SBIR-STTR Award

HIV remains a dangerous and prevalent disease globally contributing to millions of infections and deaths per year and tens of billions of dollars in healthcare costs. The use of Truvada as Pre Exposure Prophylaxis (PrEP) is 99% effective at preventing HIV infection if taken daily, but its impact on reducing HIV burden is limited by poor adherence. PrEP is recommended by the Centers for Disease Control (CDC) in populations known to have a high risk of HIV infection, which include men who have sex with men, people with a history of an STI, intravenous drug users, and people with a partner who is HIV positive. Monitoring of drug adherence is well known to improve drug compliance, but there are no commercial products for adherence monitoring of PrEP. Hence, there is a critical unmet need for a tool that will allow physicians to monitor adherence to PrEP in their patients. In order to be acceptable to patients on a routine basis and feasible in the physician workflow, this adherence test needs to be noninvasive, painless, inexpensive, and provide rapid, accurate results. The overall goal of this project is to develop a point-of-care (POC) test that will measure patient adherence with the PrEP regimen and ultimately prevent HIV infection in people known to be at risk of exposure. The POC test will be based on a laboratory-based mass spectrometry urine test for levels of tenofovir, one of the drugs in PrEP, which UrSure already developed. The POC assay will be faster (minutes to get a result) and therefore can be used during a clinic visit to measure PrEP adherence and, if appropriate, counsel patients on how to improve their compliance. The aims of this Phase I project are to: 1) synthesize four tenofovir derivatives and conjugate them to form two immunogens for polyclonal antibody production in Aim 2 and two HRP-conjugated proteins for ELISA use in Aim 3; 2) produce two affinity purified polyclonal antibodies against each of the two immunogens produced in Aim 1; and 3) develop a prototype ELISA using these derivatives and antibodies, which will be validated with urine samples positive and negative for tenofovir. The final deliverables of this Phase I project will be conjugated tenofovir derivatives, polyclonal antibodies to these derivatives, and an ELISA, all of which will be used to develop monoclonal antibodies and ultimately a Lateral Flow Immunoassay POC urine tenofovir test in a subsequent Phase II application. This Phase I project will be a critical step towards developing our POC urine test and bringing a powerful tool with the ability to drive PrEP adherence and reduce HIV infections to physicians worldwide.

Public Health Relevance Statement:
PROJECT NARRATIVE Pre-exposure prophylaxis with drugs such as tenofovir can prevent new HIV infections and end the HIV epidemic, but eligible people at high risk for HIV exposure have a very low success rate in adhering to the recommended drug regimen. Successful completion of this research program will result in a rapid, non- invasive point-of-care urine test to monitor tenofovir use. Test results will be used during clinic visits to encourage and increase drug adherence in order to prevent new HIV infections.

Project Terms:
Adherence; Affinity; AIDS prevention; AIDS/HIV problem; Antibodies; Antibody Formation; Antigens; assay development; base; Bedside Testings; Biological Assay; Blood; Centers for Disease Control and Prevention (U.S.); Cessation of life; Chemicals; Clinic; Clinic Visits; Clinical; Compliance behavior; Counseling; Dangerousness; design; Devices; Disease; Dose; Drug Combinations; Drug Monitoring; Eligibility Determination; Enzyme-Linked Immunosorbent Assay; Epidemic; Equipment; Future; Goals; Hair; Health Care Costs; high risk; HIV; HIV Infections; HIV Seropositivity; Hour; Immunoassay; immunogenic; improved; Individual; Infection; innovation; Interruption; intravenous drug user; Laboratories; Lateral; Liquid Chromatography; Mass Spectrum Analysis; Measures; medication compliance; men who have sex with men; Methods; Monitor; Monoclonal Antibodies; novel; Painless; Patients; Persons; Pharmaceutical Preparations; Phase; Physicians; point of care; polyclonal antibody; Population; pre-exposure prophylaxis; prevent; programs; Proteins; prototype; Recording of previous events; Regimen; Research; Resources; Risk; Sampling; Sensitivity and Specificity; Sexually Transmitted Diseases; Small Business Innovation Research Grant; Specimen Handling; success; tandem mass spectrometry; Tenofovir; Test Result; Testing; tool; truvada; Urine; Work

The goal of this Phase II SBIR project is to develop a point-of-care (POC) test to measure patient adherence with Pre-exposure Prophylaxis (PrEP) as a prevention against HIV infection in people known to be at risk of HIV exposure. HIV contributes to millions of infections and deaths per year and tens of billions of dollars in healthcare costs around the world. PrEP is recommended by the Centers for Disease Control and World Health Organization in people at high risk for HIV exposure. It is 99% effective at preventing HIV infection if taken daily, but its impact on reducing HIV burden is limited by poor adherence. Monitoring actual drug use by patients improves drug adherence, but there is no commercial test for PrEP. The UrSure rapid TFV POC test will measure PrEP adherence based on the UrSure laboratory-based mass spectrometry urine test for tenofovir (TFV), which is a component of both PrEP and Truvada. The POC test will be noninvasive, painless, rapid (minutes to get a result) and inexpensive. It can be used during a clinic visit to measure PrEP adherence and, if appropriate, counsel patients on how to improve compliance. Since Truvada is a common first-line therapy for confirmed HIV infection, this test also will measure adherence in known HIV-positive patients who do not have access to frequent viral load testing. Phase I of this project demonstrated the feasibility of this approach. A novel TFV derivative was synthesized and used to produce polyclonal antibodies that are specific and sensitive for TFV. A prototype ELISA was developed and validated with urine samples confirmed as positive or negative for TFV, for use in evaluating the POC test. A urine sample bank with known TFV results was established. The objective of this Phase II project is to advance development of the UrSure TFV POC test toward commercialization. The Aims are to: 1) produce up to three monoclonal antibodies with sensitivity and specificity >95% for TFV in urine; 2) use those monoclonal antibodies to develop a Lateral Flow Immunoassay (LFIA) strip; 3) integrate that strip with a self-contained assay cassette and optimize its performance; 4) verify the performance of the LFIA and prepare it for manufacture; and 5) transfer the LFIA prototype to a contract manufacturer and produce validation lots. UrSure has relationships in place with LFIA experts and organizations with deep experience in LFIA assay development, validation and assay kit production. The final deliverables of this Phase II project will be monoclonal antibodies sensitive and specific for TFV in urine, a locked prototype of a LFIA for rapid detection of TFV in urine, and a lot of 1,000 UrSure TFV POC tests with at least 85% sensitivity and specificity for urine samples positive and negative for TFV. Successful completion of this Phase II project will result in a prototype suitable for initiating the FDA regulatory process. A Class 2 de novo classification is expected and will support commercialization of the POC test.

Thesaurus Terms:
Adherence; Advanced Development; Affinity; Aids Prevention; Aids/Hiv Problem; Antibodies; Architecture; Assay Development; B-Lymphocytes; Base; Bedside Testings; Biological Assay; Blood; Centers For Disease Control And Prevention (U.S.); Cessation Of Life; Classification; Clinic; Clinic Visits; Clinical; Clinical Application; Commercialization; Compliance Behavior; Contracts; Counseling; Cross Reactivity; Custom; Dangerousness; Development; Devices; Disease; Documentation; Dose; Drug Combinations; Drug Monitoring; Drug Usage; Enzyme-Linked Immunosorbent Assay; Epidemic; Equipment; Evaluation; Experience; Future; Goals; Health Care Costs; High Risk; Hiv; Hiv Infections; Hiv Seropositivity; Hour; Immunoassay; Improved; Individual; Industry Standard; Infection; Innovation; Institutes; Interruption; Intravenous Drug User; Laboratories; Lateral; Liquid Chromatography; Manufacturer Name; Mass Spectrum Analysis; Measures; Medical Device; Medication Compliance; Methods; Monitor; Monoclonal Antibodies; Novel; Oryctolagus Cuniculus; Painless; Patients; Performance; Persons; Pharmaceutical Preparations; Phase; Point Of Care; Polyclonal Antibody; Pre-Exposure Prophylaxis; Prevent; Prevention; Procedures; Process; Production; Programs; Prototype; Quality Control; Rapid Detection; Regimen; Reporting; Research; Resources; Risk; Sampling; Sensitivity And Specificity; Sexually Transmitted Diseases; Small Business Innovation Research Grant; Specificity; Specimen Handling; Success; Tandem Mass Spectrometry; Tenofovir; Test Result; Testing; Truvada; Urine; Validation; Viral Load Result; Work; World Health Organization;