A Second-Generation Patch Test for Tuberculosis

A Second-Generation Patch Test For Tuberculosis

Carol A Nacy, Ceo
Sequella, Inc. 9610 Medical Center Dr, Ste 200 Rockville, Md 208506332

Grant 5U01AI057313-04 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1

Abstract: This proposal outlines a development program to transition a second generation Transdermal Tuberculosis (TB) Skin Test into a commercial product. The first generation of the technology, transdermal delivery of a protein of the M. tuberculosis complex (MPB/T-64) by a Torii patch, has already been tested in three clinical trials performed in the Philippines and Japan it demonstrated remarkable ability to specifically identify active cases of TB (smear positive TB patients), while it was non-reactive in symptom-free individuals PPD positive from either BCG vaccination or prior cure of TB. Sequella licensed this exciting technology from the Japan BCG Laboratory, and GMP manufacturing of recombinant MPT64 began in January 2003. We will begin a World Health Organization-supported clinical trial in South Africa in March 2003 that will test the sensitivity and specificity of this first generation test. We believe that the Transdermal TB Skin Test will be an invaluable tool in the fight against TB. Because of the incredible need for a rapid diagnostic for active TB, we hope to begin distribution in the developing world as soon as possible using the existing test format. However, there are many aspects of the product that need to be improved, and we hope to develop a more effective second generation product in this proposal. Our specific aims include Specific Aim I We will optimize expression, purification, and quality control protocols for MPT64. Specific Aim II We will complete development of a second-generation patch technology that incorporates the protein directly into the patch adhesive. Specific Aim III We will manufacture the second-generation patches for clinical studies using Good Manufacturing Practices. Specific Aim IV We will perform clinical studies of the Transdermal TB Skin Test.

Keywords: communicable disease diagnosis, diagnosis design /evaluation, diagnostic test, noninvasive diagnosis, rapid diagnosis, transdermal drug delivery, tuberculosis, Mycobacterium tuberculosis, bacterial antigen, bacterial protein, clinical trial phase I, clinical trial phase II, cooperative study, diagnosis quality /standard, recombinant protein, Peru, biopsy, bioterrorism /chemical warfare, clinical research, human subject, patient oriented research, protein purification, transfection /expression vector

Project start date: 2003-08-15

Project end date: 2007-01-31

5U01AI057313-04 (2006): $299160

Sponsored Links Excellgen http://Excellgen.com

	Recombinant Lentivirus & Adenovirus High Yield and High Titer up to 10¹⁰ (lentivirus) and 10¹³ (adenovirus) for Guaranteed Expression of GOI. ~~$3000~~, $2500
	Transient Protein Expression in CHO and HEK293 Cells Transient Expression, Truly Functional Protein, 95% purity, 1~20 mg, fast turnaround. ~~$5500~~, $3950
	Baculovirus Protein Expression Fast turn around, >95% purity functional protein. No outsourcing to China or India. ~~$5500~~, $3950

A Second-Generation Patch Test For Tuberculosis

Carol A Nacy, Ceo
Sequella, Inc. 9610 Medical Center Dr, Ste 200 Rockville, Md 208506332

Grant 5U01AI057313-03 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1

Project start date: 2003-08-15

Project end date: 2007-01-31

5U01AI057313-03 (2005): $736439

5U01AI057313-02 (2004): $621250

Grants awarded to Carol A Nacy

Transdermal Test For Active Tuberculosis

Carol A Nacy, Ceo
Sequella, Inc.
9610 Medical Center Dr, Ste 200
rockville, Md 208506332

Grant 1R43AI050273-01 from National Institute Of Allergy And Infectious Diseases IRG: ZRG1

Abstract: This proposal seeks to establish a development program to transition an experimental Transdermal Tuberculosis Skin Test into a commercial product. This technology has already demonstrated a remarkable ability to specifically identify AFB+ TB patients by adminstering a small amount of a TB protein (MPB64) to the skin and monitoring for an erythemic skin response. PPD+ symptomless individuals are non-reactive to the protein. The MPB-64 Transdermal Patch was applied to sixty-two patients, 49 with sputum-positive active disease and 13 who had completed TB chemotherapy, and 28 non-TB but tuberculin-positive controls. The results were read at 72h. The sensitivity of the Transdermal Patch was 87.8 percent, efficacy 92.9 percent, and specificity 100 percent. The thirteen TB patients who had each completed 6 months of TB chemotherapy showed different reactions to MPB64 patch test those who had completed chemotherapy less than 4 months before testing were positive; 50 percent of patients who completed chemotherapy 5 months prior were positive; and those who had completed chemotherapy 7 and 8 months before were negative. All the non-TB controls with positive tuberculin tests were negative to the MPB-64 Transdermal Patch, even at the highest protein dose tested. This test may be a useful method to distinguish active TB patients from TB-infected but asymptomatic individuals. The current version of the technology is a research grade patch. This proposal will outline a development pathway to improve product packaging, ease of use and product stability. Initial efforts will be to characterize a unique animal model for use in examining component materials and technologies. Our specific aims are 1. Validate an animal model to test variables in the formulation and transdermal delivery of the skin test antigen, 2. Establish the most cost effective source of MPB64, 3. Establish the best packaging / delivery technology for the patch, 4. Establish animal test procedures for toxicity, product release and to test product stability. Phase II aims include optimizing protein production and manufacturing scale up, and performance of clinical trials for product registration. PROPOSED COMMERCIAL APPLICATION The proposed commercial application would be to develop a patch test for diagnostic screening of active cases of tuberculosis by using transdermal delivery of MPB64 protein. The patch test could be used in place of other screening tests, such as the PPD skin test or the AFB smear, as a means of diagnosing patients with active tuberculosis

Keywords: bacterial antigen, bacterial protein, diagnosis design /evaluation, technology /technique development, transdermal drug delivery, tuberculin test, tuberculosis Mycobacterium tuberculosis, delayed hypersensitivity

Project start date: 2001-09-30

Project end date: 2002-09-29

1R43AI050273-01 (2001): $232038

A Second-Generation Patch Test For Tuberculosis

Carol A Nacy, Ceo
Sequella, Inc. 9610 Medical Center Dr, Ste 200 Rockville, Md 208506332

Grant 1U01AI057313-01 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1

Project start date: 2003-08-15

Project end date: 2007-01-31

1U01AI057313-01 (2003): $237679

Advancing A New TB Drug Through Early Clinical Trials

Carol A Nacy, Ceo
Sequella, Inc. 9610 Medical Center Dr, Ste 200 Rockville, Md 208506332

Grant 1UC1AI062534-01 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1

Abstract: This application for a Biodefense Product Development Challenge Grant requests support for the early clinical trials of a new tuberculosis drug. The new drug-SQ109-is effective against multi-drug resistant TB, which is a Category C NIAID high priority pathogen. SQ109 was developed by a small biopharmaceutical company, in collaboration with the NIH. All discovery phase research is completed, and the compound is at the final stage of pre-clinical development. The scope of work proposed would advance SQ109 through the final regulatory requirements for filing an IND, through all of the Phase I safety trials, and through two of the anticipated three Phase II clinical trials. The Phase II clinical trials to be completed will be a seven-day study of SQ109 with early bactericidal activity (EBA) as a key indicator of anti-TB efficacy, and a two-month combination-treatment study with sputum conversion as the primary outcome measure. These studies will position SQ109 for the last stages of clinical development-a final, longer-term Phase II study that will serve as a pilot study for the Phase III clinical trial, and the Phase III clinical trial itself. Development of a new TB drug that is effective against multi-drug resistant TB will have enormous public health benefits and global health impact, as well as providing defense against the possible use of TB as a bio-weapon.

Keywords: antibacterial agent, bioterrorism /chemical warfare, clinical trial, drug design /synthesis /production, tuberculosis, clinical trial phase I, clinical trial phase II, cooperative study, ethambutol, human therapy evaluation, method development, multidrug resistance, respiratory disorder chemotherapy, human subject, patient oriented research

Project start date: 2004-09-02

Project end date: 2006-08-31

1UC1AI062534-01 (2004): $1479750

GENE-BASED AND BACTERIA HIGH THROUGHOUT TB DRUG SCREEN

Carol A Nacy, Ceo
Sequella Global Tuberculosis Foundation
9610 Medical Center Dr, Ste 200
rockville, Md 208506332

Grant 1RC1AI048896-01 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1

Abstract: The search for new TB drugs has finally caught up with the pharmaceutical revolution of the last half of this century solid-matrix-based methods of chemical synthesis now create combinatorial chemistry libraries of pharmaceuticals that contain millions of unique compounds. Using robotics and sophisticated assay instrumentation, these libraries can be screened with high throughput to identify compounds which are lethal to specific organisms, or which affect specific pathways known to be critical for bacterial survival. A year-old CRADA between the NIAID and Sequella, Inc. to develop combinatorial chemistry and a high throughput screening assay succeeded in synthesizing and screening approximately 100,000 analogues of the TB drug ethambutol in a very short period of time. The gene-based screen used in these studies identified compounds that affect specific regions of the M. tuberculosis genome that are activated in response to ethambutol therapy, and are involved in the cellular response to cell wall damage. The Sequella, Inc./NIH collaboration had impressive momentum (100,000 compounds screened, 200 hits, 4 lead compounds in 9 months). The Foundation intends to provide several gene-based and whole-cell high throughput screening assays to interested pharmaceutical companies to screen their chemical libraries in a similar timeframe. One company, for example, has 500,000 well- characterized chemicals that have never been tested for TB. They are interested in providing the Foundation with these chemicals to test in a specific whole cell-screening assay as an initial screen for hits. Setting up the high throughput screen in-house, while not that expensive, represents an opportunity cost for that company and would require a BL3 facility. In addition, a validated whole-bacteria high throughput screening assay would be of benefit to the current NIAID/DAIDS drug screening program. The foundation would be interested in transferring the screen to the NIH program over the course of this challenge grant. The Foundation goal is to reduce the barriers to entry that the major pharmaceutical companies face when considering the market for new TB drugs. Thus, providing tailored screening programs that support the specific needs of pharmaceutical partners is a relatively inexpensive endeavor that may improve the likelihood that medicines will emerge to improve the treatment of this neglected disease

Keywords: Mycobacterium tuberculosis, antitubercular agent, drug screening /evaluation, high throughput technology biotechnology

Project start date: 2000-05-05

Project end date: 2000-08-31

1RC1AI048896-01 (2000): $26000

A Rapid Lateral Flow Test For TB In Nonhuman Primates

Carol A Nacy, Ceo
Sequella, Inc. 9610 Medical Center Dr, Ste 200 Rockville, Md 208506332

Grant 5R44RR019845-03 from National Center For Research Resources IRG: ZRG1

Abstract: The goal of this project is to develop and bring to market a new diagnostic test for tuberculosis (TB) in nonhuman primates. The new diagnostic is based upon the detection, in infected animals, of antibodies to a set of recombinant TB antigens. The proposed format is a proprietary lateral flow test that uses a drop of blood and gives a visual result within 15 minutes. Two small U.S. biotechnology companies with complementary areas of expertise, Sequella Inc. and Chembio Diagnostic Systems, have entered into a cooperative research and development venture to develop and market the new diagnostic. Following on the success of a Phase I feasibility study, the work proposed for Phase II is tightly focused on rapidly completing the development and validation of the test. As required by the USDA, an Outline of Production detailing process standardization, quality control, and the method for assuring equivalency of each lot of diagnostic tests will be prepared for the approval of the USDA. Test kits in the final commercial format will be manufactured and used to obtain the efficacy data for licensure. The claims to be sought are that the test will diagnose TB caused by either Mycobacterium tuberculosis (M. tb) or M. bovis in macaque and saimiri species of primates, and that the test does not give false positive results due to infections/exposures to M. avium/intracellulare, M. kansasii, M. gordonae, or M. scrofulaceum. To the extent possible, sera from naturally infected animals will used to estimate sensitivity and specificity. Experimental infections will be conducted as needed to provide the data required by the USDA. The resulting test is expected to have advantages over the present tuberculin skin test in terms of sensitivity, specificity, and operational simplicity. Many individuals and entities involved in the management of nonhuman primate colonies, including the Association of Primate Veterinarians, the NIH National Center for Research Resources, and the Centers for Disease Control have called for the development of a better, more reliable test for TB in nonhuman primates, and it has been termed an "urgent need" by the NIH SBIR program.

Project start date: 2001-09-01

Project end date: 2007-08-31

5R44RR019845-03 (2004): $986376

9R44RR019845-02A1 (2003): $986376

1R43AI050271-01 (2001): $162215

ANTIBODY BASED DIAGNOSTIC FOR TB IN NONHUMAN PRIMATES

Carol A Nacy, Ceo
Sequella, Inc.
9610 Medical Center Dr, Ste 200
rockville, Md 208506332

Grant 1R43AI043812-01 from National Institute Of Allergy And Infectious Diseases IRG: ZRG5

Abstract: Sequella will explore use of early-onset Ab produced by infection of rhesus monkeys with M. tb as a potential diagnostic for identifying or confirming absence of this disease in quarantined animals and nonhuman primate colonies. The current skin test reagent produces unacceptably high numbers of false negative and positive reactions. The purpose of this Phase I proposal is (a) to identify antigens of M. tb to which nonhuman primates (as opposed to humans) develop immunological reactivity, and (b) to determine the time course and immunoglobulin class of Ab produced to TB antigens in serum and in feces. The results of these studies will help select both the correct antigen(s) to detect early Ab and the type of Ab detection system (by Ab class) that will most appropriately form the basis of a new serologic or scatologic diagnostic. A successful Phase I study would stimulate a Phase II application to confirm Ab to the same or different antigens in M. bovis- infected rhesus monkeys, extend these observations to other nonhuman primates infected with the two etiologic agents of TB, identify the appropriate test format for highest sensitivity and specificity, and perform clinical trials to validate the serologic/scatologic diagnostic and compare it to the current skin test. PROPOSED COMMERCIAL APPLICATION Sequella will develop a proprietary position with Mycos, Inc. on antigens that are specifically useful for the diagnosis of TB in nonhuman primates; Sequella will also patent any innovative technology it develops to detect Ab, (specifically anti-mycobacterial Ab) in serum or feces. Should a sensitive and specific diagnostic be the result of these studies, and the clinical evaluation suggests that this test is superior to the currently approved skin test, Sequella and its partners will seek approval of the diagnostic by the USDA as an adjunct or replacement for the skin test. With approval, Sequella and its partners will have such the diagnostic manufactured and marketed to interested veterinary groups, in private or public institutions

Keywords: antibacterial antibody, bacterial antigen, diagnosis design /evaluation, serology /serodiagnosis, tuberculin test, tuberculosis Mycobacterium bovis, Mycobacterium tuberculosis, immunologic skin test Macaca mulatta, SDS polyacrylamide gel electrophoresis

Project start date: 1998-09-01

Project end date: 2000-02-29

1R43AI043812-01 (1998): $100000

DETECTION OF INTRACELLULAR PATHOGENS BY FLOW CYTOMETRY

Carol A Nacy, Ceo
Entremed, Inc.
9610 Medical Center Dr, Ste 200
rockville, Md 20850

Grant 1R41AI036572-01 from National Institute Of Allergy And Infectious Diseases IRG: ZRG7

Abstract: Many intracellular pathogens that infect man fail to induce detectable levels of antibodies until after disease symptoms appear. Thus, early diagnosis by classical antibody-based serological techniques is problematic. In such cases, an easy, sensitive, specific, and rapid method for identification of the pathogen itself is required. EntreMed has developed proprietary technology using flow cytometry to detect intracellular pathogens in specific and sensitive screening assays. Flow cytometry differentiates cell populations based on their size, internal complexity, and antibody-defined surface markers. Monoclonal antibodies can also be used in this system to identify pathogens inside cells. The short-term objective of this proposal is to refine and increase the sensitivity of a flow cytometric technique for diagnosis of active Leishmania-infections. The long-term objective, however, is to develop this method as a platform technology for diagnosis of infections caused by a range of intracellular pathogens. Most hospitals routinely use flow cytometric analyses for diagnosis of certain diseases (HIV, leukemia, etc.). Thus, a well defined, technologically informed commercial market presently exists for this assay. A large pharmaceutical company that participated in the concept´s early development, remains interested in possible commercial application of the technology, if feasibility is demonstrable in Phase I

Keywords: communicable disease diagnosis, diagnosis design /evaluation, flow cytometry, leishmaniasis Leishmania, antimicroorganism antibody, diagnosis quality /standard, intracellular parasitism, monoclonal antibody, monocyte, protozoal antigen, rapid diagnosis, surface antigen fluorescent dye /probe, human tissue, microorganism culture

Project start date: 1994-08-15

Project end date: 1995-08-14

1R41AI036572-01 (1994): $100000

High Throughput TB Drug Screens

Carol A Nacy, Ceo
Sequella Global Tuberculosis Foundation 9610 Medical Center Dr, Ste 200 Rockville, Md 208506332

Grant 5R01AI049497-03 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1

Abstract: adapted from applicant s ) The search for new TB drugs has finally caught up with the pharmaceutical revolution of the last half of this century solid-matrix-based methods of chemical synthesis now create combinatorial chemistry libraries of pharmaceuticals that contain millions of unique compounds. Using robotics and sophisticated assay instrumentation, these libraries can be screened with high throughput to identify compounds which are lethal to specific organisms, or which affect specific pathways know to be critical for bacterial survival. A year-old CRADA between the NIAID and Sequella, Inc. to develop combinatorial chemistry and a high throughput screening assay succeeded in synthesizing and screening approximately 100,000 analogues of the TB drug ethambutol in a very short period of time. The gene- based screen used in these studies identified compounds that affect specific regions of the M. tuberculosis genome that are activated in response to ethambutol therapy, and are involved in the cellular response to cell wall damage. The Sequella, Inc./NIH collaboration had impressive momentum (100,000 compounds screened, 200 hits, 4 lead compounds in 9 months). The Foundation intends to provide several gene-based and whole-cell high throughput screening assays to interested pharmaceutical companies to screen their chemical libraries in a similar timeframe. One company, for example, has 500,000 well- characterized chemicals taht have never been tested for TB. They are interested in providing the Foundation with these chemicals to test in a specific whole cell-screening assay as an initial screen for hits. Setting up the high throughput screen in-house, while not that expensive, represents an opportunity cost for that company and would require a BL3 facility. In addition, a validated whole-bacteria high throughput screening assay would be of benefit to the current NIAID/DAIDS drug screening program. The Foundation would be interested in transferring the screen to the NIH program over the course of this challenge grant. The Foundation goal is to reduce the barriers to entry that the major pharmaceutical companies face when considering the market for new TB drugs. Thus, providing tailored screening programs that support the specific needs of pharmaceutical partners is a relatively inexpensive endeavor that may improve the likelihood that medicines will emerge to improve the treatment of this neglected disease.

Keywords: Mycobacterium tuberculosis, antitubercular agent, drug screening /evaluation, genetic technique, high throughput technology, bacterial genetics, combinatorial chemistry, ethambutol, biotechnology, microorganism culture

Project start date: 2001-09-20

Project end date: 2004-06-30

5R01AI049497-03 (2003): $172765

Sponsored Links Excellgen http://Excellgen.com

	Recombinant Lentivirus & Adenovirus High Yield and High Titer up to 10¹⁰ (lentivirus) and 10¹³ (adenovirus) for Guaranteed Expression of GOI. ~~$3000~~, $2500
	Transient Protein Expression in CHO and HEK293 Cells Transient Expression, Truly Functional Protein, 95% purity, 1~20 mg, fast turnaround. ~~$5500~~, $3950
	Baculovirus Protein Expression Fast turn around, >95% purity functional protein. No outsourcing to China or India. ~~$5500~~, $3950

1R01AI049497-01 (2001): $150000

Carol A Nacy
Sequella, Inc.

Project start date: 2012-03-01

Project end date: 2014-02-28