U.S. patent application number 10/186914 was filed with the patent office on 2002-11-28 for microbiocidal properties of poly-substituted guanidinium salts.
This patent application is currently assigned to RhoCraft Research Development, Ltd.. Invention is credited to Fyles, Thomas M., Rowe, Robert D..
Application Number | 20020177627 10/186914 |
Document ID | / |
Family ID | 24189857 |
Filed Date | 2002-11-28 |
United States Patent
Application |
20020177627 |
Kind Code |
A1 |
Fyles, Thomas M. ; et
al. |
November 28, 2002 |
Microbiocidal properties of poly-substituted guanidinium salts
Abstract
This patent discloses the use of poly-substituted isothiouronium
salts (T1 where R.sub.1, R.sub.2, R.sub.3, R.sub.4=hydrogen, alkyl,
cycloalkyl, alkenyl, cycloalkenyl, aryl, heteroaryl, etc. and
X.sup.-=Cl.sup.-, Br.sup.-, NO.sub.3.sup.-, CH.sub.3CO.sub.2.sup.-,
or any other common anion), poly-substituted guanidinium salts (G1
where R.sub.1, R.sub.2, R.sub.3, R.sub.5, R.sub.6=hydrogen, alkyl,
cycloalkyl, alkenyl, cycloalkenyl, aryl, heteroaryl, etc. and
X.sup.-=Cl.sup.-, Br, NO.sub.3.sup.-, CH.sub.3CO.sub.2.sup.-, or
any other common anion), or mixtures of two or more of the above
compounds, as the biocidal component of microbiocidal or
anti-fouling formulations.
Inventors: |
Fyles, Thomas M.; (Victoria,
CA) ; Rowe, Robert D.; (Victoria, CA) |
Correspondence
Address: |
KLARQUIST SPARKMAN, LLP
121 SW SALMON STREET
SUITE 1600
PORTLAND
OR
97204
US
|
Assignee: |
RhoCraft Research Development,
Ltd.
|
Family ID: |
24189857 |
Appl. No.: |
10/186914 |
Filed: |
June 28, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
10186914 |
Jun 28, 2002 |
|
|
|
09548666 |
Apr 13, 2000 |
|
|
|
Current U.S.
Class: |
514/580 ;
514/353; 514/634 |
Current CPC
Class: |
C09D 5/1625 20130101;
C07C 335/08 20130101; A01N 47/42 20130101; A01N 47/44 20130101;
C07C 279/04 20130101 |
Class at
Publication: |
514/580 ;
514/634; 514/353 |
International
Class: |
A61K 031/44; A61K
031/155; A61K 031/17 |
Claims
We claim:
1. A composition comprising a compound selected from the group
consisting of compounds having a first formula 3where R.sub.4 and
at least one of the other R.sub.1, R.sub.2 or R.sub.3 groups are
selected independently from the group consisting of alkyl,
cycloalkyl, alkenyl, cycloalkenyl, and heteroaryl other than
phthalocyanine, the remaining R.sub.1, R.sub.2 or R.sub.3 groups
are independently selected from the group consisting of hydrogen,
alkyl, cycloalkyl, alkenyl, cycloalkenyl, and heteroaryl, and
X.sup.- is an anion; compounds having a second formula 4where
R.sub.4 and at least one of the other R.sub.1, R.sub.2 or R.sub.3
groups are independently selected from the group consisting of
alkyl, cycloalkyl, alkenyl, cycloalkenyl, and heteroaryl other than
phthalocyanine, and the remaining R.sub.1, R.sub.2 or R.sub.3
groups are independently selected from the group consisting of
hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, and heteroaryl;
compounds having a third formula 5where R.sub.1, R.sub.2, R.sub.3,
R.sub.5, and R.sub.6 are selected such that at least two of the
R.sub.1, R.sub.2, R.sub.3, R.sub.5 or R.sub.6 groups are
independently selected from the group consisting of alkyl,
cycloalkyl, alkenyl, cycloalkenyl, aryl, and heteroaryl, the
remaining R.sub.1, R.sub.2, R.sub.3, R.sub.5 or R.sub.6 groups are
independently selected from the group consisting of hydrogen,
alkyl, cycloalkyl, alkenyl, cycloalkenyl, aryl, and heteroaryl, and
X is an anion; compounds having a fourth formula 6where R.sub.1,
R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are selected such that at
least two of the R.sub.1, R.sub.2, R.sub.3, R.sub.5 or R.sub.6
groups are selected from the group consisting of alkyl, cycloalkyl,
alkenyl, cycloalkenyl, aryl, and heteroaryl and the remaining
R.sub.1, R.sub.2, R.sub.3, R.sub.5 or R.sub.6 groups are
independently selected from the group consisting of hydrogen,
alkyl, cycloalkyl, alkenyl, cycloalkenyl, aryl, and heteroaryl; and
mixtures thereof.
2. The composition according to claim 1 where X.sup.- is selected
from the group consisting of halide, nitrate, and acetate.
3. The composition according to claim 1 where the total number of
carbon atoms in selected compounds is between about 8 and about
30.
4. A compound selected from the group consisting of compounds
having a first formula 7where at least two of the R.sub.1, R.sub.2,
R.sub.3, R.sub.5 or R.sub.6 groups are independently selected from
the group consisting of alkyl, cycloalkyl, alkenyl, cycloalkenyl,
aryl, and heteroaryl, the remaining R.sub.1, R.sub.2, R.sub.3,
R.sub.5 or R.sub.6 groups are independently selected from the group
consisting of hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl,
aryl, and heteroaryl, and X.sup.- is an anion; and compounds having
a second formula 8where at least two of the R.sub.1, R.sub.2,
R.sub.3, R.sub.5 or R.sub.6 groups are selected from the group
consisting of alkyl, cycloalkyl, alkenyl, cycloalkenyl, aryl, and
heteroaryl and the remaining R.sub.1, R.sub.2, R.sub.3, R.sub.5 or
R.sub.6 groups are independently selected from the group consisting
of hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, aryl, and
heteroaryl; and mixtures thereof.
5. The compound according to claim 4 where X.sup.- is selected from
the group consisting of halide, nitrate, and acetate.
6. The compound according to claim 4 where the total number of
carbon atoms in selected compounds is between about 8 and about
30.
7. A composition comprising a compound selected from the group
consisting of compounds having a first formula 9where R.sub.1,
R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are selected such that at
least two of the R.sub.1, R.sub.2, R.sub.3, R.sub.5 or R.sub.6
groups are independently selected from the group consisting of
alkyl, cycloalkyl, alkenyl, cycloalkenyl, aryl, and heteroaryl, the
remaining R.sub.1, R.sub.2, R.sub.3, R.sub.5 or R.sub.6 groups are
independently selected from the group consisting of hydrogen,
alkyl, cycloalkyl, alkenyl, cycloalkenyl, aryl, and heteroaryl, and
X.sup.- is an anion; compounds having a second formula 10where
R.sub.1, R.sub.2, R.sub.3, R.sub.5, and R.sub.6 are selected such
that at least two of the R.sub.1, R.sub.2, R.sub.3, R.sub.5 or
R.sub.6 groups are selected from the group consisting of alkyl,
cycloalkyl, alkenyl, cycloalkenyl, aryl, and heteroaryl and the
remaining R.sub.1, R.sub.2, R.sub.3, R.sub.5 or R.sub.6 groups are
independently selected from the group consisting of hydrogen,
alkyl, cycloalkyl, alkenyl, cycloalkenyl, aryl, and heteroaryl; and
mixtures thereof.
8. The compound according to claim 7 where X.sup.- is selected from
the group consisting of halide, nitrate, and acetate.
9. The compound according to claim 7 where the total number of
carbon atoms in selected compounds is between about 8 and about 30.
Description
CROSS REFERENCE TO RELATED APPLICATION
[0001] This is a divisional application of currently pending U.S.
patent application Ser. No. 09/548,666 filed on Apr. 13, 2000,
which is incorporated herein by reference.
BACKGROUND
[0002] Surfaces exposed to humid and aqueous environments are
readily colonized by microorganisms and may be further colonized by
higher organisms. The resultant fouling has many adverse effects on
these surfaces and the objects they compose. Current anti-fouling
methods often involve the use of highly toxic and environmentally
stable compounds, usually including a metal ion such as cooper (as
in CuO.sub.2) or tin (as in tri-butyltin fluoride, TBTF). Research
has shown that these heavy metals remain in the environment and
retain their toxicity for many years. Furthermore, it has been
demonstrated that these compounds become concentrated in plants and
animals higher up on the food chain with many adverse effects.
These compounds while effective as anti-fouling agents are under
increasing pressure from environmental regulations that seek to
limit the concentration of heavy metals in the environment. An
effective anti-fouling agent with a short and known biological
lifetime is therefore of great interest to the industry.
[0003] This invention describes the use of poly-substituted
isothiouronium salts (T1), and poly-substituted guanidinium salts
(G1). The biological activity of these classes of compounds has
been recognized previously as variously fungicides, bactericides,
lepidoptericides, antibiotics, etc. It is also known that
isothiouronium salts (T1) and neutral isothioureas (T2) can be
interconverted. Similarly, the interconversion of guanidinium salts
(G1) and neutral guanidines (G2) is also well established. These
pairs of compounds are related as acid and conjugate base,
differing only in protonation state. Consequently, in many cases
the biological activity of isothiouronium salts can be inferred
from the known activity of isothioureas, and vice versa. Similarly
the biological activity of guanidinium salts can be inferred from
the known activity of neutral guanidines, and vice versa. This is
particularly true when the compounds are dispersed in an aqueous
environment.
[0004] Many authors have reported examples of the activity of
isothiouronium salts and isothioureas. For example, U.S. Pat. No.
4,515,813 discloses the lepidoptericidal properties of isothiourea
compounds. Fungicidal and bactericidal activity of this class of
compounds were also noted. Similarly, the use of pyridyl
thiouronium salts as fungicides are disclosed in U.S. Pat. No.
3,655,898, and related pyridyl thiouronium N-oxides are useful as
wood preservatives as described in Japan Patent No. 53109903.
German Patent No. 2637651 describes the use of
S-(p-isopropylbenzyl)thiouronium chloride as one of the biocidal
components in a water-based paint formulation. Marine anti-fouling
activity by dissolved isothioureas is disclosed in Japan Patent No.
05163105.
[0005] Similarly, there are many examples of the biological
activity of guanidinium salts and guanidines. Several naturally
occurring toxins from marine organisms contain the guanidinium
functional group, most notably tetrodotoxin. The best-known
commercial example is dodecyl guanidinium acetate (dodine), widely
used as a fungicide and bactericide to control scab on hard fruits.
It is also used as an industrial biocide and preservative. Dodine
also shows synergistic anti-fouling activity in conjunction with
other well-known anti-fouling agents such as tributyltin oxide as
reported by Evans, Callow and Wood (1986). Dodine, in conjunction
with quaternary ammonium salts, is reported by Bidwell, Farris and
Cherry (1995) to control the growth of zebra mussels and Asian
clams (moluscicidal activity). Such soluble formulations have also
been disclosed in U.S. Pat. No. 4,816,163, U.S. Pat. No. 4,906,385,
and Canadian Patent No. 1,269,927. A method to prepare an
anti-fouling coating from a mixture of dodine and additional
biocides has been disclosed in Japan Patent No. 04225945.
BRIEF SUMMARY OF THE INVENTION
[0006] Most of the previously reported isothiouronium and
guanidinium containing compounds are monosubstituted with a
relatively low carbon-number substituent. Although this is
appropriate for applications requiring soluble biocides, it is
obvious to someone skilled in the art that a successful coating
application in contact with water will require sparingly soluble
biocides. Solubility can be limited by increasing the carbon number
of a single substituent, or by increasing the number of similar
sized substituents. The present invention discloses the utility of
the second strategy.
[0007] A second issue, previously unrecognized, is the role that an
anion exchange may play in biocidal activity. Prior discussion of
the mode of action of biocidal formulations containing
isothiouronium or guanidinium salts focused on their detergent
capabilities (references cited above). Our parallel work on the
development of ion-exchange membranes for dissolved gas sensors
(U.S. patent application Ser. No. 09/444,867) showed that
guanidinium salts are effective agents for the exchange of
hydroxide ions across membranes. In the context of biocidal
activity, an anion exchanger would disrupt the normal ionic and pH
balance across a cell membrane that would prove to be fatal for
microorganisms.
[0008] This invention describes the synthesis of a series of
poly-substituted isothiouroniums of type T1 and poly-substituted
guanidinium salts of type G1, via intermediate thioureas of type
T3, their formulation in paints and their activity in limiting the
growth of marine organisms on the treated surfaces as a result of
prolonged immersion in open seawater. In addition to the biocidal
and anti-fouling activity disclosed below, compounds of types T1
and G1 possess two additional properties of significant utility.
The first is that they are colorless, that is they do not absorb
significant amounts of visible light. Thus they could be used to
inhibit fouling on windows exposed to humid or aqueous
environments. The second is that they degrade easily in a marine
environment to produce benign by-products. Thus a buildup of these
compounds in the environment will be avoided.
[0009] Structural Formulae:
[0010] General Formulae of Compounds Discussed: 1 2
DETAILED DESCRIPTION OF THE INVENTION
[0011] Synthesis of Compounds:
[0012] The synthesis of the various compounds is given in the
reaction schemes in terms of a general procedure. The counterion
produced in the synthesis is typically iodide that is subsequently
exchanged for chloride or other anions by ion exchange. Procedure A
can produce monosubstituted guanidinium salts or N,N-disubstituted
guanidinium salts, depending on the starting amine. Procedure B can
produce N,N'-disubstituted thioureas, or N,N,N'-trisubstituted
thioureas, depending on the starting amine. The first step of
procedures C and D can produce N,N',S-trisubstituted isothiouronium
salts or N,N,N',S-tetrasubstituted isothiouronium salts, depending
on the starting thiourea. Direct reaction of isothiouronium salts
with ammonia (procedure C) gives either N,N'-disubstituted
guanidinium salts or N,N,N'-trisubstituted guanidinium salts.
Alternatively, reaction of isothiouronium salts with primary or
secondary amines (procedure D) can produce N,N',N"-trisubstituted
guanidinium salts, N,N,N',N"-tetrasubstituted guanidinium salts, or
N,N,N'N',N"-pentasubstituted guanidinium salts, depending on the
starting isothiouornium salt and starting amine. A total of nine
mono-, di-, and tri-substituted guanidinium salts (G1), and three
tri-substituted isothiouronium salts (T1) were prepared by the
methods shown.
[0013] All compounds shown were characterized by NMR, MS, and IR.
By the methods disclosed below, the purity of the compounds was
high without recourse to chromatographic separation. Samples of
each compound were further purified by chromatography on silica.
The purified materials all showed UV cutoff values below 300 nm,
and showed an .epsilon. less than one at 300 nm for all
compounds.
[0014] It will be obvious to someone skilled in the art that the
nature of the substituent groups R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6 in structures T1 and G1 can be varied by
judicious choice of starting amines and isothiocyanates according
to the reaction schemes presented. The following general
procedures, given for alkyl substituents, illustrate the methods
used.
[0015] Procedure A:
[0016] (!!CAUTION!!: This procedure evolves methyl mercaptan. Use a
hood. Avoid exposure.) An alkyl amine (1 eq.) and solid S-methyl
isothiouronium iodide (2 eq.) were suspended in absolute ethanol
(5-8 mL/g salt). The mixture was stirred at reflux under a reflux
condenser. The evolution of methyl mercaptan was followed using
moistened lead acetate test paper. The reaction was usually
complete in 6 hours, but the reflux was continued overnight. The
mixture was evaporated to a solid and redissolved in water. The
water was extracted on a continuous extractor overnight using
chloroform, the extracts were dried over magnesium sulfate,
filtered and evaporated to yield the iodide salt of the product.
The iodide was converted to the chloride using Amberlite IRA400
resin in methanol.
[0017] Spectroscopic Data for Compounds Prepared by Procedure
A:
[0018] G1 where R.sub.1=C.sub.10H.sub.21,
R.sub.2=R.sub.3=R.sub.5=R.sub.6=- H, X.sup.-=Cl.sup.-: .sup.1 H NMR
(CDCl.sub.3, .delta.): 0.95 (br. t., 3H), 1.54 (br. s., 14H), 1.60
(br. m., 2H), 3.10 (t., 2H), 4.90 (br. s., >SH); .sup.3C NMR
(CDCl.sub.3, .delta.):14.2, 22.7, 26.7, 28.5, 29.3 (m), 31.9, 42.5,
156.6; MS (+LSIMS, mNBA): 200.2 (M-Cl).
[0019] G1 where R.sub.1=C.sub.14H.sub.29,
R.sub.2=R.sub.3=R.sub.5=R.sub.6=- H, X.sup.-=Cl.sup.-: .sup.1H NMR
(CDCl.sub.3, .delta.): 0.95 (br. t., 3H), 1.5 (br. s., 22H), 1.6
(br. m., 2H), 3.10 (t., 2H), 4.9 (br. s., >5H); MS (+LSIMS,
mNBA): 256.2 (M-Cl).
[0020] G1 where R.sub.1=C.sub.18H.sub.37,
R.sub.2=R.sub.3=R.sub.5=R.sub.6=- H, X.sup.-=I.sup.-: .sup.1 NMR
(DMSO-d.sub.6, .delta.): 0.95 (br. t., 3H), 1.54 (br. s., 30H),
1.60 (br. m., 2H), 3.10 (t., 2H), 4.90 (br. s., >5H); MS
(+LSIMS, mNBA): 312.2 (M-I).
[0021] Procedure B:
[0022] (!!CAUTION!!: isothiocyanates are typically lachrymators.
Use a hood. Avoid exposure). An alkyl amine (1 eq.) and an alkyl
isothiocyanate (1 eq.) were dissolved in toluene (5 mL/g amine).
The mixture was stirred at reflux for 3-5 hours. The product
precipitated in some cases. The mixture was concentrated under
reduced pressure, cooled and filtered. The precipitate was washed
with pentane and air-dried. The product is sufficiently pure for
the subsequent reaction.
[0023] Spectroscopic Data for Compounds Prepared by Procedure
B:
[0024] T3 where R.sub.1=C.sub.10H.sub.21, R.sub.2=H,
R.sub.3=C.sub.4H.sub.9: .sup.1 H NMR (CDCl.sub.3,.delta.): 0.95
(m., 6H), 1.3 (br. s., 16H), 1.60 (m., 4H), 3.4 (br. s., 4H), 5.7
(br. s., 2H); .sup.13C NMR (CDCl.sub.3, .delta.): 13.7, 14.1, 20.2,
22.7, 26.9, 29.5 (m.), 31.0, 31.9, 44.0 (br.), 139.5; MS (+LSIMS,
mNBA): 273.2 (M+H).
[0025] T3 where R.sub.1=C.sub.14H.sub.29, R.sub.2=H,
R.sub.3=C.sub.4H.sub.9: .sup.1 H NMR (CDCl.sub.3, .delta.): 0.95
(m., 6H), 1.3 (br. s., 24H), 1.60 (m., 4H), 3.4 (br. s., 4H), 5.7
(br. s., 2H); .sup.13C NMR (CDCl.sub.3, .delta.): 13.7, 14.1, 20.2,
22.7, 26.9, 29.5 (m.), 31.0, 31.9, 44.0 (br.), 139.5; MS (+LSIMS,
mNBA): 329.2 (M+H).
[0026] T3 where R.sub.1=C.sub.18H.sub.37, R.sub.2=H,
R.sub.3=C.sub.4H.sub.9: .sup.1 H NMR (CDCl.sub.3, .delta.): 0.95
(m., 6H), 1.3 (br. s., 32H), 1.60 (m., 4H), 3.4 (br. s., 4H), 5.7
(br. s., 2H); .sup.13C NMR (CDCl.sub.3, .delta.): 13.7, 14.1, 20.2,
22.7, 26.9, 29.5 (m.), 31.0, 31.9, 44.0 (br.), 139.5; MS (+LSIMS,
mNBA): 385.2 (M+H).
[0027] Procedure C:
[0028] (!!CAUTION!!: This procedure evolves methyl mercaptan. Use a
hood. Avoid exposure). A substituted thiourea (1 eq.) was suspended
in absolute ethanol (5-10 mL/g, thiourea may not dissolve) and
idomethane (3 eq.) was added. The mixture was sealed in a low
pressure hydrogenation bottle, stirred and heated to 80.degree. C.
After cooling, the vessel was opened and the unreacted idomethane
and solvent was removed by evaporation. The product at this stage
is an isothiouronium salt of type T1 and may be worked up as
indicated after the ammonia treatment below.
[0029] The product was dissolved in absolute ethanol (SmL/g) and
ammonia was added via a bubbler at a rate to allow dissolution.
Ammonia addition was continued until a large excess was assured.
The mixture was again sealed and heated at 80.degree. C. for 24
hours. After cooling the vessel was opened (!!CAUTION!!: use a fume
hood) and reheated to drive off the methyl mercaptan. The mixture
was then evaporated to a thick oil. The oil was dissolved in water,
extracted with methylene chloride, the extracts were dried over
magnesium sulfate, filtered and evaporated to yield the iodide salt
of the product.
[0030] The iodide was converted to the chloride using Amberlite
IRA400 resin in methanol.
[0031] Spectroscopic Data for Compounds Prepared by Procedure
C:
[0032] T1 where R.sub.1=C.sub.10H.sub.21, R.sub.2=H,
R.sub.3=C.sub.4H.sub.9, R.sub.4=CH.sub.3, X.sup.-=Cl.sup.-: .sup.1
H NMR (CDCl.sub.3, .delta.): 0.9 and 0.95 (2 br. t., 6H), 1.2 (br.
s., 16H), 1.6 (br. m., 4H), 2.8 (br. s., 3H), 3.75 (m., 4H), 7.8
(br. 2H); .sup.13C NMR (CDCl.sub.3, .delta.): 13.7, 14.1, 19.8,
22.6, 27.0, 29.3 (m.), 31.8, 44.7, 45.0, 50.1, 166.5; MS (+LSIMS,
mNBA): 287.4 (M-Cl).
[0033] T1 where R.sub.1=C.sub.14H.sub.29, R.sub.2=H,
R.sub.3=C.sub.4H.sub.9, R.sub.4=CH.sub.3, X.sup.-=I.sup.-: .sup.1 H
NMR (CDCl.sub.3, .delta.): 0.9 and 0.95 (2 br. t., 6H), 1.2 (br.
s., 24H), 1.6 (br. m., 4H), 2.8 (br. s., 3H), 3.75 (m., 4H), 7.85
(br. 1H), 8.4 (br. 1H); 13C NMR (CDCl.sub.3, .delta.): 13.7, 14.1,
19.7, 22.9, 27.1, 29.4 (m.), 31.8, 44.7, 45.1, 50.3, 166.5; MS
(+LSIMS, mNBA): 343.4 (M-I).
[0034] T1 where R.sub.1=C.sub.18H.sub.37, R.sub.2=H,
R.sub.3=C.sub.4H.sub.9, R.sub.4=CH.sub.3, X.sup.-=Cl.sup.-: .sup.1
H NMR (CDCl.sub.3, .delta.): 0.9 and 0.95 (2 br. t., 6H), 1.2 (br.
s., 32H), 1.6 (br. m., 4H), 2.8 (br. s., 3H), 3.75 (m., 4H); 8.0
(br. 2H); .sup.13C NMR (CDCl.sub.3, .delta.): 13.7, 14.1, 19.7,
22.6, 27.1, 29.3 (m.), 31.8, 44.7, 44.8, 50.3, 166.5; MS (+LSIMS,
mNBA): 399.4 (M-Cl).
[0035] G1 where R.sub.1=C.sub.10H.sub.21, R.sub.3=C.sub.4H.sub.9,
R.sub.2=R.sub.5=R.sub.6=H, X.sup.-=Cl.sup.-: .sup.1 H NMR
(CDCl.sub.3, .delta.): 0.9 and 0.95 (2 br. t., 6H), 1.2 and 1.4
(br. s. +m., 16H), 1.6 (br. m., 4H), 3.15 (br. m., 4H), 6.4-7.0
(br., 4H); .sup.13C NMR (CDCl.sub.3, .delta.): 13.7, 14.1, 20.0,
22.7, 26.8, 29.4 (m.), 31.9, 42.2, 42.5, 155.8; MS (+LSIMS, mNBA):
256.4 (M-Cl).
[0036] G1 where R.sub.1=C.sub.14H.sub.29, R.sub.3=C.sub.4H.sub.9,
R.sub.2=R.sub.5=R.sub.6=H, X.sup.-=Cl.sup.-: .sup.1 H NMR
(CDCl.sub.3, .delta.): 0.9 and 0.95 (2 br. t., 6H), 1.2 and 1.4
(br. s. +m., 24H), 1.6 (br. m., 4H), 3.15 (br. m., 4H), 6.4-7.0
(br., 4H); .sup.13C NMR (CDCl.sub.3, .delta.): 13.7, 14.1, 19.9,
22.7, 26.9, 29.4 (m.), 31.9, 41.8, 42.1, 156.2; MS (+LSIMS, mNBA):
312.5 (M-Cl).
[0037] G1 where R.sub.1=C.sub.18H.sub.37, R.sub.3=C.sub.4H.sub.9,
R.sub.2=R.sub.5=R.sub.6=H, X.sup.-=Cl.sup.-: .sup.1 H NMR
(CDCl.sub.3, .delta.): 0.9 and 0.95 (2 br. t., 6H), 1.2 and 1.4
(br. s. +m., 32H), 1.6 (br. m., 4H), 3.15 (br. m., 4H), 6.4-7.0
(br., 4H); MS (+LSIMS, mNBA): 368.6 (M-Cl).
[0038] Procedure D:
[0039] The procedure was identical to C with the exception of an
alkyl amine (2 eq.) was used in place of ammonia.
[0040] Spectroscopic Data for Compounds Prepared by Procedure
C:
[0041] G1 where R.sub.1=C.sub.10H.sub.21,
R.sub.3=R.sub.5=C.sub.4H.sub.9, R.sub.2=R.sub.6=H,
X.sup.-=Cl.sup.-: .sup.1 H NMR (CDCl.sub.3, .delta.): 0.95 (m.,
9H), 1.2-1.4 (m., 18H), 1.6 (m., 6H), 3.25 (br. m., 6H), 7.0-7.2
(br., 3H); .sup.3C NMR (CDCl.sub.3, .delta.): 13.7, 13.8, 14.1,
19.9, 20.0, 22.7, 26.8, 29.4 (m.), 31.8, 31.9, 42.3,42.6, 155.4; MS
(+LSIMS, mNBA): 312.2 (M-Cl).
[0042] G1 where R.sub.1=C.sub.14H.sub.29,
R.sub.3=R.sub.5=C.sub.4H.sub.9, R.sub.2=R.sub.6=H,
X.sup.-=Cl.sup.-: .sup.1 H NMR (CDCl.sub.3, .delta.): 0.95 (m.,
9H), 1.2-1.4 (m., 26H), 1.6 (m., 6H), 3.25 (br. m., 6H), 7.0-7.2
(br., 3H); .sup.13C NMR (CDCl.sub.3, .delta.): 13.7, 14.1, 19.9,
22.7, 26.8, 29.4 (m.), 31.8, 31.9, 42.5, 42.8, 155.7; MS (+LSIMS,
mNBA): 368.6 (M-Cl).
[0043] G1 where R.sub.1=C.sub.18H.sub.37,
R.sub.3=R.sub.5=C.sub.4H.sub.9, R.sub.2=R.sub.6=H, X.sup.-=I.sup.-:
.sup.1H NMR (CDCl.sub.3, .delta.): 0.95 (m., 9H), 1.2-1.4 (m.,
34H), 1.6 (m., 6H), 3.25 (br. m., 6H), 7.0-7.2 (br., 3H); .sup.13C
NMR (CDCl.sub.3, .delta.): 13.7, 14.1, 19.9, 22.7, 26.8, 29.4 (m.),
31.8, 31.9, 42.5, 42.8, 154.6; MS (+LSIMS, mNBA): 324.7 (M-I).
[0044] Assessment of Marine Anti-Fouling Activity
[0045] An experiment was designed to follow the onset and
development of algal growth on painted panels held approximately 1
m below the surface in open seawater. The apparatus consisted of a
moored floating superstructure with a set of test panels suspended
below it. The superstructure allowed the test panels to be lifted
from the water periodically to assess the extent of growth and to
photograph the panels. The apparatus was designed to hold 90 test
panels each 10 cm square. The experiment examined nine of the
compounds prepared, at two different dose levels (5 and 10 wt %) in
three different topside marine paints (9.times.2.times.3=54 primary
samples). None of the paints contained any commercial anti-fouling
agent. A total of 12 control samples were included: 6 which were
not touched, and 6 which were used for scrapes to examine the type
of organisms populating the surface fouling layer. The remaining 24
test panels were assigned to randomized replicates of the primary
samples, in sets of 8 for each paint type. The locations of the
controls were fixed on the six arrays and the remaining 78 test
panels were randomly assigned to the other locations.
[0046] The lexan test panels were sandblasted to provide a surface
for paint adhesion, cleaned in methanol, and then in
trifluorethanol immediately prior to painting. Paint samples were
prepared from a weighed amount of the test compound and a known
volume of paint using the measured paint density to arrive at the
nominal 5 and 10 wt % dose levels. In most cases the compounds were
dissolved in a few mL of methylene chloride before the paint was
added. The paint samples were mixed by hand until homogeneous to
the eye. A measured volume of the paint sample was spread on the
cleaned test panel with a silk-screen tool using a jig designed to
form a 250 .mu.m paint layer. The painted test panels were then
glued in place on the array and allowed to air dry for 72 hours.
After painting, the sole identifier for the compound and
formulation was from the array coordinates. Given that the six
arrays were virtually indistinguishable after drying, the specific
location of any particular compound was essentially hidden from the
subsequent observers.
[0047] The experiment was initiated in the summer of 1999.
Qualitatively, the panels remained clean for the first two weeks,
then rapidly fouled over the next two weeks. By the end of a
six-week period, the late-summer die-off of marine flora was
evident from the amount of plant debris in the water column and the
exposure of some previously fouled surfaces on the test panels. The
main fouling observed was filamentous algae that hung from the
frame of the apparatus, from the clean sections between the test
panels, and from some fouled panels.
[0048] The extent of fouling was assessed and scored by two
independent observers. Statistical controls establish excellent
agreement between the observers. The observers scored the control
panels as "heavily" fouled after a six-week exposure. At the same
time, a total of 8 test panels corresponding to 6
compound-dose-paint formulations showed significantly less growth
than the controls. Some test panels remained completely free of
algal growth after six-week exposure.
[0049] Formulations containing T1 (R.sub.1=C.sub.10H.sub.21,
R.sub.3=C.sub.4H.sub.9, R.sub.2=H, R.sub.4=CH.sub.3,
X.sup.-=Cl.sup.-) showed virtually no growth over the first six
weeks of the experiment in three different formulations. In two
formulations, growth on panels containing G1
(R.sub.1=C.sub.14H.sub.29, R.sub.3=C.sub.4H.sub.9,
R.sub.2=R.sub.5=R.sub.6=H, X.sup.-=Cl.sup.-) was inhibited relative
to controls during the initial growth period, but increased after
five weeks to levels that were less fouled but not statistically
significantly relative to controls. These data establish that these
compounds inhibit initial growth on the surfaces.
[0050] After a 9-month exposure all control panels and untreated
surfaces were heavily fouled with brown and green algae, and
barnacles had set in many places. Several other organisms inhabited
regions of the dense algal mat around and on the painted panels.
Several test panels were significantly less fouled than control
surfaces with a substantial portion of the surface (>90% in some
cases) free of attached algae and barnacles. All formulations
containing T1 (R.sub.1=C.sub.10H.sub.21, R.sub.3=C.sub.4H.sub.9,
R.sub.2=H, R.sub.4=CH.sub.3, X.sup.-=Cl.sup.-) showed clear dose
dependent anti-fouling activity. The majority of formulations
containing G1 (R.sub.1=C.sub.18H.sub.37, R.sub.3=C.sub.4H.sub.9,
R.sub.2=R.sub.5=R.sub.6=H, X.sup.-=Cl.sup.-) or G1
(R.sub.1=C.sub.10H.sub.21, R.sub.3=R.sub.5=C.sub.4H.sub.9,
R.sub.2=R.sub.6=H, X.sup.-=Cl.sup.-) also showed dose-dependent
anti-fouling activity. These data establish that these compounds
inhibit marine growth on treated surfaces, both during the initial
colonization phase, and over the longer term.
[0051] An experiment was designed to examine the stability of the
compounds in seawater over a period of time to determine the rate
of microbial degradation of the compound. Seawater samples (20 L)
were held at 1.degree. C. in an east-facing window and air was
bubbled for 2 hours each day to maintain saturation. Compound T1
(R.sub.1=C.sub.10H.sub.21, R.sub.2=H, R.sub.3=C.sub.4H.sub.9,
R.sub.4=CH.sub.3, X.sup.-=Cl.sup.-) was added at an initial
concentration of 100 nM. At intervals over 5 days, 100 mL samples
of seawater were withdrawn and analyzed by electrospray mass
spectrometry. A steady decline in the concentration of T1
(R.sub.1=C.sub.10H.sub.21, R.sub.2=H, R.sub.3=C.sub.4H.sub.9,
R.sub.4=CH.sub.3, X.sup.-=Cl.sup.-) was observed with an apparent
half-life of 80 hours under the experimental conditions. This
experiment establishes that the compound is likely to be degraded
in the environment. The product of the degradation is initially the
corresponding urea that is then further degraded by the
microorganisms in the seawater.
REFERENCES CITED
[0052] Patents
[0053] US Patents
[0054] U.S. Pat. No. 4,515,813 Fancher et al Application No.
426,366, Filed: Sept. 29, 1982
[0055] U.S. Pat. No. 3,655,898 Driscoll, Patrick Application No.
1969000083794 Filed: Jun. 30, 1969
[0056] U.S. Pat. No. 4,906,385 Lyons et al Application No.
1989000305231 Filed: Feb. 1, 1989
[0057] U.S. patent application Ser. No. 09/444,867 Filed: Nov. 22,
1999
[0058] Patents: Other
[0059] CA Patent No. 1269927 Lyons et al Application No. 535696
Filed: Apr. 27, 1987
[0060] JP Patent No. 53109903A2 Nishimoto Application No.
JP1977000024060 Filed: Mar. 4, 1977
[0061] DE Patent No. 2637651 Young et al Application No.
DE19762637651 Filed: Aug. 20, 1976
[0062] JP Patent No. 5163105A2 Hamachi et al Application No.
JP1991000332078 Filed: Dec. 16, 1991
[0063] JP Patent No. 225945A2Akashietal Application No.
JP1991000140999 Filed: May 15, 1991
OTHER REFERENCES
[0064] Evans, Callow and Wood, Synergism between antifouling
biocides, Stud. Env. Sci. (1986), 28 (Algal Biofouling) 55-64.
[0065] Bidwell, Farris and Cherry, Comparative response of the
zebra mussel, Dreissena polymorpha, and the Asian claim, Corbicula
fluminea, to DGH/QUAT, a non-oxidizing molluscicide. Aquat.
Toxicol. (1995), 33(3, 4) 183
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