U.S. patent application number 11/765348 was filed with the patent office on 2007-11-08 for triterpene quaternary salts as biologically active surfactants.
This patent application is currently assigned to Regents of the University of Minnesota. Invention is credited to Dmytro Avilov, Oksana Kolomitsyna, Pavel A. Krasutsky, Dmytre A. Krasutskyy, Tatiana Sergeeva.
Application Number | 20070259842 11/765348 |
Document ID | / |
Family ID | 27613290 |
Filed Date | 2007-11-08 |
United States Patent
Application |
20070259842 |
Kind Code |
A1 |
Krasutsky; Pavel A. ; et
al. |
November 8, 2007 |
TRITERPENE QUATERNARY SALTS AS BIOLOGICALLY ACTIVE SURFACTANTS
Abstract
The invention provides novel compounds that are quaternary amine
derivatives of betulin and other triterpenes. The compounds have
antibacterial, antifungal, and surfactant properties.
Inventors: |
Krasutsky; Pavel A.;
(Duluth, MN) ; Avilov; Dmytro; (Duluth, MN)
; Sergeeva; Tatiana; (Gainesville, FL) ;
Krasutskyy; Dmytre A.; (Duluth, MN) ; Kolomitsyna;
Oksana; (Duluth, MN) |
Correspondence
Address: |
Schwegman, Lundberg, Woessner & Kluth, P.A.
P.O. Box 2938
Minneapolis
MN
55402
US
|
Assignee: |
Regents of the University of
Minnesota
|
Family ID: |
27613290 |
Appl. No.: |
11/765348 |
Filed: |
June 19, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11765338 |
Jun 19, 2007 |
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11765348 |
Jun 19, 2007 |
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11760242 |
Jun 8, 2007 |
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11765338 |
Jun 19, 2007 |
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11612423 |
Dec 18, 2006 |
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11760242 |
Jun 8, 2007 |
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10893147 |
Jul 16, 2004 |
7199114 |
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11612423 |
Dec 18, 2006 |
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PCT/US03/01666 |
Jan 21, 2003 |
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10893147 |
Jul 16, 2004 |
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60349564 |
Jan 18, 2002 |
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Current U.S.
Class: |
514/176 ;
514/182; 540/47; 540/90; 564/282 |
Current CPC
Class: |
A61P 31/10 20180101;
C07J 53/00 20130101; A61P 31/04 20180101; C07J 63/008 20130101;
C07J 63/00 20130101 |
Class at
Publication: |
514/176 ;
514/182; 540/047; 540/090; 564/282 |
International
Class: |
A61K 31/56 20060101
A61K031/56; A61K 31/58 20060101 A61K031/58; A61P 31/04 20060101
A61P031/04; C07C 211/43 20060101 C07C211/43; C07J 53/00 20060101
C07J053/00 |
Claims
1-188. (canceled)
189. A compound of formula (IV) ##STR98## wherein R.sub.1, R.sub.2,
R.sub.3, and R.sub.4 are together --O--CH.sub.2--; R.sub.5 is
absent, oxy, thio, or imino; R.sub.6 is absent or alkylene; R.sub.7
is N.sup.+-containing heteroaryl, N.sup.+-containing heterocycle,
or --N.sup.+R.sub.aR.sub.bR.sub.c; R.sub.a, R.sub.b, and R.sub.c
are each independently (C.sub.1-C.sub.24)alkyl, aryl, arylalkyl,
heteroarylalkyl, heterocycle, or heterocylealkyl; any heteroaryl,
heterocycle, or R.sub.a, R.sub.b, or R.sub.c of R.sub.4 and R.sub.7
can optionally be substituted on carbon with one or more alkyl,
hydroxyalkyl, arylalkyl, heteroarylalkyl, aryl, heterocycle,
heterocyclealkyl, oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d,
--NR.sub.dR.sub.d, or cycloalkylalkyl; any cycloalkylalkyl can
optionally be substituted on carbon with one or more hydroxyl,
N.sup.+-containing heteroaryl, N.sup.+-containing heterocycle,
--N.sup.+R.sub.aR.sub.bR.sub.c, N.sup.+-containing
heteroarylalkyloxy, N.sup.+-containing heterocyclealkyloxy, or
--NR.sub.aR.sub.bR.sub.coxy; R.sub.d and R.sub.e are each
independently hydrogen or alkyl; any alkyl or alkylene of R.sub.3,
R.sub.4, R.sub.6, or R.sub.7 can be optionally substituted on
carbon with one or more oxo, hydroxy, halo, aryl, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and is optionally partially unsaturated;
or an acceptable salt thereof.
190. The compound of claim 189, wherein R.sub.5 is oxy.
191. The compound of claim 189, wherein R.sub.6 is acetyl.
192. The compound of claim 189, wherein R.sub.7 is
N-diazabicyclo[2.2.2]octyl; N-pyridinium; or
--N.sup.+(CH.sub.3).sub.3.
193. The compound of claim 192, wherein R.sub.5 is oxy, thio, or
imino; R.sub.6 is alkylene optionally substituted on carbon with
one or more oxo, hydroxy, halo, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, or --NR.sub.dR.sub.e, and optionally interrupted
on carbon with one or more oxy, imino, or thio, and is optionally
partially unsaturated; and R.sub.7 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl optionally substituted on carbon with one
or more oxo, hydroxy, halo, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d, or --NR.sub.dR.sub.e, and optionally
interrupted on carbon with one or more oxy, imino, or thio, and
optionally partially unsaturated; wherein R.sub.d and R.sub.e are
each independently hydrogen or alkyl.
194. The compound of claim 189, wherein N.sup.+-containing
heteroaryl is N-pyridinium, N-methyl-2-pyridinium,
N-methyl-3-pyridinium, N-methyl-4-pyridinium, N-ethyl-2-pyridinium,
N-ethyl-3-pyridinium, N-ethylpyridinium, 3,5-dimethylpyridinium, or
4-(dimethylamino)pyridinium; wherein any heteroaryl can optionally
be substituted on carbon with one or more alkyl, hydroxyalkyl,
arylalkyl, heteroarylalkyl, aryl, heterocycle, heterocyclealkyl,
oxo, hydroxy, halo, nitro, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d, --NR.sub.dR.sub.dR.sub.e, or
cycloalkylalkyl; wherein R.sub.d and R.sub.e are each independently
hydrogen or alkyl.
195. The compound of claim 189, wherein N.sup.+-containing
heterocycle is N-diazabicyclo[2.2.2]octyl;
N-azabicyclo[2.2.2]octyl; N-methyl-N-piperidino;
N,N-dimethyl-2-piperidino; N,N-dimethyl-3-piperidino;
N,N-dimethyl-4-piperdino; N-methyl-N-morpholino;
N,N-dimethyl-2-morpholino; or N,N-dimethyl-3-morpholino; wherein
any heterocycle can optionally be substituted on carbon with one or
more alkyl, hydroxyalkyl, arylalkyl, heteroarylalkyl, aryl,
heterocycle, heterocyclealkyl, oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d,
--NR.sub.dR.sub.e, or cycloalkylalkyl; wherein R.sub.d and R.sub.e
are each independently hydrogen or alkyl.
196. The compound of claim 189, wherein
--N.sup.+--R.sub.aR.sub.bR.sub.c is
N'-benzyl-N,N,N',N'-tetramethylethylenediamine-N-yl;
N,N,N',N'-tetramethylethylenediamine-N-yl; octyldimethylammonium;
tetradecyldimethylammonium; trimethylammonium; triethylammonium, or
tri(hydroxymethyl)ammonium.
197. The compound of claim 189, wherein R.sub.6 is alkylene
optionally substituted with one or more oxo
198. The compound of claim 189, wherein R.sub.7 is
--N+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl optionally substituted on carbon with one
or more oxo, hydroxy, halo, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d, or --NR.sub.dR.sub.e, and optionally
interrupted on carbon with one or more oxy, imino, or thio, and
optionally partially unsaturated.
199. The compound of claim 189, wherein R.sub.7 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl.
200. The compound of claim 189, wherein R.sub.7 is
--N+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl and R.sub.c is (C.sub.6-C.sub.24)alkyl.
201. The compound of claim 189, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl and R.sub.c is
(C.sub.6-C.sub.24)alkyl.
202. A compound of formula (IV) ##STR99## wherein R.sub.1, R.sub.2,
R.sub.3 and R.sub.4 are together --O--C(.dbd.X)--; wherein X is two
hydrogens, oxo, or thioxo (.dbd.S); R.sub.5 is oxy; R.sub.6 is
absent or alkylene; R.sub.7 is N.sup.+-containing heteroaryl,
N.sup.+-containing heterocycle, or --N.sup.+R.sub.aR.sub.bR.sub.c;
R.sub.a, R.sub.b, and R.sub.c are each independently
(C.sub.1-C.sub.24)alkyl, aryl, arylalkyl, heteroarylalkyl,
heterocycle, or heterocylealkyl; any heteroaryl, heterocycle, or
R.sub.a, R.sub.b, or R.sub.c of R.sub.4 and R.sub.7 can optionally
be substituted on carbon with one or more alkyl, hydroxyalkyl,
arylalkyl, heteroarylalkyl, aryl, heterocycle, heterocyclealkyl,
oxo, hydroxy, halo, nitro, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d, --NR.sub.dR.sub.e, or
cycloalkylalkyl; any cycloalkylalkyl can optionally be substituted
on carbon with one or more hydroxyl, N.sup.+-containing heteroaryl,
N.sup.+-containing heterocycle, --N.sup.+R.sub.aR.sub.bR.sub.c,
N.sup.+-containing heteroarylalkyloxy, N.sup.+-containing
heterocyclealkyloxy, or --N.sup.+R.sub.aR.sub.bR.sub.coxy; R.sub.d
and R.sub.e are each independently hydrogen or alkyl; any alkyl or
alkylene of R.sub.3, R.sub.4, R.sub.6, or R.sub.7 can be optionally
substituted on carbon with one or more oxo, hydroxy, halo, aryl,
nitro, cyano, (C.sub.1-C.sub.6)alkoxy, trifluoromethyl,
--COOR.sub.d, or --NR.sub.dR.sub.e, and optionally interrupted on
carbon with one or more oxy, imino, or thio, and is optionally
partially unsaturated; or an acceptable salt thereof.
203. The compound of claim 202, wherein R.sub.1, R.sub.2, R.sub.3,
and R.sub.4 are together --O--CH.sub.2--.
204. The compound of claim 202, wherein R.sub.6 is acetyl.
205. The compound of claim 202, wherein R.sub.7 is
N-diazabicyclo[2.2.2]octyl; N-pyridinium; or
--N.sup.+(CH.sub.3).sub.3.
206. The compound of claim 202, wherein N.sup.+-containing
heteroaryl is N-pyridinium, N-methyl-2-pyridinium,
N-methyl-3-pyridinium, N-methyl-4-pyridinium, N-ethyl-2-pyridinium,
N-ethyl-3-pyridinium, N-ethyl-4-pyridinium, 3,5-dimethylpyridinium,
or 4-(dimethylamino)pyridinium; wherein any heteroaryl can
optionally be substituted on carbon with one or more alkyl,
hydroxyalkyl, arylalkyl, heteroarylalkyl, aryl, heterocycle,
heterocyclealkyl, oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d,
--NR.sub.dR.sub.e, or cycloalkylalkyl; wherein R.sub.d and R.sub.e
are each independently hydrogen or alkyl.
207. The compound of claim 202, wherein N.sup.+-containing
heterocycle is N-diazabicyclo[2.2.2]octyl;
N-azabicyclo[2.2.2]octyl; N-methyl-N-piperidino;
N,N-dimethyl-2-piperidino; N,N-dimethyl-3-piperidino;
N,N-dimethyl-4-piperdino; N-methyl-N-morpholino;
N,N-dimethyl-2-morpholino; or N,N-dimethyl-3-morpholino; wherein
any heterocycle can optionally be substituted on carbon with one or
more alkyl, hydroxyalkyl, arylalkyl, heteroarylalkyl, aryl,
heterocycle, heterocyclealkyl, oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d,
--NR.sub.dR.sub.e, or cycloalkylalkyl; wherein R.sub.d and R.sub.e
are each independently hydrogen or alkyl.
208. The compound of claim 202, wherein --N+--R.sub.aR.sub.bR.sub.c
is N'-benzyl-N,N,N',N'-tetramethylethylenediamine-N-yl;
N,N,N',N'-tetramethylethylenediamine-N-yl; octyldimethylammonium;
tetradecyldimethylammonium; trimethylammonium; triethylammonium, or
tri(hydroxymethyl)ammonium.
209. The compound of claim 202, wherein R.sub.6 is alkylene
optionally substituted with one or more oxo
210. The compound of claim 202, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl optionally substituted on carbon with
one or more oxo, hydroxy, halo, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d, or --NR.sub.dR.sub.e, and optionally
interrupted on carbon with one or more oxy, imino, or thio, and
optionally partially unsaturated.
211. The compound of claim 202, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl.
212. The compound of claim 202, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl and R.sub.c is
(C.sub.6-C.sub.24)alkyl.
213. The compound of claim 202, wherein R.sub.7 is
--N+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl and R.sub.c is (C.sub.6-C.sub.24)alkyl.
214. A compound of formula (IV) ##STR100## wherein R.sub.1,
R.sub.2, R.sub.3 and R.sub.4 are together O--C(.dbd.X)--; wherein X
is two hydrogens, oxo, or thioxo (.dbd.S); R.sub.5 is absent, oxy,
thio, or imino; R.sub.6 is acetyl; R.sub.7 is N.sup.+-containing
heteroaryl, N.sup.+-containing heterocycle, or
--N.sup.+R.sub.aR.sub.bR.sub.c; R.sub.a, R.sub.b, and R.sub.c are
each independently (C.sub.1-C.sub.24)alkyl, aryl, arylalkyl,
heteroarylalkyl, heterocycle, or heterocylealkyl; any heteroaryl,
heterocycle, or R.sub.a, R.sub.b, or R.sub.c of R.sub.4 and R.sub.7
can optionally be substituted on carbon with one or more alkyl,
hydroxyalkyl, arylalkyl, heteroarylalkyl, aryl, heterocycle,
heterocyclealkyl, oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d,
--NR.sub.dR.sub.e, or cycloalkylalkyl; any cycloalkylalkyl can
optionally be substituted on carbon with one or more hydroxyl,
N.sup.+-containing heteroaryl, N.sup.+-containing heterocycle,
--N.sup.+R.sub.aR.sub.bR.sub.c, N.sup.+-containing
heteroarylalkyloxy, N.sup.+-containing heterocyclealkyloxy, or
--NR.sub.aR.sub.bR.sub.cR.sub.coxy; R.sub.d and R.sub.e are each
independently hydrogen or alkyl; any alkyl or alkylene of R.sub.3,
R.sub.4, R.sub.6, or R.sub.7 can be optionally substituted on
carbon with one or more oxo, hydroxy, halo, aryl, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and is optionally partially unsaturated;
or an acceptable salt thereof.
215. The compound of claim 214, wherein R.sub.1, R.sub.2, R.sub.3,
and R.sub.4 are together --O--CH.sub.2--.
216. The compound of claim 214, wherein R.sub.5 is oxy.
217. The compound of claim 214, wherein R.sub.7 is
N-diazabicyclo[2.2.2]octyl; N-pyridinium; or
--N.sup.+(CH.sub.3).sub.3.
218. The compound of claim 214, wherein N.sup.+-containing
heteroaryl is N-pyridinium, N-methyl-2-pyridinium,
N-methyl-3-pyridinium, N-methylpyridinium, N-ethyl-2-pyridinium,
N-ethyl-3-pyridinium, N-ethylpyridinium, 3,5-dimethylpyridinium, or
4-(dimethylamino)pyridinium; wherein any heteroaryl can optionally
be substituted on carbon with one or more alkyl, hydroxyalkyl,
arylalkyl, heteroarylalkyl, aryl, heterocycle, heterocyclealkyl,
oxo, hydroxy, halo, nitro, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d, --NR.sub.dR.sub.e, or
cycloalkylalkyl; wherein R.sub.d and R.sub.e are each independently
hydrogen or alkyl.
219. The compound of claim 214, wherein N.sup.+-containing
heterocycle is N-diazabicyclo[2.2.2]octyl;
N-azabicyclo[2.2.2]octyl; N-methyl-N-piperidino;
N,N-dimethyl-2-piperidino; N,N-dimethyl-3-piperidino;
N,N-dimethyl-4-piperdino; N-methyl-N-morpholino;
N,N-dimethyl-2-morpholino; or N,N-dimethyl-3-morpholino; wherein
any heterocycle can optionally be substituted on carbon with one or
more alkyl, hydroxyalkyl, arylalkyl, heteroarylalkyl, aryl,
heterocycle, heterocyclealkyl, oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d,
--NR.sub.dR.sub.e, or cycloalkylalkyl; wherein R.sub.d and R.sub.e
are each independently hydrogen or alkyl.
220. The compound of claim 214, wherein
--N.sup.+--R.sub.aR.sub.bR.sub.c is
N'-benzyl-N,N,N',N'-tetramethylethylenediamine-N-yl;
N,N,N',N'-tetramethylethylenediamine-N-yl; octyldimethylammonium;
tetradecyldimethylammonium; trimethylammonium; triethylammonium, or
tri(hydroxymethyl)ammonium.
221. The compound of claim 214, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl optionally substituted on carbon with
one or more oxo, hydroxy, halo, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d, or --NR.sub.dR.sub.e, and optionally
interrupted on carbon with one or more oxy, imino, or thio, and
optionally partially unsaturated.
222. The compound of claim 214, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl.
223. The compound of claim 214, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl and R.sub.c is
(C.sub.6-C.sub.24)alkyl.
224. The compound of claim 214, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl and R.sub.c is
(C.sub.6-C.sub.24)alkyl.
225. A compound of formula (IV) ##STR101## wherein R.sub.1,
R.sub.2, R.sub.3 and R.sub.4 are together --O--C(.dbd.X)--; wherein
X is two hydrogens, oxo, or thioxo (.dbd.S); R.sub.5 is absent,
oxy, thio, or imino; R.sub.6 is alkylene optionally substituted
with one or more oxo; R.sub.7 is N.sup.+-containing heteroaryl,
N.sup.+-containing heterocycle, or --N.sup.+R.sub.aR.sub.bR.sub.c;
R.sub.a, R.sub.b, and R.sub.c are each independently
(C.sub.1-C.sub.24)alkyl, aryl, arylalkyl, heteroarylalkyl,
heterocycle, or heterocylealkyl; any heteroaryl, heterocycle, or
R.sub.a, R.sub.b, or R.sub.c of R.sub.4 and R.sub.7 can optionally
be substituted on carbon with one or more alkyl, hydroxyalkyl,
arylalkyl, heteroarylalkyl, aryl, heterocycle, heterocyclealkyl,
oxo, hydroxy, halo, nitro, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d, --NR.sub.dR.sub.e, or
cycloalkylalkyl; any cycloalkylalkyl can optionally be substituted
on carbon with one or more hydroxyl, N.sup.+-containing heteroaryl,
N.sup.+-containing heterocycle, --N.sup.+R.sub.aR.sub.bR.sub.c,
N.sup.+-containing heteroarylalkyloxy, N.sup.+-containing
heterocyclealkyloxy, or --N.sup.+R.sub.aR.sub.bR.sub.coxy; R.sub.d
and R.sub.e are each independently hydrogen or alkyl; any alkyl or
alkylene of R.sub.3, R.sub.4, R.sub.6, or R.sub.7 can be optionally
substituted on carbon with one or more oxo, hydroxy, halo, aryl,
nitro, cyano, (C.sub.1-C.sub.6)alkoxy, trifluoromethyl,
--COOR.sub.d, or --NR.sub.dR.sub.e, and optionally interrupted on
carbon with one or more oxy, imino, or thio, and is optionally
partially unsaturated; or an acceptable salt thereof.
226. The compound of claim 225, wherein R.sub.1, R.sub.2, R.sub.3,
and R.sub.4 are together --O--CH.sub.2--.
227. The compound of claim 225, wherein R.sub.5 is oxy.
228. The compound of claim 225, wherein R.sub.6 is acetyl.
229. The compound of claim 225, wherein R.sub.7 is
N-diazabicyclo[2.2.2]octyl; N-pyridinium; or
--N.sup.+(CH.sub.3).sub.3.
230. The compound of claim 225, wherein N.sup.+-containing
heteroaryl is N-pyridinium, N-methyl-2-pyridinium,
N-methyl-3-pyridinium, N-methylpyridinium, N-ethyl-2-pyridinium,
N-ethyl-3-pyridinium, N-ethyl-4-pyridinium, 3,5-dimethylpyridinium,
or 4-(dimethylamino)pyridinium; wherein any heteroaryl can
optionally be substituted on carbon with one or more alkyl,
hydroxyalkyl, arylalkyl, heteroarylalkyl, aryl, heterocycle,
heterocyclealkyl, oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d,
--NR.sub.dR.sub.e, or cycloalkylalkyl; wherein R.sub.d and R.sub.e
are each independently hydrogen or alkyl.
231. The compound of claim 225, wherein N.sup.+-containing
heterocycle is N-diazabicyclo[2.2.2]octyl;
N-azabicyclo[2.2.2]octyl; N-methyl-N-piperidino;
N,N-dimethyl-2-piperidino; N,N-dimethyl-3-piperidino;
N,N-dimethyl-4-piperidino; N-methyl-N-morpholino;
N,N-dimethyl-2-morpholino; or N,N-dimethyl-3-morpholino; wherein
any heterocycle can optionally be substituted on carbon with one or
more alkyl, hydroxyalkyl, arylalkyl, heteroarylalkyl, aryl,
heterocycle, heterocyclealkyl, oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d,
--NR.sub.dR.sub.e, or cycloalkylalkyl; wherein R.sub.d and R.sub.e
are each independently hydrogen or alkyl.
232. The compound of claim 225, wherein
--N.sup.+R.sub.aR.sub.bR.sub.c is
N'-benzyl-N,N,N',N'-tetramethylethylenediamine-N-yl;
N,N,N',N'-tetramethylethylenediamine-N-yl; octyldimethylammonium;
tetradecyldimethylammonium; trimethylammonium; triethylammonium, or
tri(hydroxymethyl)ammonium.
233. The compound of claim 225, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl optionally substituted on carbon with
one or more oxo, hydroxy, halo, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d, or --NR.sub.dR.sub.e, and optionally
interrupted on carbon with one or more oxy, imino, or thio, and
optionally partially unsaturated.
234. The compound of claim 225, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl.
235. The compound of claim 225, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl and R.sub.c is
(C.sub.6-C.sub.24)alkyl.
236. The compound of claim 225, wherein R.sub.7 is
--N.sup.+--R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are
each (C.sub.6-C.sub.24)alkyl and R.sub.c is
(C.sub.6-C.sub.24)alkyl.
237. A pharmaceutical composition comprising a pharmaceutically
effective diluent or carrier, and a compound of any one of claims
189, 202 and 225.
238. A method of killing or inhibiting a bacterium, the method
comprising contacting the bacterium with an effective
anti-bacterial amount of a compound of any one of claims 189, 202
and 225.
239. A method of killing or inhibiting a fungus, the method
comprising contacting the fungus with an effective anti-fungal
amount of a compound of any one of claims 189, 202 and 225.
Description
BACKGROUND OF THE INVENTION
[0001] Outer layers of plants such as leaf cuticles, fruit peels,
and bark protect the plant against abrasion, prevent water loss,
and protect against pathogenic microorganisms. Breaking through the
plant protective outer layer is a prerequisite for a pathogen to
enter the plant's internal tissues. Some studies have suggested
that penetration of the protective layer involves dissolution of
the host cuticle by enzymes secreted by the pathogen Nicholson, R.
L et al., in The Fungal Spore and Disease Initiation in Plants and
Animals, eds. Cole, G. T, and Hoch, H. C., 1991, Plenum Press, New
York, pp. 3-23.
[0002] Pentacyclic triterpenes are among the most common plant
secondary metabolites, but their function in plants has not been
fully understood. They are usually concentrated in the outermost
layers such as plant cuticle, fruit peel, and bark.
[0003] Betulin is a pentacyclic triterpenoid derived from the outer
bark of paper birch trues (Betula papyrifera, B. pendula, B.
venucosa, etc.). It can be present at concentrations of up to about
24% of the bark of white birch Merck Index, twelfth edition, page
1236 (1996). Lupeol is a related compound also found in birch bark
and in other plant sources. Lupeol is present at concentrations of
about 1.5-3% of the birch bark and at up to about 8.2% in Canavalia
ensiformis, a plant widespread in the humid tropics of Asia and
Africa. Allobetulin is another triterpenoid found in birch bark. A
typical pulp mill that process birch produces enough bark waste to
allow for the inexpensive isolation of significant quantities of
these triterpenoids.
[0004] Literature supplies examples of enzymes that can be
inhibited by triterpenes, indicating the ability of triterpenes to
act broadly in a non-specific mode on multiple targets. For
example, Buchler et al. (Biochem. Biophys. Acta 1075, 206 (1991)
showed inhibition of rat renal 11.beta.-hydroxysteroid
dehydrogenase. Koch et al. (Phytother, Res. 8, 109 (1994)) showed
in vitro inhibition of adenosine deaminase. This leads to the
hypothesis that pentacyclic triterpenoids in plant protective outer
layers may protect against infection by inhibiting enzymes that
would degrade the cuticle.
[0005] Several triterpenoids have been found to have utility. For
example, betulin and related compounds have been shown to have
anti-viral activity against herpes simplex virus. Carlson et al.,
U.S. Pat. No. 5,750,578. Betulin and related compounds have also
been shown to have anti-fungal and anti-bacterial activity.
However, triterpenoids are hydrophobic compounds with relatively
low interfacial activity and water solubility. For instance, the
solubility of betulin in water is about 0.15 mg/l. The relatively
low interfacial activity and water solubility can make handling and
administration of the compounds difficult. Low interfacial activity
also limits the efficient interaction with target (fingi or
bacteria) cell membranes. It also limits accessibility to
hydrophilic biological targets or targets protected by a
hydrophilic barrier.
[0006] There is a need for new compounds with anti-fungal, and
anti-bacterial activities. Preferably the compounds would be
relatively water soluble surfactants. The water-soluble compounds
would retain their utility, e.g., antifungal, antibacterial, etc.,
activity.
SUMMARY OF THE INVENTION
[0007] The present invention provides novel compounds useful for
their anti-fungal and antibacterial properties. The compounds have
the advantage of being derived from abundant and relatively
inexpensive staring materials. They have the further advantage of
being based on natural compounds, and thus being less likely to
pose a risk to the environment, humans, or beneficial organisms.
They have the flier advantage of being relatively water soluble and
amphiphilic. This makes the compounds easier to handle and reduces
the use of hazardous organic solvents. It also increases
accessibility to hydrophilic biological targets, or to targets
present in water or protected by a hydrophilic barer. The
amphiphilic character of the compounds allows them to bind
hydrophobic targets, and may allow them to adsorb on cytoplasmic
membranes. A further advantage of the compounds is that they are
stable and do not tend to readily hydrolyze.
[0008] The present invention provides a quaternary ammonium salt of
a triterpene.
[0009] The present invention further provides a compound of formula
(I): ##STR1## wherein each R.sub.1 is independently absent, oxy,
thio, or imino. Each R.sub.2 is independently absent or alkylene.
Each R.sub.3 is independently hydrogen, N.sup.+-containing
heteroaryl, N.sup.+-containing heterocycle, or
--N.sup.+R.sub.aR.sub.bR.sub.c; provided at least one R.sub.3 is
b-containing heteroaryl, N.sup.+-containing heterocycle, or
--N.sup.+R.sub.aR.sub.bR.sub.c. Each n is independently 0-4
inclusive, provided at least one n is not 0. R.sub.a, R.sub.b, and
R.sub.c are each independently (C.sub.1-C.sub.24)alkyl, aryl,
arylalkyl, heteroaylalkly, heterocycle, or heterocylealyl. Any
heteroaryl, heterocycle, R.sub.a, R.sub.b, or R.sub.c of R.sub.3
can optionally be substituted on carbon with one or more alkyl,
hydroxyalkyl, arylalkyl, heteroarylalkyl, aryl, heterocycle,
heterocycloalkyl, oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d,
--NR.sub.dR.sub.e, or cycloalkylalkyl. R.sub.d and R.sub.e are each
independently hydrogen or alkyl. Any cycloalkylalkyl can optionally
be substituted on carbon with one or more hydroxyl,
N.sup.+-containing heteroaryl, N.sup.+-containing heterocycle, or
--N.sup.+R.sub.aR.sub.bR.sub.c N.sup.+-containing heteroarylalkoxy,
N.sup.+-containing heterocyclealkyloxy, or
--N.sup.+R.sub.aR.sub.bR.sub.coxy. Any alkyl or alkylene of R.sub.3
can optionally be substituted on carbon with one or more oxo,
hydroxy, halo, nitro, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d, or --NR.sub.dR.sub.e, and optionally
interrupted on carbon with one or more oxy, imino, or thio, and is
optionally partially unsaturated. The invention also provides
acceptable salts of a compound of formula (I).
[0010] The present invention further provides a compound of formula
(II): ##STR2##
[0011] In formula (II), R.sub.1, R.sub.4, and R.sub.7 are each
independently absent or alkylene. R.sub.3 and R.sub.6 are each
independently absent, oxy, thio, or imino. R.sub.2, R.sub.5, and
R.sub.5 are each independently hydrogen, N.sup.+-containing
heteroaryl N.sup.+-containing heterocycle, or
--N.sup.+R.sub.aR.sub.bR.sub.c; provided at least one of R.sub.2,
R.sub.5, and R.sub.8 is N.sup.+-containing heteroaryl, N-containing
heterocycle, or --N.sup.+R.sub.aR.sub.bR.sub.c, R.sub.a, R.sub.b,
and R.sub.c are each independently (C.sub.1-C.sub.24)alkyl, aryl,
arylalkyl, heteroarylalkly, heterocycle, or heterocylealkyl. Any
heteroaryl, heterocycle, or R.sub.a, R.sub.b, or R.sub.c of
R.sub.2, R.sub.5, and R.sub.8 can optionally be substituted on
carbon with one or more alkyl hydroxyalkyl, arylalkyl,
heteroarylalkyl, aryl, heterocycle heterocyclealkyl, oxo, hydroxy,
halo, nitro, cyano, (C.sub.1-C.sub.6)alkoxy, trifluoromethyl,
--COOR.sub.d, --NR.sub.dR.sub.e, or cycloalkylalkyl. R.sub.d and
R.sub.e are each independently hydrogen or alkyl. Any
cycloalkylalkyl can optionally be substituted on carbon with one or
more hydroxyl, N.sup.+-containing heteroaryl, N.sup.+-containing
heterocycle, --N.sup.+R.sub.aR.sub.bR.sub.c N.sup.+-containing
heteroarylalkyloxy, N.sup.+-containing heterocyclealkyloxy, or
--N.sup.+R.sub.aR.sub.bR.sub.coxy. Any alkyl or alkylene of
R.sub.1, R.sub.2, R.sub.4, R.sub.5, R.sub.7, or R.sub.8 can be
optionally substituted on carbon with one or more oxo, hydroxy,
halo, nitro, cyano, (C.sub.1-C.sub.6)alkoxy, trifluoromethyl,
--COOR.sub.d, or --NR.sub.dR.sub.e, and optionally interrupted on
carbon with one or more oxy, imino, or thio, and is optionally
partially unsaturated. The invention also provides acceptable salts
of a compound of formula (II).
[0012] The present invention further provides a compound of formula
(III) ##STR3## In formula (III), each R.sub.1 is independently
absent, oxy, thio, or imino. Each R.sub.2 is independently absent
or alkylene. Each R.sub.3 is independently hydrogen,
N.sup.+-containing heteroaryl, N.sup.+-containing heterocycle, or
--N.sup.+R.sub.aR.sub.bR.sub.c; provided at least one R.sub.3 is
N.sup.+-containing heteroaryl, N.sup.+-containing heterocycle, or
--N.sup.+R.sub.aR.sub.bR.sub.c. R.sub.4 is hydrogen, alkyl, or
hydroxyalkyl. Alternatively, R.sub.4 together with one
R.sub.1R.sub.2R.sub.3 forms a --OC.sub.2-- bridging carbons 19 and
17. Each n is independently 0-4 inclusive, provided at least one n
is not 0. R.sub.a, R.sub.b, and R.sub.c are each independently
(C.sub.1-C.sub.24)alkyl, aryl, arylalkyl, heteroarylalkyl,
heterocycle, or heterocylealkyl. Any heteroaryl, heterocycle,
R.sub.a, R.sub.b, or R.sub.c of R.sub.3 can optionally be
substituted on carbon with one or more alkyl hydroxyalkyl,
arylalkyl, heteroarylalkyl, aryl, heterocycle, heterocyclealkyl,
oxo, hydroxy, halo, nitro, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d--NR.sub.dR.sub.e, or cycloalkylalkyl.
R.sub.d and R.sub.e are each independently hydrogen or alkyl. Any
cycloalkylalkyl can optionally be substituted on carbon with one or
more hydroxyl, N-containing heteroaryl, N.sup.+-containing
heterocycle, --N.sup.+R.sub.aR.sub.bR.sub.c, N.sup.+-containing
heteroarylalkyloxy, N.sup.+-containing heterocyclealkyloxy, or
--N.sup.+R.sub.aR.sub.bR.sub.coxy. Any alkyl or alkylene of R.sub.3
can optionally be substituted on carton with one or more oxo,
hydroxy, halo, nitro, cyano, (C.sub.1-C.sub.6)alkoxy,
trifluoromethyl, --COOR.sub.d, or --N.sub.dR.sub.e, and optionally
interrupted on carbon with one or more oxy, imino, or thio, and is
optionally partially saturated. The invention also provides
acceptable salts of a compound of formula (III).
[0013] The present invention further provides a compound of formula
(IV): ##STR4## In formula (IV) R.sub.1 is hydrogen, alkyl, or
hydroxyalkyl. R.sub.2 is oxymethylene, thiomethylene,
iminomethylene, or methylene. R.sub.3 and R.sub.6 are each
independently absent or alkylene. R.sub.4 and R.sub.7 are each
independently hydrogen, N.sup.+-containing heteroaryl,
N.sup.+-containing heterocycle, or --N.sup.+R.sub.aR.sub.bR.sub.c;
provided at least one of R.sub.4 and R.sub.7 is N.sup.+-containing
heteroaryl, N.sup.+-containing heterocycle, or
--N.sup.+R.sub.aR.sub.bR.sub.c. Alternatively R.sub.1, R.sub.2,
R.sub.3, and R.sub.4 are together --O--C(.dbd.X)--; wherein X is
two hydrogens, oxo, or thioxo (.dbd.S). R.sub.5 is absent, oxy,
thio, or imino. R.sub.a, R.sub.b, and R.sub.c are each
independently (C.sub.1-C.sub.24)alkyl, aryl, arylalkyl,
heteroarylalkyl, heterocycle, or heterocylealkyl. Any heteroaryl,
heterocycle, R.sub.a, R.sub.b, or R.sub.c of R.sub.4 and R.sub.7
can optionally be substituted on carbon with one or more alkyl,
hydroxyalkyl, arylalkyl, heteroarylalkyl aryl, heterocycle,
heterocyclealkyl, oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d,
--NR.sub.dR.sub.e, or cycloalkylalkyl wherein any cycloalkylalkyl
can optionally be substituted on carbon with one or more hydroxyl,
N.sup.+-containing heteroaryl, N.sup.+-containing heterocycle,
--N.sup.+R.sub.aR.sub.bR.sub.c, N.sup.+-containing
heteroarylalkyloxy, N.sup.+-containing heterocyclealkyloxy, or
--N.sup.+R.sub.aR.sub.bR.sub.coxy. R.sub.d and R.sub.e each
independently hydrogen or alkyl. Any alkyl or alkylene of R.sub.3,
R.sub.4, R.sub.6, or R.sub.7 can be optionally substituted on
carbon with one or more oxo, hydroxy, halo, aryl, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and is optionally partially
unsaturated.
[0014] The present invention further provides a method of
inhibiting or killing a fungus comprising contacting the fungus
with an effective anti-fungal amount of a compound of formula
(I)-(IV).
[0015] The present invention further provides a method of
inhibiting or killing a bacterium comprising contacting the
bacterium with an effective anti-bacterial amount of a compound of
formula (I)-(IV).
DETAILED DESCRIPTION OF THE INVENTION
[0016] The following definitions are used, unless otherwise
described: halo is fluoro, chloro, bromo, or iodo. Alkyl, alkoxy,
etc. denote both straight and branched groups; but reference to an
individual radical such as "propyl" embraces only the straight
chain radical, a branched chain isomer such as "isopropyl" being
specifically referred to. Alkyl and alkylene are defined herein as
(C.sub.1-C.sub.6)alkyl or (C.sub.1-C.sub.6)alkylene unless
specified otherwise. The alkyl or alkylene portion of other groups,
e.g., alkoxy, cycloalkyl, etc., is also defined herein as
comprising 1-6 carbons unless otherwise specified. Aryl denotes a
phenyl radical or an ortho-fused bicyclic carbocyclic radical
having about nine to ten ring atoms in which at least one ring is
aromatic. Heteroaryl encompasses a radical attached via a ring atom
of a monocyclic aromatic ring containing five or six ring atoms
consisting of carbon and one to four heteroatoms each selected from
the group consisting of non-peroxide oxygen, sulfur, and N(X)
wherein X is absent or is H, O, (C.sub.1-C.sub.4)alkyl, phenyl or
benzyl, as well as a radical of an ortho-fused bicyclic heterocycle
of about eight to ten ring atoms derived therefrom, particularly a
benz-derivative or one derived by fusing a propylene, trimethylene,
or tetraethylene diradical thereto. N+-containing heteroaryl can be
attached via a nitrogen atom of the heteroaryl ring, or via another
ring atom. If attached via another ring atom, the one or more
nitrogens of the ring can optionally be derivatized with one or two
alkyl or hydroxyalkyl groups to make the nitrogen quaternary.
Heterocycle encompasses a radical attached via a ring atom, wherein
the ring can be a single ring, ortho-fused rings, or bicyclic
rings, and wherein the ring system is non-aromatic. The heterocycle
ring preferably comprises 5-20 atoms, of which preferably one to
four are heteroatoms each selected from the group consisting of
non-peroxide oxygen, sulfur, and N(X) wherein X is absent or is H,
O, (C.sub.1-C.sub.6)alkyl, phenyl or benzyl. The ring system can
optionally be substituted. The ring system can optionally be
partially unsaturated.
[0017] The term "cycloalkyl" encompasses a radical attached via a
ring atom of a non-aromatic cyclic ring system, wherein the ring
system comprises one to five ortho-fused rings, each ring
consisting of 4-10 carbon atoms. The ring system can optionally be
substituted. The ring system can optionally be partially
unsaturated.
[0018] The term "acceptable salt" refers to a salt comprising one
of the cationic compounds of the invention and an acceptable anion.
An acceptable anion is an anion that does not interfere with the
functioning of the compound. For instance, in pharmaceutical uses,
an acceptable anion is one that does not have any significant
deleterious effect on the health of the patient. In agricultural
use of the compounds of the invention as anti-fungal agents, an
acceptable anion is one that does not interfere with the antifungal
properties of the compounds and does not have a deleterious effect
on plant health. Preferred cations include chloride, bromide, and
iodide. Other preferred cations include organic cations, such as
formate, acetate, propionate, or butyrate.
[0019] The term "N-diazabicyclo[2.2.2]octyl" refers to the group
##STR5##
[0020] The term "N-pyridinium" refers to the group ##STR6##
[0021] The term "N-methyl-N-piperidino" refers to the group
##STR7##
[0022] The term "N-methyl-N-morpholino" refers to the group
##STR8##
[0023] The term "N-azabicyclo[2.2.2]octyl" refers to the group
##STR9##
[0024] The term "bacterium" or "bacteria" refers to any prokaryotic
organism. See Biology of Microorganisms, 6.sup.th ed., Brock, T.
D., and Madigan, M. T., (1991), pp. 9-11.
[0025] The term "fungus" refers to a distinct group of eukaryotic,
spore-forming organisms with absorptive nutrition and lacking
chlorophyll. The term includes mushrooms, molds, and yeasts. See
id., pp. 817-822.
[0026] The term "titerpene" refers to one of a class of compounds
having approximately 30 carbon atoms and synthesized from six
isoprene units in plants and other organisms. Triterpenes consist
of carbon, hydrogen, and optionally oxygen. Most triterpenes are
secondary metabolites in plants. Moat, but not all, triterpenes are
pentacyclic. Examples of triterpenes include betulin, allobetulin,
lupeol, friedelin, and all sterols, including lanosterol,
stigmasterol cholesterol, .beta.-sitosterol and ergosterol.
[0027] The term "quaternary ammonium salt" refers to a compound
comprising at least one positively charged nitrogen atom with four
covalent bonds to non-hydrogen atoms. Typically the four bonds will
be to carbon atoms. Two or three of the bonds can make up a double
or triple bond respectively to a single atom.
[0028] The term "quaternary ammonium salt of a triterpene" refers
to triterpene covalently attached to a group comprising at least
one positively charged nitrogen atom with four covalent bonds to
non-hydrogen atoms. Examples of quaternary ammonium salts of a
triterpene include a compound of formulas (I)-(IV).
[0029] It will be appreciated by those skilled in the art that
compounds of the invention having a chiral center may exist in and
be isolated in optically active and racemic forms. Some compounds
may exhibit polymorphism. It is to be understood that the present
invention encompasses any racemic, optically-active; polymorphic,
or stereoisomeric form, or mixtures thereof, of a compound of the
invention, which possess the useful properties described herein, it
being well known in the art how to prepare optically active forms
(for example, by resolution of the racemic form by
recrystallization techniques, by synthesis from optically-active
starting materials, by chiral synthesis, or by chromatographic
separation using a chiral stationary phase) and how to determine
anti-fungal or anti-bacterial activity using the standard tests
described herein, or using other similar tests which are well known
in the art.
[0030] Specific and preferred values listed here for radicals,
substituents, and ranges, are for illustration only, they do not
exclude other defined values or other values within defined ranges
for the radicals and substituents
[0031] Specifically, alkyl can be methyl, ethyl, propyl, isopropyl,
butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl, or hexyl. Cycloalkyl
can be cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the
triterpene ring system of betulin, allobetulin, or lupeol.
Cycloalkylalkyl can be cyclopropylmethyl, cyclobutylmethyl,
cyclopentylmethyl, cyclohexylmethyl, 2-cyclopropylethyl,
2-cyclobutylethyl, 2-cyclopentylethyl, or 2-cyclohexylethyl, or a
synthetically feasible radical derived from a compound of formula
(V) or (VI). ##STR10## The synthetically feasible radical derived
from a compound of formula (V) or (VI) can be formed by removal of
a hydrogen or methyl from any suitable carbon atom on the compound,
or by hydrolysis of the ether bridge of compound (VI) followed by
removal of a hydrogen from resultant hydroxyl, methyl, or
methylene. The point of attachment thus can be a side chain methyl,
a ring methylene, a ring methylidyne, carbons 29 or 30 of the side
chain isopropylene of compound (V), or a methylene, oxymethyl, or
oxy formed from the product of hydrolysis of the ether bridge of
compound (VI).
[0032] (C.sub.1-C.sub.6)alkoxy can be methoxy, ethoxy, propoxy,
isopropoxy, butoxy, isobutoxy, sec-butoxy, pentoxy, 3-pentoxy, or
hexyloxy. (C.sub.1-C.sub.5)alkylenecarbonyl can be acetyl,
propionyl, butyryl, pentanoyl, or hexanoyl.
[0033] Alkylene can be methylene, ethylene, propylene, butylene,
pentamethylene, or hexamethylene.
[0034] N.sup.+-containing heteroaryl can be N-pyridinium,
N-methyl-2-pyridinium, N-methyl-3-pyridinium,
N-methyl-4-pyridinium, N-ethyl-2-pyridinium, N-ethyl-3-pyridinium,
N-ethylpyridinium, 3,5-dimethylpyridinium, or
4-(dimethylamino)pyridinium.
[0035] N.sup.+-containing heterocycle can be
N-diazabicyclo[2.2.2]octyl; N-azabicyclo[2.2.2]octyl;
N-methyl-N-piperidino; N,N-dimethyl-2-piperidino;
N,N-dimethyl-3-piperidino; N,N-dimethyl-4-piperidino;
N-methyl-N-morpholino; N,N-dimethyl-2-morpholino; or
N,N-diethyl-3-morpholino.
[0036] Hydroxyalkyl can be hydroxymethyl, hydroxyethyl,
3-hydroxypropyl, 2-hydroxypropyl, 1-hydroxyisopropyl,
4-hydroxybutyl, 5-hydroxypentyl, or 6-hydroxyhexyl.
[0037] Arylalkyl can be benzyl, phenylethyl, phenylpropyl,
phenylbutyl, or phenylpentyl.
[0038] --N.sup.+R.sub.aR.sub.bR.sub.c can be
N'-benzyl-N,N,N',N'-tetramethylethylenediamine-N-yl;
N,N,N',N'-tetramethylethylenediamine-N-yl; octyldimethylammonium;
tetradecyldimethylammonium; trimethylammonium; triethylammonium, or
tri(hydroxymethyl)ammonium.
[0039] The present invention is directed to triterpenes derivatized
with N-containing groups. The compounds of the invention are found
to be rather resistant to hydrolysis. Derivatization with
N.sup.+-containing groups is also found to make the compounds of
the invention rather water soluble. For instance, the solubility of
some quaternary salts of betulin disclosed herein is 400-600
g/l.
[0040] In one specific embodiment of a compound of formula (I), at
least one R.sub.3 is --N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a
and R.sub.b are each independently (C.sub.6-C.sub.24)alkyl
optionally substituted on carbon with one or more oxo, hydroxy,
halo, cyano, (C.sub.1-C.sub.6)alkoxy, trifluoromethyl,
--COOR.sub.d, or --NR.sub.dR.sub.e, and optionally interrupted on
carbon with one or more oxy, imino, or thio, and optionally
partially unsaturated. In particular compounds of this embodiment,
one R.sub.3 is --N.sup.+R.sub.aR.sub.bR.sub.c and the other
R.sub.3s are hydrogen.
[0041] In one specific embodiment of a compound of formula (II),
R.sub.2, R.sub.5, and R.sub.8 are each independently absent,
hydroxyl, N-diazabicyclo[2.2.2]octyl, N-pyridinium,
N-alkyl-N-piperidino, N-alkyl-N-morpholino,
N-azabicyclo[2.2.2]octyl, or --N.sup.+R.sub.aR.sub.bR.sub.c;
provided at least one of R.sub.2, R.sub.5, and R.sub.8 is
N.sup.+-containing heteroaryl, N.sup.+-containing heterocycle, or
--N.sup.+R.sub.aR.sub.bR.sub.c. In this embodiment
N-diazabicyclo[2.2.2]octyl; N-pyridinium; N-alkyl-N-piperidino;
N-alkyl-N-morpholino; and N-azabicyclo[2.2.2]octyl can optionally
be substituted on one or more suitable carbon atoms with one or
more oxo, hydroxy, mercapto, alkyl, hydroxyalkyl, halo, nitro,
cyano, (C.sub.1-C.sub.6)alkoxy, --COOR.sub.d, or --NR.sub.dR.sub.e.
In this embodiment also, any alkyl or alkylene of R.sub.1, R.sub.2,
R.sub.4, R.sub.5, R.sub.7, or R.sub.8 can optionally be substituted
with one or more oxo or --NR.sub.dR.sub.e, and optionally
interrupted with one or more oxy, imino, or thio, and can
optionally be partially unsaturated.
[0042] In another specific embodiment of q compound of formula
(II), R.sub.1 is absent and R.sub.2 is hydrogen,
N-diazabicyclo[2.2.2]octyl, or N-dimethylamino-N-pyridinium.
[0043] In another specific embodiment of a compound of formula
(II), R.sub.3 and R.sub.4 are absent, and R.sub.5 is hydrogen.
[0044] In another specific embodiment of a compound of formula
(II), R.sub.3 is oxy; R.sub.4 is absent or
(C.sub.1-C.sub.5)alkylenecarbonyl; and R.sub.5 is hydrogen,
N-diazabicyclo[2.2.2]octyl; 4 dimethylamino-N-pyridinium;
4-hydroxybutyl-N-diazabicyclo[2.2.2]octyl
4-benzyl-N-diazabicyclo[2.2.2]octyl;
tetramethylethylenediamine-N-yl;
N'-benzyl-N,N,N',N'-tetramethylethylenediamine-N-yl; N-pyridinium;
4-hydroxymethyl-N-pyridinium; 2,4-dimethyl-N-pyridinium;
3,5-dimethyl-N-pyridinium; octyldimethylammonium; or
tetradecyldimethylammonium.
[0045] In another specific embodiment of a compound of formula
(II), R.sub.6 is oxy; R.sub.7 is absent or
(C.sub.1-C.sub.5)alkylenecarbonyl; and R.sub.8 is hydrogen,
N-diazabicyclo[2.2.2]octyl; 4-dimethylamino-N-pyridinium;
N'-(4-hydroxybutyl)-N-diazabicyclo[2.2.2]octyl;
N'-benzyl-N-diazabicyclo[2.2.2]octyl;
N,N,N',N'-tetramethylethylenediamine-N-yl;
N'-benzyl-N,N,N',N'-tetramethylethylenediamine-N-yl; N-pyridinium;
4-hydroxymethyl-N-pyridinium; 2,4-dimethyl-N-pyridinium;
3,5-dimethyl-N-pyridinium; octyldimethylammonium;
tetradecyldimethylammonium; 2-methyl-N-pyridinium; 4
hydroxy-N-methyl-N-piperidinium; or N-methyl-N-morpholino.
[0046] In particular embodiments of the invention, the compound of
formula (II) is: [0047]
lup-20(29)-ene-3,28-bis-(N-pyridiniumacetate); [0048]
lup-20(29)-ene-3-[N-(4-oxybutyl)-1,4-diazabicyclo[2.2.2]octyl-N'--
acetate]; [0049]
lup-20(29)-ene-3,28-bis[N-(1,4-diazabicyclo[2.2.2]octyl)acetate];
[0050]
lup-20(29)-ene-3,28-bis[N--(N'-benzyldiazabicyclo[2.2.2]octyl)acetate);
[0051]
lup-20(29)-ene-3,28-bis[N--(N'-(4-oxybutyl)diazabicyclo[2.2.2]oct-
yl)acetate]; [0052]
lup-20(29)-ene-3-[N-(1,4-diazabicyclo[2.2.2]octyl)acetate]; [0053]
lup-20(29)-ene-3,28-bis[(tetramethylethylenediamine-N-yl)acetate];
[0054]
lup-20(29)-ene-3,28-bis[N'-benzyl-N,N,N',N'-tetramethylethylenedi-
amine-N-yl)acetate]; [0055]
lup-20(29)-ene-3-[N--(N'-(benzyl)diazabicyclo[2.2.2]octyl)acetate];
[0056] bis(N,N'-pyridinium-2-ethyl)lup-20(29)-ene-3,28-dicarbamate;
[0057]
1-(3,28-diacetoxy)lup-20(29)-ene-30-yl)-4-(dimethylamino)pyridini-
um; [0058] lup-20(29)-ene-3,28-bis(N-pyridinium-2-propionate);
[0059] lup-20(29)-ene-3,28-bis(N-pyridinium-3-propionate); [0060]
lup-20(29)-ene-3,28-bis(N-pyridinium-4-butyrate); [0061]
lup-20(29)-ene-3,28-bis(N-pyridinium-4-butyrate); [0062]
lup-20(29)-ene-3,28-bis(N-pyridinium-2-butyrate); [0063]
1-[3,28-(diacetoxy)lup-20(29)-ene-30-yl]-1,4-diazabicyclo[2.2.2]octyl;
[0064] 3,28-bis[3-(1-piperidinyl)propanoyloxy]lup-20(29)-ene;
[0065]
1-(3,28-dihydroxylup-20(29)-ene-30-yl)-4-(dimethylamino)pyridinium;
[0066]
lup-20(29)-ene-3,28-bis[N-(4-dimethylaminopyridinium)-2-propionat-
e]; [0067]
lup-20(29)-ene-3,28-bis[N-(1,4-diazabicyclo[2.2.2]octyl)-2-propionate];
[0068] 1-(lup-20(29)-ene-30-yl)-1,4-diazabicyclo[2.2.2]octane;
[0069] 1-(3,28-dihydroxylup-20(29)-ene-30-yl)-pyridinium; [0070]
lup-20(29)-ene-3,28-bis[N-(1,4-diazabicyclo[2.2.2]octyl)-4-butyrate];
[0071]
1-(3,28-dihydroxylup-20(29)-ene-30-yl)-[N-3-(hydroxymethyl)pyridi-
nium]; [0072]
1-(3,28-dihydroxylup-20(29)-ene-30-yl)-[N-(3,5-dimethylpyridinium)];
[0073]
bis[N-(1,4-diazabicyclo[2.2.2]octyl)-2-ethyl]-lup-20(29)-ene-3,28-
-dicarbamate; [0074]
lup-20(29)-ene-3,28-bis[N-(3-oxymethylpyridinium)acetate]; [0075]
lup-20(29)-ene-3,29-bis[N-(2-oxymethylpyridinium)acetate]; [0076]
lup-20(29)-ene-3,28-bis[N-(2-methylureapyridinium)acetate]; [0077]
lup-20(29)-ene-3-[N-(2-oxymethylpyridinium)acetate]; [0078]
lup-20(29)-one-3,28-bis[N--(N-methylmorpholino)acetate]; [0079]
lup-20(29)-ene-3,28-bis[N-(4-hydroxyl-N-methylpiperidino)acetate];
[0080] lup-20(29)-ene-3-[N-(3-ureamethylpyridinium)acetate]; [0081]
lup-20(29)-ene-3-(N-pyrdiniumacetate); [0082]
lup-20(29)-ene-3,28-bis[N-(1,4-diazabicyclo[2.2.2]octyl)-2-butyrate];
[0083]
lup-20(29)-ene-3,28-bis[N-(4-dimethylpyridinium)-2-butyrate];
[0084]
lup-20(29)-ene-3,28-bis[N-(4-dimethylaminopyridinium)-4-butyrate]-
; [0085]
lup-20(29)-ene-3,28-bis[N-(4-dimethylaminopyridinium)-3-propion-
ate]; [0086]
1-(3,28-dihydroxylup-20(29)-ene-30-yl)-4-(hydroxymethyl)pyridinium;
[0087]
1-(3,28-dihydroxylup-20(29)-ene-30-yl)-3-hydroxy-1-azabicyclo[2.2-
.2]octane; [0088]
lup-20(29)-ene-3,28-bis[N-(2,4-dimethylpyridinium)acetate]; [0089]
lup-20(29)-ene-3,28-bis[N-(3,5-dimethylpyridinium)acetate]; [0090]
lup-20(29)-one-3,28-bis[N-(4-dimethylaminopyridinium)acetate];
[0091] lup-20(29)-ene-3-[N-(2-methylpyridinium)acetate]; [0092]
lup-20(29)-ene-3-[N-(2,4-dimethylpyridinium)acetate]; [0093]
lup-20(29)-ene-3-[N-(4-hydroxy-N-methylpiperidino)acetate]; [0094]
lup-20(29)-ene-3-[N--(N-methylmorpholino)acetate]; [0095]
lup-20(29)-ene-3-[N-(3,5-dimethylpyridinium)acetate]; [0096]
lup-20(29)-ene-3-[N-(4-dimethylaminopyridinium)acetate]; [0097]
lup-20(29)-ene-3,28-bis(octyldimethylammoniumacetate); [0098]
lup-20(29)-ene-3-octyldimethylammoniumacetate; [0099]
lup-20(29)-ene-3,28-bis(tetradecyldimethylammoniumacetate); [0100]
lup-20(29)-ene-3-tetradecyldimethylammoniumacetate; [0101]
N,N,N',N'-tetramethylethylenediamine-N,N'-bis-[lup-20(29)-ene-3-acetate];
[0102]
1-[(lup-20(29)-en-3.beta.-yl)oxycarbonylmethyl]-4-aza-1-azonia-b-
icyclo[2.2.2]octane; [0103]
1-[(lup-20(29)-en-3.beta.-yl)oxycarbonylmethyl]trimethylammonium;
or [0104]
1-[(lup-20(29)-en-3.beta.-yl)oxycarbonylmethyl]pyridinium.
[0105] A specific embodiment of the compound of formula (II) is the
compound wherein at least one of R.sub.2, R.sub.5, and R.sub.8 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
independently (C.sub.6-C.sub.24)alkyl optionally substituted on
carbon with one or more oxo, hydroxy, halo, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and optionally partially unsaturated. In
more particular embodiments, R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl, or are each (C.sub.8-C.sub.24)alkyl, or
are each (C.sub.10-C.sub.24)alkyl.
[0106] In a specific embodiment of a compound of formula (II)
wherein at least one of R.sub.2, R.sub.5, and R.sub.8 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
independently (C.sub.6-C.sub.24)alkyl optionally substituted on
carbon with one or more oxo, hydroxy, halo, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and optionally partially unsaturated,
R.sub.1 is absent and R.sub.2 is hydrogen.
[0107] In a specific embodiment of a compound of formula (O), the
cation of the cation of the compound is
betulin-3,28-bis(didecylmethylammoniumacetoxy).
[0108] In a specific embodiment of a compound of formula (II),
wherein at least one of R.sub.2, R.sub.5 and R.sub.8 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
independently (C.sub.6-C.sub.24)alkyl optionally substituted on
carbon with one or more oxo, hydroxy, halo, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and optionally partially unsaturated,
R.sub.1 is absent and R.sub.2 is hydrogen, R.sub.3 is absent or
oxy, rid is absent, and R.sub.5 is hydrogen. In another specific
embodiment, R.sub.6 is oxy, R.sub.7 is absent, and R.sub.5 is
hydrogen.
[0109] In a specific embodiment of a compound of formula (II), the
cation of the compound is
betulin-3-(didecylmethylammoniumacetoxy).
[0110] In a specific embodiment of a compound of formula (II),
R.sub.3 is absent, or oxy.
[0111] In a particular embodiment of a compound of formula (II),
R.sub.4 is absent or alkylene optionally substituted on carbon with
one or more oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and is optionally partially unsaturated.
In a specific embodiment, R.sub.4 is acetyl.
[0112] In a particular embodiment of the compound of formula (II),
R.sub.5 is --N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and
R.sub.b are each (C.sub.6-C.sub.24)alkyl optionally substituted on
carbon with one or more oxo, hydroxy, halo, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and optionally partially unsaturated. In
a more specific embodiment, R.sub.5 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl. In a more specific embodiment, R.sub.4 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl and R.sub.c is (C.sub.1-C.sub.24)alkyl. In
a more specific embodiment, R.sub.5 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl and R.sub.e is (C.sub.1-C.sub.6)alkyl.
[0113] In a specific embodiment of a compound of formula (II),
R.sub.6 is absent, or oxy.
[0114] In a particular embodiment of a compound of formula (II),
R.sub.7 is absent or alkylene optionally substituted on carbon with
one or more oxo, hydroxy, halo, nitro, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and is optionally partially unsaturated.
In a specific embodiment, R.sub.7 is acetyl.
[0115] In a particular embodiment of the compound of formula (II),
R.sub.8 is --N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and
R.sub.b are each (C.sub.6-C.sub.24)alkyl optionally substituted on
carbon with one or more oxo, hydroxy, halo, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and optionally partially unsaturated. In
amore specific embodiment, R.sub.8 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl. In a more specific embodiment, R.sub.4 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl and R.sub.c is (C.sub.1-C.sub.24)alkyl. In
a more specific embodiment, R.sub.8 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl and R.sub.c is (C.sub.1-C.sub.6)alkyl.
[0116] A specific embodiment of the compound of formula (III) is
the compound wherein at least one R.sub.3 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
independently (C.sub.6-C.sub.24)alkyl optionally substituted on
carbon with one or more oxo, hydroxy, halo, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and optionally partially unsaturated. In
specific compounds of this embodiment, R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl, are each C.sub.8-C.sub.24)allyl, or are
each (C.sub.10-C.sub.24)alkyl. In other specific compounds of this
embodiment, one R.sub.3 is --N.sup.+R.sub.aR.sub.bR.sub.c and the
other R.sub.3s are hydrogen.
[0117] A specific embodiment of the compound of formula (IV) is the
compound wherein R.sub.1 is hydrogen, alkyl, or hydroxyalkyl;
R.sub.2 is oxymethylene, thiomethylene, iminomethylene, or
methylene; R.sub.3 and R.sub.6 are each independently absent or
alkylenecarbonyl; R.sub.4 and R.sub.7 are each independently
hydrogen, N-diazabicyclo[2.2.2]octyl; N-pyridinium;
N-alkyl-N-piperidino; N-alkyl-N-morpholino;
N-azabicyclo[2.2.2]octyl; or --N.sup.+R.sub.aR.sub.bR.sub.c; or
R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are together
--O--CH.sub.2--. In this case, N-diazabicyclo[2.2.2]octyl;
N-pyridinium; N-alkyl-N-piperidino; N-alkyl-N-morpholino; and
N-azabicyclo[2.2.2]octyl can optionally be substituted on carbon
with one or more alkyl, hydroxyalkyl, hydroxy, COOR.sub.d, or
NR.sub.dR.sub.e. R.sub.a, R.sub.b, and R.sub.c are each
independently aryl or (C.sub.1-C.sub.24)alkyl; wherein R.sub.d and
R.sub.e are each independently hydrogen or alkyl. Any alkylene or
alkyl can optionally be substituted on carbon with one or more oxo,
hydroxy, halo, nitro, cyano, trifluoromethyl, COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted with one or more oxy,
imino, or thio, and where any alkyl or alkylene can optionally be
partially unsaturated.
[0118] Another specific embodiment of the compound of formula (IV)
is the compound wherein R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are
together O--CH.sub.2--.
[0119] Another specific embodiment of the compound of formula (IV)
is the compound wherein R.sub.5 is oxy.
[0120] Another specific embodiment of the compound of formula (IV)
is the compound wherein R.sub.6 is acetyl.
[0121] Another specific embodiment of the compound of formula (IV)
is the compound wherein R.sub.7 is N-diazabicyclo[2.2.2]octyl;
N-pyridinium; or --N.sup.+(CH.sub.3).sub.3.
[0122] In particular embodiments of the invention, the compound of
formula (IV) is: [0123]
1-[(19.beta.,28-epoxy-18.alpha.-oleanan-3.beta.-yl)oxycarbonylmethyl]-4-a-
za-1-azonia-bicyclo[2.2.2]octane; [0124]
[(19.beta.,28-epoxy-18.alpha.-oleanan-3.beta.-yl)oxycarbonylmethyl]trimet-
hylammonium; or [0125]
1-[(19.beta.,28-epoxy-18.alpha.-oleanan-3.beta.-yl)oxycarbonylmethyl]pyri-
dinium.
[0126] In a specific embodiment of the compound of formula (IV), at
least one of R.sub.4 and R.sub.7 is --N.sup.+R.sub.aR.sub.bR.sub.c
wherein R.sub.a and R.sub.b are each (C.sub.6-C.sub.24)alkyl
optionally substituted on carbon with one or more oxo, hydroxy,
halo, cyano, (C.sub.1-C.sub.6)alkoxy, trifluoromethyl,
--COOR.sub.d, or --NR.sub.dR.sub.e, and optionally interrupted on
carbon with one or more oxy, imino, or thio, and optionally
partially unsaturated.
[0127] In a more specific embodiment of the compound of formula
(IV), at least one of R.sub.4 and R.sub.7 is --NR.sub.dR.sub.e,
wherein R.sub.a and R.sub.b are each (C.sub.6-C.sub.24)alkyl
optionally substituted on carbon with one or more oxo, hydroxy,
halo, cyano, (C.sub.1-C.sub.6)alkoxy, trifluoromethyl,
--COOR.sub.d, or --NR.sub.dR.sub.e, and optionally interrupted on
carbon with one or more oxy, imino, or thio, and optionally
partially unsaturated, R.sub.5 is oxy, ado, or imino; R.sub.6 is
alkylene optionally substituted on carbon with one or more oxo,
hydroxy, halo, cyano, (C.sub.1-C.sub.6)alkoxy, trifluoromethyl, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and is optionally partially unsaturated;
and R.sub.7 is --N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and
R.sub.b are each (C.sub.6-C.sub.24)alkyl optionally substituted on
carbon with one or more oxo, hydroxy, halo, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and optionally partially unsaturated.
[0128] In a particular embodiment of the compound of formula (IV),
R.sub.5 is oxy.
[0129] In a particular embodiment of the compound of formula (IV),
R.sub.6, is alkylene optionally substituted on carbon with one or
more oxo, hydroxy, halo, cyano, (C.sub.1-C.sub.5)alkoxy,
trifluoromethyl, or --NR.sub.dR.sub.e, and optionally interrupted
on carbon with one or more oxy, imino, or thio, and is optionally
partially unsaturated, In a more specific embodiment, R.sub.6 is
alkylene optionally substituted with one or more oxo. In a more
specific embodiment, R.sub.6 is acetyl.
[0130] In a particular embodiment of the compound of formula (IV),
R.sub.7 is --N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and
R.sub.b are each (C.sub.6-C.sub.24)alkyl optionally substituted on
carbon with one or more oxo, hydroxy, halo, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and optionally partially unsaturated. In
a more specific embodiment R.sub.7 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl. In a more specific embodiment, R.sub.7 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl and R.sub.c is (C.sub.1-C.sub.24)alkyl. In
a more specific embodiment, R.sub.7 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl and R.sub.2 is (C.sub.1-C.sub.4)alkyl.
[0131] In a particular embodiment of a compound of formula (I),
R.sub.2 is oxymethylene, thiomethylene, iminomethylene, or
methylene or R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are together
--O--CH.sub.2--. In another particular embodiment, R.sub.2 is
oxymethylene or R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are together
--O--CH.sub.2--.
[0132] In a particular embodiment of a compound of formula (IV),
R.sub.3 is absent or alkylene optionally be substituted on carbon
with one or more oxo, hydroxy, halo, nitro, cyano, trifluoromethyl,
COOR.sub.d, or --NR.sub.dR.sub.e and optionally interrupted with
one or more oxy, imino, or thio, and optionally be partially
unsaturated.
[0133] In a particular embodiment of the compound of formula (IV),
R.sub.4 is --N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and
R.sub.b are each (C.sub.6-C.sub.24)alkyl optionally substituted on
carbon with one or more oxo, hydroxy, halo, cyano,
(C.sub.1-C.sub.6)alkoxy, trifluoromethyl, --COOR.sub.d, or
--NR.sub.dR.sub.e, and optionally interrupted on carbon with one or
more oxy, imino, or thio, and optionally partially unsaturated. In
amore specific embodiment, R.sub.4 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.9-C.sub.24)alkyl. In a more specific embodiment, R.sub.4 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl and R.sub.c is (C.sub.1-C.sub.24)alkyl. In
a more specific embodiment, R.sub.4 is
--N.sup.+R.sub.aR.sub.bR.sub.c wherein R.sub.a and R.sub.b are each
(C.sub.6-C.sub.24)alkyl and R.sub.a is (C.sub.1-C.sub.6)alkyl.
[0134] In a particular embodiment of a compound of formula (IV),
R.sub.1 is hydrogen or R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are
together --O--CH.sub.2--.
[0135] In a particular embodiment of the compound of formula (IV)
the cation of the compound is
3.beta.-[(N-methyl-N,N-didecyl)aminoacetyloxy]-19.beta.,28-epoxy-18.alpha-
.-oleanan chloride.
[0136] Another embodiment of the invention provides a method of
inhibiting or killing a fungus comprising contacting the fungus
with an effective anti-fungal amount of any of the compounds of the
invention, e.g., a compound of formulas (I)-(IV). The contacting
can be in vitro or in vivo. The contacting can, for example, be on
a plant. The contacting can be done on turf grass to kill or
inhibit a fungus growing on it. The fungus can be causing the
disease dollar spot or brown patch.
[0137] Another embodiment of the invention provides a method of
inhibiting or killing a bacterium comprising contacting the
bacterium with an effective anti-bacterial amount of any one of the
compounds of the invention, e.g., a compound of formulas (I)-(IV).
The bacterium can be Staphylococcus sp. or Enterococcus sp., for
example. The bacterium, for example, can be S. aureus or E.
faecium. In one embodiment of the invention, the bacterium is
antibiotic resistant.
[0138] Another embodiment of the invention provides a
pharmaceutical composition comprising any one of the compounds of
the invention.
[0139] The compounds of the invention comprise one triterpene
moiety derivatized with one or more quaternary ammonium group
(e.g., N.sup.+-containing group). Preferred N.sup.+-containing
groups include N-containing heteroaryl, N.sup.+-containing
heterocycle, or --N.sup.+R.sub.aR.sub.bR.sub.c, wherein R.sub.a,
R.sub.b, and R.sub.c are each independently
(C.sub.1-C.sub.24)alkyl, aryl, arylalkyl, heteroarylalkyl,
heterocycle, or heterocyclealkyl. Preferably, a single triterpene
moiety is derivatized with one, two, three, or four
N.sup.+-containing groups.
[0140] The compounds of the invention also comprise more than one
triterpene moiety derivatized to a single N.sup.+-containing group
and comprise oligomers of alternating triterpene moieties and
N.sup.+-containing groups. In these cases, the triterpene moieties
can be further derivatized with additional N.sup.+-containing
groups.
[0141] For instance, one embodiment of the invention provides a
compound of formula (VII) or (VIII): ##STR11## Each R.sub.1 is
independently (C.sub.1-C.sub.24)alkyl or is alkylcarbonyl attached
through the carbonyl the oxy at the 3 or 28 carbon of betutlin,
lupeol, or allobetulin, or to an imino or thio in place of the oxy
at the 3 or 28 carbon of betulin, lupeol, or allobetulin, wherein
if it is attached to an oxy, imino, or thio at the 28 carbon of
allobetulin, carbon 19 is a methylene. R.sub.2 is
(C.sub.1-C.sub.24)alkyl. R.sub.3 is absent or
(C.sub.1-C.sub.24)alkyl or is alkylcarbonyl attached through the
carbonyl to the oxy at the 3 or 28 carbon of betulin, lupeol, or
allobetulin, or to an imino or thio in place of the oxy at the 3 or
28 carbon of betulin, lupeol, or allobetulin, wherein if it is
attached to an oxy, imino, or thio at the 28 carbon of allobetulin,
carbon 19 is a methylene. In particular embodiments of compound
VII, two R.sub.1s are (C.sub.6-C.sub.24)alkyl. In particular
embodiments of compound VII, at least two R.sub.1s are
(C.sub.8-C.sub.24)alkyl or (C.sub.10-C.sub.24)alkyl. In particular
embodiments of compound VIII, at least two R.sub.1s attached to
either nitrogen atom are (C.sub.6-C.sub.24)alkyl. In particular
embodiments of compound VIII, at least two R.sub.1s attached to
either nitrogen atom are (C.sub.8-C.sub.24)alkyl or
C.sub.10-C.sub.24)alkyl. Any alkyl or alkylcarbonyl can optionally
be substituted with one or more oxo, hydroxy, mercapto, or
NR.sub.dR.sub.e. R.sub.d and R.sub.e are each independently
hydrogen or alkyl. The compound in this case comprises at least two
moieties selected from the group of betulin, allobetulin, and
lupeol.
[0142] In one specific embodiment of the compound of formula (VI),
the compound is
N,N,N',N'-tetramethylethylenediamine-N,N'-bis-[lup-20(29)-ene-3-acetate].
[0143] The compounds of the invention include one or more
triterpene moieties covalently attached via a linker to a
quaternary ammonium salt. The linker can attach to the triterpene
moiety at any suitable position of the triterpene. The linker can
attach to the quaternary ammonium salt at the N.sup.+ atom or at
any other suitable position. The linker can be, for instance,
alkylene, alkylcarbonyl, alkoxy, alkylimino, oxyalkylcarbonyl,
carbonylalkylcarbonyl, or carbonylalkyloxy.
[0144] The quaternary ammonium salt can also be attached directly
to the triterpene without a linker. The attachment in this case can
be at any suitable position of the triterpene and any suitable
position of the quaternary ammonium salt.
[0145] Processes for preparing compounds of formulas (I)-(IV) are
provided as further embodiments of the invention and are
illustrated by the following procedures in which the meanings of
the generic radicals are as given unless otherwise qualified.
Specifically, the compounds of formulas (I)-(IV) can be prepared
from convenient starting materials, employing procedures (e.g.,
reagents and reaction conditions) known to those of skill in the
art. For example, suitable reagents and reaction-conditions are
disclosed, e.g., in Advanced Organic Chemistry, Part B: Reactions
and Synthesis, Second Edition, Cary and Sundberg (1983); Advanced
Organic Chemistry, Reactions, Mechanisms, and Structure, Second
Edition, March (1977); Protecting Groups in Organic Synthesis,
Second Edition, Greene, T. W., and Wutz, P. G. M., John Wiley &
Sons, New York; and Comprehensive Organic Transformations, Larock,
R. C., Second Edition, John Wiley & Sons, New York (1999).
[0146] The compounds of the invention, e.g., compounds of formulas
(I)-(IV), can be formulated as compositions for the treatment or
prevention of fungal or bacterial infections of plants. Generally,
the compositions comprising compounds of the invention will be
applied topically to the plants, generally by spraying. The
compositions can also be injected into the plant or applied to the
soil for uptake into the plant through the root system. The
compositions can comprise an acceptable vehicle, such as an inert
diluent or a carrier that facilitates uptake into the plant.
Acceptable vehicles are known in the art. Methods of formulating
anti-fungal compounds for spraying onto plants are disclosed, for
instance in Glinsky et al. (U.S. Pat. No. 6,303,589 B1).
[0147] If desired, the compounds of the invention may be applied in
conjunction with one or more inert or active ingredients. Exemplary
materials include dyes, additives affecting stability of the
compositions, additives affecting physical properties of the
sprayed layer, foliar fertilizers, fungicides, and
insecticides.
[0148] The compounds of the invention, e.g. compounds of formulas
(I)-(IV), are also useful for cosmetic treatment of nails and
adjacent skin infected with fungus in humans. In cosmetic
treatment, the compounds of the invention are applied topically to
the affected nails and adjacent skin either alone or in a
composition with an acceptable carrier. The compounds of the
invention can be applied with one or more other inert or active
ingredients.
[0149] Acceptable salts may be obtained using standard procedures
well known in the art, for example by reacting a sufficiently basic
compound such as an amine with a suitable acid affording an
acceptable anion.
[0150] The compounds of formulas (I)-(IV) can be formulated as
pharmaceutical compositions and administered to a mammalian host,
such as a human patient, in a variety of forms adapted to the
chosen route of administration. Typically, the pharmaceutical
compositions will be administered topically. They may also be
administered, e.g., orally or parenterally, and by intravenous,
intramuscular, or subcutaneous routes.
[0151] Thus, the present compounds may be systemically
administered, e.g., orally, in combination with a pharmaceutically
acceptable vehicle such as an inert diluent or an assimilable
edible carrier. They may be enclosed in hard or soft shell gelatin
capsules, may be compressed into tablets, or may be incorporated
directly with the food of the patient's diet. For oral therapeutic
administration, the active compound may be combined with one or
more excipients and used in the form of ingestible tablets, buccal
tablets, troches, capsules, elixirs, suspensions, syrups, wafers,
and the like. Such compositions and preparations should contain at
least 0.1% of active compound. The percentage of the compositions
and preparations may, of course, be varied and may conveniently be
between about 2 to about 60% of the weight of a given unit dosage
form. The amount of active compound in such therapeutically useful
compositions is such that an effective dosage level will be
obtained.
[0152] The tablets, troches, pills, capsules, and the like may also
contain the following: binders such as gum tragacanth, acacia, corn
starch or gelatin; excipients such as dicalcium phosphate; a
disintegrating agent such as corn starch potato starch, alginic
acid and the like; a lubricant such as magnesia stearate; and a
sweetening agent such as sucrose, fructose, lactose or aspartame or
a flavoring agent such as peppermint, oil of wintergreen, or cherry
flavoring may be added. When the unit dosage form is a capsule, it
may contain, in addition to materials of the above type, a liquid
carrier, such as a vegetable oil or a polyethylene glycol. Various
other materials may be present as coatings or to otherwise modify
the physical form of the solid unit dosage form. For instance,
tablets, pills, or capsules may be coated with gelatin, wax,
shellac or sugar and the like. A syrup or elixir may contain the
active compound, sucrose or fructose as a sweetening agent, methyl
and propylparabens as preservatives, a dye and flavoring such as
cherry or orange flavor. Of course, any material used in preparing
any unit dosage form should be pharmaceutically acceptable and
substantially non-toxic in the amounts employed. In addition, the
active compound may be incorporated into sustained-release
preparations and devices.
[0153] The active compound may also be administered intravenously
or intraperitoneally by infusion or injection. Solutions of the
active compound or its salts can be prepared in water, optionally
mixed with a nontoxic surfactant. Dispersions can also be prepared
in glycerol, liquid polyethylene glycols, triacetin, and mixtures
thereof and in oils. Under ordinary conditions of storage and use,
these preparations contain a preservative to prevent the growth of
microorganisms.
[0154] The pharmaceutical dosage forms suitable for injection or
infusion can include sterile aqueous solutions or dispersions or
sterile powders comprising the active ingredient which are adapted
for the extemporaneous preparation of sterile injectable or
infusible solutions or dispersions, optionally encapsulated in
liposomes. In all cases, the ultimate dosage form should be
sterile, fluid and stable under the conditions of manufacture and
storage. The liquid carrier or vehicle can be a solvent or liquid
dispersion medium comprising, for example, water, ethanol, a polyol
(for example, glycerol propylene glycol, liquid polyethylene
glycols, and the like), vegetable oils, nontoxic glyceryl esters,
and suitable mixtures thereof. The proper fluidity can be
maintained, for example, by the formation of liposomes, by the
maintenance of the required particle size in the case of
dispersions or by the use of surfactants. The prevention of the
action of microorganisms can be brought about by various
antibacterial and antifungal agents for example, parabens,
chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In
many cases, it will be preferable to include isotonic agents, for
example, sugars, buffers or sodium chloride. Prolonged absorption
of the injectable compositions can be brought about by the use in
the compositions of agents delaying absorption, for example,
aluminum monostearate and gelatin.
[0155] Sterile injectable solutions are prepared by incorporating
the active compound in the required amount in the appropriate
solvent with various of the other ingredients enumerated above, as
required, followed by filter sterilization. In the case of sterile
powders for the preparation of sterile injectable solutions, the
preferred methods of preparation are vacuum drying and the freeze
drying techniques, which yield a powder of the active ingredient
plus any additional desired ingredient present in the previously
sterile-filtered solutions.
[0156] For topical administration, the present compounds may be
applied in pure form, i.e., when they are liquids. However, it will
generally be desirable to administer them to the skin as
compositions or formulations, in combination with a
dermatologically acceptable carrier, which may be a solid or a
liquid.
[0157] Useful solid carriers include finely divided solids such as
talc, clay, microcrystalline cellulose, silica, alumina and the
like. Useful liquid carriers include water, alcohols or glycols or
water-alcohol/glycol blends, in which the present compounds can be
dissolved or dispersed at effective levels, optionally with the aid
of non-toxic surfactants. Adjuvants such as fragrances and
additional antimicrobial agents can be added to optimize the
properties for a given use. The resultant liquid compositions can
be applied from absorbent pads, used to impregnate bandages and
other dressings, or sprayed onto the affected area using pump-type
or aerosol sprayers.
[0158] Thickeners such as synthetic polymers, fatty acids, fatty
acid salts and esters, fatty alcohols, modified celluloses or
modified mineral materials ca also be employed with liquid carriers
to form spreadable pastes, gels, ointments, soaps, and the like,
for application directly to the skin of the user.
[0159] Examples of useful dermatological compositions which can be
used to deliver the compounds of formula I to the skin are known to
the art; for example, see Jacquet et al. (U.S. Pat. No. 4,608,392),
Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S. Pat. No.
4,559,157) and Wortzman (U.S. Pat. No. 4,820,508).
[0160] Useful dosages of the compounds of formulas (I)-(IV) can be
determined by comparing their in vitro activity, and as vivo
activity in animal models. Methods for the extrapolation of
effective dosages in mice, and other animals, to humans are known
to the art for example, see U.S. Pat. No. 4,938,949.
[0161] The ability of a compound of the invention to act as an
antibacterial or antifungal agent may be determined using
pharmacological models which are well know to the art, including
the tests described in the Examples below.
[0162] The compounds of the invention may also be useful as
pharmacological tools for the further investigation of the
mechanism of their antibacterial or antifungal action.
[0163] The compounds of the invention can also be administered in
combination with other therapeutic agents that are effective to
treat bacterial or fungal infections, or to inhibit or kill
bacteria or fungi.
[0164] The compounds of the invention possess a rigid and
hydrophobic fused pentacyclic ring portion, and a hydrophilic
quaternary amine portion. Because of their amphiphilic character,
they are also useful as surfactants. Their activity as surfactants
may be the basis for their antibacterial and antifungal activities,
by for instance, adsorbing to cytoplasmic membranes.
[0165] The structures and carbon numbering of three exemplary
triterpenes used as starting materials in the synthesis of the
triterpene quaternary ammonium salts of the invention are shown
below. ##STR12##
[0166] The invention will now be illustrated with by the following
non-limiting Examples.
EXAMPLES
[0167] In the syntheses below, "DABCO" refers to
1,4-diazabicyclo[2.2.2]octane.
Example 1
Lup-20(29)-ene-3,28-bis-(N-pyridiniumacetate)dichloride
[0168] ##STR13##
[0169] 4.0 g (67.1 mmol) of betulin-3,28-dichloroacetate was
dissolved in 20 ml of dry pyridine and the solution kept at
80.degree. C. for 6 h. The mixture was placed then into benzene,
the precipitate filtered, washed with the brine, and dried to yield
5.0 g (98.8%) of the product NMR 1H (DMSO, TMS, 300 MHz): 9.18 (r,
2.times.2H, Pyr-H), 8.74 (T, 2.times.1H, Pyr-H) 8.27 (DD,
2.times.2H, Pyr-H), 5.80 (P, 4H, --CO--CH.sub.2), 4.64 (D, 1H,
29-H), 4.53 (DD, 1-H, 3H), 4.36 (D, 1H, 28-H), 3.91 (D, 1H, 3-H),
1.91 (D, 1H, 19-H), 1.65, 0.97, 0.95, 0.84, 0.79, 0.67 (all S,
6.times.3H, 30-, 27-, 26-, 25-, 24-, 23-Me), 1.01-1.64 (complex
CH--, CH.sub.2, 24H); NMR 13C (DMSO): 167.36, 166.58, 150.55,
147.65, 146.84, 128.72, 110.88, 84.35, 65.24, 61.23, 55.46, 50.34,
48.98, 47.84, 46.74, 43.02, 41.17, 38.50, 38.25, 37.99, 37.27,
34.63, 29.77, 28.47, 27.34, 25.49, 23.82, 21.15, 19.53, 18.42,
16.88, 16.51, 16.37, 15.22 IR (KBr): 3426, 2944, 2872, 1742, 1631,
1272, 1222.
Example 2
Lup-20(29)-ene-3-[N-(4-oxybutyl)-1,4-diazabicyclo[2.2.2]octyl-N'-2-acetate-
]
[0170] ##STR14##
[0171] To 3.0 g of betulin-3-chloroacetate (5.85 mmol) in 100 ml of
1-butanol, 1.29 g (5.85 mmol) of N-(4-oxybutyl) DABCO chloride was
added and the mixture was refluxed overnight. The solvent was
evaporated until a viscous liquid formed. Dry toluene was added and
mixed until a solid material appeared. The latter was filtered and
dried to yield 4.03 g (93%) of the product. 1H NMR (DMSO, TMS):
3.28-4.53 (complex CH.sub.2, 21H), 3.07 (D, 1H, 28-H), 0.83-1.87
(complex CH, CH.sub.2, CH.sub.3).
Example 3
[0172]
Lup-20(29)-ene-3,28-bis[N-(1,4-diaza[2,2,2]bicycloocyl)acetate]dic-
hloride ##STR15##
[0173] To a solution of 8.0 g (13.4 mmol) of
betulin-3,28-dichloroacetate in 40 ml of dry dimethylformamide, 6.0
g (4.times.13.4 mmol) of DABCO (diazabicyclooctane) in 20 ml of
DMFA was added at once. In a few minutes, a white precipitate was
formed. The mixture was kept at room temperature for 24 hr. The
precipitate filtered off, washed with DMFA then with dry benzene,
and dried at 90.degree. C. in vacuum. Yield 9.1 g (8.28%) IR (KBr)
cm.sup.-1.
[0174] 1H NMR (DMSO, TMS) 4.72 (S, 1H, 29-H), 4.58 (S, 1H, 29-H),
4.47 (S, 2.times.2H, CH.sub.2--CO), 4.42 DD, 1H, 29-H), 3.92 (DD,
1H, 29-H), 3.51 (T, 12H, DABCO H), 3.08 (T, 12H, DABCO H), 1.93 (M,
1H, 19-H), 1.66, 1.01, 0.97, 0.84, 0.82 (all S, 5.times.3H, 27-,
26-, 25-, 24-, 23-Me), 1.09-1.45 (complex CH--, CH.sub.2, 25H); 13C
NMR (DMSO, TMS) 165.06, 164.53, 149.80, 110.22, 82.65, 63.59,
60.66, 54.40, 52.23, 49.42, 48.25, 46.99, 46.17, 44.52, 42.37,
37.64, 37.12, 36.62, 33.61, 29.16, 27.77, 26.68, 24.75, 23.33,
20.34, 18.86, 17.76, 16.43, 15.90, 15.69, 14.58.
Example 4
Lup-20(29)-ene-3,28-bis[N--(N'-benzyldiazabicyclo[2.2.2]octyl)acetate]tetr-
achloride
[0175] ##STR16##
[0176] To a solution of 5.0 g (6.0 mmol) of
betulin-3,28-bis[N-(diaza[2,2,2]bicycloocyl)acetate] in 70 ml of
dry ethanol, 1.5 ml of benzylchloride was added and the solution
was refluxed overnight. The resulting clear solution was evaporated
in vacuum to 30 ml and then 100 ml of toluene added. A white
precipitate was formed, then filtered, washed with toluene and
dried in vacuum to yield 6.50 g of the product (99%).
[0177] IR (KBr):
[0178] 1H NMR (DMSO, TMS) 7.16-7.57 (complex, 2.times.5H, Ph--H),
491 (S, 2.times.2H, PhCH.sub.2), 3.53-4.79 (complex, 33H, 29-, 28-,
COCH.sub.2--, DABCO H), 1.65, 0.99, 0.96, 0.83, 0.82 (all S,
5.times.3H, 27-, 26-, 25-, 24-, 23-Me), 1.05-1.5 (complex CH--,
CH.sub.2, 25H); 13C NMR (DMSO, TMS) 164.58, 164.06, 133.22, 130.83,
129.32, 128.31, 125.42, 110.25, 83.12, 66.37, 64.02, 60.79, 54.36,
51.90, 51.43, 50.85, 49.91, 49.41, 48.26, 46.99, 46.16, 43.77,
42.38, 37.67, 37.13, 36.62, 33.96, 33.56, 29.11, 27.76, 26.70,
24.76, 23.35, 20.35, 18.86, 17.76, 16.40, 15.90, 15.69, 14.59.
Example 5
Lup-20(29)-ene-3,28-bis[N--(N'-(4-oxybutyl)diazabicyclo[2.2.2]octyl)acetat-
e]tetrachloride
[0179] ##STR17##
[0180] To a solution of 8.68 g (14.60 mmol) of
betulin-3,28-bis(chloroacetate) in 120 ml of 1-butanol, 6.50 g
(29.2 mmol) of N-(4-oxybutyl)diazabicyclo[2.2.2]octane was added.
The mixture was refluxed for 24 hours. After that, the mixture was
cooled down and evaporated in vacuum to yield 15.0 g (98.8%) of the
product.
[0181] 1H NMR (DMSO, TMS) 7.20 (M, 2.times.1H, OH), 3.21-5.15
(complex CH.sub.2, CH), 0.83-1.95 (complex CH.sub.3, CH.sub.2, CH).
13C NMR (DMSO, TMS) 54.39, 51.39, 51.12, 50.41, 50.06, 49.44,
48.27, 46.99, 46.19, 44.19, 42.38, 37.70, 37.12, 36.63, 33.59,
28.96, 27.77, 26.69, 24.77, 23.36, 20.35, 18.87, 18.47, 18.24,
17.74, 16.41, 15.89, 15.70, 14.59.
Example 6
Lup-20(29)-ene-3-[N-(1,4-diazabicyclo[2.2.2]octyl)acetate]chloride
[0182] ##STR18##
[0183] To a solution of 2.90 g (5.58 mmol) of
betulin-3-chloroacetate in 20 ml of toluene, 1.25 g (5.58.times.2
mmol) of DABCO in 20 ml of toluene was added and the mixture was
kept at 80.degree. C. for 6 hr. The precipitate was filtered,
washed with toluene and dried to yield 3.50 g (99%) of the
product.
[0184] 1H NMR (DMSO, TMS): 4.3-4.69 (complex CH, CH.sub.2), 3.54
(T, 6H, DABCO CH.sub.2), 3.11 (T, 6H, DABCO CH.sub.2), 1.9 (M, 1H,
19-H), 0.85-1.67 (complex, CH, CH.sub.2, CH.sub.3).
Example 7
Lup-20(29)-ene-3,28-bis[(tetramethylethylenediamine-N-yl)acetate]dichlorid-
e
[0185] ##STR19##
[0186] To a solution of 8.0 g (0.0134 mol) of
betulin-3,28-bis(chloroacetate) in 40 ml of dry DMFA, 12 ml of
tetramethylethylenediamine was added. The mixture was kept at room
temperature overnight then diluted with benzene, the precipitate
filtered, washed with benzene and dried to yield 11.0 g (99% Y) of
the product.
[0187] 1H NMR (DMSO, TMS): 4.53-4.72 (complex CH.sub.2, 4H), 4.35
(D, 1H, 28-n), 3.89 (D, 1H, 28-H), 2.5-3.65 (complex NCH.sub.2,
NCH.sub.3), 0.82-1.66 (complex CH, CH.sub.2, CH.sub.3.
Example 8
Lup-20(29)-ene-3,29-bis[N-benzyl-N,N,N',N'-tetramethylethylenediamine-N-yl-
)acetate]tetrachloride
[0188] ##STR20##
[0189] To a solution of 4.5 g (5.43 mmol) of
Lup-20(29)-ene-3,28-bis[(tetramethylethylenediamine-N-yl)acetate]dichlori-
de (7) in 70 ml of ethanol 1.8 ml of benzylchloride was added and
the solution was refluxed overnight, then evaporated in vacuum to
yield 5.76 g (97.0%) of the product.
[0190] 1H NMR (DMSO, TMS): 7.1-7.6 (complex benzene H), 2.51-4.85
(complex CH, CH.sub.2, CH.sub.3).
Example 9
Lup-20(29)-ene-3-[N--(N'-(benzyl)diazabicyclo[2.2.2]octyl)acetate]dichlori-
de
[0191] ##STR21##
[0192] To a solution of 4.5 g (5.43 mmol) of Lup-20
(29)-ene-3-[(DABCO-N'-yl)acetate]chloride (6) in 70 ml of ethanol,
1.8 ml of benzylchloride was added and the solution was refluxed
overnight then evaporated in vacuum to yield 5.70 g (97.0%) of the
product.
[0193] 1H NMR (DMSO, TMS): 7.1-7.6 (complex benzene H), 2.51-4.85
(complex CH, CH.sub.2, CH.sub.3).
Example 10
N,N,N',N'-tetramethylethylenediamine-N,N'-bis-[lup-20(29)-ene-3-acetate]
[0194] ##STR22##
[0195] To a solution of 0.72 g (1.38 mmol) of
betulin-3-chloroacetate in 10 ml of dry DMFA, 0.08 g (0.69 mmol) of
tetramethylethylenediamine was added and the mixture was kept at
80.degree. C. for 6 hr then the solvent evaporated in vacuum. The
residue was washed with toluene and filtered to yield 0.23 g (28%)
of the product.
[0196] 1H NMR (DMSO, TMS): 4.29-4.66 (complex CH.sub.2, CH),
3.09-3.68 (complex CH, CH.sub.3, CH.sub.2), 0.84-2.70 (complex CH,
CH.sub.2, CH.sub.3).
Example 11
Bis(N,N'-pyridinium-2-ethyl)lup-20(29)-ene-3,28-dicarbamate
dichloride
[0197] ##STR23##
[0198] 4.0 g (6.1 mmol) of lup-20(29)-ene 3,28-di(ethyl) carbamate
was dissolved in 5 mL of dry pyridine and the solution kept at
80.degree. C. for 12 h. The mixture was then placed into benzene,
the precipitate filtered, washed with the brine, and dried to yield
4.6 g (93%) of the product.
[0199] 1HNMR (CDCl.sub.3, d): 9.2 (m, 4H), 8.6 (m, 2H), 8.2 (m,
4H), 7.5 (m, 2H), 4.8 (m, 5H), 4.6 (s, 1H), 4.1 (m, 2H), 3.6 (m,
5H), 2.4 (m, 1H), 2.0-0.7 (m, 42H).
Example 12
1-(3,28-(diacetoxy)lup-20(29)-ene-30-yl)-4-(dimethylamino)pyridinium
bromide
[0200] ##STR24##
[0201] To a solution of 3,28-diacetyl-30-bromobetulin (1 g, 1.669
mmol) in 10 ml of toluene was added a solution of 4-(dimethylamino)
pyridine (DMAP) (0.204 g, 1.669 mmol) in 10 ml of toluene under
nitrogen atmosphere. The solution was heated overnight at
70.degree. C. Then the solvent was evaporated and the residue was
washed with hexane. The solids were dried to yield 1.18 g (97%) of
the 1-(3,28-diacetoxylup-20-en-30-yl)-4-(dimethylamino)pyridinium
bromide. M.p. 119-123.degree. C. 1H NMR (CDCl.sub.3, TMS):
0.76-1.90 (40H, m), 2.04 (3H, s), 2.06 (3H, s), 3.32 (6H, s), 3.73,
4.24 (2H, AB, J=11.1 Hz), 4.46 (1H, m), 4.64 (1H, s), 4.89, 5.03
(2H, AB, J=15.9 Hz), 5.09 (1H, s), 7.08 (2H, m), 8.32 (2H, m). 13C
NMR (CDCl.sub.3 TMS): 14.70, 15.93, 16.08, 16.40, 18.04, 20.82,
20.96, 21.25, 23.57, 26.82, 27.28, 27.85, 29.57, 31.44, 33.99,
34.17, 36.95, 37.23, 37.68, 38.31, 40.63, 40.79, 42.58, 43.25,
46.26, 50.02, 55.23, 62.11, 80.75, 108.22, 112.00, 125.19, 128.12,
128.93, 142.79, 149.15, 156.27, 170.91, 171.41, IR (KBr): 3414.17,
2941.70, 2872.27, 1728.88, 1647.69, 1568.23, 1447.32, 1367.09,
1240.59, 1167.30, 1030.57, 978.51, 839.85, 732.24, 514.70 Calcd: C,
67.66; H, 8.72. Found: C, 66.50; H, 8.69.
Example 13
Lup-20(29)-ene-3,28-bis(N-pyridinium-2-propionate)dichloride
[0202] ##STR25##
[0203] 4.0 g (6.4 mmol) of
lup-20(29)-ene-3,28-di(2'-chloropropionate) was dissolved in 5 mL
of dry pyridine. The solution was kept at 80.degree. C. for 12 h.
The mixture was then placed into benzene, the precipitate filtered,
washed with the brine, and dried to yield 4.6 g (92%) of the
product.
Example 14
Lup-20(29)-ene-3,28-bis(N-pyridinium-3-propionate)dichloride
[0204] ##STR26##
[0205] 4.0 g (6.4 mmol) of
lup-20(29)-ene-3,28-di(3'-chloropropionate) was dissolved in 5 ml
of dry pyridine. The solution kept at 80.degree. C. for 12 h. The
mixture was placed then into benzene, the precipitate filtered,
washed with the brine, and dried to yield 4.6 g (92%) of the
product.
Example 15
Lup-20(29)-ene-3,28-bis(N-pyridinium-4-butyrate)dichloride
[0206] ##STR27##
[0207] 4.0 g (6.1 mmol) of
lup-20(29)-ene-3,28-di(2'-chlorobutyrate) was dissolved in 5 ml of
dry pyridine. The solution kept at 80.degree. C. for 12 h. The
mixture was then placed into benzene, the precipitate filtered,
washed with the brine, and dried to yield 4.5 g (90%) of the
product.
Example 16
Lup-20(29)-ene-3,28-bis(N-pyridinium-4-butyrate)dibromide
[0208] ##STR28##
[0209] 4.0 g (5.4 mmol) of lup-20(29)-ene-3,28-di(4'-bromobutyrate)
was dissolved in 5 ml of dry pyridine. The solution was kept at
80.degree. C. for 12 h. The mixture was placed then into benzene
the precipitate filtered, washed with brine, and dried to yield 4.3
g (89%) of the product.
Example 17
Lup-20(29)-ene 3,28-bis(N-pyridinium-2-butyrate) dibromide
[0210] ##STR29##
[0211] 4.0 g (5.4 mmol) of lup-20(29)-ene-3,28-di(2'-bromobutyrate)
was dissolved in 5 ml of dry pyridine. The solution was kept at
80.degree. C. for 12 h. The mixture was placed then into benzene,
the precipitate filtered, washed with brine, and dried to yield 4.3
g (89%) of the product.
Example 18
1-[3,28-(diacetoxy)lup-20(29)-ene-30-yl]-1,4-diazabicyclo[2.2.2]octane
bromide
[0212] ##STR30##
[0213] To a solution of 30-bromo-3,28-diacetylbetulin (21.9 g,
0.03616 mol) in dry toluene (100 ml) was added a solution of DABCO
(12.2 g, 0.1085 mol) in dry toluene (250 ml) with stirring under
nitrogen atmosphere. The mixture was refluxed for 3 hours. White
solids were formed. The solids were filtered off and washed with
toluene and hexane to give the pure product (20.2 g, 78%). M.p.
254-257.degree. C. (dec.) 1H NMR (CDCl.sub.3, TMS): 0.76-1.95 (39H,
m), 3.26 (6H, m), 3.73 (7H, m), 4.17 (1H, d, J=13 Hz), 4.25 (1H, d,
J=11 Hz), 4.47 (2H, m), 5.71 (2H, m). 13C NMR (CDCl.sub.3, TMS):
14.51, 15.95, 16.10, 16.45, 18.08, 20.87, 21.01, 21.29, 23.63,
26.72, 27.88, 28.56, 29.62, 34.04, 34.47, 36.97, 37.72, 38.25,
40.79, 42.64, 45.52, 46.71, 49.90, 52.69, 55.22, 56.48, 62.01,
71.56, 80.73, 125.89, 144.03, 170.94, 171.33. IR (KBr): 3410.70,
2947.87, 1729.26, 1463.90, 1371.72, 1246.37, 1031.54, 979.26,
921.04, 846.02, 646.61 cm.sup.-1. Calcd.: C, 66.93; H, 9.13. Found:
C, 66.10; H, 9.51.
Example 19
3,28-bis[3-(1-piperidinyl)propanoyloxy]lup-20(29)-ene
dihydrochloride
[0214] ##STR31##
[0215] To a solution of 5.74 g (0.0104 mol) of
3,28-diacryloylbetulin in 300 ml MeOH was added 4.44 g (0.0521 mol)
of piperidine. The mixture was refluxed for 2 h. The solvent was
evaporated and the product was dried in vacuum. Yield 7.51 g
(100%). 1H NMR (free base) (CDCl.sub.3, TMS): 0.7-2.00 (43H, m),
2.30-2.80 (10H, m), 2.98 (2H, m), 3.85, 4.28 (2H, AB, J=11.0 Hz),
4.49 (1H, m), 4.55-4.70 (2H, m). 13C NMR (free base) CDCl.sub.3,
TMS): 14.67, 15.95, 16.07, 16.48, 18.09, 19.05, 23.62, 24.26,
24.67, 25.07, 25.90, 26.97, 27.85, 29.50, 29.69, 32.52, 32.82,
34.04, 34.50, 36.97, 37.48, 37.79, 38.29, 40.81, 42.61, 45.77,
46.35, 47.66, 48.70, 50.18, 54.17, 54.36, 54.40, 55.31, 62.57,
80.64, 109.79, 150.07, 172.45, 173.13.
[0216] This product was dissolved in 50 ml of aqueous HCl (1%), and
the solution was evaporated to give 7.86 g of salt M.p.
211-215.degree. C. IR (hydrochloride) (KBr): 3420.73, 2957.51,
2637.19, 2539.99, 1727.91, 1454.84, 1391.01, 1198.93, 975.23,
881.70, 548.45, 445.47 cm.sup.-1. Calcd.: C, 69.58; H, 9.90. Found:
C, 68.97; H, 10.06.
Example 20
1-(3,28-dihydroxylup-20(29)-ene-30-yl)-4-(dimethylamino)pyridinium
bromide
[0217] ##STR32##
[0218] To a mixture of 30-bromobetulin (501 mg, 0.961 mmol) and
4-dimethylaminopyridine (117 mg, 0.961 mmol), ethanol (40 ml) and
THF (10 ml) were added under nitrogen. The solution was refluxed
for 3 h. The solvent was evaporated, and the solids were
recrystallized from methanol/EtOAc (1:5) twice to yield 469 mg
(76%) of the product. M.p. 220-223.degree. C. 1H NMR (CD3OD, TMS):
0.69-2.38 (m, 40H), 3.12 (dd, J1=10.8 Hz, J2=5.4 Hz, 1H),
3.201-3.28 (m, 7H), 3.72 (d, J=11.1 Hz, 1H), 4.59 (s, 1H), 4.79 (s,
1H), 5.12 (s, 1H), 7.03, 8.11 (AB, J=7.8 Hz, 4H). 1H NMR
(CDCl.sub.3, TMS): 0.65-2.20 (m, 39H), 2.34 (m, 1H), 3.20 (m, 1H),
3.30 (s, 6H), 3.40, 3.68 (AB, J=9.6 Hz, 2H), 4.72 (s, 1H), 4.91 (s,
2H), 5.11 (s, 1H), 7.02 (m, 2H), 8.34 (m, 2H). 1H NMR (DMSO-d6,
TMS): 0.60-1.76 (m, 36H), 1.85 (m, 3H), 2.31 (m, 1H), 3.01 (m, 2H),
3.50 (m, 1H), 4.67 (br s, 1H), 4.79 (dd, J1=17.6 Hz, J2=17.6 Hz,
2H), 5.01 (s, 1H), 7.06 (d, J=7.4 Hz, 2H), 8.23 (d, J=7.4 Hz, 2H).
13C NMR (DMSO-d6, TMS): 13.98, 14.46, 15.59, 15.72, 15.80, 17.85,
20.42, 20.66, 26.36, 26.49, 27.06, 27.99, 30.95, 33.34, 33.69,
36.49, 36.57, 42.09, 47.22, 49.23, 49.62, 54.74, 57.53, 59.64,
76.67, 107.51, 110.29, 142.46, 142.54, 150.95, 155.79, 170.21, IR
(KBr): 3362.11, 2932.25, 2866.10, 1648.07, 1567.85, 1445.78,
1400.65, 1237.11, 1168.27, 1036.94, 819.02, 517.41 cm.sup.-1.
Calcd.: C, 69.03; H, 9.24; Br, 12.41.
Example 21
Lup-20(29)-ene-3,28-bis[N-(4-dimethylaminopyridinium)-2-propionate]dichlor-
ide
[0219] ##STR33##
[0220] To a solution of 1.76 g (2.8 mmol) of
betulin-3,28-di(2'-chloropropionate) in 10 ml of dimethylacetamide,
1.38 g (4.times.2.8 mmol) of 4-(dimethylamino)pyridine (DMAP) in 10
ml of dimethylacetamide was added at once. The mixture was kept at
room temperature for 24 hr. Then ether was added, the precipitate
filtered off, washed with ether, and dried at 90.degree. C. in
vacuum. Yield 2 g (82%) 1H NMR (CDCl.sub.3, d): 8.6 (m, 3H), 7.0
(m, 3H), 6.0-5.8 (m, 2H), 4.7 (s, 1H), 4.6 (s, 1H), 4.5-4.3 (m,
2H), 4.1-3.8 (m, 1H), 3.4 (s, 12H), 2.4 (m, 1H), 22-0.7 (m, 50H)
13C NMR (CDCl.sub.3, d): 169.6, 168.9, 156.5, 149.5, 142.0, 116.2,
109.9, 107.8, 84.0, 83.3, 70.4, 64.9, 63.9, 63.7, 60.0, 55.1, 52.6,
50.0, 58.6, 47.4, 463, 42.5, 40.7, 40.4, 37.8, 37.7, 37.4, 36.8,
35.0, 34.2, 33.8, 29.2, 28.0, 26.8, 24.9, 23.3, 21.4, 20.5, 18.9,
18.2, 17.9, 16.3, 15.9, 14.5.
Example 22
Lup-20(29)-ene-3,28-bis[N-(1,4-diazabicyclo[2.2.2]octyl)-2-propionate]-dic-
hloride
[0221] ##STR34##
[0222] To a solution of 1.61 g (2.6 mmol) of
betulin-3,28-di(2'-chloropropionate) in 10 ml of dimethylacetamide,
1.16 g (4.times.2.6 mmol) of DABCO in 20 ml of dimethylacetamide
was added at once. The mixture was kept at room temperature for 24
hr. Then ether was added, the precipitate filtered off, washed with
ether and dried at 90.degree. C. in vacuum. Yield 1.75 g (80%).
[0223] 1H NMR (DMSO, d): 4.9 (s, 1H), 4.8 (s, 1H), 4.7 (m, 1H), 4.6
(d, 1H), 4.2 (d, 1H), 3.7 (m, 6H), 3.6 (m, 6H), 3.7 (m, 12H), 2.6
(m, 1H), 2.0-0.9 (m, 50H).
Example 23
1-(lup-20(29)-ene-30-yl)-1,4-diazabicyclo[2.2.2]octane bromide
[0224] ##STR35##
[0225] A mixture of 30-bromobetulin (503 mg, 0.964 mmol) and
1,4-diazabicyclo[2.2.2]octane (130 mg, 1.16 mmol) in methanol (25
ml) was refluxed for 3 hours under nitrogen. The solvent was
evaporated; THF was added, and solids were filtered off and washed
with THF. The solids were recrystallized from isopropanol to yield
310 mg (51%) of the product. M.p. 317-319.degree. C. (dec.) 1H NMR
(DMSO-d6, TMS): 0.60-1.76 (m, 34H), 1.91 (m, 2H), 2.21 (m, 3H),
2.95-3.45 (m, 14H), 3.51 (m, 1H), 3.94 (dd, J1=35.4 Hz, J2=12.3 Hz,
2H), 5.40 (s, 1H), 5.62 (s, 1H). 13C NMR (DMSO-d6, TMS): 14.22,
15.54, 15.71, 15.77, 17.86, 20.50, 26.47, 27.55, 27.97, 28.64,
28.67, 33.40, 33.71, 34.07, 36.27, 36.53, 38.10, 38.37, 42.07,
44.63, 47.54, 49.39, 51.75, 54.65, 56.52, 57.66, 70.54, 76.60,
108.23, 124.02, 145.42, IR KBr): 3373.68, 2925.50, 2862.63,
1466.02, 1374.62, 1040.99, 844.28, 797.61, 651.44 cm.sup.-1.
Calcd.: C, 68.22; H, 9.70; Br, 12.61. Found: C, 67.87; H, 9.74; Br,
12.51.
Example 24
1-(3,28-dihydroxylup-20(29)-ene-30-yl)-pyridinium bromide
[0226] ##STR36##
[0227] 30-bromobetulin (505 mg, 0968 mmol) was dissolved in
pyridine (2.5 ml, 30.9 mmol) under nitrogen. The mixture was kept
at 80.degree. C. for 2 hours. Benzene was added; solids were
filtered off and recrystallized from MeCN to yield 395 mg (68%) of
product M.p. 280-282.degree. C. 1H NMR (CD3OD, TMS): 0.68-2.06 (m,
39H), 2.34 (m, 1H), 3.12 (dd, J1=10.8 Hz, J2=5.1 Hz, 1H), 3.24,
3.72 (AB, 1=11.3 Hz, 2H), 4.65 (s, 1H), 5.24 (s, 1H), 5.29 (dd,
J1=15.9 Hz, J2-15.9 Hz, 2H), 8.18 (m, 2H), 8.67 (m, 1H), 9.01 (m,
2H). 13C NMR (CD3OD, TMS): 15.20, 16.16, 16.55, 16.71, 19.46,
22.11, 28.07, 28.16, 28.64, 30.35, 32.71, 34.84, 35.47, 38.30,
38.50, 39.98, 40.10, 42.17, 43.84, 43.93, 45.09, 51.59, 51.70,
56.80, 60.08, 66.91, 66.93, 66.95, 79.61, 113.65, 129.66, 146.74,
147.59, 151.42. IR (KBr): 3382.74, 2936.88, 2867.64, 1629.37,
1481.26, 1388.89, 1150.53, 1040.80, 769.46, 689.04. Calcd.: C,
69.98; H, 9.06; Br, 13.30. Found: C, 69.90; H, 8.95; Br, 13.50.
Example 25
Lup-20(29)-ene-3,28-bis[(N-(1,4-diazabicyclo[2.2.2]octyl)-4-butyrate)]dibr-
omide
[0228] ##STR37##
[0229] To a solution of 1.5 g (2.0 mmol)
lup-20(29)-ene-3,28-di(4'-bromobutyrate) in 10 ml of
dimethylacetamide, 0.9 g (4.times.2.0 mmol) of DABCO in 5 ml of
dimethylacetamide was added at once. The mixture was kept at room
temperature for 24 hr. Then ether was added, the precipitate
filtered off, washed with ether and dried at 90.degree. C. in
vacuum. Yield 1.73 g (78%).
[0230] 1H NMR DMSO, d): 4.75 (s, 1H), 4.6 (s, 1H), 4.5 (m, 1H),
4.35 (d, 1H), 3.8 (d, 1H), 3.5-3.0 (m, 30H), 2.5 (m, 7H), 2.0-0.9
(m, 42H).
Example 26
Lup-20(29)-ene-3,28-bis[N-(1,4-diazabicyclo[2.2.2]octyl)-4-butyrate]dichlo-
ride
[0231] ##STR38##
[0232] To a solution of 1.5 g (2.3 mmol)
lup-20(29)-ene-3,28-di(4-chlorobutyrate) in 10 ml of
dimethylacetamide, 1.03 g (4.times.2.3 mmol) of DABCO in 5 ml of
dimethylacetamide was added at once. The mixture was kept at room
temperature for 24 hr. Then ether was added, the precipitate
filtered off, washed with ether and dried at 90.degree. C. in
vacuum. Yield 1.75 g (87%).
[0233] 1H NMR (DMSO, d): 4.75 (s, 1H), 4.6 (s, 1H), 4.5 (m, 1H),
4.35 (d, 1H), 3.8 (d, 1H), 3.5-3.0 (m, 30H), 2.5 (m, 7H), 2.0-0.9
(m, 42H).
Example 27
1-(3,28-dihydroxylup-20(29)-ene-30-yl)-[N-3-(hydroxymethyl)pyridinium]brom-
ide
[0234] ##STR39##
[0235] A mixture of 30-bromobetulin (1 g, 1.917 mmol) and
3-hydroxymethylpyridine (418 mg, 3.83 mmol) in THF (10 ml) was
refluxed for 17 hours under nitrogen. The solids were filtered off,
washed with THF, and recrystallized from MeCN to yield 719 g (59%)
of product M.p. 244-247.degree. C. 1H NMR (CD3OD, TMS): 0.70-2.10
(m, 39H), 2.40 (m, 1H), 3.14 (dd, J1=10.8 Hz; J2=5.1 Hz, 1H), 3.23,
3.72 (AB, J=11.0 Hz, 2H), 4.66 (s, 1H), 5.24-536 (m, 3H), 8.12 (m,
1H), 8.59 (m, 1H), 8.88 (m, 1H), 8.93 (br s, 1H). 13C NMR (CD3OD,
TMS): 15.19, 16.16, 16.55, 16.71, 19.46, 22.13, 28.07, 28.64,
28.71, 30.35, 32.70, 34.85, 35.49, 38.31, 38.52, 39.99, 40.10,
42.19, 43.86, 45.08, 51.59, 51.73, 56.81, 60.10, 61.07, 66.95,
79.65, 113.75, 129.06, 144.16, 145.08, 145.65, 151.35, IR (KBr):
3356.51, 2938.61, 2866.68, 1458.89, 1390.04, 1032.70, 987.38,
921.04, 688.27, 545.37 cm.sup.-1. Calcd.: C, 68.55; H, 8.95; Br,
12.67. Found: C, 66.65; H, 8.76; Br, 12.48.
Example 28
1-(3,28-dihydroxylup-20(29)-ene-30-yl)-(N-3,5-dimethylpyridinium)bromide
[0236] ##STR40##
[0237] A mixture of 30-bromobetulin (1.004 g, 1.925 mmol) and
3,5-dimethylpyridine (412 mg, 3.85 mmol) in THF (10 ml) and MeOH (5
ml) was refluxed for 20 hours under nitrogen. The solvent was
evaporated, and the solids were recrystallized from MeCN to give
993 mg (82%) of the product. M.p. 293-296.degree. C. 1H NMR (CD3OD,
TMS): 0.68-2.10 (m, 39H), 2.38 (m, 1H), 2.55 (s, 6H), 3.12 (dd,
J1=10.8 Hz, J2=5.4 Hz, 1H), 3.24, 3.72 (AB, J=11.4 Hz, 2H), 4.66
(s, 1H), 5.16 (m, 2H), 5.22 (s, 1H), 8.33 (s, 1H), 8.67 (s, 2H).
13C NMR (CD3OD, TMS): 15.19, 16.15, 16.54, 16.71, 16.75, 18.25,
19.44, 19.47, 22.14, 28.04, 28.08, 28.64, 30.37, 30.39, 32.70,
32.71, 34.87, 35.47, 38.31, 38.50, 39.99, 40.11, 40.13, 42.18,
43.84, 45.01, 51.42, 51.44, 51.73, 56.81, 60.11, 79.63, 113.57,
140.66, 143.43, 148.50, 151.14, IR (KBr): 3327.10, 3075.15,
2930.51, 2867.45, 1631.10, 1453.10, 1388.50, 1308.28, 1036.94,
930.10, 659.92, 559.64 cm.sup.-. Calcd.: C, 70.68; H, 9.30; Br,
12.71. Found: C, 70.47; H, 929; Br, 12.48.
Example 29
Bis[N-(1,4-diazabicyclo[2.2.2]octyl)-2-ethyl]-lup-20(29)-ene-3,28-dicarbam-
ate dichloride
[0238] ##STR41##
[0239] To a solution of 0.75 g (1.15 mmol)
120(29)-ene-3,28-di-ethylcarbamate in 10 ml of dimethylacetamide,
0.51 g (4.times.1.15 mmol) of DABCO in 5 ml of dimethylacetamide
was added at once. The mixture was kept at boiling temperature for
24 hr. Then ether was added, the precipitate filtered oil; washed
with ether and dried at 90.degree. C. in vacuum. Yield 0.75 g (56%)
1H NMR (DMSO, TMS, d): 4.8 (s, 1H), 4.6 (s, 1H), 4.4-4.0 (m, 5H),
3.9-3.1 (m, 32H), 2.6 (m, 1H), 2.0-0.9 (m, 42H).
Example 30
Lup-20(29)-ene-3,28-bis[N-(3-oxymethylpyridinium)acetate)dichloride
[0240] ##STR42##
[0241] 3 g of betulin-3,28-di(chloroacetate) (0.005 mol) was
dissolved in 7 ml of 1-methyl-2-pyrrolidinone, and 2.2 g of
3-pyridylcarbinol (0.02 mol) was added. The reaction mixture was
heated up to 70.degree. C. for 20 hours. Reaction was monitored by
TLC analysis. The reaction mixture then was diluted with TIE and
filtrated. The precipitate was washed with THF and dried with an
oil pump. Yield 3.55 g.
Example 31
Lup-20(29)-ene-3,28-bis[N-(2-oxymethylpyridinium)acetate]dichloride
[0242] ##STR43##
[0243] 3 g of betulin-3,28-di(chloroacetate) (0.005 mol) was
dissolved in 7 ml of 1-methyl-2-pyrrolidinone, and 2.2 g of
2-pyridylcarbinol (0.02 mol) was added. The reaction mixture was
heated up to 70.degree. C. for 20 hours. Reaction was monitored by
TLC analysis. The reaction mixture then was diluted with THF and
filtrated. The precipitate was washed with THF and dried on oil
pump. Yield 1.75 g.
Example 32
Lup-20(29)-ene-3,28-bis[N-(2-methylureapyridinium)acetate]dichloride
[0244] ##STR44##
[0245] 5 g of betulin-3,28-di(chloroacetate) (0.008 mol) was
dissolved in 35 ml of 1-methyl-2-pyrrolidinone, and 4 g of
1-(3-pyridylmethyl)urea (0.0265 mol) was added. The reaction
mixture was heated up to 70.degree. C. for 20 hours Reaction was
monitored by TLC analysis. The reaction mixture then was diluted
with THF and filtrated. The precipitate was washed with THF and
dried with an oil pump. Yield 5.8 g (85%).
Example 33
Lup-20(29)-ene-3-[N-(2-oxymethylpyridinium)acetate]chloride
[0246] ##STR45##
[0247] 3 g of betulin-3<chloroacetate) (0.006 mol) was dissolved
in 15 ml of 1-methyl-2-pyrrolidinone, and 2.56 g of
2-pyridylcarbinol (0.024 mol) was added. The reaction mixture was
heated up to 70.degree. C. for 20 hours. Reaction was monitored by
TLC analysis. The reaction mixture then was diluted with THF and
filtered. Precipitate was washed with CHCl.sub.3 and dried with an
oil pump. Yield 1.66 g.
Example 34
Lup-20(29)-ene-3,28-bis[N--(N-methylmorpholino)acetate]dichloride
[0248] ##STR46##
[0249] 3 g of betulin-3,28-bis-(chloroacetate)dichloride (0.005
mol) was dissolved in 15 ml of 1-methyl-2-pyrrolidinone, and 2.02 g
of 4-methylmorpholine (0.024 mol) was added. The reaction mixture
was heated up to 70.degree. C. for 12.0 hours. Reaction was
monitored by TLC analysis. The reaction mixture then was diluted
with THF and filtered. The precipitate was washed with CHCl.sub.3
and dried with an oil pump. Yield 2.95 g.
Example 35
Lup-20(29)-ene-3,28-bis[(4-hydroxyl-N-methylpiperidino)acetate]dichloride
[0250] ##STR47##
[0251] 3 g of betulin-3,28-bis(chloroacetate)dichloride (0.005 mol)
was dissolved in 15 ml of 1-methyl-2-pyrrolidinone, and 4.6 g of
4-hydroxy-1-methylpiperidine (0.04 mol) was added. The reaction
mixture was heated up to 70.degree. C. for 20 hours. Reaction was
monitored by TLC analysis. The reaction mixture then was diluted
with THF and filtered. The precipitate was washed with CHCl.sub.3
and dried with an oil pump. Yield is 2.01 g.
Example 36
Lup-20(29)-ene-3-[N-(3-(ureamethyl)pyridinium)acetate]chloride
[0252] ##STR48##
[0253] 3 g of betulin-3-chloroacetate (0.006 mol) was dissolved in
15 ml of 1-methyl-2-pyrrolidinone, and 1.8 g of
1-(3-pyridylmethyl)urea (0.012 mol) was added. The reaction mixture
was heated up to 70.degree. C. for 20 hours. Reaction was monitored
by TLC analysis. The reaction mixture then was diluted with THF and
filtered. The precipitate was washed with CHCl.sub.3 and benzene
and dried with an oil pump. Yield 2 g.
Example 37
Lup-20(29)-ene-3-(N-pyridiniumacetate)chloride
[0254] ##STR49##
[0255] 8 g of betulin-3-(chloroacetate) were dissolved in 50 ml of
pyridine and heated up to 70.degree. C. for 3 hours. The
precipitate was filtered and washed with THF and then with benzene
twice, and dried with an oil pump. Yield is 7.5 g.
Example 38
Lup-20(29)-ene-3,28-bis[N-(1,4-diazabicyclo[2.2.2]octyl)-2-butyrate]dibrom-
ide
[0256] ##STR50##
[0257] To a solution of 1.5 g (2.0 mmol)
lup-20(29)-ene-3,28-di(2'-bromobutyrate) in 10 ml of
dimethylacetamide, 0.9 g (4.times.2.0 mmol) of DABCO in 5 ml of
dimethylacetamide was added at once. The mixture was kept at room
temperature for 24 hr. Then ether was added, the precipitate
filtered off, washed with ether, dried at 90.degree. C. in vacuum
Yield 1.84 g (94%). 1H NMR (CD3OD, d): 8.0 (m 2H), 4.9-4.7 (dd,
2H), 4.6 (m, 2H), 4.2 (m, 2H), 3.8 (m, 4H), 3.5 (m, 5H), 3.2 (m,
1H), 2.5 (m, 1H), 2.3 (m, 2H), 2.0-0.7 (m, 42H).
Example 39
Lup-20(29)-ene-3,28-bis[N-(4-dimethylaminopyridinium)-2-butyrate]dibromide
[0258] ##STR51##
[0259] To a solution of 4 g (5.4 mmol) of
betulin-3,28-di(2'-bromobutyrate) in 10 ml of benzene, 2.7 g
(4.times.5.4 mmol) of 4-(dimethylamino)pyridine (DMAP) in 10 ml of
benzene was added at once. The mixture was kept at room temperature
for 24 hr. Then ether was added, the precipitate filtered off,
washed with ether and dried at 90.degree. C. in vacuum. Yield is
4.8 g (90%) 1H NMR (DMSO, d): 8.4 (m, 4H), 7.1 (m, 4H), 5.4 (m,
2H), 4.7-4.5 (dd, 2H), 4.5-4.2 (m, 2H), 3.8 (m, 1H), 3.4 (m, 4H),
3.2 (m, 12H), 2.4 (m, 5H), 2.0-0.7 (m, 4H+2H).
Example 40
Lup-20(29)-ene-3,28-bis[N-(4-dimethylaminopyridinium)butyrate]dibromide
[0260] ##STR52##
[0261] To a solution of 3 g (4.1 mmol) of
betulin-3,28-di(4'-bromobutyrate) in 10 ml of benzene, 2 g
(4.times.4.1 mmol) of 4-(dimethylamino) pyridine (DMAP) in 10 ml of
benzene was added at once. The mixture was kept at room temperature
for 24 hr. Then ether was added, the precipitate filtered off,
washed with ether and dried at 90.degree. C. in vacuum. Yield 3.7 g
(93%) 1H NMR (DMSO, d): 8.4 (m, 4H), 7.1 (m, 4H), 4.7-4.5 (dd, 2H),
4.4 (m, 1H), 4.2 (m, 5H), 3.8 (d, 1H), 3.2 (m, 12H), 2.4 (m, 5H),
2.1 (m, 4H), 2.0-0.7 (m, 42H).
Example 41
Lup-20(29)-ene-3,28-bis[N-(4-dimethylaminopyridinium)-3-propionate]dichlor-
ide
[0262] ##STR53##
[0263] To a solution of 4 g (6.1 mmol) of
betulin-3,28-di(3'-chloropropionate) in 10 ml of benzene 3 g
(4.times.6.1 mmol) of 4-(dimethylamino) pyridine (DMAP) in 10 ml of
benzene was added at once. The mixture was kept at room temperature
for 24 hr. Then the ether was added, the precipitate filtered off,
washed with ether and dried at 90.degree. C. in vacuum. Yield 4.3 g
(77%).
[0264] 1H NMR (DMSO, d): 8.4 (m, 4H), 7.1 (m, 4H), 4.7-4.5 (dd,
2H), 4.4 (m, 4H), 4.3 (m, 1H), 3.8 (d, 1H), 3.5 (d, 1H), 3.2 (m,
12H), 3.1 (m, 4H), 2.4 (m, 1H), 2.2-0.7 (m, 42H).
Example 42
1-(3,2-dihydroxylup-20(29)-ene-30-yl(hydroxymethyl pyridinium
bromide
[0265] ##STR54##
[0266] A mixture of 30-bromobetulin (0.23, 0.441 mmol) and
4-pyridinemethanol (200 mg, 1.83 mmol) in MeOH (10 ml) was refluxed
for 17 hours under nitrogen. The solution was evaporated to a small
volume and poured into ether (40 ml) to yield a precipitate.
Example 43
1-(3,28-dihydroxylup-20(29)-ene-30-yl)-3-hydroxy-1-azabicyclo[2.2.2]octane
bromide
[0267] ##STR55##
[0268] A mixture of 30-Bromobetulin (498 mg 0.9547 mmol) and
3-quinuclidinol (121.4 mg, 0.9547 mmol) in MeOH (10 ml) was
refluxed for 16 hours under nitrogen. The solvent was evaporated,
and the residue was recrystallized from EtOAc/MeOH to give 398.8 mg
(64%) of product. M.p. 308-310.degree. C. 1H NMR (CD3OD, TMS):
0.68-2.40 (45H, m), 3.11 (1H, dd, J1=10.4 Hz, J2=5.3 Hz), 3.23,
3.72 (2H, AB, J=10.8 Hz), 3.33-3.56 (6H, m), 3.80-3.95 (2H, m),
4.20 (1H, m), 5.46 (1H, m), 5.70 (1H, d, J=6 Hz). 13C NMR (CD3OD,
TMS): 9.07, 10.23, 10.60, 10.77, 13.04, 13.54, 16.25, 16.67, 22.11,
22.28, 22.71, 23.95, 24.41, 29.19, 29.57, 29.80, 32.37, 32.43,
34.06, 34.14, 36.21, 37.90, 41.02, 45.65, 49.40, 49.72, 50.12,
50.30, 50.86, 52.57, 52.90, 54.36, 58.83, 59.57, 67.04, 73.69,
119.56, 119.76, 141.16, 141.39, IR (KBr): 3219.96, 2941.50,
2866.29, 1459.85, 1377.12, 1037.13, 920.27. Calcd: C, 68.50; H,
9.63. Found: C, 65.04; H, 9.23.
Example 44
Lup-20(29)-ene-3,28-bis[N-(2,4-dimethylpyridinium)acetate]dichloride
[0269] ##STR56##
[0270] 3 g betulin-3,28-bis-(chloroacetate) (0.005 mol) was
dissolved in 5 ml of 1-methyl-2-pyrrolidinone, and 2.15 g of
2,4-lutidine (0.02 mol) was added. The reaction mixture was heated
up to 65.degree. C. overnight. Reaction mixture was added in ethyl
ether dropwise with stirring. The precipitate was filtrated and
dissolved in a small volume of CHCl.sub.3. The CHCl.sub.3 solution
was added to ethyl ether and the filtered precipitate was washed
twice with ethyl ether. 3.151 g Yield.
Example 45
Lup-20(29)-ene-3,28-bis[N-(3,5-dimethylpyridinium)acetate]dichloride
[0271] ##STR57##
[0272] 3 g of betulin-3,28-bis(chloroacetate) (0.005 mol) was
dissolved in 5 ml of 1-methyl-2-pyrrolidinone and 2.15 g of
3,5-lutidine (0.02 mol) was added. The reaction mixture was heated
up to 65.degree. C. overnight. The reaction mixture was added in
ethyl ether dropwise with stirring. The precipitate was filtered
and dissolved in a small volume of CHCl.sub.3. The CHCl.sub.3
solution was added to ethyl ether and the filtered precipitate was
washed twice with ethyl ether. 3.44 g Yield.
Example 46
Lup-20(29)-ene-3,28-bis[N-(4-dimethylaminopyridinium)acetate]dichloride
[0273] ##STR58##
[0274] 1 g of betulin-3,28-bis-(chloroacetate) (0.0017 mol) was
dissolved in 5 ml of DMFA, and 0.75 g of 4-(dimethylamino)pyridine
(0.06 mmol) was added. The reaction mixture was heated up to
65.degree. C. overnight. The reaction mixture was added to ethyl
ether dropwise with stirring. The precipitate was filtered and
washed twice with ethyl ether.
Example 47
Lup-20(29)-ene-3-[N-(2-methylpyridinium)acetate]chloride
[0275] ##STR59##
[0276] 1 g of Betulin 3-(chloroacetate) (0.0019 mol) was dissolved
in 5 ml of DMFA and 0.36 g of 2-methylpyridine (0.004 mol) was
added. The reaction mixture was heated up to 65.degree. C.
overnight. The reaction mixture was added in ethyl ether dropwise
with stirring. The precipitate was filtered and washed twice with
ethyl ether. 0.8 g Yield.
Example 48
Lup-20(29)-ene-3-[N-(2,4-dimethylpyridinium)acetate]chloride
[0277] ##STR60##
[0278] 1 g of betulin 3 (chloroacetate) (0.0019 mol) was dissolved
in 5 ml of DMFA, and 0.41 g of 2,4-lutidine (0.004 mol) was added.
The reaction mixture was heated up to 65.degree. C. overnight. The
reaction mixture was added to ethyl ether dropwise with stirring.
The precipitate was filtered and washed twice with ethyl ether.
1.02 g Yield.
Example 49
Lup-20(29)-ene-3-[N-(4-hydroxy-N-methylpiperidino)acetate]chloride
[0279] ##STR61##
[0280] 2 g of betulin-3-(chloroacetate) (0.0039 mol) was dissolved
in 5 ml of dimethylacetamide and placed into a 50-ml flask. A
solution of 1.78 g of 4-hydroxy-1-methylpiperidine (0.0156 mol) in
3 ml of dimethylacetamide was added into the same flash. The
reaction mixture was heated up to 60.degree. C. for 2 hours. The
dimethylacetamide with the precipitate of the product of reaction
was cooled down to room temperature and diluted with ethyl ether.
Solid part was filtrated and washed twice with Ethyl Ether. Then
the precipitate was dissolved in a minimal volume of isopropanol
and reprecipitated with ethyl ether. Traces of solvents were
evaporated with an oil pump.
Example 50
Lup-20(29)-ene-3-[N--(N-methylmorpholino)acetate)chloride
[0281] ##STR62##
[0282] 2 g of betulin-3-(chloroacetate) (0.0039 mol) was dissolved
in 5 ml of dimethylacetamide and placed into a 50-ml flask. A
solution of 0.8 g of 4-methylmorpholine (0.0078 mol) in 3 ml of
dimethylacetamide was added into the same flask. The reaction
mixture was heated up to 60.degree. C. for 2 hours. The
dimethylacetamide with the precipitate of the product of reaction
was cooled down to room temperature and diluted with ethyl ether.
The solid part was filtered and washed twice with ethyl ether. Then
the precipitate was dissolved in a minimal volume of isopropanol
and reprecipitated with ethyl ether. Traces of solvents were
evaporated with an oil pump.
Example 51
Lup-20(29)-ene-3-[N-(3,5-dimethylpyridinium)acetate]chloride
[0283] ##STR63##
[0284] 2 g of betulin-3-(chloroacetate) (0.0039 mol) was dissolved
in 5 ml of dimethylacetamide and placed into a 50-ml flask. A
solution of 0.85 g of 3,5-Lutidine (0.0078 mol) in 3 ml of
dimethylacetamide was added into the same flask. Reaction mixture
was heated up to 60.degree. C. for 2 hours. The dimethylacetamide
with the precipitate of the product of reaction was cooled down to
room temperature and diluted with ethyl ether. The solid part was
filtered and washed twice with ethyl ether. Then the precipitate
was dissolved in a minimal volume of isopropanol and reprecipitated
with ethyl ether. Traces of solvents were evaporated with an oil
pump.
Example 52
Lup-20(29)-ene-3-[N-(4-dimethylaminopyridinium)acetate]chloride
[0285] ##STR64##
[0286] 2 g of betulin-3-(chloroacetate) (0.0039 mol) was dissolved
in 5 ml of dimethylacetamide and placed into a 50-ml flask. A
solution of 0.95 g of 4-(dimethylamino) pyridine (0.0078 mol) in 3
ml of dimethylacetamide was added into the same flask. The reaction
mixture was heated up to 60.degree. C. for 2 hours.
Dimethylacetamide with the precipitate of the product of reaction
was cooled down to room temperature and diluted with ethyl ether.
The solid part was filtered and washed twice with ethyl ether. Then
the precipitate was dissolved in a minimal volume of isopropanol
and reprecipitated with ethyl ether. Traces of solvents were
evaporated with an oil pump.
Example 53
Lup-20(29)-ene-3,28-bis(octyldimethylammoniumacetate)dichloride
[0287] ##STR65##
[0288] 5 g of betulin-3,28-bis(chloroacetate) (0.0084 mol) was
dissolved in 15 ml of DMFA, and 2.9 g of ADMA 8 Amine
(Octyldimethylamine) (0.0185 mol) was added. The reaction mixture
was heated up to 65.degree. C. for 7 hours. The solid part of
reaction mixture was filtered and washed with ethyl ether once.
Then the precipitate was dissolved in a minimal volume of
isopropanol (15 ml) and diluted with ethyl ether. The precipitate
was filtered and dried with an oil pump.
Example 54
Lup-20(29)-ene-3,28-bis(octyldimethylammoniumacetate)dichloride
[0289] ##STR66##
[0290] 5 g of betulin 3(chloroacetate) (0.0096 mol) was dissolved
in 15 ml of DMFA, and 1.67 g of ADMA 8 Amine (Octyldimethylamine)
(0.0106 mol) was added. The reaction mixture was heated up to
65.degree. C. for 7 hours. The solid part of the reaction mixture
was filtered and washed with ethyl ether once. Then the precipitate
was dissolved in a minimal volume of isopropanol (15 ml) and
diluted with ethyl ether. The precipitate was filtered and dried
with an oil pump.
Example 55
Lup-20(29)-ene-3,28-bis(tetradecyldimethylammoniumacetate)dichloride
[0291] ##STR67##
[0292] 5 g of betulin-3,28-bis(chloroacetate) (0.0084 mol) was
dissolved in 15 ml of DMFA, and 4.46 g of ADMA 14 Amine
(tetradecyldimethylamine) (0.0185 mol) was added. The reaction
mixture was heated up to 65.degree. C. for 7 hours. The solid part
of the reaction mixture was filtered and washed with ethyl ether
once. Then the precipitate was dissolved in a minimal volume of
isopropanol (15 ml) and diluted with ethyl ether. The precipitate
was filtered and dried with an oil pump.
Example 56
Lup-20(29)-ene-3-tetradecyldimethylammoniumacetate chloride
[0293] ##STR68##
[0294] 5 g of betulin-3(chloroacetate) (0.0096 mol) was dissolved
in 15 ml of DMFA, and 2.55 g of ADMA 14 Amine
(tetracyldimethylamine) (0.0106 mol) was added. The reaction
mixture was heated up to 65.degree. C. for 7 hours. The solid part
of the reaction mixture was filtered and washed with ethyl ether
once. Then the precipitate was dissolved in a minimal volume of
isopropanol (15 ml) and diluted with ethyl ether. The precipitate
was filtered and dried with an oil pump.
Example 57
1-[(19.beta.,28-epoxy-18.alpha.-oleanan-3.beta.-yl)oxycarbonylmethyl]-4-az-
a-1-azonia-bicyclo[2.2.2]octane chloride
[0295] ##STR69##
[0296] To a solution of 2.0 g (3.8 mmol) of
3.beta.-chloroacetoxy-19.beta.,28-epoxy-18.alpha.-oleanan in 20 mL
of toluene, 0.86 g (7.6 mmol) of DABCO in 20 mL of toluene was
added and the mixture was kept at 80.degree. C. for 6 h. The
precipitate was filtered, washed with toluene, and dried to yield
2.24 g (92%) of the product .sup.1H NMR (CDCl.sub.3, TMS):
0.76-1.95 (45H, m), 3.11 (6H, m), 3.48, 3.81 (2H, AB, J=7.8 Hz),
3.54 (6H, m), 3.56 (1H, br s), 4.58 (1H, m), 5.30 (2H, s).
Example 58
[(19.beta.,28-epoxy-18.alpha.-oleanan-3.beta.-yl)oxycarbonylmethyl]trimeth-
ylammonium chloride
[0297] ##STR70##
[0298] To the solution of 2.0 g (3.8 mmol) of
3.beta.-chloroacetoxy-19.beta.,28-epoxy-18.alpha.-oleanan in 10 mL
of dry DMFA, trimethylamine was bubbled at room temperature
overnight. The mixture was diluted with benzene; the precipitate
was filtered, washed with benzene, and dried to yield 2.1 g (94%)
of the product .sup.1H NMR (CDCl.sub.3, TMS): 0.70-1.90 (45H, m),
3.40 (9H, s), 3.47, 3.81 (2H, AB, J=7.8 Hz), 3.57 (1H, br s), 4.57
(1H, m), 5.10 (2H, s).
Example 59
1-[(19.beta.,
28-epoxy-18.alpha.-oleanan-3.beta.-yl)oxycarbonylmethyl]pyridinium
chloride
[0299] ##STR71##
[0300] A solution of 2.0 g (3.8 mmol) of
3.beta.-chloroacetoxy-19.beta.,28-epoxy-18.alpha.-oleanan in 10 mL
of dry pyridine was kept at 80.degree. C. for 6 h. Benzene (30 ml)
was added; the solids were filtered off, washed with benzene, and
dried to give 2.24 g (97%) of the product. .sup.1H NMR (CDCl.sub.3,
TMS): 0.65-1.87 (45H, m), 3.46, 3.80 (2H, AB, J=7.8 Hz), 3.54 (1H,
br s), 4.55 (1H, m), 5.95 (2H, s), 8.27 (2H, m), 8.76 (1H, m), 9.20
(2H, m).
Example 60
1-[(lup-20(29)-en-3.beta.-yl)oxycarbonylmethyl]-4-aza-1-azonia-bicyclo[2.2-
.2]octane chloride
[0301] ##STR72##
[0302] To a solution of 2.0 g (4.0 mmol) of
3.beta.-chloroacetoxylup-20(29)-ene in 20 mL of toluene 0.89 g (7.9
mmol) of DABCO in 20 mL of toluene was added, and the mixture was
kept at 80.degree. C. for 6 b. The precipitate was filtered, washed
with toluene, and dried to yield 2.24 g (92%) of the product
.sup.1H NMR (CDCl.sub.3, TMS): 0.75-1.80 (46H, m), 3.10 (6H, m),
3.55 (6H, m), 4.51 (1H, m), 4.57 (1H, s), 4.70 (1H, s), 5.25 (2H,
s).
Example 61
1-[(lup-20(29)-en-3.beta.-yl)oxycarbonylmethyl]trimethylammonium
chloride
[0303] ##STR73##
[0304] To the solution of 2.0 g (4.0 mmol) of
3.beta.-chloroacetoxylup-20(29)-ene in 10 mL of dry DMFA
trimethylamine was bubbled at room temperature overnight. The
mixture was diluted with benzene; the precipitate filtered, washed
with benzene and dried to yield 2.03 g (91%) of the product. 1H NMR
(CDCl.sub.3, TMS): 0.77-1.89 (46H, m), 3.43 (9H, s), 4.47 (1H, m),
4.59 (1H, s), 4.72 (1H, s), 5.15 (2H, s).
Example 62
1-[(lup-20(29)-en-3.beta.-yl)oxycarbonylmethyl]pyridinium
chloride
[0305] ##STR74##
[0306] A solution of 2.0 g (4.0 mmol) of
3.beta.-chloroacetoxylup-20(29)-ene in 10 mL of dry pyridine was
kept at 80.degree. C. for 6 h. Benzene (30 ml) was added; the
solids were filtered off, washed with benzene, and dried to give
2.29 g (99%) of the product. .sup.1H NMR (CDCl.sub.3, TMS):
0.80-1.90 (461, m), 4.50 (1H, m), 4.55 (1H, s), 4.68 (1H, s), 5.90
(2H, s), 8.25 (2H, m), 8.78 (1H, m), 9.21 (2H, m).
Example 63
Betulin-3,28-bis(N,N-dimethylethanolacetoxyammonium)dibromide
[0307] ##STR75##
[0308] Betulin-3,28-dichloroacetate (3 g., 4.4 mmol) and
N,N-dimethylethanolamine (2.34 g., 0.0264 mol) were dissolved in 10
ml. of dimethylacetamide. The reaction mixture was kept at room
temperature for one hour. The solid part was filtered, washed twice
with ethyl ether, dissolved in methanol (3 ml.), and diluted with
ethyl ether again. 3.60 g. of final product was obtained after
filtration and drying in vacuum.
[0309] 1H NMR (300 MHz, CDCl.sub.3+(CD.sub.3).sub.2SO): 4.71 s,
1H), 4.61 (m, 6H), 4.39 (d, J=10.4, 1H), 3.95 (m, 5H), 3.78 (bs,
4H), 3.41 (m, 12H), 2.46 (m, 1H), 1.96-0.65 (m, 42H).
Example 64
Betulin-3,28-bis(N,N-dimethylethanolbutiroxyammonium)dibromide
(64)
[0310] ##STR76##
[0311] Betulin-3,28-di(4-bromoacetate) (2.85 g, 3.1 mmol) and
N,N-dimethylethanolamine (2.0 g, 18.0 mmol) were dissolved in 10 ml
of dimethylacetamide. The reaction mature was kept at room
temperature overnight (16 hours). The solid part was filtered,
washed twice with ethyl ether, dissolved in methanol (3 ml), and
diluted with ethyl ether. 3.24 g of final product was obtained
after filtration and drying in vacuum.
[0312] 1H NMR (300 MHz, CDCl.sub.3+(CD.sub.3).sub.2SO): 4.72 (s,
1H), 4.57 (s, 1H), 4.48 (dd, J=6.8, J.sub.2=6.0 Hz, 1H), 4.29 (d,
J=10.4, 1H), 4.10 (bs, 4H), 3.86 (d, J=10.5 Hz, 1H), 3.69 (m, 8H),
3.36 (m, 12H), 2.35 (m, 5H), 2.10 (m, 4H), 1.96-0.65 (m, 42H).
Example 65
Betulin-3,28-bis(didecylmethylammoniumacetoxy)dichloride
[0313] ##STR77##
[0314] Betulin-3,28-di(chloroacetate) (5 g, 8.4 mmol) and
didecylmethylamine (5.75 g, 18.5 mmol) were dissolved in 25 ml of
tetrahydrofurane. The reaction mixture was kept at temperature
70.degree. C. for 6 hours. THF was evaporated on rotary evaporator.
The sold part was washed with hexane. 8.1 g of final product was
obtained after filtration and drying in vacuum
Example 66
Betulin-3,28-bis(didecylmethylammoniumacetoxy) dibromide (66)
[0315] ##STR78##
[0316] Betulin-3,28-di(bromoacetate) (5 g., 7.3 mmol) and
didecylmethylamine (5.03 g., 16 mmol) were dissolved in 25 ml of
tetrahydrofurane. The reaction mixture was kept at 70.degree. C.
for 6 hours. THF was evaporated on rotary evaporator. The solid
part was washed with hexane. 9.2 g of final product was obtained
after filtration and drying in vacuum
Example 67
Betulin-3-(didecylmethylammoniumacetoxy)chloride
[0317] ##STR79##
[0318] Betulin-3-bromoacetate (3 g, 9.6 mmol) and
didecylmethylamine (2.0 g, 10.6 mmol) were dissolved in 25 ml of
tetrahydrofurane. The reaction mixture was kept at 70.degree. C.
for 6 hours THF was evaporated on rotary evaporator. The solid part
was washed with hexane. 9.2 g of final product was obtained after
filtration and drying in vacuum.
Example 68
3.beta.-(N-diazabicyclo[2.2.2]octylacetyloxy)-19.beta.,28-epoxy-18.alpha.--
oleanan bromide
[0319] ##STR80##
[0320] A solution of 0.82 g (0.0074 M) of diazabicyclo[2.2.2]octane
(DABCO) in 15 ml of DMFA was added to a suspension of
3-bromoacetylallobetulin (2.064 g, 0.0037 M) in 20 ml of DMFA. The
mixture was stirred at room temperature overnight diethyl ether and
benzene were added to the mixture. The white precipitate was
filtered off, washed with diethyl ether and hexane, and dried
furnishing the title compound (2.3 g, 0.0034 M, 92%) as a white
solid.
[0321] M.p.=339-340.degree. C. (Decomp.)
[0322] .sup.1H NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 300 MHz): 4.72
(dd, 1H, 3H), 4.55-4.14 (complex, 7.times.2H, CH2), 3.98 (d,
J=10.3, 1H, 28H), 3-9 (s, 1H, 19H), 3.66 (d, J=10.3, 1H, 28H),
1.84-1.43 (complex, CH, CH2), 1.0-0.86 (7.times.3H, CH3).
[0323] .sup.13C NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 75 MHz): 163.11,
89.70, 87.11, 71.12, 61.94, 55.51, 51.80, 50.93, 46.62, 44.37,
41.70, 40.74, 40.60, 38.35, 37.90, 37.07, 36.20, 35.96, 34.08,
33.70, 32.44, 28.30, 27.58, 26.18, 25.75, 24.22, 23.31, 20.88,
18.01, 16.13, 15.76, 15.29, 13.37
[0324] IR (KBr); 3433.6, 3239.3, 2938.3, 2860.6, 1735.9, 1221.2
Example 69
3.beta.-(N-pyridiniumacetyloxy)-19.beta.,28-epoxy-18.alpha.-oleanan
bromide
[0325] ##STR81##
[0326] A suspension of 3-bromoacetylallobetulin (2.064 g, 0.0037 M)
in 10 ml of pyridine was stirred at room temperature overnight.
Diethyl ether was added to the suspension. The white precipitate
was filtered of X; washed with diethyl ether and hexane, and dried,
furnishing the title compound (2.3 g, 97%) as a yellowish
solid.
[0327] M.p. 327-329.degree. C. (Decomp.)
[0328] .sup.1H NMR(CDCl.sub.3+CF.sub.3CO.sub.2D, 300 MHz): 8.93
(d), 8.61 (t) and 8.17 (t) (all pyridine H, 5H), 5.77 and 5.67 (dd,
2H, CH2), 4.68 (t, 1H, 3H), 3.99 (d, J=10.3, 1H, 28H), 3.89 (s, 1H,
19H), 3.65 (d, J=10.3, 28H), 1.82-1.19 (complex, CH CH2),
0.995-0.82 (7.times.3H, CH3).
[0329] .sup.13C NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 75 MHz): 165.99,
146.83, 145.99, 128.52, 89.87, 87.43, 71.19, 61.79, 55.57, 50.99,
46.67, 41.76, 40.79, 40.65, 38.41, 38.03, 37.12, 36.22, 35.99,
34.15, 33.73, 32.47, 28.27, 27.77, 26.21, 25.75, 24.23, 23.44,
20.93, 18.02, 16.16, 15.99, 15.30, 13.39
[0330] IR (KBr): 3417.1, 2922.6, 2867.6, 1742.4, 1637.1, 1490.6,
1444.8, 1376.1, 1234.2
Example 70
3.beta.-[-(N',N'-dimethylaminopyridinium)-N-acetyloxy]-19.beta.,28-epoxy-1-
8.alpha.-oleanan bromide
[0331] ##STR82##
[0332] A mixture of 3-bromoacetylallobetulin (2.064 g, 0.0037 M)
and N,N-dimethylaminopyridine (0.9, 0.0074 in) in 50 ml
dimethylacetamide was stirred at room temperature over 3 h, and
treated with diethyl ether. The precipitate was filtered off,
washed with ether and dried, furnishing the title compound (23 g,
91) as a yellowish solid.
[0333] M.p. 323-326.degree. C. (Decomp. subl.)
[0334] .sup.1H NMR (CDCl.sub.3, 300 MHz): 8.54 (t) and 6.99 (d)
(all pyridine H, 4H), 5.5 (d, 2H, CH2), 4.57 (t, 1H, 3H), 3.77 (d,
J=10.3, 1H, 28H), 3.52 (s, 1H, 19H), 3.44 (d, J=10.3, 28H), 3.3 (s,
6H, 3.times.CH3), 1.76-1.12 (complex, CH, CH2), 0.97-0.79
(7.times.3H, CH3).
[0335] .sup.13C NMR (CDCl.sub.3, 75 MHz): 166.13, 155.93, 143.30,
107.14, 87.36, 83.76, 70.70, 57.32, 54.97, 50.40, 46.25, 40.91,
40.17, 40.11, 40.06, 37.99, 37.41, 36.57, 36.18, 35.72, 33.57,
33.42, 32.14, 28.27, 27.70, 25.86, 25.70, 24.01, 23.15, 20.49,
17.53, 16.0, 15.15, 12.96
[0336] IR (KBr): 3427.4, 2934.3, 2859.9, 1736.8, 1648.4, 1448.3,
1392.5, 1211.1
Example 71
3.beta.-[-(N',N'-dimethylaminopyridinium)-N-acetyloxy]-19.beta.,28-epoxy-1-
8.alpha.-oleanan chloride
[0337] ##STR83##
[0338] A mixture of 3-chloroacetylallobetulin (1.91 g, 0.0037 M)
and N,N-dimethylaminopyridine (0.9 g, 0.0074 M) in 50 ml
dimethylacetamide was stirred at room temperature overnight, and
treated with diethyl ether and methanol. The precipitate was
filtered off, washed with ether and dried furnishing the title
compound (2.03 g, 86%) as a white solid.
[0339] M.p. 325-327.degree. C. (decomp, subl.)
[0340] 1H NMR (CDCl.sub.3, 300 MHz): 8.57 (t) and 6.94 (d) (all
pyridine H, 4H), 5.51 (a, 2H, CH2), 4.52 (m, 1H, 3H), 3.73 (d,
J=10.3, 1H, 28H), 3.47 (s, 1H, 19H), 3.40 (d, J=10.3, 28H), 3.25
(s, 6H, 3.times.CH3), 1.71-1.07 (complex, CH, CH2), 0.3-0.73
(7.times.3H, CH3).
[0341] .sup.13C NMR (CDCl.sub.3, 75 MHz): 166.62, 156.25, 149.60,
143.74, 107.44, 87.69, 83.95, 71.02, 57.48, 55.29, 50.71, 46.22,
42.46, 41.23, 40.49, 40.38, 38.30, 37.71, 36.88, 36.76, 36.50,
36.04, 33.89, 33.56, 32.47, 28.60, 28.00, 26.18, 24.33, 23.44,
20.80, 17.86, 16.33, 15.47, 13.29
[0342] IR (KBr): 3390.2, 2934.3, 2859.9, 1736.8, 1648.4, 1564.6,
1448.3, 1387.9, 1215.7
Example 72
3.beta.-(N-octyldimethylaminoacetyloxy)-19.beta.,28-epoxy-18.alpha.-oleana-
n bromide
[0343] ##STR84##
[0344] A mixture of 3-bromoacetylallobetulin (2.064 g, 0.0037 M)
and octyldimethylamine (1.16 g, 0.0074 M) in 30 ml
dimethylacetamide was stirred at room temperature overnight and
treated with diethyl ether/benzene mixture. The precipitate was
filtered off, washed with ether and dried, furnishing the title
compound (1.7 g, 64%) as a white solid.
[0345] M.p. 275-277.degree. C. (decomp.)
[0346] .sup.1H NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 300 MHz): 4.67
(dd, 1H, 3H), 4.36*m, 2H, CH2), 3.97 (d, J=10.3, 1, 28H), 3.87 (s,
1H. 19H), 3.64 (m, 3H, 28H and N--CH2), 3.43 (s, 6H, 2.times.3H,
CH3), 1.83-1.15 (complex, CH, CH2), 1.00-0.87 (7.times.3H,
CH3).
[0347] .sup.13C NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 75 MHz): 163.80,
89.0, 85.25, 70.91, 6530, 61.43, 55.48, 52.56, 52.44, 50.91, 46.57,
41.53, 40.66, 40.55, 38.43, 37.87, 37.07, 36.25, 35.96, 33.94,
33.69, 32.46, 31.49, 28.84, 28.46, 27.86, 26.18, 25.91, 25.8,
24.37, 23.48, 22.81, 22.47, 20.85, 18.0, 16.32, 16.16, 15.39, 1392,
13.45
[0348] IR (KBr): 3436.7, 2925.0, 2850.6, 1727.5, 1462.3, 1262.3,
1201.8
Example 73
3.beta.-[N-(2-hydroxyethyl)aminoacetyloxy]-19.beta.,28-epoxy-18.alpha.-ole-
anan bromide
[0349] ##STR85##
[0350] A mixture of 3-bromoacetylallobetulin (2.064 g, 0.0037 M)
and ethanolamine (0.45 g, 0.0074 M) in 30 ml of DMFA was sired at
room temperature for 24 hours and treated with methanol. The
precipitate was filtered off, washed with methanol and dried,
furnishing the title compound (1.1 g, 48%) as a white solid.
[0351] M.p. 319-322.degree. C. (decomp.)
[0352] .sup.1H NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 300 Hz): 4.71 (m,
1H, 3H), 4.13-3.39 (complex, 9H, CH2, 19H and 28H), 1.83-1.19
(complex, CH, CH2), 0.99-0.86 (7.times.3H, CH3).
[0353] .sup.13C NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 300 Hz): 165.92,
89.16, 86.06, 70.94, 57.05, 55.43, 50.88, 49.85, 48.12, 46.57,
41.56, 40.67, 40.55, 3835, 38.11, 37.87, 37.05, 36.25, 35.96,
33.98, 33.67, 32.47, 28.43, 27.58, 26.18, 25.81, 24.35, 23.34,
20.85, 17.99, 16.32, 16.16, 15.38, 13.41
[0354] IR (KBr): 3213.4, 2934.3, 2859.9, 1732.1, 1453.0, 1387.9,
1220.4, 1197.1, 1146.0
Example 74
3.beta.-[N,N-dimethyl-N-(2-hydroxyethyl)aminoacetyloxy]-19.beta.,28-epoxy--
18.alpha.-oleanan bromide
[0355] ##STR86##
[0356] A mixture of 3-bromoacetylallobetulin (2.064 g, 0.0037 M)
and dimethylaminoethanol (0.67 g, 0.0074 M) in 50 ml of DMA was
stirred at room temperature overnight and treated with diethyl
ether/hexane. The precipitate was filtered off, washed with
methanol/hexane and dried, furnishing the title compound (2.27 g,
94%) as a white solid.
[0357] M.p. 316-318.degree. C. (decomp.)
[0358] .sup.1H NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 300 Hz): 4.70 (m,
1H, 3H), 4.48-3.64 (complex, 9H, CH2, 19H and 28H), 3.49 (s, 6H,
2.times.3H, CH3), 1.83-1.20 (complex, CH, CH2), 1.00-0.86
(7.times.3, CH3).
[0359] .sup.13C NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 300 Hz): 163.91,
89.32, 86.18, 85.70, 71.00, 66.36, 63.15, 62.87, 6236, 60.88,
55.99, 55.50, 54.20, 53.49, 50.92, 46.59, 41.615, 40.71, 40.58,
38.39, 37.87, 37.08, 36.23, 35.97, 34.03, 33.70, 32.46, 28.39,
27.76, 26.18, 25.79, 24.32, 23.45, 20.87, 18.01, 16.24, 16.04,
15.35, 13.43
[0360] IR (KBr): 3283.6, 2934.3, 1732.8, 1643.2, 1199.9
Example 75
3.beta.-[N,N-dimethyl-N-(2-hydroxyethyl)aminoacetyloxy]-19.beta.,28-epoxy--
18.alpha.-oleanan chloride
[0361] ##STR87##
[0362] A mixture of 3-chloroacetylallobetulin (1.91 g, 0.0037 M)
and dimethylaminoethanol (0.67 g, 0.0074 M in 50 ml of DMA was
stirred at room temperature for 24 hours and treated with diethyl
ether/methanol. The precipitate was filtered off, washed with
hexane and dried furnishing the title compound (1.7 g, 76%) as a
white solid.
[0363] M.p. 271-273.degree. C. (decomp.)
[0364] .sup.1NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 300 Hz): 4.71 (m,
1H, 3H), 4.48-3.24 (complex, 9H, CH2, 19H and 28H), 3.47 (s, 6,
2.times.3H, CH3), 1.82-1.19 (complex, CH, CH2), 1.00-0.86
(7.times.3H, CH3).
[0365] .sup.13C NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 300 Hz): 163.86,
89.28, 85.67, 70.98, 66.34, 62.27, 56.06, 55.50, 53.39, 50.90,
46.59, 41.60, 40.69, 40.56, 38.38, 37.84, 37.06, 36.23, 35.96,
34.02, 33.69, 32.47, 28.39, 27.71, 26.18, 25.80, 24.32, 23.42,
20.86, 18.0, 16.24, 16.0, 15.34, 13.43
[0366] IR (KBr): 3390.2, 3287.8, 2934.3, 2869.2, 2357.4, 1736.8,
1634.4, 1457.6
Example 76
3.beta.-[N-(3-hydroxymethylpyridinium)acetyloxy]-19.beta.,28-epoxy-18.alph-
a.-oleanan bromide
[0367] ##STR88##
[0368] A mixture of 3-bromoacetylallobetulin (2.064 g, 0.0037 M)
and 3-pyridylcarbinol (0.82 g, 0.0075 M) in 50 ml of DMA was
stirred at room tenure overnight and treated with diethyl
ether/hexane. The precipitate was filtered off, washed with hexane
and dried, furnishing the title compound (2.26 g, 94%) as a white
solid.
[0369] M.p. 325-327.degree. C. (decomp.)
[0370] .sup.1H NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 300 Hz): 9.0 (s),
8.65 and 8.58 (dd), 8.09 (m) (4H, pyridine), 5.6, (m, 2H, CH2),
5.12 (s, 2H, CH2), 4.690 (m, in, 3H), 4.02 (d, J=10.3, 1H, 28H),
3.89 (s, 1H, 19H), 3.66 (d. J=10.3, 1H, 28H), 1.83-1.20 (complex,
at CH2), 0.99-0.84 (7.times.3H, CH3).
[0371] .sup.13C NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 300 Hz): 165.30,
145.99, 144.84, 143.99, 142.57, 135.55, 128.44, 127.67, 89.27,
86.90, 70.97, 64.24, 61.80, 61.49, 60.10, 55.46, 50.87, 46.57,
41.58, 40.69, 40.55, 38.35, 37.92, 37.03, 36.22, 35.96, 34.00,
33.67, 32.44, 28.38, 27.77, 26.16, 25.78, 24.31, 23.36, 20.85,
17.96, 16.22, 16.05, 15.33, 13.40
[0372] IR (Br): 3265.7, 2943.3, 2862.7, 1737.3, 1638.7, 1468.6,
1379.0, 1280.5, 1240.2
Example 77
3.beta.-[(N,N,N',N'-tetramethylethylenediamino)acetyloxy]-19.beta.,28-epox-
y-18.alpha.-oleanan bromide
[0373] ##STR89##
[0374] A mixture of 3-bromoacetylallobetulin (2.064 g, 0.0037 M)
and tetramethylethylenediamine (TMEDA) (0.86 g, 0.0075 M) in 30 ml
of DMFA was sired at room temperature overnight and treated with
diethyl ether/benzene. The precipitate was filtered off, washed
with ether/benzene and dried, furnishing the title compound (2.35
g, 93%) as a white solid.
[0375] M.p. 324-326.degree. C. (decomp.)
[0376] .sup.1H NMR (CDCl.sub.3+CF.sub.3CO.sub.2D, 300 Hz): 4.7 (m,
1H, 3H), 4.3-3.6 (complex, 9H, CH, CH2), 3A6 (s, 2.times.3H, CH3),
3.1 (s (6H, CH3), 1.89-1.20 (complex, CH, CH2), 1.00-0.86
(7.times.3H, CH3).
[0377] .sup.13C NMR (CDCl.sub.3+CF.sub.3CO.sub.2), 300 Hz) 163.31,
89.25, 86.42, 70.98, 62.77, 58.72, 55.47, 52.37, 52.25, 50.91,
50.0, 46.59, 44.15, 41.59, 40.69, 40.57, 38.35, 37.82, 37.05,
36.24, 35.97, 34.02, 33.68, 32.46, 28.39, 27.62, 26.18, 25.80,
24.31, 23.30, 20.86, 18.01, 16.22, 15.84, 15.34, 13.42
[0378] (KBr): 3426.2, 2931.7, 2867.6, 1733.3, 1467.7, 1449.4,
1266.2, 1220.4
Example 78
3.beta.-[(N,N-dimethyl-N-octyl)aminoacetyloxy]-19.beta.,28-epoxy-18.alpha.-
-oleanan chloride
[0379] ##STR90##
[0380] A mixture of 3-chloroacetylallobetulin (30 g, 0.058 mol) and
octyldimethylamine (18.2 g, 0.116 mol) in 300 ml of
dimethylacetamide was stirred at room temperature over 48 h,
treated with diethyl ether afterwards. The precipitate was filtered
off, washed with ether/hexane and dried, furnishing the title
compound (32 g, 81%) as a white solid.
[0381] M.p. 245-247.degree. C. (decomp.)
[0382] .sup.1H NMR (CDCl.sub.3, 300 MHz): 4.89 (s, 2H, CH2), 4.64
(m, 1H, 3H), 3.8-3.48 (1, 15H, CH, CH2, CH3), 1.81-1.29 (m, CH,
CH2), 1.00-0.82 (s, 7.times.3H, CH3).
[0383] .sup.13C NMR(CDCl.sub.3, 75 MHz): 164.59, 87.86, 84.45,
71.20, 64.31, 61.00, 55.49, 51.97, 50.28, 46.75, 41.40, 40.66,
40.55, 38.42, 37.92, 37.06, 36.67, 36.20, 34.05, 33.71, 32.63,
31.53, 29.0, 28.75, 28.07, 26.18, 24.49, 23.64, 22.88, 22.50,
20.97, 18.03, 16.49, 15.63, 13.99, 13.42
[0384] IR (KBr): 3422.3, 2923.5, 2844.8, 1735.5, 1634.9, 1464.2,
1389.9, 1254.2, 1201.5
Example 79
3.beta.-[(N,N-dimethyl-N-tetradecyl)aminoacetyloxy]-19.beta.,28-epoxy-18.a-
lpha.-oleanan chloride
[0385] ##STR91##
[0386] A mixture of 3-chloroacetylallobetulin (2.59 g, 0.005 mol)
and tetradecyldimethylamine (2.35 g, 0.01 mol) in 50 ml
dimethylacetamide was stirred at room temperature over 48 h,
treated with diethyl ether/hexane afterwards. The precipitate was
filtered off, washed with ether/hexane and dried, furnishing the
title compound (3.5 g, 90%) as a white solid.
[0387] M.p. 244-246.degree. C. (decomp.)
[0388] .sup.1H NMR (CDCl.sub.3, 300 MHz): 4.89 (s, 2H, CH2), 4.63
(m, 1H, 3-H), 3.8-3.7 (m, 9H, CH, CH2, CH3), 3.54 (s, 1H, 19-H),
3.45 (d, J=10.3, 28-H), 1.8-1.22 (m, CH, CH2), 0.99-0.81
(7.times.3H, CH3).
[0389] .sup.13C NMR (CDCl.sub.3, 75 MHz): add data
[0390] IR (KBr): 34135, 2923.5, 28.53.5, 1735.5, 1468.6, 1254.2,
1201.7
Example 80
3.beta.-[(N-methyl-N,N-didecyl)aminoacetyloxy]-19.beta.,28-epoxy-18.alpha.-
-oleanan chloride
[0391] ##STR92##
[0392] A mixture of 3-chloroacetylallobetulin (31 g, 0.06 mol.) and
methyldidecylamine (37.1 g, 0.12 mol.) in 300 ml dimethylacetamide
was stirred at 60.degree. C. over 3 days. The reaction mixture was
poured into large volume of diethyl ether. The precipitate was
filtered off, washed with ether and dried, furnishing the title
compound (37.3 g, 75%) as a colorless solid.
[0393] M.p. 203-204.degree. C. (decomp.)
[0394] .sup.1H NMR (CDCl.sub.3, 300 MHz): 4.77 (s, 2H, CH2), 4.62
(m, 1H, 3-H), 3.86-3.45 (m, 10H, CH, CH2, CH3), 1.79-1.22 (m, CH,
CH2), 1.00-0.82 (7.times.3H, CH3).
[0395] .sup.13C NMR (CDCl.sub.3, 75 MHz): 164.59, 87.87, 84.53,
83.29, 71.20, 61.98, 59.63, 55.52, 50.94, 49.41, 46.76, 41.41,
41.19, 40.66, 38.48, 37.92, 37.09, 36.68, 36.20, 34.07, 32.65,
31.77, 29.32, 29.16, 28.74, 26.36, 26.18, 24.48, 23.63, 22.59,
20.97, 18.04, 16.46, 15.64, 14.04, 13.42
[0396] IR (KBr): 3396, 2923.5, 2853.5, 1735.5, 14643, 1376.7,
1228.0
Example 81
3.beta.-[N,N-di(2-hydroxyethyl)aminoacetyloxy]-19.beta.,28-epoxy-18.alpha.-
-oleanan bromide
[0397] ##STR93##
[0398] A mixture of 3-bromoacetyl allobetulin (3.1 g, 0.0055 mol)
and diethanol amine (1.16 g, 0.011 mol) in 50 ml dimethylacetamide
was stirred at room temperature overnight, treated with
ethanol\hexane mixture. The precipitate was filtered off, washed
with hexane and dried, furnishing the title compound (3.96 g, 99%)
as a colorless solid.
[0399] M.p. 240-244.degree. C. (decomp.)
[0400] .sup.1H NMR (CDCl.sub.3, 300 MHz): 4.61 (m, 1H, 3-M),
3.83-3.48 (m, 10H, CH, CH2), 2.82 (m, 4H, CH2), 1.79-123 (m, CH,
CH2), 1.02-0.84 (7.times.3H, CH3).
[0401] .sup.13C NMR (CDCl.sub.3, 75 MHz): 172.98, 87.93, 82.15,
71.25, 60.95, 59.89, 57.67, 56.10, 55.53, 50.94, 50.76, 46.80,
41.55, 40.71, 38.53, 37.85, 37.14, 36.72, 36.25, 34.12, 33.79,
32.69, 28.79, 28.02, 26.40, 24.52, 23.71, 21.00, 18.10, 16.59,
15.68, 13.48
[0402] IR (KBr): 3374.4, 2932.7, 1729.9, 1714.5, 1451.4, 1197.6,
1070.7
Example 82
Betulin-3,28-bis(N,N-dimethylethanolacetoxyammonium)dichloride
(62)
[0403] ##STR94##
[0404] Betulin-3,28-dichloroacetate (3 g, 5 mmol) and
N,N-dimethylethanolamine (2.6 g, 30 mmol) were dissolved in 50 ml
of dimethylacertamide. The reaction mixture was kept at room
temperature overnight (16 hours). The solid part was filtered,
washed twice with ethyl ether, dissolved in methanol (4 ml.) and
diluted with ethyl ether again 3.45 g of final product was obtained
after filtration and drying in vacuum.
Example 83
[0405] Several uncharged or negatively charged triterpenes, and
several positively charged triterpenes of the invention were tested
for anti-bacterial activity against antibiotic-susceptible and
antibiotic resistant strains of four types of bacteria of concern
in clinics and hospitals. The triterpenes were tested in broth
culture in triplicate for ability to inhibit bacterial growth. The
compounds tested are those listed below.
[0406] Compounds 3, 6, 12, 18, 25, and 40 are from the
correspondingly numbered examples.
[0407] Compound 83 is lupeol.
[0408] Compound 84 is lupenone, the compound of formula (II)
wherein R.sub.1 and R.sub.2 together are hydrogen; R.sub.3,
R.sub.4, and R.sub.5 together are hydrogen; and R.sub.6, R.sub.7,
and R.sub.8 together are oxo.
[0409] Compound 85 is lupeneno-1-ene-2-ol, with the structure below
##STR95##
[0410] Compound 86 is lupeol-3-maleate, the compound of formula
(II) wherein R.sub.1 and R.sub.2 together are hydrogen; R.sub.3,
R.sub.4, and R.sub.5 together are hydrogen; and R.sub.6, R.sub.7,
and R.sub.8 together are --OCOCH.dbd.CHCOOH.
[0411] Compound 87 is lupeol-3-phosphate, the compound of formula
(II) wherein wherein R.sub.1 and R.sub.2 together are hydrogen;
R.sub.3, R.sub.4, and R.sub.5 together are hydrogen; and R.sub.6,
R.sub.7, and R.sub.8 together are phosphate.
[0412] Compound 88 is allobetulone-1-ene-2-ol, with the structure
below ##STR96##
[0413] Compound 89 is betulin-3,28,30-triol, the compound of
formula (II) wherein wherein R.sub.1 and R.sub.2 together are
hydroxyl; R.sub.3, R.sub.4, and R.sub.5 together are hydroxyl; and
R.sub.6, R.sub.7, and R.sub.8 together are hydroxyl.
[0414] Compound 90 is betulon-1-ene-2-ol, with the structure below
##STR97##
[0415] Compound 91 is betulin-3,28-disuccinate, the compound of
formula (II) wherein wherein R.sub.1 and R.sub.2 together are
hydrogen; R.sub.3, R.sub.4, and R.sub.5 together are
--OCOCH.sub.2CH.sub.2COOH; and R.sub.6, R.sub.7, and R.sub.8
together are --OCOCH.sub.2CH.sub.2COOH.
[0416] Compound 92 is betulin-28-succinate, the compound of formula
(II) wherein wherein R.sub.1 and R.sub.2 together are hydrogen;
R.sub.3, R.sub.4, and R.sub.5 together are --OCOCH.sub.2
CH.sub.2COOH; and R.sub.6, R.sub.7, and R.sub.8 together are
hydroxyl.
[0417] Compound 93 is betulin-3,28-dimaleate, the compound of
formula (II) wherein wherein R.sub.1 and R.sub.2 together are
hydrogen R.sub.3, R.sub.4 and R.sub.5 together are
--OCOCH.dbd.CHCOOH; and R.sub.6, R.sub.7, and R.sub.8 together are
--OCOCH.dbd.CHCOOH.
[0418] Compound 94 is betulin-28-maleate, the compound of formula
(II) wherein wherein R.sub.1 and R.sub.2 together are hydrogen;
R.sub.3, R.sub.4, and R.sub.5 together are --OCOCH.dbd.CHCOOH; and
R.sub.6, R.sub.7, and R.sub.8 together are hydroxyl.
[0419] Compound 95 is betulin-3,28-bisdiacetyltartrate, the
compound of formula (II) wherein R.sub.1 and R.sub.2 together are
hydrogen; R.sub.3, R.sub.4, and R.sub.5 together are
--COCHOHCHOHCOCH.sub.2COCH.sub.2COOH; and R.sub.6, R.sub.7, and
R.sub.8 together are --COCHOHCHOHCOCH.sub.2COCH.sub.2COOH.
Results
[0420] The lowest concentration which inhibits bacterial growth is
referred to as the minimum inhibitory concentration (MIC), the
standard measurement of anti-microbial activity. Table 1 gives
approximate MIC values for the 19 triterpenes tested. The standard
for antibiotic development in the pharmaceutical industry is that
compounds under study must be active at 10 .mu.g/ml or less to be
considered for further development. By this criterion, all five of
the quaternary amine triterpene derivatives tested were effective
against both S. aureus and E. faecium. In addition, the compounds
were approximately equally effective against both
antibiotic-resistant and antibiotic-sensitive strains of the two
bacteria. In contrast, of the triterpenes not derivatized with a
quaternary amine, only lupeol-3-maleate had an MIC of 10 .mu.g/ml
or less, and that against only one strain of bacteria.
TABLE-US-00001 TABLE 1 Anti-Bacterial Activity of Betulin and
Derivatives (Approximate Minimum Inhibitory Concentration, MIC,
.mu.g/ml) Compound number Test Organism 3 6 12 18 25 40 83 84 85 86
87 88 89 90 91 92 93 94 95 Staphylococcus aureus 5 2 5 5 2 5 -- --
100 -- -- -- -- -- -- -- 200 -- -- Methicillin-resistant
Staphylococcus aureus 5 2 2 5 2 5 -- -- 50 -- -- -- -- -- 200 50
200 50 -- Methicillin-sensitive Enterococcus faecium 5 5 2 5 5 2 --
-- 20 10 -- -- -- -- -- -- -- 200 -- Vancomycin-sensitive
Enterococcus faecium 5 10 2 5 10 5 -- -- 50 50 -- -- -- -- -- -- --
200 -- Vancomycin-sensitive Pseudomonas aeruginosa -- -- -- -- --
200 -- -- -- -- -- -- -- -- -- -- -- -- -- Multi-drug resistant
Pseudomonas aeruginosa -- -- -- -- -- 200 -- -- -- -- -- -- -- --
-- -- -- -- -- Antibiotic sensitive Eschericia coli -- -- 50 -- 100
50 -- -- -- -- -- -- -- -- -- -- -- -- -- Ampicillin-resistant
Escherichia coli -- -- -- -- 50 20 -- -- -- -- -- -- -- -- -- -- --
-- -- Ampicillin sensitive --= no inhibition of bacterial
growth.
Example 84
Methods.
[0421] Plots of creeping bentgrass, cv. "Pennlinks," were treated
with water based sprays of the test compounds. Sprays were
delivered by CO.sub.2-charged sprayer to deliver the equivalent of
2 gallons of spray per 1000 sq. ft of plot area. Plots were 15 sq.
ft. each, and treatments were mixed to apply material to 75 sq. ft
(5 replicates), although only 4 replicates were treated. The
quaternary salts of triterpenes were applied at 17.8 g/1000 sq. ft.
for 14 days. Daconil Ultrex.RTM. was applied at 3.2 oz/1000 sq. ft.
for 14 days. The experiment used a randomized complete block
design, with 4 replicates per treatment. Disease pressure was high.
Pennlinks bluegrass is moderately to highly susceptible to both
dollar spot and brown patch. For brown patch, caused by Rhozoctonia
sofani, a visual estimate of the percent of the plot area affected
was recorded. Turf quality was estimated on a 1 to 9 scale, with 1
being worst and 9 being perfect A rating of 6 is considered
acceptable.
Results.
[0422] Table 2 and Table 3 report the results of two experiments.
The compound numbers in the Tables correspond to the Example
numbers above. In Table 3, compounds 15 and 24 suppressed the
development of brown patch after the first spray was applied. The
other treatment suppressed brown patch also, but to a lesser extent
on Jun. 25, 2001, one week after the first treatment was applied.
At Jul. 2, 2001, compounds 1, 15, and 24 suppressed dollar spot,
but not as well as the chemical standard, Daconil Ultrex.RTM.. By
Jul. 9, 2001, none of the treatments differed significantly from
the control for dollar spot.
[0423] On June 25 and Jul. 2, 2001 in Table 3, all of the test
compounds apparently deceased brown patch, but only compound 15 (on
June 25) and Daconil Ultrex.RTM. achieved statically significant
suppression. By Jul. 9, 2001, compounds 7, 18, and 24 were judged
to significantly suppress brown patch. On July 16, there were no
significant differences regarding brown patch.
[0424] In summary, for the first two weeks, all of the tested
quaternary amine salts of triterpenes suppressed both dollar spot
and brown patch for the first two weeks of treatment in one of the
studies. In the study of Table 2, for die compounds appeared to be
less effective. Many reasons might account for this, in particular
weather and soil conditions. TABLE-US-00002 TABLE 2 Antifungal
Treatment on Turf Grass Date Jun. 25, 2001 Jul. 2, 2001 Jul. 9,
2001 Jul. 16, 2001 Dollar Dollar Brown Dollar Spot Brown Spot Brown
Patch Spot Brown # of Patch # of Patch % # of Patch spots % of
spots % of of spots % of per plot Turf per plot Turf plot Turf per
plot Turf Sample plot infected Quality plot infected Quality
infeted Quality plot infected Quality Control 17.0 23.8 4.63 15.8
30.5 4.75 27.5 5.00 2.5 5.0 5.72 Comp. 1 21.0 18.8 4.50 22.3 35.0
4.50 13.8 6.00 2.8 3.8 5.57 Comp. 7 18.3 24.5 4.50 22.3 20.5 4.25
20.0 4.50 2.3 3.8 5.60 Comp. 14 22.3 25.8 4.38 21.8 36.8 4.25 25.0
4.63 3.5 5.0 5.60 Comp. 17 17.8 20.0 4.63 19.3 33.8 4.25 22.8 5.13
2.0 5.0 5.60 Comp. 24 17.0 15.8 4.75 16.3 30.0 4.50 14.3 5.00 2.8
2.5 5.65 Daconil 11.3 17.5 5.25 8.3 5.8 5.75 13.3 6.00 1.3 3.8
5.85
[0425] TABLE-US-00003 TABLE 3 Antifungal Treatment on Turf Grass
Date Jun. 25, 2001 Jul. 2, 2001 Jul. 9, 2001 Jul. 16, 2001 Dollar
Dollar Dollar Dollar Spot Brown Spot Brown Spot Brown Spot Brown #
of Patch # of Patch # of Patch # of Patch spots % of spots % of
spots % of spots % of per plot Turf per plot Turf per plot Turf per
plot Sample plot infected Quality plot infected Quality plot
infeted Quality plot infected Control 21.3 24.5 4.00 27.0 41.3 4.38
17.8 27.5 4.88 2.0 7.5 Comp. 1 11.5 15.8 4.88 15.8 24.0 4.50 13.0
20.5 5.38 1.5 8.6 Comp. 7 14.8 17.5 4.36 21.5 25.5 4.50 14.5 10.8
5.88 2.0 8.8 Comp. 14 16.3 16.3 4.88 20.8 31.3 4.75 14.5 16.5 5.00
2.8 7.5 Comp. 15 10.5 13.8 4.88 16.0 26.8 4.75 16.3 14.5 5.50 1.5
10.0 Comp. 18 16.8 14.8 4.63 20.8 37.5 4.63 14.3 12.0 5.50 1.8 6.3
Comp. 24 10.8 17.0 4.75 14.0 29.5 4.50 11.3 12.5 5.50 1.5 7.5
Daconil 11.0 7.5 5.63 10.5 8.3 6.13 11.0 7.3 6.20 1.5 8.8
Example 85
Methods
[0426] Plots of creeping bentgrass, cv. Pennlinks, were treated
with water-based sprays as in Example 84 at Clemson University.
Treatment application dates were May 7, May 14, May 28, and June
11. Plots were rated visually, estimating the percent of the area
affected by dollar spot fungus (Agrotis stolonifera). Turf quality
was rated on a 1 to 9 scale, with 9 being perfect turf, as in
Example 84. Disease pressure was moderately high to high when the
trial was initiated, with existing dollar spot present. Plots did
not receive additional innoculum, as it was not needed.
Results
[0427] Table 4 shows the amount of active ingredient applied for
each tested compound. TABLE-US-00004 TABLE 4 Product Rate Treatment
(grams active ingredient Name per 1000 square feet) Comp. 15 50
Comp. 17 50 Comp. 22 50 Comp. 67 50 Comp. 56 50 Comp. 53 50 Comp.
46 50 Comp. 55 50
[0428] Tables 5 and 6 show the turf quality and percent area of the
plots infected with dollar spot after treatment with the test
compounds in duplicate experiments. In both cases all of the tested
compounds somewhat reduced the extent of dollar spot infection and
somewhat improved turf quality by the end of the treatment trial,
compared to the no treatment control. Compound 67 gave the best
results. TABLE-US-00005 TABLE 5 % of area % of area Turf quality
infected Turf quality infected Turf quality May 27, 2002 Jun. 03,
2002 Jun. 03, 2002 Jun. 03, 2002 Jun. 17, 2002 Treatment Name
Control 2.95 b 28.3 a 3.95 c 30.5 a 4.70 ab Comp. 15 3.13 b 24.3 a
4.95 abc 23.0 a 3.60 b Comp. 17 3.10 b 29.0 a 4.47 bc 26.0 a 4.70
ab Comp. 22 3.05 b 24.5 a 4.80 abc 17.3 ab 5.07 ab Comp. 67 4.85 a
13.0 b 6.00 a 9.0 b 5.80 a Comp. 56 4.47 ab 22.0 a 5.30 ab 19.3 ab
5.38 ab Comp. 53 3.60 ab 22.5 a 5.18 ab 19.5 ab 5.38 ab Comp. 46
4.00 ab 23.5 a 5.40 ab 24.0 a 4.52 ab Comp. 55 3.38 ab 28.8 a 5.10
ab 24.3 a 5.2 ab LSD (P = .05) 0.941 5.38 0.773 8.49 1.072 Standard
Deviation 0.658 3.77 0.541 5.94 0.750 CV 17.83 15.44 10.56 25.24
15.49 Bartlett's X2 41.594 3.327 11.42 14.151 7.6 P(Bartlett's X2)
0.001* 0.998 0.653 0.439 0.909 Replicate F 3.632 5.654 4.409 2.304
2.843 Replicate Prob(F) 0.0204 0.0024 0.0088 0.0907 0.0491
Treatment F 3.335 4.371 3.225 3.741 2.065 Treatment Prob(F) 0.0013
0.0001 0.0017 0.0005 0.0354 Means followed by same letter do not
significantly differ (P = .05, Student-Newman-Keuls)
[0429] TABLE-US-00006 TABLE 6 % of area % of area % of area % of
area infected Turf quality infected Turf quality infected Turf
quality infected May 08, 2002 May 08, 2002 May 13, 2002 May 13,
2002 May 20, 2002 May 20, 2002 May 27, 2002 Treatment Name Control
30.0 a 3.13 a 28.0 a 3.15 a 38.3 a 2.75 b 36.3 a Comp. 15 30.3 a
3.00 a 27.5 a 3.13 a 27.3 bc 3.13 ab 29.3 a-d Comp. 17 31.5 a 3.00
a 28.0 a 3.13 a 32.0 bc 3.00 ab 31.5 ab Comp. 22 34.3 a 3.00 a 28.5
a 3.13 a 31.5 bc 2.95 ab 32.0 ab Comp. 67 28.3 a 3.20 a 26.0 a 3.17
a 24.3 c 3.55 a 21.0 d Comp. 56 31.0 a 3.13 a 26.5 a 3.00 a 32.0 bc
3.03 ab 21.8 cd Comp. 53 33.5 a 3.00 a 28.5 a 3.05 a 27.3 bc 3.25
ab 26.8 bcd Comp. 46 30.5 a 3.25 a 28.0 a 3.38 a 25.0 bc 3.50 a
26.8 bcd Comp. 55 33.5 a 3.05 a 30.0 a 3.00 a 32.8 b 2.97 ab 29.3
a-d LSD (P = .05) 4.64 0.253 4.08 0.273 4.67 0.418 5.45 Standard
Deviation 3.25 0.177 2.86 0.191 3.27 0.293 3.82 CV 10.47 5.74 10.23
6.15 11.37 9.08 13.6 Bartlett's X2 8.725 10.65 17.52 6.974 7.938
21.133 14.183 P(Bartlett's X2) 0.848 0.222 0.23 0.801 0.893 0.07
0.436 Replicate F 21.790 10.976 6.502 5.058 5.755 3.248 6.185
Replicate Prob(F) 0.0001 0.0001 0.0010 0.0044 0.0022 0.0311 0.0014
Treatment F 1.152 1.113 0.505 1.421 5.743 3.277 4.520 Treatment
Prob(F) 0.3456 0.3750 0.9167 0.1860 0.0001 0.0014 0.0001 Means
followed by same letter do not significantly differ (P = .05,
Student-Newman-Keuls)
[0430] Similar experiments were performed at Rutgers University to
test for treatment of bentgrass infected with dollar spot. Test
compounds were mixed in aqueous solution with the adjuvant Latron
B-1956.RTM. (Rohm and Haas) and applied to the grass at a rate of
4.18 fluid ounces per 1000 square feet and an interval of every 7
days. The number of lesions per plot were then counted and averaged
over the replicate plots. Treatment with fertilizers or other
fungicides were also performed. The results are presented in Table
7. Compound 80 and to a lesser extent compound 65 were particularly
effective. TABLE-US-00007 TABLE 7 Spray Number of Lesion Interval
Centers/Plot Treatment and Rate/1000 sq. ft. (days).sup.2 18 Jun 27
Jun 3A. Comp. 24 11.8 fl oz -- -- -- B. +Latron B-1956 .TM. 4.18 fl
oz 7 161.0 112.5 4A. Comp. 52 11.8 fl oz -- -- -- B. +Latron B-1956
.TM. 4.18 fl oz 7 118.8 74.0 5A. Comp. 54 11.8 fl oz -- -- -- B.
+Latron B-1956 .TM. 4.18 fl oz 7 158.0 125.0 6A. Comp. 65 11.8 fl
oz -- -- -- B. +Latron B-1956 .TM. 4.18 fl oz 7 22.5 17.5 7A. Comp.
1 11.8 fl oz -- -- -- B. +Latron B-1956 .TM. 4.18 fl oz 7 115.2
143.8 8A. Comp. 3 11.8 fl oz -- -- -- B. +Latron B-1956 .TM. 4.18
fl oz 7 200.0 187.5 10A. Comp. 71 14.6 fl oz -- -- -- B. +Latron
B-1956 .TM. 4.18 fl oz 7 98.0 162.5 11A. Comp. 75 14.6 fl oz -- --
-- B. +Latron B-1956 .TM. 4.18 fl oz 7 160.0 162.5 12A. Comp. 78
14.6 fl oz -- -- -- B. +Latron B-1956 .TM. 4.18 fl oz 7 159.5 107.8
13A. Comp. 79 14.6 fl oz -- -- -- B. +Latron B-1956 .TM. 4.18 fl oz
7 182.5 142.5 14A. Comp. 80 14.6 fl oz -- -- -- B. +Latron B-1956
.TM. 4.18 fl oz 7 6.2 0.0 15. FNX-100 .TM. 8.0 fl oz 14 159.5 187.5
16. FNX-100 .TM. 12.0 fl oz 14 162.0 200.0 17. FNX-100 .TM. 16.0 fl
oz 14 158.0 200.0 18. PS00KP .TM. 2.0 fl oz 7 200.0 175.0 19.
710-140 .TM. 10 fl oz 7 125.2 200.0 20. 710-140 .TM. 15 fl oz 7
84.2 125.2 21. 710-140 .TM. 20 fl oz 7 46.8 125.0 22. 710-140 .TM.
15 fl oz 14 133.5 162.5 23. 710-140 .TM. 20 fl oz 14 120.8 160.0
24. 710-140 .TM. 25 fl oz 14 119.8 112.5 25A. 710-140 .TM. 20 fl oz
-- -- -- 25B. /Daconil Ultrex .TM. 82.5SDG 14 57.8 2.0 3.25 oz 26A.
710-140 .TM. 20 fl oz -- -- -- 26B. /Chipco 26GT 2SC .TM. 3.0 fl oz
14 159.5 1.8 27A. 710-140 .TM. 20 fl oz -- -- -- 27B. /Chipco 26GT
2SC .TM. 3.0 fl oz 14 156.8 2.8 28A. 710-140 .TM. 20 fl oz -- -- --
28B. /Daconil Ultrex .TM. 82.5SDG 14 200.0 6.5 3.25 oz 29A. 710-140
.TM. 20 fl oz -- -- -- 29B. /710-140 .TM. 20 fl oz 14 116.8 131.25
30. 710-150 .TM. 20 fl oz 14 99.8 141.25 31. Daconil Ultrex .TM.
82.5SDG 1.8 oz 7 1.5 0.0 32. Daconil Ultrex .TM. 82.5SDG 3.25 oz 14
25.2 0.8 33. Fertilizer 18-6-12 1.6 oz N + -- -- -- 0.048 oz Fe 7
110.0 175.0 34. Fertilizer 18-6-12 1.6 oz N + -- -- -- 0.048 oz Fe
14 95.5 165.0 35. GE-2 Plus .TM. 4 oz 14 156.5 175.0 36. Untreated
Control -- 200.0 180.0
Example 86
[0431] The following illustrate representative pharmaceutical
dosage forms, containing a compound of formulas (I)-(IV) (`Compound
X`), for therapeutic or prophylactic use in humans or other
mammals. TABLE-US-00008 (i) Tablet 1 mg/tablet `Compound X` 100.0
Lactose 77.5 Povidone 1 Croscarmellose sodium 12.0 Microcrystalline
cellulose 92.5 Magnesium stearate 3.0 300.0 (ii) Tablet 2 mg/tablet
`Compound X` 20.0 Microcrystalline cellulose 410.0 Starch 50.0
Sodium starch glycolate 15.0 Magnesium stearate 5.0 500.0 (iii)
Capsule mg/capsule `Compound X` 10.0 Colloidal silicon dioxide 1.5
Lactose 465.5 Pregelatinized starch 120.0 Magnesium stearate 3.0
600.0 (iv) Injection 1 (1 mg/ml) mg/ml `Compound X` 1.0 Dibasic
sodium phosphate 12.0 Monobasic sodium phosphate 0.7 Sodium
chloride 4.5 1.0 N Sodium hydroxide solution (pH adjustment to
7.0-7.5) q.s. Water for injection q.s. ad 1 mL (v) Injection 2 (10
mg/ml) mg/ml `Compound X` 10.0 Monobasic sodium phosphate 0.3
Dibasic sodium phosphate 1.1 Polyethylene glycol 400 200.0 01 N
Sodium hydroxide solution (pH adjustment to 7.0-7.5) q.s. Water for
injection q.s. ad 1 mL (vi) Aerosol mg/can `Compound X` 20.0 Oleic
acid 10.0 Trichloromonofluoromethane 5,000.0
Dichlorodifluoromethane 10,000.0 Dichlorotetrafluoroethane
5,000.0
[0432] The above formulations may be obtained by conventional
procedures well known in the pharmaceutical art.
[0433] The compounds of the invention, e.g., compounds of formulas
(I)-(IV), may also be formulated into fungicidal compositions or
bacteriacidal compositions for use on plants, the compositions
comprising at least one compound of the invention and conventional
excipients.
[0434] All publications, patents, and patent documents are
incorporated by reference herein, as though individually
incorporated by reference. The invention has been described with
reference to various specific and preferred embodiments and
techniques. However, it should be understood that many variations
and modifications may be made while remaining within the spirit and
scope of the invention.
* * * * *