. 78(10): p. 2251-64. 

23. Pasqual, G., et al., Old world arenaviruses enter the host cell via the multivesicular body and depend on 

the endosomal sorting complex required for transport. PLoS Pathog, 2011. 7(9): p. e1002232. 

24. Perez, M., R.C. Craven, and J.C. de la Torre, The small RING finger protein Z drives arenavirus budding: 

implications for antiviral strategies. Proc Natl Acad Sci U S A, 2003. 100(22): p. 12978-83. 

25. Pornillos, O., et al., Structure of the Tsg101 UEV domain in complex with the PTAP motif of the HIV-1 p6 

protein. Nat Struct Biol, 2002. 9(11): p. 812-7. 

26. Pornillos, O., et al., Structure and functional interactions of the Tsg101 UEV domain. EMBO J, 2002. 

21(10): p. 2397-406. 

27. Timmins, J., et al., Ebola virus matrix protein VP40 interaction with human cellular factors Tsg101 and 

Nedd4. J Mol Biol, 2003. 326(2): p. 493-502. 

References Cited Page 175 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

28. Urata, S., et al., Interaction of Tsg101 with Marburg virus VP40 depends on the PPPY motif, but not the 

PT/SAP motif as in the case of Ebola virus, and Tsg101 plays a critical role in the budding of Marburg 

virus-like particles induced by VP40, NP, and GP. J Virol, 2007. 81(9): p. 4895-9. 

29. Urata, S., et al., Cellular factors required for Lassa virus budding. J Virol, 2006. 80(8): p. 4191-5. 

30. Loughran, H.M., et al., Quinoxaline-based inhibitors of Ebola and Marburg VP40 egress. Bioorg Med 

Chem Lett, 2016. 26(15): p. 3429-35. 

31. Han, Z., et al., ITCH E3 Ubiquitin Ligase Interacts with Ebola Virus VP40 to Regulate Budding. J Virol, 

2016. 

32. Madara, J.J., et al., The multifunctional Ebola virus VP40 matrix protein is a promising therapeutic target. 

Future Virol, 2015. 10(5): p. 537-546. 

33. Han, Z., et al., ALIX Rescues Budding of a Double PTAP/PPEY L-Domain Deletion Mutant of Ebola 

VP40: A Role for ALIX in Ebola Virus Egress. J Infect Dis, 2015. 212 Suppl 2: p. S138-45. 

34. Han, Z., et al., Calcium Regulation of Hemorrhagic Fever Virus Budding: Mechanistic Implications for 

Host-Oriented Therapeutic Intervention. PLoS Pathog, 2015. 11(10): p. e1005220. 

35. Lu, J., et al., A host-oriented inhibitor of Junin Argentine hemorrhagic fever virus egress. J Virol, 2014. 

88(9): p. 4736-43. 

36. Han, Z., et al., Small-molecule probes targeting the viral PPxY-host Nedd4 interface block egress of a 

broad range of RNA viruses. J Virol, 2014. 88(13): p. 7294-306. 

37. Liu, Y., et al., Bimolecular Complementation to Visualize Filovirus VP40-Host Complexes in Live 

Mammalian Cells: Toward the Identification of Budding Inhibitors. Adv Virol, 2011: 341816. 

38. Lu, J., et al., Host IQGAP1 and Ebola virus VP40 interactions facilitate virus-like particle egress. J Virol, 

2013. 87(13): p. 7777-80. 

39. Okumura, A., et al., Suppressor of Cytokine Signaling 3 Is an Inducible Host Factor That Regulates Virus 

Egress during Ebola Virus Infection. J Virol, 2015. 89(20): p. 10399-406. 

40. Han, Z., et al., Small-Molecule Probes Targeting the Viral PPxY-Host Nedd4 Interface Block Egress of a 

Broad Range of RNA Viruses. J Virol, 2014. 88(13): p. 7294-7306. 

41. Irie, T., et al., In Vivo Replication and Pathogenesis of Vesicular Stomatitis Virus Recombinant M40 

Containing Ebola Virus L-Domain Sequences. Infect Dis (Auckl), 2012. 5: p. 59-64. 

42. Cai, Z.W., et al., Synthesis, SAR, and Evaluation of 4-[2,4-Difluoro-5-

(cyclopropylcarbamoyl)phenylamino]pyrrolo[2,1-f][1,2,4]triazine -based VEGFR-2 kinase inhibitors. 

Bioorg Med Chem Lett, 2008. 18(4): p. 1354-8. 

43. Oikonomakos, N.G., et al., Crystallographic studies on acyl ureas, a new class of glycogen 

phosphorylase inhibitors, as potential antidiabetic drugs. Protein Sci, 2005. 14(7): p. 1760-71. 

44. Bray, M. and T.W. Geisbert, Ebola virus: The role of macrophages and dendritic cells in the pathogenesis 

of Ebola hemorrhagic fever. International Journal of Biochemistry & Cell Biology, 2005. 37(8): p. 1560-

1566. 

45. Hensley, L.E., et al., Pathogenesis of Marburg Hemorrhagic Fever in Cynomolgus Macaques. Journal of 

Infectious Diseases, 2011. 204: p. S1021-S1031. 

46. Maurya, C.K., A. Mazumder, and P.K. Gupta, Phosphorus pentasulfide mediated conversion of organic 

thiocyanates to thiols. Beilstein J Org Chem, 2017. 13: p. 1184-1188. 

47. Heinrich, M.R. and S.Z. Zard, Generation and intermolecular capture of cyclopropylacyl radicals. Org 

Lett, 2004. 6(26): p. 4969-72. 

48. Liu, L.Q., et al., Design, Synthesis, and Activity Study of Water-Soluble, Rapid-Release Propofol 

Prodrugs. J Med Chem, 2020. 63(14): p. 7857-7866. 

49. Stella, V.J. and K.W. Nti-Addae, Prodrug strategies to overcome poor water solubility. Adv Drug Deliv 

Rev, 2007. 59(7): p. 677-94. 

50. Varia, S.A. and V.J. Stella, Phenytoin prodrugs V: In vivo evaluation of some water-soluble phenytoin 

prodrugs in dogs. J Pharm Sci, 1984. 73(8): p. 1080-7. 

51. McDonnell, M.E., et al., Anilino-monoindolylmaleimides as potent and selective JAK3 inhibitors. Bioorg 

Med Chem Lett, 2014. 24(4): p. 1116-21. 

52. Pelletier, J.C., et al., Dipeptide Prodrugs of the Glutamate Modulator Riluzole. ACS Med Chem Lett, 

2018. 9(7): p. 752-756. 

53. Lindell, S.D., et al., The design and synthesis of novel inhibitors of NADH:ubiquinone oxidoreductase. 

Bioorg Med Chem Lett, 2004. 14(2): p. 511-4. 

54. Lu, Q., et al., TSG101 interaction with HRS mediates endosomal trafficking and receptor down-regulation. 

Proc Natl Acad Sci U S A, 2003. 100(13): p. 7626-31. 

References Cited Page 176 
 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

55. Morris, C.R., et al., A Knockout of the Tsg101 Gene Leads to Decreased Expression of ErbB Receptor 

Tyrosine Kinases and Induction of Autophagy Prior to Cell Death. PLoS One, 2012. 7(3): p. e34308. 

56. Carlton, J.G. and J. Martin-Serrano, Parallels between cytokinesis and retroviral budding: a role for the 

ESCRT machinery. Science, 2007. 316(5833): p. 1908-12. 

57. Elia, N., et al., Dynamics of endosomal sorting complex required for transport (ESCRT) machinery during 

cytokinesis and its role in abscission. Proc Natl Acad Sci U S A, 2011. 108(12): p. 4846-51. 

58. Horgan, C.P., et al., Tumor susceptibility gene 101 (TSG101) is a novel binding-partner for the class II 

Rab11-FIPs. PLoS One, 2012. 7(2): p. e32030. 

59. Baldys, A. and J.R. Raymond, Critical role of ESCRT machinery in EGFR recycling. Biochemistry, 2009. 

48(40): p. 9321-3. 

60. Chen, J., et al., Transferrin-directed internalization and cycling of transferrin receptor 2. Traffic, 2009. 

10(10): p. 1488-501. 

61. Stuffers, S., et al., Multivesicular endosome biogenesis in the absence of ESCRTs. Traffic, 2009. 10(7): 

p. 925-37. 

62. Anantpadma, M., et al., Large-Scale Screening and Identification of Novel Ebola Virus and Marburg Virus 

Entry Inhibitors. Antimicrobial Agents and Chemotherapy, 2016. 60(8): p. 4471-4481. 

63. Shtanko, O., et al., Retro-2 and its dihydroquinazolinone derivatives inhibit filovirus infection. Antiviral 

Res, 2018. 149: p. 154-163. 

64. Rodrigo, W.W.S.I., J.C. de la Torre, and L. Martinez-Sobrido, Use of Single-Cycle Infectious Lymphocytic 

Choriomeningitis Virus To Study Hemorrhagic Fever Arenaviruses. Journal of Virology, 2011. 85(4): p. 

1684-1695. 

65. Lukashevich, I.S., et al., Lassa and Mopeia virus replication in human monocytes/macrophages and in 

endothelial cells: Different effects on IL-8 and TNF-alpha gene expression. Journal of Medical Virology, 

1999. 59(4): p. 552-560. 

66. Batra, J., et al., Protein Interaction Mapping Identifies RBBP6 as a Negative Regulator of Ebola Virus 

Replication. Cell, 2018. 175(7): p. 1917- 1930.e13. 

67. Rogers, K.J., et al., Frontline Science: CD40 signaling restricts RNA virus replication in M phi s, leading 

to rapid innate immune control of acute virus infection. Journal of Leukocyte Biology, 2020. 

68. Qiu, X.G., et al., Establishment and Characterization of a Lethal Mouse Model for the Angola Strain of 

Marburg Virus. Journal of Virology, 2014. 88(21): p. 12703-12714. 

69. Rogers, K.J., et al., Acute Plasmodium Infection Promotes Interferon-Gamma-Dependent Resistance to 

Ebola Virus Infection. Cell Reports, 2020. 30(12): p. 4041-51. 

References Cited Page 177 

 

 

 

Contact PD/PI: Harty, Ronald N. 

RESOURCE SHARING PLAN 

Fox Chase Chemical Diversity Center Inc. is committed to rapidly sharing data, following NIH guidelines as 

noted in the document “NIH Grants Policy Statement and the Principles and Guidelines for Recipients of NIH 

Research Grants and Contracts on Obtaining and Disseminating Biomedical Research Resources”. After the 

filing of patents if appropriate, data will be prepared and submitted for publication in scholarly journals. Peer 

reviewed publications will be submitted to publicaccess@nih.gov according to and within the time parameters 

required. Project resources will also be made available as per NIH policy. For example, we will distribute any 

materials requested to collaborators and will fill requests in a timely fashion. 

Sharing model organisms: No model organisms will be generated. 

Genome-wide association studies: No genome-wide association studies will be performed. 

Resource Sharing Plan(s) Page 189 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

AUTHENTICATION OF KEY BIOLOGICAL AND CHEMICAL RESOURCES 

FOX CHASE CHEMICAL DIVERSITY CENTER, INC. 

Medicinal Chemistry. FCCDC will synthesize all new chemical entities and scale-ups at its Doylestown, PA or 

King of Prussia, PA facilities using modern methods of synthetic organic chemistry and established method of 

analysis for chemical identity and purity (e.g. ≥95% by 1H NMR and LC/MS). Record keeping and compound 

registration and management are delineated by our standard operating procedures described below. 

Laboratory Notebook Maintenance and Records Retention: 

1. Ideas, calculations and experimental results are entered into the notebook as soon as possible, preferably 

the same date they occur, so that the laboratory notebook becomes a daily record of the inventor's 

activities. 

2. All notebook pages are signed and dated by the researcher on a daily basis. 

3. All entries are made in the notebook in permanent ink to be as legible and complete as possible. 

4. Entries are made in the notebook without skipping pages or leaving empty spaces at the bottom of a page. 

5. NMR spectral data is archived onto a primary server, and periodically downloaded onto a back-up server. 

6. ACD software is used when printing out NMR data for patent applications and publications, with chemical 

shifts and coupling contents added in. 

Compound Registration and Management: 

1. We require ≥95% purity by one electronic measure (e.g. NMR, LC/MS). If purity is <95%, then the structural 

assignment and data package is reviewed by FCCDC management, and the apparent purity is indicated 

in a separate comments field. 

2. Once approved, the structure, notebook page number, comments, and project number are entered into an 

electronic database for registration and assignment of a unique FCCDC number in Instant J Chem. 

3. Upon receipt of FCCDC number, this number is recorded in the laboratory notebook on the specified page. 

4. Compounds are stored under refrigeration in well-marked vials in a separate compound repository. 

5. Compounds prepared for one project are not cross-screened for a different project, without the express 

written consent of the original collaborator(s). 

CROs and Vendors, We will seek authentication of reagents, starting materials, antibodies, and biological results 

from well-established vendors via their product specifications, work descriptions and statistical analyses, and we 

will independently check or verify these if issues arise. 

Authentication of Key Biological and/or Chemical Resources Page 190 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

University of Pennsylvania - Authentication of Key Biological Resources 

We will use the following antibodies in this proposal: anti-eVP40, anti-Flag (to detect Flag-tagged 

mVP40, and LAFV-Z proteins), anti-Nedd4, and anti-VSV M (Abcam, Cell Signaling Technology, 

Santa Cruz Biotechnology, Cocalico). Each manufacturer will authenticate these antibodies and 

detailed specification sheets will be provided upon receipt or available online. Upon receipt and 

when we use them after a long period of storage (>6 months), we will authenticate these 

antibodies by Western blot using HEK293T cells overexpressing the corresponding gene product 

or HEKs in which corresponding genes have been silenced by gene-specific shRNAs. 

The following cell lines will be used in this proposal: human embryonic kidney epithelial 293T cells 

(HEK293T), baby hamster kidney 21 cells (BHK-21), Vero-E6 cells, HeLa cells. These cells were 

either purchased from ATCC or from the Univ. of Penn. High Throughput Screening (HTS) Core 

where they have been certified to be free of mycoplasma. Cells purchased from ATCC have been 

authenticated by ATCC using Short Tandem Repeat (STR) profiling. We will continuously monitor 

every 10th passages or 12-month of use, whichever comes first for mycoplasma contamination. 

The following BSL-2 viruses will be used in this proposal: vesicular stomatitis virus (VSV, Indiana 

serotype) wild type and various recombinants expressing foreign viral sequences (L-domains). 

These viruses will be authenticated by plaque purification and subsequent sequencing to confirm 

their identity and insertions. 

The other key reagents to be used in this proposal will include expression plasmids. We will 

authenticate all expression plasmids in this proposal by direct sequencing. In subsequent 

amplifications and when we distribute them to the interested parties, we will verify the identity of 

each construct by appropriate restriction enzyme digest, followed by gel electrophoresis, or direct 

DNA sequencing. 

Scientific Rigor and Statistical Analysis . We will statistically evaluate data from all immunoblot 

assays and virus titration experiments (above) as follows. P-values less than 0.05 will be 

interpreted as statistically significant. Briefly, the normality of the data will be assessed by 

visualizing the distribution of each dataset using probability plots and carrying out Shapiro-Wilks 

tests. Data that are not normally distributed will be compared using Mann-Whitney U test, and 

the results of this test will be interpreted in combination with evaluation of the distribution of 

scores for the groups of interest. Data that are not normally distributed (Shapiro Wilks t-test pvalue <0.05) will be log transformed and reassessed for normality. If raw data or transformed 

data are normally distributed, then we will use an F-test to determine equality of variance. For 

those comparisons in which equality of variance is not met (F-test p-value <0.05), we will 

compare data using Welch's t-test. Data that do not violate the assumption of homoscedasticity 

will be compared using Student t-test. 

Authentication of Key Biological and/or Chemical Resources Page 191 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

AUTHENTICATION OF KEY RESOURCES PLAN 

Texas Biomedical Research Institute 

Virus. This project will use low passage, replication competent wild-type EBOV variant Mayinga, MARV strain 

Musoke, and LASV strain Josiah, all available in Texas Biomed’s repository. The virus stocks have been purified 

by ultracentrifugation through a sucrose cushion, and their titer is 107 focus-forming units (FFUs) per mL on 

Vero cells using the fluorescent focus assay. The mouse-adapted MARV strain Angola at a titer of 107 plaqueforming units (PFUs) per mL on Vero cells is also available in the repository. All virus stocks are currently stored 

at ≤-65°C in the ABSL4 laboratory. The stocks have been confirmed mycoplasma- and endotoxin-free by 

MycoSEQ mycoplasma detection and LAL endotoxin kits, respectively. Virus genome of the stocks was either 

deep- sequenced by Illumina sequencing technology or sequenced by PCR to determine that the stocks were 

identical to those reported for NCBI accession numbers: 

EBOV variant Mayinga NC_002549 

MARV strain Musoke NC_001608 

LASV strain Josiah HQ_688672, HQ_688674 

Mouse-adapted MARV strain Angola KM_261523 

Cell lines. Vero cell line will be used for generation of virus stocks and virus stock titering. HeLa cell line will 

be used to test whether small-molecule compounds affect EBOV, MARV, and LASV replication and spread. 

The cells will be obtained from ATCC where authentication of cells is now routinely performed. Cells will be 

routinely checked for mycoplasma contamination. 

Human macrophages. The proposed research will use macrophages to study EBOV, MARV, and LASV spread 

in the presence of small-molecule compounds. Macrophages will be derived from peripheral blood mononuclear 

cells isolated from human subjects by venipuncture. Subjects will be limited to normal healthy adult (18-50 years 

old) volunteers without symptoms of recent infections. Both genders will be studied and to ensure that the 

gender distribution is equal, recruitment will be directed at acquiring a 50% female:male ratio. We will obtain 

venous blood samples from 6 volunteers/year for a total pool of 18 individuals. 

Peripheral blood cells will be obtained from venipuncture, following standard sterile procedures from healthy 

subjects, according to the approved UT Health IRB protocol #HSC20180013H. The date on which blood was 

given and the amount of blood withdrawn will be recorded in a notebook to be kept locked in the medical 

personnel’s (phlebotomist’s) research office. This information and the peripheral blood are the only things 

collected from the human subjects. Specimens and data will be labeled with a numerical code. The subjects’ 

name will not appear on any specimens and participation will remain confidential. 

Monocyte-derived macrophages (MDMs) will be prepared at Texas Biomed as we described. Briefly, heparinized 

human peripheral blood will be layered on a Ficoll-Paque cushion to collect peripheral blood mononuclear cells. 

Subsequently, the cells will be cultured in suspension in medium supplemented with autologous serum to 

differentiate monocytes into macrophages. Macrophages will be harvested and adhered to tissue culture dishes 

while lymphocytes will be washed away. Such macrophage monolayers are 99% pure, viable and highly 

susceptible to infection with viruses to be used in this study. 

A typical MDM yield is over 15-20 million cells/pint of blood from each donor, which is sufficient to complete the 

proposed studies. We will utilize MDMs differentiated in autologous serum to more closely mimic the 

physiological environment. This eliminates artifactual or well-established inflammatory cellular responses that 

may arise from exposure to human AB or bovine sera. 

Mouse macrophages. The proposed studies will use primary peritoneal macrophages isolated from wild-type 

Balb/c mice. These cells will be isolated using well-established, published protocols. Isolated cell populations 

will be characterized visually for the morphology and by immunofluorescence for surface staining of antigens 

specific for macrophages. 

Authentication of Key Biological and/or Chemical Resources Page 192 
 

 

 

 

 

 

Contact PD/PI: Harty, Ronald N. 

Antibodies, Chemicals, siRNAs, PCR primers. The reagents will be selected based on published literature 

and then purchased from reputable commercial vendors in quantities sufficient to complete the proposed studies. 

If it becomes necessary to obtain additional batches, we will purchase only products with identical specifications 

from the same vendors. 

Lymphocytic choriomeningitis virus (LCMV) mouse monoclonal anti-GP antibody (83.6) which cross-reacts with 

LASV GP2 will be provided by Dr. Luis Martinez-Sobrido (Texas Biomed; letter of collaboration is attached). 

Rigor and Reproducibility. For each virus infection experiment, ≥3 independent tests in triplicate will be

performed. Experiments with human MDMs will be performed from ≥6 different healthy adult donors (50:50

female:male ratio) per year. We expect the same outcome trend among donors, although there may be a 

different magnitude of response. In vivo studies will be repeated at least once and will use ≥5 mice/experimental 

condition for both sexes. All data will be analyzed for statistical significance using either a Student t-test to 

compare averages of 2 groups or one-way ANOVA to compare averages of >2 groups, with p<=0.05 being 

considered significant. We will inspect data for normality. Where deviations from normality are seen, we will 

use appropriate transformations, or non-parametric tests. 

Authentication of Key Biological and/or Chemical Resources Page 193