ABSTRACT
The phase I SBIR application, from which this phase II application is a continuation, was in response to the
NIEHS solicitation for “Improved Test Systems for Prioritization and Safety Evaluation.” Specifically, the
application was written to address the need for a nonanimal ocular safety test to differentiate ocular nonirritant
substances from irritant and corrosive substances. Ocular safety tests are routinely conducted on live animals
with a time to results of up to 21 days and are considered by many to be inhumane. Recent government
legislation and consumer pressure are directed at reducing or eliminating the use of live animals for the routine
testing of consumer products and cosmetics. However, limited options exist for nonanimal testing. Currently,
accepted nonanimal test methods require the advanced purchase (up to three weeks) of live tissues and a
technician skilled in aseptic technique; additionally, such tests are unable to classify compounds that cause
reversible irritation, do not have a long shelf life, often misclassify ocular corrosives as nonirritants, take a long
time (both hands-on and incubation), and require other advanced laboratory techniques including cell culture or
dissection and subjective interpretation of ocular lesions. The subject of this proposal is the production of a
rapid, irritancy-predictive, quantitative, standardized test kit with a one-year shelf life to provide standardized
results more accurately and with higher sensitivity than any other available nonanimal ocular irritation test.
SUCCESSFUL COMPLETION OF ALL SBIR PHASE I AIMS
Using chemicals selected and coded by a government agency (NICEATM), and evaluated by representatives
from U.S. governmental agencies [ICCVAM Ocular Toxicology Working Group (OTWG)], the phase I SBIR
demonstrated that the test kit has a transferability of 92%, according to results from a blind transferability
study. For the detection of nonirritants, out of the 90 chemicals evaluated, no GHS false-negative results were
obtained (100% sensitivity). The overall sensitivity and accuracy outperformed all other available nonanimal
ocular irritation test methods. In addition to the test being rapid and shelf stable, test kit performance for the
detection of nonirritant compounds was excellent.
Only a limited number of ocular corrosives were included in the validation study. While the overall sensitivity for
ocular corrosives was 87%, only 16 ocular corrosives were tested in total. Additional validation with an
increased number of ocular corrosives will better define the sensitivity of the method and improve confidence in
the results.
AIMS OF THE PHASE II APPLICATION
During the first year of the phase II project, we propose to further optimize the test kit and convert
manufacturing to full GMPs.
During the second year of the project, we propose a second, larger and more complete validation study with
90–140 substances, tested in triplicate. As selected by NICEATM and the OTWG, we will test a large number
of coded ocular corrosives, irritants, and nonirritants, representative of different chemical and physical classes.
NICEATM has already agreed to select, purchase, and provide these substances to us in coded form without
charge. Concurrently, since the test is already in kit form, we will initiate low-volume kit sales, scale up GMP
production operations, and seek regulatory approval through the U.S. government (NICEATM and ICCVAM)
and worldwide (OECD).
Additional development and validation studies are required to better define accuracy (EPA classes I, II, III, and
IV, and GHS categories 1, 2, and NC) and determine whether the test is able to detect compounds in all
classes of irritants, ranging from slight irritants to corrosives. In addition, we will scale up and convert
production operations to full GMP production. Concomitantly, we will seek worldwide regulatory acceptance
through the OECD. An accurate, multiclass, rapid, stable, nonanimal eye safety test kit represents a significant
business opportunity in the current market of consumer products and cosmetics, and will make available an
accurate, validated, humane, and standardized test kit that will provide a needed solution to both private
industry and regulatory agencies.
