A MODE OF SELECTIVE ACTION OF

THIADIAZOLYL UREA HERBICIDES

ILLE NAMME LEE and KOZO ISHIZUKA

The Institute of Physical and Chemical Research

Wako-Shi, Saitama 351, Japan

1,l-Dimethyl-3-(5-tert-butyl-l,3,4-thiadiazol-2-yl)urea(I) was newly synthesized

by the authors. (I) was found to have the strongest herbicidal activities among the
thiadiazolylurea derivatives and selectivity of (I) was found between barley (tolerant)
and wheat (susceptible) plants. Neither absorption by roots nor translocation from roots
to shoots of 35S-labeled (I) in barley and wheat correlated with selectivity between
these species of plants. (I) was metabolized in both species of plants by
N-demethylation to 1-methyt-3-(5-tert-butyl-1,3,4-thiadiazol-2-yl)urea(II), and further
to non-phytotoxic 3-(5-tert-butyl-l,3,4-thiadiazol-2-yl)urea(III).

The rates of the N-demethylating reactions from (I) to (II) and from (II) to (III) were
much greater in barley shoots than in wheat shoots, especially in the second
demethylation.

It was concluded that the selective activity of (I) between barley and wheat plants
was mainly due to the difference in rates of N-demethylation of (I). In addition, it was
found that cotton plants, well known as tolerant to phenylurea herbicides, were
remarkably susceptible to (I). It was indicated that different rates of the
N-demethylating reaction between the phenyl and thiadiazolyl herbicides correlated
with susceptibility to them.

C o m p o u n d s containing the 1,3,4-thiadiazolyl group in their chemical structures

have been extensively synthesized for new active herbicides by Kubo et al. (1970)
and F a r b e n f a r b r i k e n B a y e r ' s groups (1968) recently. A m o n g this series o f
compounds several urea derivatives were reported by both groups to have strong
phytotoxicities.

On the basis o f the work o f Kubo (1969) and Kubo et al. (1970), the authors have
newly synthesized 1,1-dimethyl-3-(5-tert-butyl- 1,3,4-thiadiazol-2-yl)urea(l)
(Ishizuka et al. 1971), which shows the strongest herbicidal activity among the
thiadiazolylurea derivatives tested.

On the other hand, it was reported that N , N - d i m e t h y l substituted phenylurea

h e r b i c i d e s s u c h as 3 - ( 3 , 4 - d i c h l o r o p h e n y l ) - l , l - d i m e t h y l u r e a ( D C M U ) and
3 - ( 4 - c h l o r o p h e n y l ) - l , l - d i m e t h y l u r e a (CMU) were inactivated in plant tissues
through N-demethylating reactions o f these compounds (Smith and Sheets 1967,
Swanson and Swanson 1968, Frear 1968). But no research was reported on the
t h i a d i a z o l y l u r e a d e r i v a t i v e s . In a d d i t i o n to i n v e s t i g a t i n g e f f e c t s o f the

Archives of Environmental Contamination 155

and Toxicology Vol. 4, 155-165 (1976)
9 1976 by Springer-Verlag New York Inc.
156 I. N. Lee and K. Ishizuka

newly-discovered herbicide on growth of plants and to looking for a distinct

selectivity, a fate of the herbicide is to be investigated with consideration of
N-demethylating rates of the thiadiazolylurea herbicides.

Discovery of a new synthetic method

A new type of urea derivatives containing 1,3,4-thiadiazol has been developed as
herbicide by Kubo et al. (1970). These compounds showed herbicidal properties
quite similar to those of phenylurea herbicides such as DCMU and CMU. Among
this series of compounds, (I) and (II) have been reported to have a strong herbicidal
activity. Especially (II) has been evaluated by Kubo et al. (1970) as the most
promising herbicide among the thiadiazolylurea derivatives.

Actually however, the authors found that the chemical structure of (I), which was
reported by them to have amp of 80 to 82~ should be correctly 5-tert-butyl.3-N,N
dimethylcarbamoyl-2-N,N-dimethylcarbamoylimino-A4-1,3,4-thiadiazoline(V),
while the structure of (II) was confirmed to be correct. The physico-chemical proper-
ties of these compounds are shown in Table I.

It was discovered that 5-tert-butyl-2-arnino-l,3,4-thiadiazol (IV) reacted with

N-methyl isocyanate to produce (II) and reacted with N,N-dimethyl-carbamoyl
chloride to produce (I). It was clarified that, in the latter reaction, substitution of the
thiadiazolyl ring of (IV) by the N,N-dimethylcarbamoyl group occurred non-
selectively both at 2- and 3- positions. Although (I) was expected to be produced by
the reaction of (IV) with N,N-dimethylcarbamoyl chloride by Kubo (1969) and

Table I. Physico-chemical properties of compounds ([), (II), and ( V)

(I) (II) (V)

mp ~ 130-131 210 g0-82

Mass spectrum m/e 228 (M§ 214 (M§ 299 (M+)

UV k MeOH, m/.t(e) 253(9420) 253(10000) 283(17000)

max

KBr cm-I 3150(NH) 3340 ~ 1715(shoulder)~

IR 3' max' 3240 j (NH) 1702 ~ (C=0)
3170 1615(C=N)
1670(C-0)
NMR ppm, C D C I a , 3.18(6H,CHa
) 2.97(3H,CH3) 2.95(3H))
TMS 11.60(1H,NH) 6.30(1H,NH) 3"06(3H)~:N CH )
11.55(1H,NH) 3.10(3H)(t - 3
3.18(3H) j
 Selective Action of Thiadiazolyl Herbicides 157

, /CH3
N--r~ CH3 N--N--CON_ N~N
II 11 II / 5% KOH. MeOH CH311 rl
CH3 ~ " S ' ~ N H 2 ' "3CH~"S'%NCON'~r, u3 CH "~J"S'J~'NHCON~ CH3
CH3 (IV) ~ CH3 (V) "n 3 3 ~CH3 (I) CH3

CH3_N=C=O N~N
,,,gH3JJ ~, .,CH3
-- 9 ',..,n3"~ ~'S NHCON
CH3 (11) \H

Fig. 1. Scheme of the synthetic method.

Farbenfarbriken Bayer's groups (1969), the authors obtained only (V) as the final
reaction product under various conditions tested.
In order to obtain the real (I), the authors hydrolyzed (V) to (I) with 5% KOH in
methyl alcohol solution in consistently high yields. The scheme of the synthetic
method is shown in Figure 1. Other 1,l-dimethyl-3-(5-alkyl-substituted-l,3,4-
thiadiazol-2-yl)ureas were synthesized similarly by the alkali hydrolytic reaction.

Herbicidal activities of newly synthesized (I) and its derivatives

Herbicidal activities of a series of thiadiazolylurea derivatives obtained by the
newly proposed synthetic method have been surveyed with several species of plants.
Wheat and barley seedlings of the two-to-three leaf stage of development were at
first used for examination of the herbicidal activities of (I), (II) and (V). Solutions
of the compounds at various concentrations in 20% methyl alcohol containing a
surfactant were sprayed onto shoots of both plants. Growth expressed as fresh
weight of the plants twenty days after treatment is shown in Figure 2.

1.2~/~ Barley 2.4~, Wheat

0.8
(v)
= i" (v) ~ 1.2'- (.)
== (") o8-
"~ 0 " 2 - ~ X ~ x (I) u. 0.4 -
" - '= X ~ x (I)
I I I I 1 I I I
0 5 10 20 30 0 5 10 20 30

Fig. 2. Effects of 5-tert-butyl-l,3,4-thiadiazolyl urea derivatives on the growth of barley and

wheat seedlings.
 158 I. N. Lee and K. Ishizuka

The growth of both species of plants was actually depressed by (I) more markedly
than by (II) and (V). From the results with a wide spectrum of plant species in
addition to wheat and barley, it was concluded that the herbicidal activity of (I) was
remarkably stronger than (II), which was evaluated by Kubo et al. (1970) as the
most promising herbicide.

Herbicidal activities of a series of l,l-dimethyl-3-(5-alkyl-substituted-1,3,4-

thiadiazol-2-yl)ureas toward wheat seedlings were surveyed. The results are shown
in Figure 3. Among these compounds 5-tert-butyl and 5-isopropyl derivatives
showed strong phytotoxities. 5-isopropyl derivatives showed phytotoxities to about
same extent as (II). On the other hand, 5-methyl, 5-ethyl and 5-propyl derivatives
showed little phytotoxic effects.

Selective activity of (I)

For the purpose o f discovering the selective activity of (I), solanaceous,
graminaceous, cucurbitaceous and leguminous plants, such as barley, wheat, egg-
plant, cucumber, soybean, and pea seedlings of the three-to-four-leaf stage were
prepared. Plants were grown in pots (8.6 cm in diameter x 11.6 cm high) in a
controlled phytotron. Twenty ml of solutions of serial concentrations of 200, 400,
600,800, and 1000 ppm of(I) in 5% ethyl alcohol containing a surfactant were either
sprayed onto shoots of these plants, or applied to the soil surface.

1.4

A CH3
1.o
c
\ \ ----o-c.2c.2c.3
" 0.8-- X
v

0.6

t-

0.4

0.2
--CH 3 CH 3
I
I I I CH3
0 1000 2000 3000
Concentration (ppm)

Fig. 3. Effects of 1,t-dimethyl-3-(5-alkyl-substituted-t,3,4-thiadiazol-2-yl) ureas on the

growth of wheat seedlings.
 Selective Action of Thiadiazolyl Herbicides 159

Ten days after the treatment these plants, except barley, were completely with-
ered by both methods of application at 200 ppm of (I). Effects on the growth of
barley by shoot application of (I) were so strong that a refined selectivity between
barley and wheat plants was not obtained. Solutions of much lower concentrations
than used before were applied to roots for the purpose of obtaining a refined
selectivity. Most of the herbicides were used by application to roots. Development
o f new leaves was observed in barley seven days after treatment, although growth
was retarded.

A survey for selectivity between barley and wheat plants was made. Barley plant
was observed a little more tolerant for (I) than other species of plants tested. Twenty
ml each of solutions of 20, 40, 100, 200, 400, and 1000 ppm concentrations was
supplied to the soil surface of a culture-pot where five seedlings of barley or wheat
were grown. Growth expressed as fresh weight of the plants fourteen days after
treatment are shown in Figure 4. Growth of wheat plants was remarkably depressed
even by 20 ppm of (I), while growth of barley plants was depressed by concentra-
tions higher than 200 ppm of (I), while barley plants were not completely withered
even by 400 ppm. A difference in susceptibility toward (I) was found between
barley and wheat plants, especially in the treatments with rather low concentrations.

Mechanism on selective a c t i o n o f (I)

To clarify the selectivity between barley (tolerant) and wheat (susceptible) plants
obtained by root application, absorption by roots, translocation from roots to shoots,
and chemical transformation in plants of (I) were investigated. Wheat and barley

120

I I I I
0 20 40 100 200 400
Concentration (ppm)

Fig. 4. Selective herbicidal effects of (I) on barley and wheat plants.

 160 I. N. Lee and K. Ishizuka

seedlings of the three-to-four-leaf stage of development were grown in water culture

and 35S-labeled (I), (II), (III) and (IV) were synthesized and applied directly to the
roots.

Rates of translocation and absorption of (I) by both species o f p l a n t s were

determined after 2, 14, 26, 38, and 50 hr. exposure of (I) to roots. The results are
shown in Table H.

The rate of absorption of (I) by barley roots (tolerant) was much more rapid than
by wheat roots (susceptible). It was not shown that a plant susceptible to (I)
absorbed (I) much more than a tolerant plant, and that the rate of the absorption was
one of the main factors which determined the susceptibility. Little difference in
translocation from roots to shoots of both species of plants was observed.

To examine the chemical transformation of (I) in plant tissues, plants were grown
in test tubes in solutions containing aSS-labeled (I) for two hr and 2, 4, 7 and 10
days. The plants, removed from the test tube, were cut to root and shoot parts after
the roots were washed with water. Each part of the plant tissues was extracted with
200 ml of 80% ethyl alcohol solution.

Very little aSS-radioactivity remained in residual fraction. These extracts were

further fractionated into water- and acetone-soluble fractions. Less than 10% of the
incorporated aSS-radioactivity was in the water-soluble fraction from both species.

Table II. Absorption and translocation of 1,1-dimethyl-3-(5-tert-butyl-l,3,4-thiadiazol-

2-yl)urea by wheat and barley plants(xlO s dpm/dry wt. g)

Absorption Translocation
Wheat Barley Wheat Barley
Exposure
Hrs dpm a %b dpm % dpm c %d dpm %

2 1924 27 2132 28 86 7 163 13

14 2581 31 3255 41 360 24 829 46
26 2866 40 3898 45 787 48 1109 50
38 3280 44 4692 60 1200 57 1603 59
50 4515 50 5662 70 1533 63 1704 58

a. Dpm/g. dry wt. of roots.

b. Indicates as percentages of the initially applied amount of (I).
c. Values indicate as S radioactivities translocated to shoot parts.
[ 3s s in sh~176 ]
d. Values indicate . X I O0
total as S in plants
 Selective Action of Thiadiazolyl Herbicides 161

After concentration in vacuo, the fractions were thin-layer chromatographed and the
radioactivities on chromatography plates were measured. 35S-determination was
conducted by a liquid scintillation spectrometer (Beckman, LS-150). The metabo-
lites in acetone fraction were identified by mean of co-chromatography.

Percentages of 35S-labeled (I) and its metabolic products detected in shoots were
measured in time course. The root parts were exposed to 35S-labeled (I) solution
continuously during the application periods. Results are shown in Figure 5. (II),
(III), (IV), and three more unidentified compounds were found as metabolic pro-
ducts of (I) in both species of plants.

It was shown that (I) was metabolized in both species of plants to (II), which still
possessed a herbicidal activity, and furthermore to non-phytotoxic (III). But the rate
of the metabolic N-demethylating reaction from (I) to (II) was much greater in
barley shoots (tolerant) than in wheat shoots (susceptible).

100

8O

"0

6o B
v

v
"0
c

0
O.
E
o 4o

Er
"o

20 m

ol I I I I
2h 2 4 7 10
Exposure time (days)

Fig. 5. Change in percentages of I, t-dimethyl-3-(5-tert-butyl- 1,3,4-thiadiazol-2-yl) urea in

wheat and barley shoots.
 162 I. N. Lee and K. Ishizuka

Since (II), possessing a herbicidal activity, was present at higher percentages in

plant tissues than other non-phytotoxic metabolites, study on metabolism of (II) was
done under same conditions as (I). (II) was metabolized in both species of plants to
(III) and (IV). The rate of the metabolic N-demethylating reaction from (II) to (III)
was obviously much more rapid in barley shoots than in wheat shoots. 9The differ-
ence in metabolic N-demethylating rates of both species of plants was apparently
much faster in the second demethylating reaction than the first. It is important to
emphasize that the second demethylation is a reaction from the phytotoxic com-
pound to the completely non-phytotoxic compound toward both wheat and barley
plants. Results are shown in Figure 6.

It is concluded that the selective activity of (I) between barley and wheat plants is
mainly due to the remarkable difference in the rates of N-demethylation of (I) and
(II).

IO0

80

E 0
e~

20

[. I I I I
2h. 2 4 7 10

Exposure time (days)

Fig. 6. Change in percentages of l-methyl-3-(5-tert-butyl-l,3,4-thiadiazol-2-yl)urea in

wheat and barley shoots.
 Selective Action of Thiadiazolyl Herbicides 163

C o m p a r i s o n o f herbicidal activities and N - d e m e t h y l a t i o n

o f phenyl- and thiadiazolyl-urea herbicides
DCMU and CMU have been used extensively as soil-applied herbicides. It was
reported that these phenylurea herbicides exhibited no herbicidal activity toward
cotton and plantain, but were very active toward corn (Swanson and Swanson 1968).
This selectivity was correlated with the N-demethylating reaction of the herbicides.
DCMU was N-demethylated as a great rate in cotton and plantain plants. It was of
interest to determine whether the thiadiazolylurea herbicides had no herbicidal
activity toward these species of plants. Therefore, herbicidal activities of phenyl-
urea herbicides such as DCMU, 3-(3-chloro-4-difiuorochloromethylthiophenyl)-
1,1-dimethylurea (fluothiuron), recently developed as a herbicide for rice-fields, and
thiadiazolylurea herbicides such as (I), (II) were surveyed toward cotton plants. It
was found that cotton plants were also tolerant to the phenylurea herbicides,
coinciding with the results obtained by Frear (1968). But they were observed, as
shown in Figure 7, to be very susceptible to the thiadiazolylurea herbicides.

Rates of N-demethylation of (I) were examined in cotton and barley shoots under
the same conditions as mentioned before, sSS-radioactivities were found in several
metabolic products in shoots of barley plants. Within two hr, more than 70% of the
parent compound was transformed chemically. But in cotton plants it was mostly
detected in (I) and some in other metabolic products. The thiadiazolylurea herbicide
(I) was only slightly N-demethylated in cotton plants, while phenylurea herbicides

'4o r
DCMU

_~ 100 ................................

\
80 -- X Fluothiuron

60

x ~ x
--X,
40 - X
(I)

20 -

I . I I I I
0 20 40 100 200 400
Concentration (pprn)

Fig. 7. Effects of phenyl- and thiadiazolyl-urea herbicides on the growth of cotton plants.
164 I. N. Lee and K. Ishizuka

80
0

60- ~,~rley
"O
c
.-i
o 40--
E
8
~ 20--
E
0.

I I I .I
0.5 1.0 1.5 2.0
Time (hr)
Fig. 8. Metabolism of 1,1-dimethyl-3-(5-tert-butyl- 1,3,4-thiadiazol-2-yl) urea herbicide in
shoots of cotton and barley plants.

such as DCMU were reported by Swanson et al. (1968) to be remarkably

N-demethylated. Results are shown in Figure 8.

It was concluded that the great difference in the herbicidal activity of (I) and
DCMU on cotton plants was derived mainly from the difference in rates of
N-demethylation of both compounds. A substrate specificity o f N-demethylating
enzymes in cotton plants is to be investigated.

Acknowledgment

The authors owe much to Dr. T. Tatsuno and Dr. Y. Kubota, The Institute of
Physical and Chemical Research for this synthetic work.

References

Farbenfarbriken Bayer Aktiengesellschaft, Netherland Patent 6813438(1969).

Frear, D. S.: Microsomal N-demethylation, by a cotton leaf oxidase system, of
3-(4-chlorophenyl)-l,l-dimethylurea (Monuron). Sci., 162, 674 (1968).
Ishizuka, K., I. N. Lee, T. Tatsuno, and Y. Kubota: Mechanisms of activation and
inactivation of physiologically active substances in plants (part 13). "On
herbicidal activities of 1,3,4-thiadiazolylurea derivatives." Agr. Biol. Chem.,
35, 964 (1971).
Kubo, H., R. Sato, and T. Ohi: Herbicidal activity of 1,3,4-thiadiazole derivatives.
J. Agr. Food Chem., 18, 60 (1970).
 Selective Action of Thiadiazolyt Herbicides 165

Kubo, H.: Herbicidal activity of 1,3,4-thiadiazole derivatives. Chemical Regulation

of Plants (Japan), 4, 146 (1969).
Swanson, C. R., and H. R. Swanson: Metabolic fate of Monuron and Diuron in
isolated leaf discs. Weed Sci., 16, 137 (1968).
Smith, J. W., and T. J. Sheets: Uptake, distribution, and metabolism of Monuron
and Diuron by several plants. J. Agr. Food Chem., 15, 577 (1967).

Manuscript received September 28, 1974; accepted November 23, 1974.