RESPONSE OP TUMORIGENIC VIRUSES AND OP CELLS TO
BIOLOGICALLYACTIVE
COMPOUNDS.
I.
METHODS FOR DETERMINING RESPONSE AND APPlICATION OP
TX1
Gustave
Freeman,
Audrey
Kuehn,
and Ishkhan
Sultanian2
ABSTRkCT
Methods
i. e. ; threshold,
were
applied
for assessing
concentrations
the responses
of 65 biologically
of cell-virus
active
systems
compounds.
The
in culture
systems
to subtox.ic,
were
a pair
of tu
morigenic viruses,
Rous sarcoma (RSV) and polyoma (PYV), aixi a pair of lytic viruses,
encephalomyocarditis
(EMCV) and vaccinia
(W) . RSV and EMCV are ribonucleic
acid viruses;
PYV and SN are deoxyribonucleic
acid viruses.
RSV and W were assayed in chick cells and PYV and EMCV in mouse cells.
The viral species
had a major influence
on response,
the type of viral nucleic acid was of secondary
importance,
and the
host species
had a negligible
influence.
INTRODUCTION
New compounds,
synthesized
or extracted
by investigators
in antibiotic
research
and cancer chemo
therapy,
are being used effectively
(1, 3-6, 8) to study the biochemistry
of viral synthesis,
1. e. , to in
hibit specific
biochemical
steps in cellular
and cell-viral
metabolism.
These compounds
are generally
classified
according
to the type of biosynthesis,
ribonucleic
acid (RNA) or deoxyribonucleic
acid (DNA),
with which they interfere in normal and infected cell cultures.
However,
the conceniration
of a compound
required to achieve maximum inhibition
of a biochemical
step has been reversibly
injurious,
at the least,
or lethal to the cells.
This report discusses
the chemical control of viral propagation
and/or tumor cell induction
and rep
lication without apparent jeopardy to the cellular homeostasis,
or, more particularly,
the control of cellu
lar activities
caused by viruses without disruption of activities
essential
to cell growth.
Our main con
siderations
are (1) methods of determining
the effectiveness
of compounds
and (2) the kinds of information
that can be derived by the methods.
The 65 biologically
active
compounds
chosen
for the study
have
mechanisms
of action
that
are rea
sonably well understood.
Initial tests were done with the Rous sarcoma
virus (RSV) , in chick cells,
and the polyoma virus
(PYv) , in mouse cells.
They were chosen to form the pair of tumorigenic
cell-virus
complexes
because:
1.
RSV (Bryan strain) and PYV are being studied as etiologic
agents in great detail
in several laboratories.
The infected
cells can be studied directly.
When RSV and PYV are used, a background
of unirifected
cells can be observed.
RSV and PYV replicate
as RNA and DNA products,
respectively,
and take on cell
derived or specific
viral proteins.
RSV and PYV lend themselves
to quantitative
analyses
of normal and infected
cells and viruses.
The effect on the results
of infection
with RSV and PYV can be defined as a dose
response
relationship.
2.
3.
4.
5.
6.
7.
1 Supported
Transfiguration3
by
contract
can be observed
PH-43-65-43
Cancer Institute,
National
Education,
and Welfare.
Institutes
from
the
directly.
Cancer
of Health,
Chemotherapy
Public
Health
National
Service,
Service
Center,
U. S. Department
National
of Health,
2 Department
of Medical Sciences,
Stanford Research Institute,
Menlo Park, California.
3 We prefer “transfiguration―to “transfcrmation@because the changes are morphologic
and behavioral
are not yet known to be genetic.
1609
and
1610
Cancer Research
Later a pair of nontumorigenic
(W) was selected
for use in chick
cells.
The reason
for their
Vol. 25, October 1965, Part 2
cell-virus
systems
was added to the
cells,
and the RNA encephalomyocarditis
addition
is discussed
study.
virus
The DNA vaccinia
virus
(EMCV) for use in mouse
in the Results.
MATERIALS
The secondary
chick
embryos
The
cell
cultures
(Kimber Farms)
RSV (standard
were
derived
from
embryos
of randombred
known to have a low incidence
Bryan
strain)
was
obtained
from
Dr.
The PYV was that used at the California
Institute
of Technology
Laboratories,
where it was prepared from infected hens' eggs.
Type
Culture
Swiss
mice
and
from
a line
of
of leukosis.
W.
Ray Bryan,
National
Cancer
Institute.
(2). The VV was obtained
from the Lederle
The EMCV was obtained
from the American
Collection.
The 65 compounds
in other
systems,
Service
Center
tested
including
(Table
tumors
1) included
in man.
(CCNSC) , National
several
The agents
Cancer
were
of each
supplied
type that has
shown
by the
Chemotherapy
Cancer
biologic
activity
National
Institute.
METHODS
Toxicity
in Mouse and Chick Cells. --The least concentration
of the compound that inhibited
cell
division
during exponential
growth was determined
in unin.fected
cells to indicate
the dose range for the
cell-virus
assay.
Experiments
were done on 24-hour secondary
cultures
in 35-mm plastic
Petri dishes.
All cultures were grown initially in modified Eagle' s medium supplemented
with 10 per cent calf serum
before their use in experiments.
Then, for all mouse cell assays,
medium and serum described
by Dulbecco
and Freeman (2) was used.
For all chick cell assays,
medium, serum, and tryptose
phosphate
was used
according
to Temin and Rubin (7).
Cells were counted on sample pairs of plates by a Coulter Counter,
calibrated
for each cell type.
Then serial dilutions
of the compound were added to the cultures.
The compound was dissolved
in the
appropriate
solvent (see Table 1) , generally
at a concentration
of 2 x 102 gm/mI, and five tenfold dilu
tions were prepared in medium to give final concentrations
of l03 to io7 gm/mi.
Each of five pairs of
Petri dishes,
with equivalent
numbers of cells in the monolayers,
was covered with 2 ml of one tenfold
concentration;
concentrations
were shifted when indicated.
For the control,
a sixth pair of dishes
(con
taming monolayers)
was covered with medium without compound.
Also the concenirations
of solvent used
to dissolve
the compound were added to control monolayers
if the solvent was nonaqueous.
The cultures
were incubated
for 48 to 72 hours at 37° C in a humidified
atmosphere
containing
CO2.
After
and
incubation
1 ml of buffered
the medium
trypsin
solution
was
removed,
was
added
the monolayers
to each
washed
were washed
plate
to suspend
with
the
serum-free
cells.
medium,
The
paired
cultures
were pooled,
diluted to fit the linear range of the counter,
and counts were made at the correct
setting for the cell type.
The 95 per cent confidence
level of significant
difference
between
mean counts
of treated cultures
and controls was taken as ±15 per cent deviation
from controls,
because
the standard
deviation
(SD) from the mean count of 20 consecutive
cultures not treated with a compound was determined
to be ±6per cent for mouse and chick cells on several occasions.
(Cell counts were consonant
with those
done by a hemocytometer.)
were
Cell-Virus
essentially
Assays
Inhibition
of Virus and Cytotoxicity.
those described
previously
(2, 7).
--Plaque-
and focus-fcrmation
methods
Culture plates were prepared as for the toxicity
tests (cell count) , except that 60-mm Petri dishes
were used for plaque formation with Ply, w, or EMCV, and 35-mm dishes for RSV. The medium differed
from that used in the toxicity
tests only by the use of 0. 9 per cent agar in mouse cell cultures
and 0. 6
per cent agar in chick cell cultures
for overlaying
the infected
monolayers.
After removal of the medium from 24with Eagle' s medium, and 0. 1 ml of virus
two dilutions
(one about twice the other) of
one dilution of RSV (focus-forming)
per pair
or 48-hour
secondary
cultures,
the monolayers
were washed
suspension
was allowed to spread over the surface.
Each of
the plaque-forming
viruses
was placed on one pair of plates;
of plates was sufficient.
RSV cultures
were incubated
for 45
FREEMAN
minutes
et at.
Cancer
and the others
fcr 60 minutes,
Che,motherapy
at 370 C.
was then flowed on and allowed to gel.
Screening
Medium
Data
containing
1611
one tenfold
dilution
of the compound
Cultures were stained with neutral red the day before reading
the
assays.
EMCV plaques
were counted
on the fourth day, RSV foci and W plaques
on the fifth, and PYV
plaques on the eleventh.
Plaques were counted on either cc both pairs of plates,
depending
on which had
the largest clearly countable
number.
Foci on either or both plates of RSV cultures
were counted.
Counts
were compared
medium
with those
contained
Variance
cultures
only
studies
of the control
the solvent
plates,
in which
which were prepared
the compound
were done to determine
significant
differences,
treated with compounds and control cultures.
cells,
prepared as
tively.
The SD' s
of plaques
or foci
iently,
and 50 to
in the same way,
in plaque
Twenty consecutive
and focus
plates
centrations
SD' s.
studies,
Another
done as working conditions
indication
of drug activity
above the minimum causing
In addition
scopically
between
described
for each of several doses of virus, were infected
with PYV and RSV, respec
varied between
10 and 1 5 per cent from the means,
inversely
with the average
number
per series of plates.
(Between 40 and 150 plaques
per plate can be counted conven
300 foci per plate.)
even though new variance
reduced
counts,
each of mouse and chick
On the basis of an average SD of about 12 per cent, a value of 25 per cent was
of significant
difference
(@ 0. 05) from control counts.
This value was retained
what
except that the
was dissolved.
to counting
for quality
plaques
and compared
significant
and techniques
was that
improved,
continued
on the control
or morphologically
Toxic
effects
No stained
Few
++
Moderate
number
size or shape.
of stained
+
No obvious
cells
±
change in pattern of the monolayer, or an alteration in size,
configuration,
cc internal morphology.
Uncertain changes but suggestive of the type considered in
regardless
dead
of
but
with
were examined
were
+++
cells,
intact
cells on all plates
plates.
+1±1-
stained
showed some
to decrease
con
reduction.
or foci,. the background
with those
counts
selected
as the level
throughout
the study
graded
micro
as follows:
cells.
changes
in
cells , regardless
a reduction
size
or
shape.
of changes
in concentration,
in
a
the + category.
-
Treated
and
at higher
Results are expressed
occurred
significant
changes
fold.
Cell-Virus
the compound,
Ratio. --The
control
monolayers
identical,
but
gradual
changes
concentrations.
as ranges of minimum concentrations
(p.g/ml) of the compounds
within
from controls
(@ = 0. 05). Ranges were usually tenfold and sometimes
concept
of cell-@virus ratio
(CV?.) was used
to define
the relationship
the host (cell) , and the virus (plaque or focus formation) . Theoretically,
which
five
between
the CVR is the
ratio of the minimum amount of compound causing
threshold
injury tocells
to the minimum causing
sig
nificant reduction in plaques or foci.
Operationally,
the “amounts―
were ranges of concentrations
(@g/ml).
The amount of compound that produces threshold
injury (numerator)
lies between
the highest
concentration
showing no injury and the lowest showing at least ±injury. The amount of compound that produces sig
nificant reduction of plaques or foci (denominator) lies betweeTi the highest concentration
causing less
than a significant reduction and the lowest concentration causing a significant reduction.
Numerators and
denominators
ample:
were absolute
Highest
Lowest
Highest
values
concentration
concentration
concentration
and were
neither
interpolated
showing no cellular
showing
least
causing
in plaques
or foci
Lowest concentration
causing
in plaques or foci
cellular
change
change
ncr derived
from plotted
1 @.@g/ml
10 p@g/ml
<2 5 per cent reduction
. 1
@g/ml
>25 per cent reduction
1 @g/ml
—
1-10
.
p.g/ml
1-1
p.g/ml
points;
for ex
1612
•
Cancer Research
Vol. 25, October 1965, Part 2
RESULTS
Toxicity in Mouse and Chick Cells. --Dose-response
slopes varied considerably.
Figures
1 and 2
are examples
of typical responses.
As the concentration
of 2' -deoxy-5-fluorouridine
was increased,
in
hibition
of growth (cell count) of mouse cells gradually
increased.
@-Anisaldehyde thiosemicarbazone
produced an abrupt increase in inhibition of chick cells at doses higher than 10 @g/ml. Curves were fitted
by eye.
Table 2 compares
mouse and chick
well within a tenfold range of concentration
responded
at the same concentrations.
cell responses
to a series of compounds.
Reproducibility
was
for a particular
cell species;
mouse and chick cells generally
However,
striking exceptions
occurred
with N-
@-([(2, 4-diamino
6-pteridinyl)methyl)amino]benzoyl]glutamlc
acid (aminoptermn)
and purine-6-thiol
hydrate
(6-mercapto
purine);
mouse cells were much more sensitive
than chick cells to both.
The reverse
was true with 5bromo-2' -deoxyuridine;
chick cells were more sensitive.
Concentrations
of the last four compounds
listed
in Table 2 were not high enough •
to achieve
significant
inhibition;
with several
other compounds,
con
centrations
greater than 500 p.g/ml were necessary
-- a level unreasonably
high for practical
purposes.
In such cases,
cell-virus
assays were made at subtoxic levels to determine
whether plaques
or foci might
be inhibited.
Cell-Virus
Assays
Inhibition
of Virus and Cytotoxicity.
azolin-2-yl-2-nitroterephthalanilide
dthydrochloride
sponse that occurred in cell-virus
assays.
--The response
is an example
As the concentration
of PYVto 4' , 4' ‘
-dl-2-imid
of the most common
type
of dose-re
of the compound increased,
cell damage
in the monolayer
was roughly parallel to reduction@ in the number of plaques
(Fig. 3 and Table 3). Thus,
the CVR was approximately
1 . The response
of EMCV to 9-@-D-arabinofuranosyladenine
represents
a
second type of dose-response.
The cell monolayer
showed injury before there was any reduction
in the
number of plaques
(Fig. 4 and Table 3) . This effect in any of the four systems
was called relative
en
hancement.
The third type of dose-response
is represented
by the response
of W to 9-@-@-arabinofuran
osyladen.tne.
A significant
reduction
in plaques occurred at a concentration
of the compound that did not
apparently
affect the background
cells (Fig. 5).
Comparison
of Tests
Cells in Monolayers.
for Toxicity
--The sensitivity
Growth
Inhibition
(Cell
Count
of cell growth and morphologic
Versus
changes
the cell-virus
monolayers
were compared as indicators
of toxicity
to determine
calculating
the CVR. Table 4 compares
the results
for each cell species.
Morpholoqy
of Uninfected
in the background
which
should
cells of
be used
in
Monolayers
of chick cells infected with RSV or W appeared to be equally sensitive
to treatment
with
a given compound, the difference being generally within a tenfold range.
The duration
of both these assays
was 5 days.
The assays with mouse cells lasted 1 1 days for PYV and 4 days for EMCV; when there was a
difference
in sensitivit'y,
the assays of longer duration (PYV) were usually
more sensitive,
i. e. , the mono
layer showed toxicity
at lower concentrations
of the compound.
Generally
the morphology of the background
cells in chick and mouse monolayers
appeared
equal to
or more sensitive
than cell growth inhibition
as an indicator
of toxicity.
The morphology
was less sensi
tive than inhibition
of growth (cell count) in only 1 (W) of 32 chick cell-virus
assays
and in only 1 PYV
assay and in 5 EMCV assays of 26 mouse cell-virus
assays.
The cell count was more sensitive
in assays
that had a relatively short duration.
On the basis of these data, the morphology of cells was selected as
the criterion of toxicity for determining the CVR. There was an additional advantage:
the environment of
the baekground cells was the same as the environment in which plaque and focus formation took place.
However,
viewer.
judging
Influence
the degree
of cell
of the Cell Species
the host cell species
in determining
damage
adequately
depends
on the training
and experience
of the
on Cell-Virus
Responses
to Compounds.
--The
question
of influence
of
cell-virus
responses
to compounds
was important.
Therefore,
we
compared
the sensitivity
of the tumorigenic
virus systems
in terms of their CVR' s with 14 compounds,
i. e. , a ratio was formed of the CVR for the mouse-PYV system and the CVR for the chick-REV system.
Then
these ratios were compared to the ratios of toxicity for the same compounds
in uninfected
mouse and chick
cells.
The ratio of toxicity was the minimum amount (range) of compound causing
significant
growth in
hibition of mouse cells divided by the minimum amount (range) causing significant
inhibition
of chick cells.
FREEMAN
et at.
Cancer
Chenwtherapy
Screening
Data
1613
The results are given in Table 5; the ratios of toxicity and of the CVR' s are ranked according
to their mag
nitude.
If the effect of the compound on plaque and focus formation depended
on the response
of the cell
species
to the compound,
there should be some correlation
between the two orders of magnitude;
in fact,
there is none.
This suggests
that cell-virus
behavior was not primarily a function
of cell species
sensi
tivity to the compounds.
This conclusion
was substantiated
later by the observation
that there was no
significant
difference
in CVR' 5 when 5-bromo-2' -deoxyuridine
was added to cultures
of established
embry
onic human lung or kidney cells or to secondary embryonic mouse or chick cells infected
with VV (Freeman,
Kuehn, and Sultanian:
unpublished
data).
Influence
of T@@eof Viral Nucleic
viral nucleic acid in determining
Acid on Response
cell-virus
to Compounds.
susceptibility
--The
significance
to compounds was investigated.
of the type
of
The first tests
were done with the tumorigenic
viruses,
PYV (DNA virus) and RSV (RNA virus).
It became obvious that the
compounds were not similarly active in both systems4
and that the differences
were not attributable
to the
host species (see Table 5). Therefore, W (DNA virus) in chick cells and EMCV (RNA virus) in mouse cells,
both nontumorigenic
viruses,
were added to serve as controls.
Table
5
6 shows
The
the effects
compounds
There appeared
to be
efficacy
against
the
equally susceptible
to
ly active in W would
are
of 29 compounds,
listed
in
order
of
randomly
their
selected
efficacy
from the agents
(ascending
CVR'
s)
in
tested,
the
on all four
W-chick
system.
a lack of correlation
between
the efficacy
of a compound against
W (DNA) and its
tumorigenic
PYV (DNA) in mouse cells.
Therefore the two DNA systems
were not
the same compounds.
Nevertheless,
the probability
was high that compounds
clear
have borderline
efficacy in PYV (Table 7).
Examination of the results in the RNA-virus systems (RSV in chick cells and the lytic EMCV in mouse
cells) failed to reveal a consistent
relationship.
Neither was there an apparent
correlation
between
the
tumorigenic
and lytic viruses
(Tables
6 and 7). On the other hand, the occasional
similarities
of re
sponses
to particular
compounds
and may have
(see Table 7) may indicate
reacted
at sites
common
that these
cell-virus
are active
in multiple
species
divided
Relative Sensitivities
Among the Four Virus Systems. --A group of 12 1 randomly
selected
CVR' s was
into three categories:
@highsI(active),
both values of the CVR at least 10; ‘1low'1(inactive) , one
value less than 1; “borderline,“
both values at least
equal number of CVR's for each of the four systems.
to different
compounds
viral
systems.
1 and one value less than 10. There were a nearly
Almost half of the CVR's were about 1, and almost
half were below 1. The rest (7 per cent, nine CVR' s) were >10; most of these (eight) were in the W-chick
cell system (Table 8). In the two tumorigenic
viruses
none of the compounds
had high CVR' s but 17 and
19 had borderline
CVR' s in the PYV-mouse system and the RSV-chick
system,
respectively.
DISCUSSION
These experiments
dependent
were designed
upon tumorigenic
to explore
viral infection
a possible
cleavage
and essential
between
homeostatic
cellular
cellular
biochemical
events.
events
To do this,
we
observed the effects of a series of compouzxis of various types in the four cell-virus
systems
(a DNA-RNA
pair of tumorigen.tc viruses and a DNA-RNA pair of lytic viruses);
the efficacies
of the compounds
against
viral-determined
events appear to support the following:
1. The outstanding
sensitivity
RNA viruses,
to drug control of plaque
properties
of the DNA-VV, and the lack of sensitivity
of the DNA-PYV or the
or focus formation
imply that the viral species
and its individual
have a major role in determining
such sensitivity.
It is especially
interesting
that the corn
pounds effective
against W -- 2' -deoxy-5-iodouridine;
5-bromo-2'
-deoxyurithne.;
l-rnethylindole-2,
3dione, 3-(thiosemicarbazone);
statolon;
and 9-p-D-arabinofuranosyladenine-represent
different
types.
The possibility
of a common responsive
factor in the cell-virus
complex to the variety of compounds
is
being studied.
2.
indicated
activity
A second but less important
determinant
of response
is the type of viral nucleic acid.
This is
by the fact that the compounds
with borderline
activity
against
the DNA-PYV had outstanding
against
the DNA-VV (Table 7).
3. The cell species
has a negligible
influence
compounds,
proved both by comparisons
of host cell
4
In
the
assays
the
lytic
action
5 Assays with the tumorigenic
in the lytic virus.
of
PYV
In
mouse
on the sensitivity
sensitivities
with
cells
viruses were often repeated
was
measured,
so that tests
of the cell-virus
plaque and focus
not
would
Its
tumorigenic
be simultaneous
system to the
responses
and
activity.
with
assays
1614
Cancer Research
by study
however
efficacy
of 5-bromo-2'
-deoxyuridine
against
extensive,
do not deny the occasional
of a compound.
Vol. 25, October 1965, Part 2
VV in three different
cell species.
These observations,
strong influence
of host specificity
in determining
the
4. The majority of responses
to the compounds
imply that plaque and focus formation were reduced
only by concentrations
that injured uninfected
cells.
This suggests
that the ability of the cell to propagate
virus and to become transfigured
usually depended
on the capacity
of the host cell to function
normally.
However,
the several
clear instances
of antiplaque
responses,
without toxic responses,
suggest
significant
cleavage
took place between
normal cellular
functions
and cellular
activities
caused
virus.
This favors potential therapeutic
efficacy.
On the other hand the effect produced by certain
icals indicates
that viral propagation
was carried
on, and possibly
favored,
despite
interference
cellularhomeostasis.
that
by the
chem
with
In the investigation
reported
here, total viral propagation,
as reflected
by plaque formation,
was
the criterion of efficd'cy of a compound against the tumorigen.tc and lytic PYV (DNA) and in the pair of con
trol lytic viruses,
EMCV (RNA) and VV (DNA); cellular
transfiguration,
as reflected
by focus formation,
was the criterion
of effectiveness
against
the tumorigenic
RSV (RNA). The mechanism
of drug control of
both types of viral effect is now being studied by using the more active compounds.
Preliminary
obser
vations
indisate
that the selective
inhibition
of virus is not achieved
by a reaction
of the drug with free
virus.
Control of propagation
may occur at any a! the multiple steps between
attachment
of virus to cells
and the discharge
of newly synthesized
virus.
Agents like actinomycin
D (5), whose mechanism
of action
with respect to DNA is known, may reflect the nature of the cell-virus
relation.
In addition,
investigation
of the biologic
effects
of less well-understood
compounds
might throw light on their modes of activity.
Our results
were based upon the effects
of single “doses―of the compound put into the medium
immediately
after infection
of the cells.
Other methods of administering
the agents are being studied in
an effort to enhance
the activity
of subtoxic
concentrations
of the compounds
that were more selective
against
plaque and focus formation,
particularly
by the tumorigenic
viruses.
ACKNOWLEDGMENT
virus
We are very grateful to Dr. W. Ray Bryan of the National
and to Lederle Laboratories
for the vaccinia
virus.
Institutes
of Health
for the Rous sarcoma
REFERENCES
1.
Cohen, S. S. ; Flaks,
J. G. ; Barner,
of 5-Fluorouracil
and Its Derivatives.
2.
Dulbecco,
R. , and Freeman,
3.
Nathans,
Ribosomes
D. , and Lipmann,
of @.coli.
Proc.
4.
Reich, E. , and Franklin,
R. M. Effect of Mitomycin
Natl. Acad. Sci., US, 47:1212—17, 1961.
5.
Reich,
E. ; Franklin,
R. M. ; Shatkin,
A. J. ; and Tatum, E. L. Effect of Actinomycin
Nucleic Acid Synthesis and Virus Production. Science, 134:556-57, 1961.
6.
Simon, E. H.
18, 1961.
7.
Temin, H. M. , and Rubin, H. Characteristics
of an Assay
Cells in Tissue Culture. Virology, 6:669-88, 1958.
for Rous Sarcoma
8.
Wecker,
E. , and Richter, A. Conditions
tativeBiology, 27:137—48,
1962.
of Infectious
Evidence
C.
H. D. ; Loeb,
Proc. Natl.
Plaque
Production
M. R. ; and Lichtenstein,
J.
Acad. Sci. , US, 44: 1004-12,
by the Polyoma
F. Amino Acid Transfer
Natl. Acad. Sci. , US,
for the Non-participation
Virus.
Virology,
for Arninoacyribonucleic
45:1721-29,
1959.
C on the Growth
The Mode
1958.
8:396-97,
Acids
on Some Animal
of DNA in Viral RNA Synthesis.
for the Replication
of Action
1959.
to Protein
Viruses.
on
Proc.
D on Cellular
Virology,
13:105-
Virus and Rous Sarcoma
Viral RNA.
Sympos
Quanti
,
FREEMAN
Cancer Chemotherapy &reening Data
et (ii.
U,
-a
@
U,
U
I
1615
I
I
I
I
0.1
I
10
00
.15
“a
U
I
0
U
-IS
—
I-'..
@
-‘5
OW
@z
0
z
0
>
‘a
0
>
C
0.1
I
10
100
COMPOUND CONCENTRATION-@jg/mI
Figure
‘a
@
I
@IOOO
C
Tooo
COMPOUND
CONCENTRATION-Mg/mI
1
Figure 2
I
I
I
U,
+25
@
I
I
I
I-.,
@
U
-25
@
INJURY
@
TO MONOLAYER
INJURY
—
@
@
I
C
5
-I-
\4N@.
I
.55.
f
10
TO
z
2
MONOLAYER
4
f
50
I
C
100
I
01
I
COMPOUND CONcENTRATION-Fig/mI
COMPOUND CONCENTRATION -,.tg/mI
.c-s.u -?4
. Fig@re
Figure 4
3
U,
‘a
I
I
I
I
::±s@@:i
‘1
TO MONOLAYER
>
I
C
I
0.001
I
aoi
01
0
COMPOUND CONCENTRATION-Fig/mI
.c-)'sz-,.
Figure
5
Figure 1. --@Typic*1 mouse CYtc*CXiCLtytest inhibition ct cell growth, compared to control, th@eased
gradually as dose of 2' -decxy-5-flucrourldine
was Increased; cells counted 48 to 72 hours eft@
addition of compound..
Figure 2. --Typical chick cytotcxiclty
test; inhibition of cell growth. compared
abruptly at doses higher then 10 @g/m1of @-anisa1dthyde thiosemicarbazone;
72 hours after addition of compound.
@
@
to control. in@eased
cells counted 48 to
Figure 3. --Most common dose-response
in cefl-viru.s assays:
cell damage In the monolayer roughly
poMMel to reduction in number of plaques.
comj@red to costrols,
as concen@at1on of compound
is increased; PYV on mouse cells
@eatedwith 4' , 4' ‘
anilide dthydrochloride;
duration of assay,
1 1 days.
Figure 4. --Second type ot dose-@response In cell-virus
assays:
injury to cell monolayer before re
duction in number of plaques,
compared to con@o1s; EMCV on mouse cells
@eated with 9-@-@arabinofuranosyladenine;
duration of assay,
4 days.
Figure 5. --ThIrd type of dose—response in cell-virus
assays:
reduction in number of plaques at a
concentration
of compound that did not apparently
damage background cells; W on chick cells
@eatedwith 9-@-@-arabinofuranosyladenine;
duration of assay. 5 days.
100
CancerResearch
1616
1Listof
NO.NSC
66635
TABLE
Studied(solvent
Compounds
andsourcecodes
explained
table)ENTRY
Vol. 25, October 1965, Part 2
at the
end of the
NO.COMPOUNDNAMEMOLECULAR
3069
FORMULASOLVENTSOURCE
2, 2-dich1oro-@-Q3-hydroxy-a-(hydroxy
2
27P
2
4p
1
39K
3
20
chloramphenicol21123053Actinomycin
-Acetamide, methyl)-@-nitrophenethyl]-;
66636
@
66637
66638
66639
66640
DN12O16C62H86836531-Adamantanamine,
hydrochloride1
HC1404241Adenine,
17
9-@3-@-arabinofuranosyl
N504C10H133056Adenosine,
3' -amino-3'
83
505 1@-Alanine,
81@42
-deoxy-@,
229A
@-dimethyl
3-(@-(bis(2-thloroethyl)amtho)phenyl)-;
2
4p
@-sarcolysinN2O2C12C
@
66641
66642
66643
66644
66645
66646
66647
66648
10@-Alanine,
3- [@-Ibis(2-chloroethyl)amlnojHC168984Q-Anisaldehyde,
phenyl]-, hydrochloride; sarcolysin221
.-
8
thiosemicarbazone11712@-Anisaldehyde,
thiosemicarbazoneN305C9H11401
575Benzimidazole,
5, 6-dichloro- l-@-@-rthofuranosyl
12H124052-Benzimidazolemethanol,
N204C12C
a-phenyl
N20C14H1215506Benzimidazole,
2-(octylthio)-N2SC15H223088Butyric
19746Carbamic acid, 4-[@-lbis(2-chloroethyl)amino]phenyl]
NO2C12C
esterN02C3H7757ColchicineNO6C22H2551954
acid, ethyl
66649
66650
66651
antibiotic
5 19 15
M259
Cyclohexanecarboxamide,
@, @‘
-[3, 6-bis(1-aziri
dinyl)-@-benzoquinon-2
66652
@
66653
@
66654
1026
, 5-yleneJbis
Cyclopentanecarboxylic
acid,
2, 2' -dichloro-@-methyl-,
HC13051Formamide,
2
57A
2
20
2
74R
2
79
2
2
3
37p
1
49
43
1
302P
2
5A
233D
1
l7A
3•
1
35D
1
1
4
6
20
1
49
3
229A
43
3
32B
1
20
2
37
2
27
1-amino
63878Crystalline
Cytosine, 1-@-@-arabinofuranosyl-,
HC1762Diethylamine,
36
2
N404C24H32
261
hydrochlorideUnknown
4
hydrochlcride1
66655
@-methy1-NOC2H5758u-Glucose,
66656
HC1739
66657
2-amino-2-deoxy-,
.Glutamic
hydrochlorideN05C6H13.
acid, N-1,@-[((2,4-diamino-6-pteridinyl)-
aminopterinN805C20H2259407Glutamic
methyl]amino]benzoyl]-;
66658
66659
66660
acid, N—(,@—([(2,
4—diamino—5,
6, 7, 8—te@aH2O185Glutarimide,
hydro-6-quinazolinyl)methyl]amino]benzoyl]-,
dihydrateN6O5C21H26.
5H236981
3-(2-(3, 5-dimethyl-2-oxocyclohexyl)2-hydroxyethylJ-;
1Indole-2,
3-dione,
2
actidione1
1-methyl-,
3-(thiosemi
carbazone)
66661
13875
Melamine,
hexamethyl-@N4SC10H10
N6C9H18
FREEMAN
Cancer Chemotherapy Screening Data
et al.
1617
TABLE1(Continued)ENTRY
NO.NSC
66662
NO.COMPOUND
750
NAMEMOLECULAR
Methanesulfonic
acid , tetramethylene
FORMULASOLVENTSOURCE
ester;
14
myleran
66663
26980
Mitomycin
66664
28693
Monocrotaline
66665
52141
Nonactin
66666
26271
2@-1 , 3, 2-Oxazaphosphorine,
2-[bis(2-chloroethyl)amino]tetrahydro-,
2-oxide,
hydrate;
cyclo
phosphamide
C
N06C1 6H23
012C40H64
66667
6396
Phosphine
66668
25154
Piperazine,
66669
753
Purine
66670
743
Purine,
66671
752
Purine-6-thiol,
66672
406021
66673
755
66674
3055
N405C15H18
sulfide,
75
612
1 , 4-bis(3-bromopropionyl)-
26
2-amino-;
9H-Purine-6-thiol,
9-@3-@-arabinofuranosy1-
Purine-6-thiol,
Puromycin,
6-thioguanine
7@@-Pyrro1o[2, 3-@jpyrimithne,
ribofuranosyl-;
tubercidin
66676
46401
a-Sarcin
66677
742
L-Serine,
66678
10123
Serine,
diazoacetate;
71901
Statolon
37917
Streptozotocin
66681
35847
Terephthalanilide,
14574
1
229A
4
35C
4
26
3
27
3
21C
1
3 65D
1
35
2
254
1
1
1
32B
3
50
4
32B
3
371?
2
66683
68929
66684
749
Thymine,
HC1
N5012C14H27
1-[[2-(diethylamino)ethyl]hydrochloride;
miradil
1-@-@-arabinofuranosyl
v-Triazolo[4,
5-djpyrimidin-7-ol,
N704C26H23
HC1
N20SC20H24. HC1
D
N206C10H14
5-amino-;
5
37
Unknown
Thioxanthen-9-one,
amino)-4-methyl-,
2
3
. 2
66682
1
32
1
4' , 4' ‘
-di-2-imidazolin-2-yl-2-
1
3
N3O4C5H7
dihydrochioride
243
50
3-phenyl
66679
1
3
N404C1 1H14
azaserine
52C
N5SC5H5
Unknown
66680
nitro-,
4-amino-7-@-@-
2
37
43
.
56408
5
1
N7O5C22H29
66675
2
N6C5H6. H20
N4SC5H4. H2O
dihydrochioride
6
317?
43
N404SC10H12
hydrate; 6-mercaptopurine
2
1
N4C5H4
hydrate
37
. H2O
tris(1-aziridinyl)-
2, 6-diamino-,
5
N60C4H4
8-azaguanine
66685
62403
@-Tryptophan, 5-@bis(2-chloroethy1)amino]-
66686
57695
@-Tryptophan,
66687
68928
Uracil,
1-I3-@-arabinofuranosy1-
66688
406444
Uracil,
1-@-@-arabinofuranosy1-5-f1uoro
Uracil,
5-bromo
66689
19940
5-methyl
19
N2O2C12H14
912
1
N2O2BrC4H3
1
32B
3
32
3
7P
1618
Cancer Research
Vol. 25, October 1965, Part 2
TABLE 1
(Continued)
NAMESOLVENTSOURCE6669082222Uracil,
NO.COMPOUND
ENTRYNO.NSC
N2O6BrC9H11332B6669123519Uracil,
1-@3-@-arabthofuranosy1-5-bromo
5-diazo-,
H20163P6669219893Uracil,
hydrateN4O2C4H3.
5—fluoro
N202FC4H33217?6669357848Uradil,
N2021C4H337?6669482221Uracil, 5—iodo
l-@-@-arabthofuranosyl-5-iodo
N206IC9H11332B6669532065Urea,
N202CH41526669638297Uridine, hydroxy
N2O5BrC9H1137?6669739661Uridine,5—bromo—2'—deoxy
.N2051C9H1137P6669827640Uridi.ne,
V—deoxy—5—iodo—
2'—deoxy—5—flucro
N2O5FC9H111217P6669949842Vincaleukoblastine,
sulfate,
hydrateN409C46H56.
H2S04
. H201365k
D@LA@ION@Cf@
CODES
SOLVENTS
Cede
Solvent
Water
Ethyl alcohol
Sodium hydroxide
Hydrochloric acid
Acetone
2
3
4
5
6
Miscible with water
SOURCE OF COMPOUNDS
Soui'ce
Chemical-Biological
Center
5k
National Research Council
2101 Constitutioji
Avenue
North Chicago,
6
D@ C.
Research
Rahway,
and
7P
Dohme
Research
Laboratories
17k
Merck, Sharp, and Dohme
Department
“P.
following a source cede number indicates
California
Prof. Clarence
I. Noll
2 1 1 Whitmore
20
Abbott Laboratories
North Chicago, Illinois
Los Angeles,
College of Science
The Pennsylvania State University
New Jersey
Dr. JohnA. Carbon
Biochemistry Research
Calbiochem
3625 MedfOrd Street
Research Laboratories
Rahway, New Jersey
5
of Laboratories
Syracuse 1, New York
Institute
New York
Section Head, Research Data and Sample Register
Sharp,
Illinois
Bristol Laboratories
Dr. Anthony H. Land
Merck,
Laboratories
Dr. H. Leo Dickison
Director
Dr. Maurice L. Tainter, Director
Sterling-Winthrop
4?
Dr. Walton E. Grundy
Abbott
Rennselaer,
4
Coordination
National Academy of Science
Washington,
2
Source
Laboratory
University Park, Pennsylvania
Dr. Frank M. Schabel, Jr.
Director, Chemotherapy Research
Southern Research Institute
2000 NinthAvenue, South
Birmingham, Alabama
that the compound was purchased by CCNSC.
FREEMAN
Cancer Chemotherapy Screening Data
et at.
1619
TABLE 1
(Continued)
SOURCE OF COMPOUNDS
Source
Source
21C
39K
Dr. Gordon N. Walker
Chemical Research Department
Ciba Pharmaceutical Products, Inc.
E. I. du Pont de Nemours
Dr. Birger H. Olson, Chief
Antibiotic and Fermentation
Section
Division of Laboratories
43
49
Parke, Davis and Company
2800 Plymouth Road
Chairman
California
Dr. Douglas
A. Shepard
Francis Earle Laboratories,
Peekskill,
New York
52
Miss Barbara Stearns
52C
30 1 Henrietta Street
35D
57A
Biological
Screening
Office
Institute
of Antibiotics
of Medical
Plrogovskaia
Sciences
11
Nutritional Biochemicals
21010 Miles Avenue
Cleveland 28, Ohio
74R
George Luttermoser
Corp.
Laboratory of Parasite Chemotherapy
National Institute of Allergy and
Infectious Diseases
National Institutes of Health
Bethesda, Maryland
79P
Moscow, U. S. S. R.
37
Prescott
63P
30 1 HenrIetta Street
Kalamazoo, Michigan
Academy
Bolshaia
Dr. Benjamin
Bethesda, Maryland
The Upjohn Company
36
Dr. Ralph E. Bennett
Science Information Section
National Institute of Allergy and
Infectious Diseases
Room 207, Building 5
National Institutes of Health
Michigan
Dr. Paul W. 0' Connell
Inc.
The Squibb Institute for IG@edical Research
New Brunswick, New Jersey
Dr. Charles G. Smith
The Upjohn Company
Kalamazoo,
Industries
Organic Chemistry Section
Squibb Institute for ?4.edical Research
New Brunswick, New Jersey
Bio-Organic Chemistry Department
Stanford Research Institute
Menlo Park,
Products
50
Michigan
Dr. Leon Goodman,
Distillation
Rochester 3, New York
Upjohn Company
Kalamazoo, Michigan
35C
Dr. Carl J. Wessel
National Academy of Sciences
National Research Council
2101 Constitution
Avenue, N. W.
Dr. George Hitchings
Washington
Research Director
Chemotherapy Division
Welicome Research Laboratories
2 17P
Scarsdale Road
Tuckahoe 7, New York
229A
25,
D.
Hoffmann-LaRoche,
Nutley,
New Jersey
C.
Inc.
07110
Dr. J. M. Ruegsegger
Clinical Research Section
37P
Service
Dr. JohnR. Dice
Ann Arbor,
35
National
National Institutes
of Health
Room B1-04A, Wiscon Building
Bethesda, Maryland
Research Laboratories
Parke, Davis and Company
2800 Plymouth Road
Ann Arbor, Michigan
32
32B
Chemotherapy
Center
National Cancer Institute
Michigan Department of Health
27P
Dr. D. Jane Taylor
Cancer
Highway M-l74
Lansing 4, Michigan
27
& Company
Experimental Station
Wilmington, Delaware
556 MorrisAvenue
Summit, New Jersey
26
Dr. Edward C. Herznann, Manager
Medicinal Chemistry Section
Welcome
Research Laboratories
Scarsdale
Road
Tuckahoe
7, New York
“P@followinga source cede number indicates
Led@le Laboratories
Division
American Cyanamid Company
Pearl Rivei@ New York
that the compound was purchased by CCNSC.
.
Cancer Research
1620
Vol. 25, October 1965, Part 2
TABLE 1
(Concluded)
SOURCE OF COMPOUNDS
Code
233D
Dr. ProsperLoustalot
Ciba, Limited
Basle, Switzerland
243
Dr. Kenneth N. Campbell
Director,
Medicinal Chemistry
Research Laboratories
Mead Johnson and Company
Evansville,
Indiana
254
Dr.
Fred
H.
Schultz,
Jr.
3 17?
Cyclo Chemical Corporation
1930 East 64th Street
Los Angeles,
California
365k
Dr. Koert Gerzon
Organic Chemical Division
The Lilly Research Laboratories
Eli Lilly and Company
Indianapolis
6, Indiana
365D
Dr. Otto
Research
Chemical
The Lilly
Eli Lilly
Director of Pharmacology
Dorsey Laboratories
P. 0. Box 1113
Lincoln 1, Nebraska
302
‘IpI@
a
source
cede
number
indicates
K. Behrens
Advisor
Research Division
Research Laboratories
and Company
Indianapolis,
Fisher Scientific Company
7722 Woodbury Drive
Silver Spring, Maryland
following
Source
Code
Source
37 1P
that
the
compound
was
purchased
Indiana
Regis Chemical Company
12 19 North Wells Street
Chicago 10, Illinois
by
CCNSC.
FREEMAN
et at.
Cancer
Chemotherapy
Screening
Data
1621
TABLE 2
MINIMUM
CONCENTRATIONS
GIVING SIGNIFICANTINHIBITION OF GROWTH
(CELL cOUNT)OF UNINFECTEDCELLS IN 72 HOURS
COMPOUND/Lg/.1.001
10,000.
-
.01
0.1
1
.
Glutainic
N-[p-[[(2,4-diaatino-6-pteridinyl)methyllaminolbenzoylltPurine-6-thiol,
acid,
hydrateL.Serine,
diazoacetate———-Diethylamine,
hydrochloride'—Uracil,
2.2'-dichloro-N-uaethyl-,
5-fluoro
Purine::::Methanesulfonic
esterD-Glucose, acid, tetramethylene
hydrochloride-@jCyclopentanecarboxylic
2-amino-2-deoxy,
acid, 1-amino
@-Colchicine—
—Phosphine
— —
tris(l-aziridinyl)-————v-Triazolo[4,5-dlpyrimidin-7-ol,
sulfide,
5-amino
>2-Benzimidazolemethanol
, a-phenyl
thioaemicarbazoneUracil,
p-Anisaldehyde,
1-,8-D-arabinofuranosyl-5-fluoro
—Uridine,
LDL-Tryptophan,
2'-deoxy-S-iodo
L___Thioxanthen-9-one,
5- [bis(2-chloroethyl)aminol
hydrochlorideLLLBenzimidazole,
l-[[2-(diethyla.ino)ethyl]amino]-4-methyl-,
5,6-dichloro-1-fl-D.rihofuranosyl
Terephthalanilide, 4'.4―-di-2-imidazolin-2-yl-2-nitro-,
dihydrochiorideGlutarimide,
3-[2_(3,5-dimethyl-2-oxocyclohexyl)-2-hydroxyethyl]-“<Adenine,
9-,8-D-arabinofuranoayl
——7H-Pyrrolo[2,3-djpyrimidine,
—
4-amino-7-@8-D-ribofuranosyl
—(Purine-6-thiol,
2-amino
>—Uridine,
———Uracil,
5-bromo-2'-deoxy
.@__tjracil,
5-iodo-
5-bromo
@:@:_:Formamide,
N-methyl
9H-Purine-6-thiol, 9-fi-D-arabinofuranoayl
S
t
Compared
The
linea
to
controls.
show
the
Cells
actual
that the concentration
counted
conceatratioaa
was nontozic
by
above
Coulter
and
Counter.
below
(>) or toxic (<).
the
points
Bracketing
of
.igaificaat
change.
Arrow
is used in lieu of interpolation
of retainingconservativevalues. C—) ouse cells, (---.) chick cells.
tail
indicates
as a .esns
10
100
1000
1622
Cancer Research
Vol. 25, October 1965, Part 2
TABLE 3
Representative
COMPOUNDDOSE
Responses
in Cell-Virus
Assays
per
pair of
MONOLAYER*Number
(@ig/ml)PLAQUESINJURY
(%)PyV
control
TO CELL
from
platesDifference
CELLSTerephthalanilide,4'.
ON
MOUSE
4―—di—2—imidazolin—2—1000—100++++yl-2-nitro-,
dihydrochloride-100++++10168—34+5252—2—Control0257--------EMCV
ON MOUSE
CELLSAdenine,
9-@-D-arabinofuranosyl
10062-21++1095+22+175-4±.177—1—Control078--------*See
Methods
for grades
of toxicity.TABLE
4Comparison
andMorphology
of Sensitivity
of Growth Inhibition Assay
of UninfectedToxicityNUMBER
Cells in Monolayers as Estimates
COMPOUNDSDEGREEOF
SENSITIVITY*DEGREE
OF COMPOUNDS CAUSINGPERCENT
OF SENSITIVITY/TOTALCAUSING
OFCOMPOUNDS
TESTtSENSITIVITYCHICK
FOR EACH
CELLSRSV
monolayersNearly
vs W
equal5/1436RSV
sensitive5/1436RSV
less
4/1428Cell
more sensitive.
count (CC)
monolayersNearly
vS
RSV
equal9/1560CC
sensitive6/1540CC
less
sensitive0/150Cell
more
monolayersNearly
count vs VV
equal10/1759CC
sensitive6/1735CC
less
more sensitive1/176MOUSE
CELlSPYV
monolayersNearly
vs EMCV
equal8/1362I'YV
sensitive0/130PYV
less
more
of
sensitive5/1338(Concluded
on following
page)
OF
DEGREE
FREEMAN
Cancer Chemotherapy Screening Data
et al.
1623
TABLE 4
(Concluded)
OF COMPOUNDS
CAUSING DEGREEOF
OF COMPOUNDS CAUSING
DEGREEOF SENSITIVITY/TOTAL
COMPOUNDS FOR EACH TESTtPERCENT
DEGREEOF SENSITIVITY*NUMBER
SENSITIVITYCell
count vs PYVmonolayers
Nearly equal
CC less
sensitive
CC more sensitive
Cell count vs EMCV monolayers
Nearly equal
CC less sensitive
CC more sensitive6/13
*
The
sensitivities
of
the
two
6/13
46
1/13
8
5/1 3
3/13
38
24
38
5/1346
systems
being
compared
were
considered
nearly
equal
if
about
the
same
amount
of
compound
produced the same degree of toxicity in both systems.
The amount of compound was expressed
as a range of minimum
concentrations;
the toxicity was between - and ±(or greater) injury to the monolayers or significant
inhibition of growth
(cell count).
The duration of assays was 5 days for RSV and VV, 11 days for PYV, 4 days for EMCV, and 48 to 72 hours
for the growth inhibition assay.
t Total of 59 compounds were used in all tests.
TABLE 5
Influence
of Cell
Species
On Drug
Inhibition
of Plaque
or Focus
-Mouse
Formation
OF
vs Chick CellsPolyoma
RousRank
.
vs
COMPOUNDSENSITIVITY
of ratio
(descending order
RousGlutamic
of Polyoma/CVR of
of_magnitude)P;:I@0ofCVR
810006300yl)methyl]amino]benzoyl]
acid, N—(,@—(((2,
4—diamino—6—pteridin
hydrate73002<1£—Serine,
Purthe-6-thiol,
diazoacetate610031Phosphine
tris(1-azlridinyl)-5102<1Uridine,
sulfide,
5104>1Diethylamine,
5-bromo-2'-deoxy
-dichloro-li-methyl-,4>531hydrochlorideUracil,
2, 2'
354>1Glutarimide,
5-fluoro
211.
3-(2@-(3, 5-dimethyl-2-oxocyclo
15hexyl)-2-hydroxyethyl]-@-Anlsaldehyde,
thiosemicarbazone2131Colchicine212<1Thioxanthen-9-one,
1-([2-(diethylamino)ethyl]-212<1amino]-4-methyl-,
hydrochlorideUridine,
2155Benzimidazole,
V -deoxy-5-iedo
1<1319B-Purine-6-thiol,
5, 6-dichloro-l-@—D-ribo
furanosyl
9-@-D-arabinofuranosyl
*
Ratio
of
causing
the
minimum
significant
concentration
growth inhibition
(range)
1<131
causing
of chick cells.
significant
growth
inhibition
of
mouse
cells
to
minimum
(range)
1624
Cancer Research
Comparative
Cell-Virus
ratios
of Compounds
Vol. 25, October 1965, Part 2
in Four Cell-Virus
Systems
VIRUSES(Chick)PYV
VIRUSESRNA
(Mouse)OrderCVR*OrderCVR*OrderCVR*OrderCVR*
(Mouse)RSV
(Chick)EMCV
COMPOUNDDNA
Purine-6-thiol,
Uradil,
2-amino-
5-fluoro-
Actinomycin
D
1
<. 1
3
. 01-. 1
--
6
<1
1
<. 1
3
<. 1
4
. 1—10
1
<. 01
2
-@. 1
4
-@.1
2
@.
5
3
@.1
7
. 1—10
2
2
<. 1
Benzimidazole,
5, 6-dichloro-1—@3-D-rthofuranosyl—
3
. 1-1
Carbamicacid,
ethylester
3
<1
10
3
<1
7
. 2-1
Glutarimlde,
3—[2@(3,5—dimethyl-2—oxocyclohexyl)-2-hydroxyethyl]-
4
. 1—10
10
2—10
Colchicine
4
. 1—10
4
-@. 1
1
@.1
5
‘@.
2
4
. 1—10
7
. 1—10
3
. 1—10
3
.01—1
4
. 1-10
7
. 1-10
--
6
4
. 1- 10
7
. 1—
10
2
. 1—
1
—-
6
. 1-1
1
. 01-1
--
@-Tryptophan, 5-fbis(2-chloroethyl)amino]-
Thioxanthen-9-one,
l-ff2-(diethylamino)ethylj-
amino]-4-methyl-,
Terephthalanilide,
Uradil,
9
—10
--
11
8
1—10
4
<.2
5—100
——
hydrochloride
2-Benzimidazolemethanol,
2 -nitro-,
—5
1. —1
a-phenyl-
4' , 4' ‘
-di-2-imidazolin—2—yl-
. 1-1
dthydrochloride
1-@3-@-arabinofuranosyl-5-fluoro-
5
>1
@-Anisaldehyde,
thiosemicarbazone
6
1—10
9
.1-10
4
.1-10
7
6
>5
2
<. 04
2
. 2-1
4
Purine-6—thiol, hydrate
7
@10
1
—.01
9
-lO
7
‘@l
Indole—2,3—dione, 1—methyl—,
3—(thiosemi—
8
>10
——
3
1—10
9
Diethylamine,
2, 2' -dichloro-N-methyl-,
.1—10
<. 2
hydrochloride
10
1—10
1-100
1
@.
01
9
1-100
9
9
1—100
5
9
1—100
6
1—100
9
carbazone)
L-Sermne, diazoacetate
Acetamide,
-
2, 2-dichloro-N-[@-hydroxy-a-
. 1-10
2
10
<1
--
1-100
3
. 01-1
10
100
(hydroxymethyl)-@-nitrophenethyl]
2H—l,3,2—Oxazaphosphorine,
ethyl)amino)tetrahydro-,
2—[bis(2—chloro—
2-oxide,
Uridine, 2'—deoxy—S—fluoro—
9H-Purine-6-thiol,9-@-@-arabinofuranosyl—
Glutamic
acid,
1—1
9
hydrate
N—(@—(((2,
4—diamino-6-pteridinyl)—
9
<1
12
.1-10
10
l0—l@0
8
>1
Uracil,5—bromo—
10
10—100
3
.01—1
Uridine,
5—bromo—V—deoxy—
10
10—100
11
—‘5
Adenine,
9—@3—@-arabinofuranosyl-
11
>100
7
>1000?
2
<1
2
1.-1
6
.1-1
9
>1
3
@.1
methyl]amino]benzoyl]
Uridine, 2'—deoxy—S—iodo—
12
200—l0@
12
Glutamic acid, N—[@—[((2,
4—d.tamino—5,
6, 7, 8—
——
7
.1—10
2
. 1—1
.1-10
4
.1-10
1—100
4
. 1—10
4
. 1—1
7
. 1-10
6
. 1—1
6
<1
6
<1
9
. 1-10
——
5
.04-1
——
. 2—5
tetrahydro-6-quinazolinyl)methyl]amino]-
benzoylj-,
Benzimidazole,
Uracil,
*
When
dihydrate
2-(octylthio)-
5—iodo—
the
CVR
is
one
10
number,
a
complete
dose-response
curve
was
not
obtained.
4
1—10
(--)
2
compound
<1
not
tested.
FREEMAN
et at.
Cancer
Chenwtherapy
Screening
Data
1625
TABLE7
Relative Sensitivities
of Systems to Compounds
--DNA
virusesvVPYvEMCVRSVUridine,
COMPOUNDSENSITIVITY
OF
virusesRNA
+*±--Uridine,
2' -deoxy-5-iodo
+±--Indole-2,
5-bromo-2'-deoxy
3-(thiosemicarbazone)+±--Statolon+±--Adenine,
3-dione,
1-methyl-,
+±--Uridine,
9-@3-D-arabinofuranosyl
2' -deoxy-5-fluoro
±--±2@j-l,
2-[bis(2-chloroethyl)-±-±amino]tetrahydro-,
3, 2-Oxazaphosphorine,
hydrateGlutamic
2-oxide,
4-diamino-6-pteridinyl)-±±-±methyl)amino]benzoyl]
acid, N-[@@-[((2,
Glutarimide,
5-dimethyl-2-oxocyclohexyl)-±±-±2-hydroxyethyl)-2-Benzimidazolemethanol,
3-(2-(3,
a-phenyl
±-±-Carbamic
ester-±±-* acid,
valueless
CVR
values
ethyl
(range
between
two
limiting
than 10; (-) one value less
values):
(÷)
both
values
at
least
10;
(±J
both
values
at
least
1
and
one
than 1.TABLE
8Relative
TestedCVRCOMPOUNDS
Susceptibilities
Systemsto
of the Four Cell-Virus
all Compounds
WITH SPECIFIC CVR AGAINST--TOTAL
CVR.
virusesVVPYV@@@RSVEMCVNumber
DNAvirusesRNA
(%)Both
values
(7)Both
at least
108
(49)value
values at least 1 and one14
COMPOUNDS WITH
SPECIFIC
(%)Number
(%)Number
(%)Number (%)Number
(28)0
(0)0
(0)1
(4)9
(48)17
(50)19
(61)9
(33)59
(24)17
(50)12
(39)17
(63)53
(100)34
(100)31
(100)27
(100)121
<10One
value (44)TOTAL29
<17
(100)