Note: Descriptions are shown in the official language in which they were submitted.
~0S04Z9
This application is a divisional of our co-pending
Canadian Patent Application Serial No. 215,984 filed December
13, 1974.
The present invention is concerned with a novel
therapeutic composition.
Many diseases caused by certain living micro-organisms
may ~e treated very effectively by administering a therapeutic-
ally effective amount of an antibiotic. Eowever, antibiotics
are not effective in the treatment of virus infections insofar
as suppressing the proliferation of the virus particles and -~
reducing cell damage are concerned.
`:
:`
,, .
: 2 -
,,
~050~29
A number of substances other than antibiotics
have been pronosed heretofore ~or use in treating virus
infections, but they have not been used extensively for
a nu~er of reasons. For instance, the previously
proposed antiviral agents usually exhibit very low anti-
viral activity and have little positive effect on the
course of the viral infection. The previously proposed
antiviral drugs also have not been e~fective in the
treatment of a wide spectrum of virus infections and
this has been a ma~or disadvantage. The therapeutic
treatment of a large number of patients having undeter-
mined viral infections of widely differing types is
impractical with the narrow spectrum prior art antiviral
drugs, as it is too difficult to determine the exact
virus causing the infection and then select an effective
drug. Many potentially effective antiviral drugs are
toxic and cannot be safely administered to patients,
and still other antiviral drugs have undesirable side
effec~s. As a result of the foregoing and other
deficiencies, it is apparent that an entirely satisfâctory
wide spectrum nontoxic antiviral drug has not been
available heretofore for routinely administerin~ tG
patlents having a viral infection caused by many of the
common viruses.
25~ It has been discovered that certain ethereal
monosu~stitutions of monosaccharide derivatives provide
important biological signals which allow living cells , q
to resist virus infections. As will be described in ~ -
.
~- ~ greater detail hereinafter, the signals also provide
other types of control in cell chemistry.
-
~ ~ ~ 3
-- . . .. .. . .
~lSQ4Z9
The therapeutic compositions of the invention overcome
the disadvantages of the prior art antiviral agents noted above,
and also produce other unusual and unexpected results. For
example, the ethereal monosubstitutions of monosaccharide
- derivatives described hereinafter are therapeutically effecti~e
at very low concentrations~ while at the same time they exhibit
..
no side effects and are very nontoxic. The ratio of the mini-
mum toxic to therapeutic dose is greater than 50 in both tissue
culture and animalq.
The antiviral compounds and the therapeutic composi-
tions containing the same disclosed and claimed in our earlier
filed copending application Serial No. 192,134 filed February 8,
1974 possess striking antiviral properties and are highly effec-
tive in the treatment of a wide variety of viral infections in
warm blooded animals. Nevertheless, the earlier described
antiviral compounds and therapeutic compositions do have certain
limitations and disadvantages which are overcome by the present
in~ention. For example, the earlier described c~mpounds possess
strong hydrophilic properties due to the presence of a plurality
of free hydro~yl groups which result in several limitations on
the effective use thereof in situ including short shelf life,
t --_
variability of the stability in solution as a function of
temperature, and extreme hy~droscopicity.
The compounds of the present invention have been found
to be more desirable with respect to the properties lis~Rd above.
This is presently believed to be the direct result of a de-
crea e in the hydrophilic properties and, therehy, an increase
in the solubility of the compounds in the fatty tissues of a
- 4 -
~05~)4~29
warm-blooded animal which has been therapeutically treated
therewith. This technique also aids in the transportation of
the drug to a desired tissue or the concentration thereof in a
desired tissue. In the practice of one variant of the invention
an organic substituent is selected which blocks at least one
free monosaccharide hydroxyl group and thereby decreases the
hydrophilic properties and allows the compound to be transported
or concentrated in fatty tissues. In a further variant, a
libile organic substituent is selected which may be removed
in vivo during treatment of the warm-blooded animal after it has
served its purpose, thereby unblocking the hydroxyl group(s).
It is an object of the present invention to provide a
therapeutic composition including a pharmaceutically acceptable
carrier and a therapeutically effective amount of certain
ethereal monosubstitutions of monosaccharide derivatives to be
described more fully hereinafter.
Detailed Description of the Invention
Including Preferred Variants Thereof
The novel therapeutic composition of the invention
comprises a pharmaceutically acceptable carrier and a thera-
peutically effective amount of an ethereal monosubstitution of a
monosaccharide derivative having the general formula S-O-Y and
organic and inorganic acid salts thereof, wherein S is the
residue of a monosaccharide selected from the group consisting
of pentoses, hexoses and heptoses which has been derivatized
with (a) one or more aliphatic alcohols containing 1-18 carbon
~ atoms and preferably 1-4 car~on atoms to produce an acetal group
'i at one or more available hydroxyl residu~, (b) one or moxe
, aldehydes c~ntaining 1-18 carbon atoms and preferably 1-4
: , ~
- 5 -
ij,, . , ~ . . . . .
lQSV4Z9
carbon atoms to produce single or m~ltiple acetal groups at one
ormore availa~le hydroxyl residua, tc) one or more ketones
containing 1-18 carbon atoms and preferably 1-4 carbon atoms
to produce single or multiple ketal groups at one or more avail-
a~le hydroxyl residua, or (d) one or more organic acid residua
containing 1-18 carbon atoms and preferably 1-4 carbon atoms to
produce ester groups at one or more available hydroxyl residua,
and Y is selected from the group consisting of cyclic monovalent -
nitrogen-containing organic radicals and residua and monovalent
organic radicals and residua having the general formula
~R3 ::
-Rl-~ , wherein Rl is a divalent organic radical having a
, R2
: linear car~on chain length of about 1-7 car~on atoms and R2 and
R3 are selected from the group consisting of -H, OH, -SH,
halogen and monovalent organic radicals and residua having a
linear carbon chain length of about 1-7 carbon atoms, but
excluding a compositio.n containing
3-0-3`-(~ `-dimethylamino-n-propyl)-1,2-o-isopro-
pylideneglucofuranose,
3-0-4'-(N'-methylpiperidyl)-1,2-0-isopropylidene-
glucofuranose,
3-0-2'-(~ dimethylaminoethyl)-1,2-0-isopropyl-
~ ~ .
ideneglucofuranose,
l 3-0-3'-(2'~ '-trimethylami~o-n-propyl)-1,2-o-
:
~: isopropylideneglucofuranose,
3-_-2'-(~ '-aimethylaminopropyl)-1,2-0-isopro-
`' pylideneglucofuranose,
6-0-3'-(N',N'-dimethylamino-n-propyl)-1,2-0-isopro-
pylidenegalactopyranosa,
: - 6 -
,- - . . . . . .
~' . : , ' ."' ' ' . .
lOS0429
6-0-2'-(~',~'-dimethylaminopropyl)-1,2-0-isopro-
pylidenegalactopyranose,
3-o-3~-t~ -dimethylamino-n-propyl)-1,2:5,6-di-_-
isopropylideneglucofuranose,
3-0-~ '-methylpiperidyl)-1,2:5,6-di-0-isopropyl.
ideneglucofuranose,
3-0-2'-(~ dimethylaminoethyl)-1,2:5,6-di-0-
isopropylideneglucofuranose,
3-0-3''-(2',~',N'-trimethylamino~n-propyl)-1,2:5,6-
10 di-0-isopropylideneglucofuranose,
3-0-2l-(N',~'-dimethylaminopropyl)-1,2:5,6-di-o~
isopropylideneglucofuranose,
6-0-3'-(~ '-dimethylamino-n-propyl)-1,2:3,4-di-0-
isopropylidenegalactopyranose,
6 0-2'-(~ '-dimethylaminopropyl)-1,2:3,4-di-0-
t isopropylidenegalactopyranose,
cC -N',N'-dimethylamino-isopropyl-2,3:5,6-di-0-iso-
propylideneglucofuranoside, and pharmaceutically acceptable
., organic and inorganic acid salts thereof. When R2 or R3 is
20 halogen, the halogen may be F, Cl, Br or I, of which Cl o~ Br
is usually preferred. The organic radical Rl, and R2 and R3
when they are organic radicals, may be branched or unbranched
t linear carbon chains and may be saturated or unsaturated,
and, when saturated, the linear and/or branched carbon chains
may contain one or more double or triple carbon-to-carbon ~.
bonds. The linear and/or branched carbon chains of Rl, R2
and R3 may be substituted or unsubstituted and, when substi-
tuted, one or more substituents may be present, such as
~ - 7- - -
., ~ .
1~50429
-OH, -SE, halogen (F, Cl, Br and/or I), branched or unbranched
and saturated or unsaturated hydrocar~on radicals containing
1-7 and preferably 1-3 carbon atoms, ~ R4 and/or -SR4 radicals
wherein R4 is a branched or unbranched and saturated or un-
saturated hydrocarbon radical containing 1-7 and preferably ~:
1-3 car~on atoms, carboxylic acid residua contain.ing 1-7 and
preferably 1-3 carhon atoms, and amino groups and aminohydro-
carbon radicals containing 1-7 and preferably 1-3 carbon atoms.
Preferably Rl is a hydrocarbon radical having a linear carbon
chain length of 1-3 or 1-4 carbon atoms and R2 and R3 are indi-
vidually selected from the group consisting of hydrogen and/or
hydrocarbon radicals having linear carbon chain lengths of
1-3 or 1~4 carbon atoms.
Examples of compounds from which cyclic organic
, radicals and residua are derived include (a) monovalent
nitrogen containing saturated, un~aturated or aromatic
',
:'j .
:;
.".
- 7
-- lOS04Z9
carkocyclic compounds containing about 4-8 carbon atoms
in the ring and preferably about 5-6 carbon ato~s in the
ring and at least one nitrogen atom attached thereto or
to an organo substituent thereon, (b) heterocyclic
S organic compounds containing about '-8 carbon atoms in
the ring and at least one ring nitrogen atom and (c)
dexivatives of the foregoing compounds wherein at least
one substituent is presert, such as -OH, -S~, halogen
(F, Cl, Br and/or I), branched or unbranched and saturated
or unsaturated hydrocarbon radicals containing 1-7 and
preferably 1-3 carbon atoms, -ORs and/or -SRs radicals
.
wherein Rs is a branched or unbranchéd and saturated or
unsaturated hydrocarbon racical containing 1-7 and
~referably 1-3 carbon atoms, carbocyclic acid residua
containing 1-7 and preferably 1-3 carbon atoms, ar.d
~ . amino groups and aminohydrocarbon radicals containing
; 1-7 and preferably 1-3 carbon atoms.-
The derivatized monosaccharide residue ~ ~ay
exist in an open chain or cyclic form having the general
; .20 formulao:
` (a) O = C - Z
,~ C = (~I, OW) i~:
.'~ ' C = (~1, OW)
Y-O
C = (E1, OW)
X
~ ~ ' ' , q
. , .
,.' . ~ :
- 8 - ~:
,~
. ~, .
.
'. ' ~ ' . :
~` ' ' ' . ' . " ' .
1050429
(b) X O z
H ~ H
(H, O~) ~ (H, OW)
/
O-Y
.
5 (c) X
H ) O Z
(H, OW) ~ / ~ H . .
(H, OW) ~ / ~ (H, OW)
' / .,
: . ~Y ~ :.
:~ 10 wherein X and Z are H, OH, hydrGxyalkyl, alkoxyl and./or
;
alkoxyalkyl containing up to 3 carbon atoms, ~ is El,
alkyl, alkenyl, cyclic alkane or cyclic aromatic containing
,~ . .
18 and preferably 1-6 carbon ~toms or acyl containin~
'~ . 1-18 and preferably 1-4 carbon atoms, and Y represQrts the
, 15 same organic radicals and residua as aforementioned for
,~ the general formula S-O-Y. The above general formulae
illustrate the various isomers of.the pentoses, hexoses :-
~, .
and heptoses, the relative spatial confisuration of the
and -OH groups a~out the ring, an~ the monosubstitution
!` :
~ 20 ~ :there~ in accordance with one presently preferred variant
, ;~ :
~ cf;the invention. The hydroxyl or alkoxyl residue of
:, . .
i; the hemiacetal or hemiketal linkage ma~ assume an ~ or a
con:figuratio~. The compounds of the invention may be
in the ~form o~ anomers or miY.tures of ano~ers.
'~ , ' : .
.~ . ' .
,
~aso42s
The configurations of the various ~erivatives
of isomers of the pentoses, hexoses and he~toses are
well known to those skilled in this art and ~umerous
reference books are available or the sub~ect, the
teachings of which are incoxporated herein by reference.
For example, see Textkook of Biochemistry, 4th Edition,
by West et al tl966) and The Monosaccharides by Stanek,
- , _
Cerny, Kocourek and Pacak (1963). The ~rior art discloses,
for example, a total of eight open chain isomers for the
reducing hexoses, and an even larger number of open ~-
chain isomers for the reducing heptoses. Either the D-
series or the L-series of the pentoses, hexoses and
heptoses may be used in practicing the invention, but
~; it is usually preferred to use the D-series. The hexoses
1~ often give the best results and especially D-talose, D-
galactose, L-galactose, D-i~ose, D-~ulo~e, D-mc,nnose,
D-glucose, L-glucose, D-altrose and D-allose. The afore-
mentioned pentoses, hexoses and heptoses may be et~ereally
monosubstituted in any available position and derl~atized
at one or n.ore of the remaining hydro~yl groups.
Nevertheless, it is understood that substitution of certain
positions of specific monosaccharide derivatives results
.: ,
in more therapeutically active or less tox~c compounds. -
For instance, substitution o f the 3-O- position o' 1,2-
O-isopropylidene-D-glucofuranose or 1,2:5,6-di O-isopro-
pylidene-D-glucofuranose and the 6-O- position of 1,2-O-
isopropylidene-D-galactopyranose ox 1,2:3,4-di-O-isopro~
pylidene-D-galactopyranose results in especially valuable
-- 1 0 -- .
., ~ ' '
: - . , ~, . ~ :- . ,:
1050429
compounds.
The following substituents may be ethereally
substituted on any of the available positions of the
: various isomers of the pentoses, hexoses and heptoses to
produce nonto~ic compounds having exceptional therapeutic
activity:
-(n-propylamino),
-(N',N'-dimethylamino-n-propyl),
-(N',N'-dimethylaminoisopropyl),
-(N'-methylpiperidyl), ~
-(N',N'-dimethylaminoethyl), ::
-(N',N'-diethylaminoethyl),
-(2',N',N'-trimethylamino-n-propyl),
-dimethylamino,
-(N',N'-dimethylaminomethyl), -
-(N',N'-dimethylaminopropyl),
-(N',~'-dimethylamino-iso-butyl),
.. . .
. -(N',~3'~ .ethylamino-n-butyl),
-(N',N'-dimethylamino-iso-pentyl), .
-~N',N'-dimethylaminopentyl),
N~-methylamino-n-propyl)~
l'-methyl-~l'-ethylamino-r.-propyl~,
-(N',N'-diethylamino-n-propyl),
(amino-n-propyl),
~: 25 -~N'-ethyla~ino-n-propyl),
; -lN~-propylamino-n-propyl)~
N',~'-iso-propylamino-n-propyl),
1',2'-ethylimino-n-propyl),
. . . :
:. : .
'' , ~ :
:
''` . ' - ~ ~ . ' ':
" ~ ` '., ' . , ' '`. ~ ' . :
105()4Z~
n-propylpyrrolidyl),
~ n-propylpiperidyl),
-piperidyl,
-(N',N'-dimethylamino-sec-butyl).
Of the foregoing substituents, -(~',N'-dimethylamino-n-
propyl) is presently preferred and especially when
: su~stituted in the 3-0- position of 1,2-0-isopropylider.e-
D-glucofuranose or 1,2:5,6-di-O~isopropvlidene-D-gluco-
furanose or the 6-0- position of 1,2-0-isopropylidene-D-
: 10 galactopyranose or 1,2:3,4-di-0-isopropylidene-D-galacto-
pyranose.
The following compounds have been found to have
exceptional wide spectrum antiviral activity and other
therapeutically valuable properties and are presently
preferred for use in the composition and method of the
invention:
3-0-3l-(n-propylamino)-1,2-0-isopropylidene-
D-glucofuranose ,
3-0-3'-(N',N'-dimethylamino-n-propyl)-1,2- -
0-isopropylidene-D-glucofuranose ,
3-0-4'-(N'-methylpiperidyl)-1,2-0-isopropyliden~-
. D-glucofuranose , ~ :
3-0-2'-~N',N'-dimethylaminoethyl~-1,2-0-iso-
~ propylidene-D-glucofuranose ,
:~ 25 3-0-2'-(N',N'-diethylaminoethyl)-1,2-0-iso-
propylidene-D-glucofuranose ,
3-0-3'-(2',N',NI-trimethylamino-n-propyl)-1,2- .
~ 0-isopropylid~ne-D-gluco~uranose ;
i; :
~ . - I2 -
., .
;,, :
:~ :
,
lOS04Z9
. 3-0-2'-(N',N'-dimethylaminopropyl)-1,2-0-iso-
propylidene-D-glucofuranose ,
6-0-3'-(N',N'-dimethylamino-n-propyl)-1,2-0-
isopropylidene-D-galactopyranose ,
6-0-2'-(N',N'-dimethylaminopropyl)-1,2-0-iso-
propylidene-D-galactopyranose ,
3-0-3'-(n-propylamiro)-1,2:5,6-di-0-isc~rcpyli-
dene-D-glucofuranose ,
3-0-3'-(N',N'-di~ethylamino-n-propyl)-1,2:5,6-
di-0-is~propylidene-D-glucofuranose,
3-0-4'-(N'-methylpiperidyl)-1,2:5,6-di-0-
isopropylidene-D-ylucofuranose ,
t 3-0-2'-(N',N'-dimethylaminoethyl)-1,2:5,6-di-
0-isopropylidene-D-glucofuranose,
3-0-2'-(N',N'-diethylaminoethyl)-1,2:5,6-di-0-
~ isopropylidene-D-glucofuranose ,
; 3-0-3'-(2',N',N'-trimethylamino-n-propyl)-
1,2:5,6-di-0-iscpropylidene-D-slucofuranose ,
3-0-2'-(N',N'-dimethylaminopropyl)-1,2:5,6-di-
. 20 0-isopropylidene-D-glucofuranose ,
: 6-0-3'-(N',N'-dimethylamino-n-pro~yl)-1,2:3,4-
6 di-0-isopropylidene-D-galactopyranose ,
: 6-0-2'-tN',N'-dimethylaminopropyl)-1,2:3,g-di-
0-isopropylidene-D-galactopyranose ,
-N',N'-dimethylamino-iso-propyl-2,3:5,6-di-0-
isopropylidene-D-glucofuranoside ,
and organic and inorganic acid salts thereof. .-
Of the foregoing compounds, 3-0-3'-tNl,N'-
,
: - 13 -
.
- . .
10504;~9
dimethylamino-n-propyl)-1,2 O-isopropylidene-D-gluco-
furanose is presently preferred.
Additional compounds of the general formula
S-O-Y, wherein Y is -R~ <R3; which may be used in
practicing the invention are listed belo~
Monosaccharide Residue 5ubstituent
~S) (~)
RIR2 R3
3-O-1,2-O-isopropylidene
lO-D-glucofuranose 3'-n-propyl II methyl
" " ethyl "
" " H ethyl
2'-iso-propyl methyl methyl
" 3'-1,2-propenyl
" sec-butyl " "
" 3'-~utyl "
2'-ethyl H ~I
" methyl H I~
6-O-1,2-O-isopropylidene
20-D-galactopyranose 3'-n-propyl H ~ethyl-
" " ethyl " - :
" " H ethyl
" 3'-1,2-pro~enyl methyl methyl
" 2'-iso-propyl
" sec-butyl " " ~
" 3'-butyl 'I ll ~-
2'-ethyl H H
" methyl H
: 3-O-1,2:5,Ç-di-O-iso-
~0propylidene-D-gluco-
furanose 3'-n-Propyl }I methyl
l ~ " ethyl "
" H ethyl
. " 2'-iso-propyl methyl methyl
" 3'-1,2-propenyl
sec-butyl
" 3'-butyl '
" 2'-ethyl ~ H
. " methyl ~ H
6-O-1,2:3,4-di-O-iso-
propylidene-D-
galactopyranose 3'-n-propyl H methyl
" " ethyl
'~ " " H ethyl
: 45 " 3'-1,2-propenyl methyl methyl
" 2'-iso-propyl " "
" . sec-butyl
: . : " 3'-butyl " "
: " 2'-ethyl ~ II
" ~ethyl H
.;
- 14 -.
,
.,
., " . . .
. . .: . ; .. : . . ~.
... . .
105042~
Still other compounds of the general formula S-0-Y,
wherein Y is a cyclic monovalent nitrogen-containing
organic radical or residue, wh ch may ~e used in
practicing the invention, are as follows: .
5Monosaccharide Residue Substituent
(S) (Y)
Substituent
Cyclic Radical on the
_ Cyclic Radical
103-0-1,2-0-isopropylidene
-~-gluco~uranose 4'-piperidyl H
~ " 3'-piperidyl me~hyl, H
" 2'-piperidyl " "
" 3'-pyrrolidyl " "
" 2'-pyrrolidyl " "
6-0-1,2-0-isopropylidene
-D-galac~opyranose 4'-piperidyl H
" 3'-piperidyl methyl, H
" 2'-piperidyl " "
" 3'-pyrrolidyl " "
" 2'-pyrrolidyl " "
3-0-1,2:5,6-~i-0-is~-
propylidene~
glucofuranose 4'-piperidyl H
2; " 3'-piperidyl methyl, ~I
.. " 2'-piperidyl " "
" 3 ' -pyrrolidyl " "
2 '-pyrrolidyl
6-0-1,2:3,4-di-0-iso-
30~sQpropylidene-D-
galactopyranose 4'-piperidyl H
: " 3 '-piperidyl methyl, H
: " 2'-piperidyl
" 3'-~yrrolidyl " "
.35 " 2'-pyrrolidyl " "
- The present invention al~o prcvides certain novel
compounds which have wide spectrum antiviral activity.
The novel compounds may be defined generically as follo~s:
3-0-3'-(N',N'-dimethylamino-n-propyl)-1,2-0-
isopropylidene~lucofuranose,
~; . 3-0-4'~(N'-methylpiperidyl)-1,2-0-isoprapylidene-
,~ glucofuranose,
.
:~ - 15 -
,
.. .
. .
.~ . . . . . . . .
~(~50429
3-0-2'-(N' ,N' -dimethylaminoethyl)-1,2-0-isopropyli-
deneglucofuranose,
3-0-3'-(2',N',N'-trimethylamino-n-propyl)-1,2-0-
isopropylideneglucofuranose,
3-0-2'-(N',N'-dimethylaminopropyl)-1,2-0-isopro-
pylideneglucofuranose,
6-0-3'-(~',N'-dimethylamino-n-propyl)-1,2-0-
~¦ isopropylidenegalactopyranose,
6-0-2'-(~',N'-dimethylaminopropyl)-1,2-0-isopro-
pylidenegalactopyranose,
3-0-3'-(N',N'-dimethylamino-n-propyl)-1,2:5,6-di-
0-isopropylideneglucofuranose,
. 3-0-4'-(N'-methylpiperidyl)-1,2:5,6-di-0-isopro-
pylideneglucofuranose,
3-0-2'-(N',N'-dimethylaminoethyl)-1,2:5,6-di-0-
isopropylideneglucofuranose,
3-0-3'-(2',N',N'-trimethylamino-n-propyl)-1,2:5,6-
di-0-isopropylidenegluco~uranose,
3-0-2'-(N',N'-dimethylaminopropyl)-1,2:5,6-di-0-
isopropylideneglucofuranose,
6-0-3'-(~',~'-dimethylamino-n-propyl)-1,2:3,4-di-
, .
~ 0-isopropylidenegalactopyranose,
'`''I
6-0 2'-(N',N'-dimethylaminopropyl)-1,2:3,4-di-0-
isopropylidenegalactopyranose,
~.. ! ' .
.. 25 -N',N'-dimethylamino-iso-propyl-2,3:5,6-di-0-
,.j
isopropylideneglucofuranoside,
and organic and inorganic acid salts thereof.
,,:
.,
~ - 16
'' .
lOS04Z9
.Species of the foregoing novel compounds which
possess striking wide spectrum antiviral activity are
as follows:
3-0-3'-(N',N'-dimethylamino-n-propyl)-1,2-0-
isopropylidene-D-glucofuranose,
3-0-4'-(N'-methylpiperidyl)-1,2-0-isopropylidene-
D-glucofuranose,
3-0-2'-(N',N'-dimethylaminoethyl)-1,2-0-isopropyli-
dene-~-gluco.uranose,
3-0-3'-(2',N',NI-trimethylamino-n-propyl)-1,2-O-
i isopropylidene-D-glucofuranose,
3-0-2'-(N',~'-dimetnylaminopropyl)-1,2-0-isopro-
: ~ pylidene-D-glucofuranose ,
6-0-3'-(N',~'-2imethylamino-n-propyl)-1,2-0-iso-
propylidene-D-galactopyranose,
6-0-2'-(~',N'-dimethylaminopropyl)-1,2-0-isopro-
: pylidene-D-galactopyranose,.
3-0-3'-(N',N'-dimethylamino-n-propyl)-1,2:5,6-di-
.
0-isopropylide~e-D-glucofuranose ,
20 3-0-4'-(N'-methylpiperidyl)-1,2:5,6-di-0-isopro-
pylidene-D-glucofuranose ,
3-o-3'-(2',~ '-trimet~ylamino-n-prapyl)-1,2:~,6-
. di-0-isopropylidene-D-glucofuranose ,
3-0-2~-(N',N'-dimethylaminoethyl)-1,2:5,6-di-0-
isopropylidene-D-glucofuranose ,
3-0-2'-(N',N'-dimethylaminopropyl)-1,2;5,6-di-0-
,; : .
; isopropylidene-D-glucofuranose , .s
'
~,
- 17 -
.~ .
~(~S(~29
6-0-3 I r (~ -dimethylamino-n-propyl)-1,2:3,4-di-
O-isopropylidene-D-galactopyranose,
6-0-2~ '-dimethylaminopropyl)1,2:3,4-di-0-
isopropylidene-D-galactopyranose,
oC -N',N'-dimethylamino-iso-propyl-2,3:5,6~di-0-
isopropylidene-D-glucofuranoside, and organic and inorganic
acid salts thereof.
In general, the preparation of ~he monosubs~ituted
compounds described herein involves the formation of alkyl
ethers or substituted alkyl ethers at selected positions on
the desired monosaccharide derivative, such as at position
3-0- of 1,2-0-isopropylidene-D-glucofuranose or 1,2:5,6-di-o-
isopropylidene-D-glucofuranose, position 6-0- of 1,2-0-isopro-
pylidene-D-galactopyranose or 1,2:3,4-di-0-isopropylidene-D-
galactopyranose, and position 3-Q- of 1,2-0-isopropylidene-D-
fructopyranose or lo 2:5,6-di-0-isopropylidene-D-fructopyranose.
The condensation of the substituent substrate with the mono-
saccharide derivative at the desired position may ~e achieved by
various prior art techniques.` One method is described in
United States patent #2,715,121, issued August 9, 1955 to
; GLEN et al, The method described in this patent requires
extreme reackion conditions and often gives low yields. The
I product purity is also less than satisfactory~
¦ The preferred methoa of preparation involves much
milder reaction conditions than employed in patent #2,715,121.
The side reactions are minimized, the purity
..
..
- 18 -
1050429
of the final product is greatly improved and the method
may be adapted to a series of solvents having varying
properties such as dioxane, tetrahydrofuran and benzene.
Briefly, the improved method involves the reaction of a
monosaccharide derivative which is blocked with
one or more organo groups in the hydroxyl group positions
adjacent the desired position to be substituted. The
blocked monosaccharide is dissolved in one of
the foregoing solvents and is reacted ~7ith a halogenated
organo amino compound having the desired carbon chain
length and configuration in the presence of a base such
as sodium hydroxide. Selective removal of one or more
; blocking groupsmay be accomplished by hydrolysis under
specific conaitions resulting in a new product which is
to ~e considered a compound of this invention. The
reaction of either the blocked compound or the hydroly~ed
,: .
compound with any organic or inorganic acid to form a
salt thereof or with any organic or inorganic base .o
` form a salt thereof results in a compound of this
invention.
It is understood that simple derivati~es of the
compounds described ~erei~ are embraced by the invention.
' Such derivatives may be prepared by prior art techniques
~- and procedures and used as an ingredient in the thera-
peutic co~position and method of the invention.
, . . .
For example, the free amine compounds are basic
and form organic acid salts and inorganic acid salts,
and the resulting salts are use~ul in the therapeutic
'' ~ .
.
.~ . - 19 -. ....
' ', ' . , .
'
~: ' . , ' :
~C)50429
composition and method of the invention. The salts may
be prepared by the usual prior art techniques, such as
by adding the free amine compound to water and then
adding the desired organic acid or mineral acid thereto
in an amount sufficient to neutralize the free amine.
Examples of suitable acids include ~.Cl, H~r, H2S~4,
HNO3, benzoic acid, p-aminobenzoic acid, p-acetamido-
benzoic acid, p-hydroxybenzoic acid, alkane sulfonic
acid, p-toluene solfonic acid, acetic acid, alkylcarboxylic
acids, oxalic acid, tartaric acid, lactic acid, pyruvic
acid, malic acid, succinic acid, gluconic acid and
glucuronic acid. The aqueous solution of the resulting
salt is evaporated to the volume necessary to assure
precipitation of the salt upon cooling. The precipitated
salt is recovered by filtration, washed and dried to
obt~in a final amine salt product. The amine salts are ~ ;
often preferred for use in formulating the therapeutic
compositions of the invention as they are crystallire
ana relatively nonhygroscopic. The amine salts are also
better adapted for intramuscular injection than are the
. .
free amir.es.
Prior art blocking techniques may be e~plGyed such
as acetonization and acetylation. Suitable prior art
blocking methods are described in the aforementioned
Ur.ited States patent ~2,715,121 and are described in
the specific e~amples appearing hereinafter. In instances
:, ~
where an aldehyde or ketone is reacted with hydroxyl `!
;~. groups on ad~acent carbon atoms, the initial compound
~ 20 - ~
.... , ~ ,. . . - . . . . . .. .~
lOS0429
may be dissolved in the desired aldehyde or ketone under
anhydrous conditions and a Lewis acid catalyst is added
in a catalytic quantity, such as 1% zinc chloride or
anhydrous phosphoric acid. Often acetone is the preferred
blocking agent, but aldehydes or ketones of much higher
molecular ~eight may be used when desired such as those
containing up to 25 carbon atoms. The reaction mixture
is agitated at room temperature for a prolonged reaction
period, such as 24-48 hours. The compound may be
blocked in a plurality of positions, such as the 1,2-
and 5,6- positions. It is usually preferred to block
positions such as the 1,2- positions as the resulting
partially blocked compound is much less toxic than
compounds blocked in all available hydroxyl groups.
It is also possible to block one or more free
hydroxyl positions o~ the compound with an ester group,
wherein the carboxy~ic acid residue contains 1-18 and
preferably 1-3 carbon atoms. The ester derivatives
likewise may be prepared following prior art techni~ues
such as by reacting a carboxylic acid anhydride with the
; compound following prior art practices. Additionally,
the ~ or ~ alkyl derivatives of monosaccharide derivatives
such as 2,3:5,6-di-O-isopropylidene-D-glucofuranoside
~ay be prepared following prior art techniques. In this
latter instance, the compound is dissolved in~ dry ~;
^ alcohol having the desired carbon chain length with i
aforementioned residua and reacted with the compound in
the presence ~f a catalyst such as hydrogen chloride
of Do~.Yex 50 H+ resi~. While the above discussed
- 21 -
'~ ~
; .
10504Z9
derivatives are presently preferred, it is understood
that still other simple derivatives may be prepared
following prior art techni~ues and then used in prac~icing
the present invention. In addition to the foregoing, . -.
the compounds may also include monosu~stitutions of
monosaccharide derivatives in ~hich the substrate
{-O-Rl-NcR3}may be replaced by a substituent R7, ~Jhere
R7 is a deoxymonosaccha~ide derivative o~ halogen, keto,
amino, lower al~yl, mercapto, alkenyl, alkynyl, aromatic,
heterocyclic or alkylcarboxylic acid and its derivatives.
R~ may also represent the same groups as the above
substrate of the monosaccharide derivative ethers. Still
othor antiviral agents have a ~eneral formula S-o-~i
wherein Y is -R~-S-Rs where Ra is a saturated or
unsaturated hydrocarbon radical containing 1-7 carbon
atoms and Rg is a monovalent saturated or unsaturate~
hyarocarbon radical containing 1-7 carbon atoms an~
hydro~en.
~ The compounds of the invention are especialiy ~;
useful as wide spectrum antiviral agents for the thera-
peutic treatment of warm-blooded animals. They exhLbit
potent antiviral activity against both RNA and D~A
viruses, contrary to the prior art antiviral ager.ts.
The compounds of the present invention exhi~it mar~ed
, ~ .
suppression of virus particle multiplication and virus-
induced cell injury in animal and human cell tissue
`~ ~ culture systems against such widely varying viruses as ~:-
g~ herpes simplex, ~nfluenza A, mumps, poliovirus and
-~2 -
. ~ '
. ~,~ .,
,. . . .
.
,. , . . . . . . . . ; ~ ,
105U4Z9
rhinovirus. In tests in the whole animal, the compounds
can reduce mortality and morbidity manifestations of
influenza A infection by from 50% to 85%.
The compounds of the present invention may be
, . .
administered to human patient or animal to be treated
either orally or by parenteral administration. When
the therapeutic composition is to be administered
orally, the compound may be admixed with a prior art
filler and/or binder such as starch and a disintegrator,
and the admixture is pressed into a tablet of a size
convenient for oral administration. Capsules also may
be filled with the powd~_ed therapeutic composition
and administered orally. Alternatively, a wa.er solution
or suspension of the therapeutic composition may be
is admixed with a flavored syrup such as cherry syrup and
administered orally. ~en the therapeutic composition
is administered by intramuscular injection, the compound
is usually dissolved in a physiolcgical saline solution
which contains sodium chloride in sufflcient concentration
to make the overall solution to be injected isotonic to
body fluids. A salt of the free amine compound is
usually preferred in instances whe*e the compound is
administexed by intramuscular injection. In treating
upper respiratory viral infections, the salt form in
;~ 25 aqueous solution may also be administered by nasopharyngeal
` spray. Administration also may be by means of a
suppository in patients unable to retain medication
administered by mouth.
'
23 - -
.
.
;,
,
1050429
The dosage may be varied over extremely wide limits,
as the compounds are effective at low dosage levels
and are relatively free of ~oxicity and adverse side
effects. The compounds may be administered in the
minimum quantity which is therapeutically effective, and
the dosage may be increased as desired up to the
maximum dosage tolerated by the patient. Animal toxicity
data indicate that the limiting nontoxic dosage may be
up to 100-200 times the minimum effective dosage. Also,
`~ 10 it is not necessary to carefully control the dosage
for patients sensitive to the prior art antiviral drugs.
As a general rule, the compound may be administered in
an amount of about 1-4~ milligrams per kilogram of body
; weight per day, and preferably in an amount of about
2-20 miligrams per kilogram per day, over the peri~
re~uired for treatment of the viral infection.
Surprisingly, the compounds described herein have
still other unusual and unexpected therapeutically
valuable properties. The learning of new tasks is
enhanced. ~he therapeutic composition also protects
against death due to water deprivation. ~he buffer
! capacity of the compounds over a pH range of approxi-
mately 7-9.8 is very good and they may be used for this
purpose. Certain compounds also exhibit lubricative
properties and may be used as a specialized lubricant.
Compounds such as 3~0-D-glucopropionitxile and its
derivatives are photosensitive and may be used in
photographic applications.
~' .
~ - 2~ -
.
~1~50429
It has been further discovered that the normal
life span of tissue culture cells bein~ grown in a
prior art tissue culture medium may be extended very
substantially and often by several fold by growin~ the
cells in a tissue culture medium which contains an
effective amount of one or more of the antiviral
com~ounds described herein. This variant o~ the present
invention is especiall~ useful in the growing of human
embryo cells and it has been observed that such cells
survive for a significantly longer period of time in a
tissue culture medium in the presence of as little as 2
micro~rams per milliliter of one or more of the antiviral
compounds described herein. However, much larger quantities
of the compound may be present, such as the quantity
described herein in tlle treatment of tissue culture
cells infected with a virus, or in amounts up to the
tolerance level of the cells. Preferably, the compound
is 3-0-3'(N',N'-dimethylamino-n-propyl)~ 2-0-
isopropylidene glucofuranose. This variant of the
invention reduces research costs due to the longer life
span of the tissue culture cells. It also has other
positive effects as the surviving cells tend to be more
; vigorous and in a better state of preservation and
more consistent research results are obtained.
It has also bean discovered that the antiviral
compounds disclosed herein, the pharmaceutical compositions
containing the same, and the method of the invention are
.. .. ~ ` .
l~Sf~429
effective in the treatment of metastatic cancer in warm
blooded animals. In practicing this variant of the invention,
a warm blooded animal having a metastatic cancer, as carcinoma,
is treated by administering thereto a therapeutically ef~ective
amount of at least one antiviral compound disclosed herein.
The compound may be administered at the dosage level described
herein for common ~iral infections such as influenza. Often
much larger quantities are more effective such as 10-50 times
this amount~ or quantities within the tolerance level of the
wa~m blooded animal regardless of the amount. Death from
cancer in warm blooded animals is caused by metas~atic spread of
the tumor cells, which in turn results from a greater freedom
to translocate that individual cells develop when they become
cancerous (Leighton, J., The Spread of Cancer, Academic Press,
~ew York and London~ 1967)o The compounds used in the treat-
ment of cancers specifically prevent this tendency to metastasis,
thereby rendering the cancer non-malignant and allowing the
warm blooded animal to live.
The compounds used in the treatment of metastatic
cancer and/ox in the tissue culture medium in the growing of
tissue culture cells are preferably the antiviral compounds ais-
closed herein. However, it is understood that the antiviral
compounds disclosed in my earlier filed application Serial No.
192,134 also are useful for this purpose and may be substituted
in equal quantities.
The f~llowing specific~examples further illustrate
the present invention~
. :.
- 26 -
1050429
- Example 1
T~ a solu~ion of 104 ~ (0~4 male) of 1,2:5,6-di-
0-isopropylidene-D-glucofuranose in 550 ml of 1,4-dioxane
~ias added 189~7 g (1.2 mole) of 3-chloro-N,~-dimethyl-
amino propane in the form of the hydrochloride salt and
144 g ~3.6 mole) of sodium hydroxide. The suspension
was mechanically stirred and heated to reflux for 18
hours. The reaction miYture thus prepared was filtered
the solids were washed with 1,4-dioxane and the washings
were combined with the filtered liquid. The solvent
was removed under reduced pxessure and an amber-colored
viscous oil was obtained.
The oil was distilled under high vacuum ~less than
I mm Hg) while using a very slight dry nitrogen purge to
obtain high and low bolling fractions. Th~ low boiling
fraction was identified as unreacted 3-chloro-N,~-di-
methylamino propane. ~he high ~oiling fraction had a
boiling point of 148-154C at 2.5 mm Hg and w~s a clear
viscous oil with an optical rotation of {~}Ds a -19 ~ 3
2 nea~ ~100 mm) and a density of 0.95 g/cc. The refractive ~ -
index was nD6 - 1.4576. Gas chromatQgraphy showe~ a
purity greater than 99~. An elemental analysis showed:
C, 59.13; H, 8.99; N, 4.12; 0, 27.7. The yield was 80~
of the novel compound 1,2:5,6-di-0-isopropylidene-3-0-3'-
j 25 (N',N'-dimethylamino-n-propyl)-D-glllcofuranose.
A portion of the above oil (10 g) was hydrolyzed
in aqueous sulfuric acid at a pH value of 1.9-2.1 for
,:,
10 hours with reflu~ingO The resulting solution was
~ 27 -
.
., ~
51)42g
adjusted to a pH value ~f 4.5 with saturated ~a(OII) 2
solution, centrifuged, and filtered through an ultrafine
filter. The filtrate was lyophillized to a white-to-
slightly yellow solid having a melting point of 78-80C.
S Gas chromatography data indicated above 99~ purity of
the novel co~pound 3-0-3'-~N',N'-dimethylamino-n-propyl)-
D-glucopyranose. In thin-layer chromatography, the flow
rate on silica gel with a solvent mixture composed of
n-propanol, ethyl acetate, H2O and NH3 in the ratio by
volume of 60:10:30:10, respectively, was Rf = 0.356.
A portion of the oil is partially hydrolyzed to
1,2-O-isopropylidene-3-O-3'-(N',N'-dimethylamino-n-
propyl~-D-glucofuranose by dissolving it in distilled
water and adjusting the pH of the approximatley lM
solution to 3;0 + 0.2 with 6~ HCi. The solution is
extracted twice with chloroform and the clear aqueous
solution is refluxed for about two hours. Completion of
partial hydrolysis reaction was monitored by gas
chromatography from disappearance of the peak of Farent
.20 compound and appearance of a new peak with larger
retention time. The solution is then cooled, made
alXaline with 30% soaium hydroxide to p~ 10.5 and then
extracted with chloroform. The chloroform phase is
separated, dried over anhydrous magnesium sulfate and
vac~um distilled to remove the solve~t. The resulting
colorless viscous oil has o~tical~rotation of ~neat = ,;
-12 and refractive~index of 1.4687 at 25C. Alter- -
`~ natively, the compound can be obtained as hydrochloride
_ 28 _
`. `~' , `
- '
lOS04Z9
salt by lyophillizing the aqueous solution after partial
hydrolysis at p~ 4.0-4.5. A white crystalline material
is obtained ~hich is recrystallized from methanol. The
crystalline hydxochloride of 1,2-0-isopropylidene-3-0-3'-
(N',~'-dimethylamino-n-propyl)-D-glucofuranose has a
melting point of 181-183C and purity as indicated by gas
chromatography is 98+~. Infrared spectrophotometry
indicates presence of strong -OH band which is not present
in parent oil. The elemental analysis for the hydro-
chloride salt in a typical batch showed: C, 49.09; H, .
8.40; N, 4.14; Cl, 10.32; O, 28.12. Theoretical values
are as follows: C, 49.19; H, 8.19; ~, 4.09; C1, 10.39;
O, 28.11.
; The gas-liquid chromatograms for the above intermediate
and final novel compounds were run on a Beckman GC, Model
72-5 with a hydrogen flame detector. The column used for
the intermediate novel compound was a commercially
available SE-52 column, wherein methyl phenyl resins act
.~ as stationary phases supported on Chromosorb ~ (H.P.)
which is made by Johns-Manville Corporation. The ~inal
~; novel compound wa.s chromatographed on a Chromosorb 103
glass co1umn, which is packed with porous resins. The
foregoing materials are commercially available.
.
Examule 2
;~ ~25 Starting with 51 g (0.3 mole) of 4-chloro-N-methyl-
piperidine~hydrochloride and 26 g (0.1 mole) o, 1,2:5,6-
di-O-isopropylidene-D-gluco uranose and 36 g of NaOH in
~e~,l( - 29 ~
,, .
. . .
~,
. . . ,
. .: : : ~
~OS()~29
150 ml 1,4-dioxane, condensation was accomplished using
the general procedure outlined in Example 1. The residue
remaining following vacuum distillation was dissolved
and recrystallized from hot methanol. The melting point
was 106-107.5C (sharp).
Hydrolysis of the above product in ~2S04 at a pH
value of 2.1 yielded 3-0~4'-(N'-methylpiperidyl)-D-gluco-
pyranose having an optical rotation Of ra~D~ = +38.42 in
H20. A gas chromatography analysis in accordance with
Example 1 indicated that the purity of the product was
in excess of 96~. The melting point was 62-65C.
, .
Example 3
A solution of 0.1 mole of 1,2:5,6-di-0-isopropylidene-
~-glucofuranose in 50 ml of tetrahydrofu~an W25 added to
a suspension of 0.3 mole of 2-chloro-N,N-diethyla~ino-
~thane hydrochloride and 36 g of sodium hydroxide in 100
ml of tetrahydrofuran. ~he suspension ~as mechanically
stirred and refluY.ed overnight and the- reaction mi~.ture
was treated as set out in Example 1. The desired product,
1,2:5,6-di-0-isopropylidene-3-0-2'-(N',N'-diethylamino-
ethyl)-D-glucofuranose was obtained as z clear yellQw
liquid (boiling point 144-150C/0.15 mm HgJ having an
optical rotation of {~}D8 = -20.6 neat and a refractive
i index of nDS = 1.4532. The liquid solidified on eY.posure
to air, probably due to formation of the carbonate salt.
. . 7
The yield was 85%~
' : `
.
:
~ ~ - 30 -
. . ~ . .
,. ,.,... ,..~
- 10504i~9
Ten grams of the above product were hydrolvzed with
aqueous sulfuric acid at a pH value of 1.9-2.1 for ten
hours under reflux. The resulting solution was adjusted
to a pH value of 4-5 with saturated barium hydroxide
solution and then centrifuged and filtered. Lyophillization
of the filtrate yielded 6.55 g of light brown crystalline
3-0-2'-~N',N'-diethylaminoethyl)-D-glucop~ranose. The
optical rotation in water was {a}D5 = ~36.33. A gas
chromatography analysis in accordance with Example 1
indicated that the purity was in excess of 99%.
., :
Example 4
To 26 g (0.1 mo~e) of 1,2:5,6-di-0-isopropylidene-
~-slucofuranose and 36 g (0.9 mole) of sodium hydroxide
; in 150 ml of refluxing tetrahydrofuran was added dropwise
over cne hour 0.3 mole of 3-bromopropionitrile in 50 ml
of tetrahydrofuran. The reaction mixture was refluxed
for an additionai six hours and then filtered. The solids
-,. :
were washed with tetrahydrofuran and the washings were
combined with the filtrate. The solvent was removed
under reduced pressure and solid 1,2:5,6-dl-0-isopro-
; pylidene-3-0-3'-propionitrile-D-glucofuranose was obtained.
The decomposition point was 165C and it was light
sensitive indicating utility in photographic applications.
Five grams (0.016 mole) of the above product was
dissolved in anhydrous ether and added dropwise to a
,.~" ~ " .
suspension of 0.76 g (0.02 mole) of lithium aluminum
hydride in ether. ~he resulting complex was dissolved
.. ~, ~; ~; '
; _ 3~
. ,~. .
'
1050429
in cold hydrochloric acid and neutralized rapidly with
sodiu~ bicarbonate. The suspension thus produced was
extraoted with chloroform and the solvent ~as removed to
obtain a yellow oil in a yield of 250 mg. Gas chromato-
graphy in accordance with Example 1 indicated a purity
of 98~ and there was a sharp infrared band at 3400 cm~~.
The oil was hydrolyzed at a pH value of 2.1 in sulfuric
acid and lyophillized to dryness. The yield was 85 mg
of 3 0-3'-(n-propylamino)-D-glucopyranose.
Example 5
The 3-0-2'-~N' ,N'-dimethylaminopropyl) derivative
of 1,2:5,6-di-0-isopropylidene-~-glucofuranose was
prepared by condensing 0.1 mole of 1,2:5,6-di-O-isopro-
pylidene-D-glucofuranose with 0.3 mole of 2-chloro-N,N-
dimethylamino propane hydrochloride in the presence of
0.9 mole of sodium hydroxide in 150 ml of 1,4-aioxane.
The reaction mixture was fractionally distilled under
reduced pressure to obtain a yellow viscous oil (~oiling
point 142-145C/0.07 mm Hg) in 81% yield. The optical
..
rotation was {~}Ds = -21.5 neat and the refractive index
~as nnS = 1. 4549. Gas chromatography in accorda~ce witn
Example 1 indicated only one component.
The above prepared yellow viscous oil (lC g) ~as
hydrolyzed with aqueous sulfuric acid at a pH value of
al~ 25 2.0 by refluxing for 10 hours. The p~I value of the
hydrolysate was adjusted to 4-5 with saturated barium
~ ydroxide solution, filtered and lyophillized to obtain
,,';~ ~
, , ~ : . .
... : ,
: ' . .
1050429
10~5~g of light yellow crystals o 3-0-2'-(N',N'-dimethyl-
aminopropyl)-D-glucopyranose. The optical rotation in
water was l~}DS = +37.86. Gas chromatography in accor~- -
ance with Example 1 indicatea a purity in excess of 82%.
A portion of the oil, 1,2:5,6-di-0-isopropylidene-
3-0-2'-(N',N'-dimethylaminopropyl)-D-glucofuranose, is
partially hydrolyzed at pX 3.0 + 0.2 as indicated in
Example 1. A white crystalline hydrochloride salt is
obtained on lyophillization. The salt obtained is highly
hygroscopic, with gas chromatographic purity being of the
order of 80~.
Example 6
To 0.1 mole of 1,2:5,6-di-0-isopropylidene-D-
glucofuranose wac added 0.3 mole of 2,N,N-trimethylamino-
lS propyl chloride hydrochloride along with 36 g of sodium
hydroxide. ~he general reaction procedure was in accord-
ance with Example 1. The oil resulting from the reaCtiGn
had a boiling point of 144-146C at 0.6 mm Hg and an
optical ro~ation of {~}D = -20.05 neat.
The above product was hydrolyzed according to the
general method outlined in Example 1 to obtain the
desired 3-0-3'-(2',N',N'-trimethylamino-n-propyl)-D-
glucopyranose. ~he optical rotation of the product in
water was {~}Dq = ~38Ø
A portion of the oil, 1,2:5,6-di-0-isopropylidene-
3-0-3'-(2',N',N'-trimethylamino-n-propyl)-D-glucofuranose,
is partially~hydrolyzed at pY 3.0 + 0.2 accordiny to
3 3 -
. .
~(~S0429
the ~rocedure mentioned in Example 1. A white crystalline
1,2-O-isopropylidene-3-0-3'-(2',~',N'-trimethylamino-n-
propyl)-D-gluco~uranose hydrochloride ~as obtained which
is highly hygroscopic in nature. Optical rotation of
S the hydrochloride salt at pH 7.0 and 25C is -21.38.
Gas chromatography analysis indicated ~etter than 99
pure major component.
Example 7
Using the general method outlined in Example 1,
0.02 mole of 1,2:5,6-di-O-isopropylidene-D glucofuranose
in 1,4-dIoxarle was reacted with 0.0225 mole of 2-(2-
chloroethyl~-N-methylpyrrolidine hydrochloride and 0.0675
mole of sodium hydroxide. After 18 hours the solver.t
was removed and the resulting orange oil was vacuum
distilled under nitrogen. The residue consisted of the
: desired product, 1,2:5,6-di-O-isopropylidene-3-0-2'-
{2"-(N"-methyl)-pyrrolidyl}-ethyl-D-glucofuranose having
; an opiic~1 rotation of {a}DS = -22.95 in chloroform.
Example 8
1,2~5,6-di~O-isopropylidene-D-glucofuranose (0.1
mQle) and N-(2-chloroethyl)-pyrrolidine hydrochloride
(0.15 mole) are mechanically stirred and refluxed with
0.~5 mole of sadium hydroxide in 150 ml of tetrahydrofuran
for 18 hours. The tetrahydrofuran is removed from the
~i
reaction products and the resulting oil is vacuum dis-
tilled under nitrogen. The 3-0-2'-{N'-pyrroiidyl)-ethyl}-
; ~
34
.. ~ . . . .
., .
: ~ ,. ; . .
10504Z9
1,2;5,6-di-O-isopropylidene-D-glucofuranose derivative
has a boiling point of 165-171C/0.15 ~m Hg. Gas chrom2to-
graphy-indicates a purity o~ 99%. Using the hydrolysis
procedure outlined in Example 1, 10 g of the blocked
oil was hydrolyzed and lyophillized giving a white
hygroscopic crystalline solid. ;'~
Example 9
The N',N'-dimethylamino-n-pentyl derivative of
1,2:5,6-di-O-isopropylidene-D-glucofuranose is made by
condensing ~,~-dimethylamino-n-pentyl-5-chloride ;
hydrochloride with 1,2:5,6-~i-O-isopropvlidene-D-
glucofuranose in the presence of pulveri'zed so~ium
hydroxide in freshly puri_ied, dry 1,4-dioxane as
described in procedure in Example 1. The product was
confirmed by gas chromatography and infrared spectra.
N,N-dimethylamino-n-pentyl chloride hydrochloride ' ''
is made from commercially available sample of N,N-
dimethylamino-n-pentyl alcohol by treatment with thionyl
chloride (SOCl2). Specifically, 10.7 g of thionyl
chloride in a 250 ml three neck round bottom flask is
cooled in a salt-ice water bath and stirred vigorou~ly.
To the cooled solution is added, dropwise,'10 g of ~,N-
dimethylamino-n-pentyl alcohol. The reaction is exo-
thermic and temperature is carefully controlled. The
2S mixture is stirred for one hour after the evolution of
2 and HCl subsides. The mixture is brought to room.
temperature and allowed to stir overnig~t. Absolute
!: .
'- _ 35 _
.
- ~ .
. . ,
,;,
. , .. . ~ . , ,.. : . - . .
105042g ' .
alcohol is added to destroy excess thionyl chloride.
Ten grams of crude N,N-dimethylamino-n-pentyl chloride
hydrochloride is obtained as a ~Ihite solid. This is
used directly for the condensation reaction with 1,2:
5,6-di-O-isopropylidene-D-glucofuranose without further
purification. The alcohol and chloride can be resolved
on Chromosorb 103 gas chromatography column.
Example 10
Bromine (9.8 g) was added slowly and drop~ise to a
mechanically stirred mixture of 50 g cracked ice and a
chilled aqueous sodium hydroxide solution (7 g/20 ml water).
After the addition of bromine is complete, 15 g of 1,2:
5,6-di-0-isopropylidene-3-O-acetamido-D-glucofuranose
(prepared by the general procedure outlined in Example 1
...~
by the condensatlon of 1,2:5,6-di-O-isopropylidene-~-
glucofuranose with 2-chloroacetamide in the presence of
sodium hydroxide) is added in four portions 15 minutes
-
apart. The reaction mixture is heated for one hour in
a water bath. After this time an additional aqueous
solution of sodium hydroxide (20 g~20 ml) is added and
- heating is continued for another hour. The mixture is
cooled and extracted three times with ether. The ether
extract is dried over anhydrous ~.agnesium sulfate. he
` yellow hygroscopic solid remaining after evaporating OfL
; 25 the ether is the desired 1,2:5,6-di-0-isopropyli~ene-3-
. .
. . .
O-aminomethyl-D-glucofuranose derivative. The product
was identifie~ by the disappearance of the carbonyl
- 36 - ,
.
.
1050~29
stretching at 1670 cm~l found in the parent acetamido
compound.
E~am~le 11
Well established methodology of prior art was
employed to determine the antiviral potency of derivatives
of 1,2-0-isopropylidene-D-glucofuranose E~Cl against
poliovirus, type 1, and rhinovirus, type lA, in tissue
culture at 37C, employing HeLa cells with an agar over-
lay and WI-38 cells respectively. tSee ~allis, C., F. -
Morales, J. Powell~ and J. L. Melnick, Plaque enhancement
of enteroviruses by magnesium chloride, c~steine, and
pancreatin. J. Bacteriol. 91 1932-1935, 1966.) Polio-
virus cell injury was determined by the study of plaque
formation ard rhinovirus ~las examined for cytopathic
effect. In Table I, the virus inhibiting effects of
three concentrations of the 3-0-3'-(N',~'-dimethylamino-
n-propyl) derivative are depicted. The results are given
as the degree of inhibition of infectivity, identiLied as
plaque formation in the polio~irus system and as cytopatnic
effect in the system studying rhinovirus. Our results
indicate that, at the appropriate dose, drug can ccmpletely
inhi~it 1000 plaque forming units lPFU) of-poliovirus
and a 1000 TCIDso dose of rhinovirus-lA, a virus dose 1000
times that amount required to kill 50% of the tissue
; 25 cultured cells.
.
J ' : ,
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- 37 -
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- 38-
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- Example 12
Derivatives of 1, 2-0-isopropylidene-D- _
glucofuranos~ hydrochloride were examined for
their capacity to suppress influenza A 2 disease
in mice and for their capacity to suppress death
and nonlethal nervous system disease produced by the
encephalomyocarditis virus in mice. In these studies,
drug effect on lung pathology produced by a 15
ID50 dose of influenza virus was examined. This dose
is 15 times the dose that produces disease in 50% of
the animals. Disease and drug effect on disease were
; determined by lung weight increase and reduction thereof.
In the encephalomyocarditis study, 10 times the dose
capable of Xilling 50% of the animals was given, and the
degree of nonlethal disease and death were determined,
as well as drug inhibition of both of these parameters.
The results for these experiments are summarized in
'~ Table II, and indicate the product1on of significant
reduction in lung weight increase by drug, as well as a
~20 significant inhibition of death and nonlethal disease ~ ~ -
produced by encephalomyocarditis virus. These effects
were more potent for the 3-0-3'-(~ '-dimethylamino-n-
propyl) derivative than for the other two derivatives
studied.
'7, ~ ;` . ql
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e
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- 40 - .
.
.
5(~4;~9
Example 13
Human embryonic fi~roblast cells, WI-38 type, were
obtained from Microbiological Associates and grown in
. .
maintenance medium 199 with 1% fetal calf serum in the
presence and absence o~ either 2, 20, or 40 ug/ml of
1,2-0-isopropylidene-3-0-3'-(N',~'-dimethylamino-n-propyl)-
D-glucofuranose HCl at 37C and 34C. For cells grown at
37C, survival o~ tissue cultured cells in the absence of
drug diminished progressively through day 15, at which time
95% of cells failed to metabolize and no longer ~ormed a
monolayer. At 34C, by 72 hours 9S% of cells without
drug were no longer actively metabolizing as determined
by pH and a monolayer was no longer evident under micro-
scopic examination. Addition of drug in the concentrations
described above reduced this loss of viability by 75% at
15 days and 72 hours at 37C and 34C respectively.
Example 14
C57Bl/6J mice bearing transplanted melanoma B-16
were obtained from The Jackson Laboratories. This tumor
was transplanted into recipient C57BL/6J mice at 10 days
and the behavior of tumor growth in recîpient mice ~as
observed o~er two months. During this period, 90% of
oontrol animals died manifesting metastatic spread to
the liver, bowel and kidney. Animals treated with 80 ms/Kg
~5 per day 1, 2-0-isopropylidene~3-0-3"-(N',N'-dimethylamino- ,
n-propyl)-D-glucofuranose-~Cl, injected subcutaneously, `
' ~
exhibited significant central necrosis of primary tumors and
only 15% succumbed to such cancer metastasis.
41-
