SEC Company Response from NANOVIRICIDES, INC. (NNVC) (CIK 0001379006) (NNVC) — Feb 14, 2007
NANOVIRICIDES, INC. (NNVC) (CIK 0001379006)
Date: Feb. 14, 2007 · CIK: 0001379006 · Accession: 0001140361-07-003550
AI Filing Summary & Sentiment
Referenced dates: February 2, 2007
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CORRESP
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filename1.htm
NanoViricides, Inc. Corresp 02-14-2007
LEVY
& BOONSHOFT, P.C.
ATTORNEYS
AT LAW
477
Madison
Avenue
New
York,
New York 10022
(212)
751-1414
Facsimile:
(212) 751-6943
February
14, 2006
Via
EDGAR and OVERNIGHT DELIVERY
Mr.
Jeffrey Riedler
Assistant
Director
Securities
and Exchange Commission
Division
of Corporate Finance
100
F
Street, N.E.
Mail
Stop
6010
Washington
DC 20549
Re:
NanoViricides,
Inc.
Amendment
No. 1 to the Form 10-SB12G filed on January 17, 2007
File
No
0-52318
Dear
Mr.
Riedler:
Please
be
advised that this firm is special securities counsel to NanoViricides, Inc.,
the
above-referenced issuer (the “Issuer”). This letter is in response to the
comments of the Staff of the U.S. Securities and Exchange Commission (the
“Commission”) with respect to the above-referenced filing provided in your
letter dated February 2, 2007 (the “Comment Letter”). The purpose of this
correspondence is to illustrate these changes to the Commission and provide
explanation, where necessary. Revisions and additions to the Registration
Statement will be shown in strikethrough and underlined format for convenience.
Our responses follow the text of each Staff comment reproduced consecutively
for
your convenience.
Form
10-SB12G / Amendment No. 1
General
1. In
your
response letter, please state our comment and then explain each change that
has
been made in response to a comment. In addition, you should also reference
each
page number in which disclosure has been revised in response to a comment
so
that we can easily place your revised disclosure in its proper
context.
The
Issuer will indicate the appropriate revisions within this
correspondence.
Securities
and Exchange Commission
February
14, 2007
Page
2
of
21
2. We
note your response to comment 43 relating to your intention to submit a
confidential treatment application for Exhibit A pursuant to Rule 406. Please
note that because you have filed Exhibit A as part of an Exchange Act document,
you should seek confidential treatment under Rule 24b-2 of the Exchange Act.
Additionally, please note that comments related to your request for confidential
treatment will be delivered under separate cover. Please be advised that
we will
not be in a position to consider notifying you that we have no further comments
on your Form 10-SB until we resolve all issues concerning the confidential
treatment request.
The
Issuer has sought confidential treatment under separate cover.
Preliminary
Efficacy Study, page 7
3. We
note your response to comment 11 and your revised disclosure and reissue
the
comment in part. Our comment also sought for you to quantify the results
to the
extent possible for studies where you indicate that preclinical testing has
shown efficacy. For example, in this section, we note you provide the efficacy
results related to a drug related to common influenza. Please revise your
disclosure to provide quantified disclosure such as the number of test subjects
you used as well as p value or other statistical analysis you conducted to
determine the efficacy of the particular study. Similarly, we note your
discussion in the section entitled “Preliminary Cell Culture Studies against
H5N1 Avian Influenza,” on page 7 where you discuss your preclinical testing of
various nanoviricides related to the treatment of H5N1. You do not provide
any
quantified disclosure in terms of the number of subjects you used or whether
you
used any statistical analysis. Please revise the above referenced sections
as
well as throughout the document where you discuss preclinical testing showing
efficacy to disclose the results of the preclinical studies, including the
number of test subjects, what your test group comprised of and whether your
results were subject to any statistical analysis.
Preliminary
Efficacy Study
A
nanoviricide drug was made using a well-known compound that has known efficacy
against common influenza. This nanoviricide and the free compound were tested
along with appropriate controls in mice infected with very high levels of
a
common influenza virus called H1N1. The results indicated that the efficacy
of
the nanoviricide was approximately 50 Times (5,000%) better than that of
the
free compound.
This
study indicated that the efficacy of a known virus-binding compound can be
enhanced to extremely high levels by using the nanoviricides technology.
We had
predicted such strong efficacy improvements theoretically. Further work will
be
necessary to substantiate that such an effect is seen across a wide range
of
compounds.
Securities
and Exchange Commission
February
14, 2007
Page 3
of
21
The
preclinical animal testing, done to study the efficacy (effectiveness) of
the
test nanoviricide (anti-human influenza, H1N1) compound, revealed potential
for
development for the reasons delineated below. Three separate and distinct
sets of experiments were performed to address different questions regarding
efficacy.
The
first
set of experiments was conducted in five different groups of ten mice to
test
efficacy of the nanoviricide compounds against H1N1 strain influenza with
ten
animals per group for statistical significance. All animals were given known
lethal doses of H1N1 human influenza virus. In all of these groups, at
least 60% or more of the animals survived when treated with the active
anti-H1N1 nanoviricide compound compared to those animals which
were administered only a saline (non-therapeutic, inert)
solution.
The next
set of experiments was conducted to determine the destruction/protection
of the
animal organs. There were also ten animals in each of five groups. Lethal
doses
of H1N1 were also administered. The active compound protected the organs so
that there were no histological (microscopic tissue) changes to the internal
organs of the treated animals. Highly significant tissue damage was found
in the
internal organs of the unprotected groups.
A
third
set of experiments was performed, again on five separate groups each containing
ten animals where the viral load was determined in the animals. The findings
revealed that the viral load (number of viral particles per cubic millimeter)
in
the treated animals was significantly lower than that found in the control
animals.
These
initial animal findings suggested that the test nanoviricide compound was
an
effective treatment for human influenza in mice and that the concept of using
a
nanoviricide as a treatment for certain viral illnesses was a valid one and
was
deserving of further study. In more scientific terms, the statistical test
was
met for validity of the findings and these findings could be considered
statistically significant. Thus, in statistical terms, one could say that
the
null hypothesis, that is the statistical likelihood that the observed results
were not due to chance but to the effect of the drug, was proven.
The
section entitled “Preliminary Cell Culture Studies Against H5N1 Avian Influenza”
on page 8 was amended to quantify the number of subjects used and statistical
analysis, as follows:
The
pre-clinical, laboratory testing of various test nanoviricides against H5N1
in
Vietnam was conducted in the following manner. Five "plates" each containing
96
culture wells were utilized. Different amounts of virus were placed in the
wells
along with differing concentrations of the 12 test compounds and their
positive and negative controls.
Securities
and Exchange Commission
February
14, 2007
Page 4
of
21
As
a test case, we have developed and evaluated the safety and efficacy of
nanoviricides against common influenza and H5N1, one of the highly pathogenic
avian influenzas and a current pandemic threat.
In vitro
(laboratory)
evaluation of 14 substances, including controls, was performed to evaluate
protection of mammalian cells against infection by the H5N1 subtype. These
assays were conducted in Vietnam under the auspices of the National Institute
of
Hygiene and Epidemiology, Hanoi (NIHE) under the Vietnam Ministry of Health.
We
identified four different nanoviricides as being highly effective against
H5N1
using two different assays, both involving cell culture, one using the
plaque
reduction method and the other involving microscopic examination, to determine
the extent of cytopathic events (CPE) reduction. All of these nanoviricides
were
effective at extremely low concentrations and many of them are considered
drug
candidates.
The
four different nanoviricides were selected on the basis of the statistical
test
called the p-value.(explained below). The p-values for these four compounds
were
p<.003 which meant that there was a high statistical probability that these
results were due to the effect of the test nanoviricides and not to chance.
Thus
the "null hypothesis" is proven and the results can be considered statistically
significant
The
most
successful of these was a nanoviricide based on an antibody fragment as
the
targeting ligand, which led to substantial suppression of CPE at an
extraordinarily low concentration level. This is being developed as
AviFluCide-I™, a drug highly specific to H5N1 that is being developed against
the Vietnam strain. We currently believe that it is very likely to work
against
the Indonesian strain although further studies will be required to determine
its
efficacy against various highly pathogenic strains of influenza. If it fails to
work against the Indonesia 2006 strain, further development may become
necessary.
Another
nanoviricide which is based on a ligand that we designed in-house to be
specific
to the group of all or a majority of highly pathogenic avian influenza
(HPAI)
viruses, also showed a very high efficacy. This is being developed as
“AviFluCide-HP™”, a drug designed to be group-specific against emergent and
existing highly pathogenic influenza viruses (including H5N1, H7N3 and
others).
Non-H5N1 HPAI (non-pathogenic avian influenza) strains could also become
a
pandemic threat as can all influenza A viruses since they all have the
ability
to mutate. It is well known that influenza strains drift constantly due
to
mutation, ressortment or recombination events leading to failure of
vaccines.
Preliminary
analysis of the H5N1 preclinical in vitro studies completed in Vietnam
showed
that many nanoviricide™ candidates were effective at as low as 5-nanomolar
concentration levels in cell culture experiments. Typically, an early
developmental drug that proves effective at concentrations less than 500
nanomolars is considered a strong candidate for FDA approval as an
“Investigational New Drug (IND)” applicant.
A
third
nanoviricide is based on a ligand that we designed for attacking all influenza
A
viruses (type-level specificity) has shown strong efficacy against H5N1
as well.
This is being developed as “FluCide-I™”, a drug designed primarily for use
against serious cases of human influenza.
Securities
and Exchange Commission
February
14, 2007
Page 5
of
21
Preliminary
Efficacy Studies in Vivo, page 8
4. We
note your
response to comment 12 and your revised disclosure to this section where
you
disclose the p value related to common influenza strain H1N1 and your reference
to the wikipedia.org website where investors may obtain the definition of
the p
value. The definition of what a p value measures should be discussed in the
text
of your document. Additionally, please further explain what it means to have
p
value less than .003. Please revise your disclosure
accordingly.
The
Registration Statement was revised to include the definition of “p-value” in the
Glossary on page 4 as follows:
P-Value:
In
statistical hypothesis testing, the p-value
is
the
probability of obtaining a result at least as extreme as that obtained, assuming
the truth of the null hypothesis that the finding was the result of chance
alone. The
fact that p-values are based on this assumption is crucial to their correct
interpretation.
More
technically, the p-value of an observed value observed of some random variable
T
used as a test statistic is the probability that, given that the null hypothesis
true, T will assume a value as or more unfavorable to the null hypothesis
as the
observed value observed. “More unfavorable to the null hypothesis can in some
cases mean greater than, in some cases less than and in some cases further
away
from a specified center.
Additionally,
the Registration Statement was revised to describe the significance of a
p-Value
of less than .003, on page 8 Preliminary Efficacy Studies in Vivo, as
follows:
In
statistics, a result is called significant if it is unlikely to have occurred
by
chance. "A statistically significant difference" simply means there is
statistical evidence that there is a difference; it does not mean the difference
is necessarily large, important or significant in the usual sense of the
word.
In
traditional frequentist statistical hypothesis testing, the significance
level
of a test is the maximum probability, assuming the null hypothesis, that
the
statistic would be observed. Hence, the significance level