CA1050429A - Therapeutic composition, novel compounds useful therein and method of using the same - Google Patents

Abstract

THERAPEUTIC COMPOSITION, NOVEL COMPOUNDS USEFUL
THEREIN AND METHOD OF USING THE SAME

ABSTRACT OF THE DISCLOSURE
The invention provides a novel therapeutic composition comprising a pharmaceutically acceptable carrier containing a therapeutically effective amount of an ethereal monosubstitution of a monosaccharide derivative having the general formula S-O-Y, wherein S is the residue of the monosaccharide derivative se-lected from the group consisting of pentoses, hexoses and heptoses as single or polysubstituted acetals or esters 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

Description

~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 ~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 : , ~

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 .
:;

.".

-- 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 - ~:
,~

. ~, .
.

'. ' ~ ' . :
~` ' ' ' . ' . " ' .

(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 -- .
., ~ ' ' : - . , ~, . ~ :- . ,:

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) (~) 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 ' ~, .~ .

~(~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 ..

..

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 '~ ~
; .

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 -. ~ ' . ~,~ ., ,. . . .

.

,. , . . . . . . . . ; ~ , 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.
' .

.

;, , 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~

. :.

- 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~

. ,~. .
' 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 ,,';~ ~
, , ~ : . .

... : , : ' . .

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 . .

~(~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}-; ~

.. ~ . . . .
., .

: ~ ,. ; . .

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 ' : , ` ' ' .~" ' ' -, . .... . .
`'`. ~: ' ;' . ~

.. , 1(~50429 C
o .o .
e~
, , - -.
U~ C . C
C
~ ~ ID
= V

~ , - . .
V ~ ~, ~
. <, . C . V
, V
. , ~ ., ~ ~ ,, ~ ,"
C ~1 G 1~ ~ ~ _ a: ~
~ ' z ~ ~
.~ G~ LL ~L ~ ~ ~ ~
. ~ V~ ~ ~ ~ CL ~ ~. ~
o _ o o o o o o 'z C ~ l ,~ 'O o _ ~ _ ~
,Co , O

1 ~- ~
c :~ ~ ~ ~
o C~: C~
L~l C _ _ _ -- -- O
E E E E E E . v ; ~:

-- O O ~l o o , C C
N ~

:~ ~ O - O E V
_- E - E- ~ ~ :
2 ~ Z ~D--- c ~ , ~ >~ - -O ~ ~ O ~ U~
oi E Q oi E Q _ ~-- ;
. . _~

' . , ~' ' -.-- .. ~ . , - -. .
:

~0504Z9 - 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 ,`.,~ ' . .
',,`-' ' "' -', ' , ' .

` ~ _3~_ ~

~ ~ ' . ' .

- 105()4Z9 , Ul va,~
I ~ ~ , ~I _ C ~ C ~ ~
V ,, ~ ~ _ . V _ J v 1-- v ~ ,~: D C1.
.; ~ C C V ~ G~ - oC _ O, V -- V C _ .-- V ~ .--.-- C -- O C ~ V C I V
'-- C ~ W C ~0 D ~.~ C >
~ c ~ a~ c -- al -- -- c --C~ ~ V ~ .VI ~ c .~n c --_C
v ~ 3 ~ ,c ~ ~ -- c v e O v v u '~ c C C
_ ~ c7 a~
L-~ c ._ ~ ~
_ ~ E 3 c~ > z ~ ~ ~ - ~
L~ ~ c- ~ o ~ v 2 V C V > ~
O _ Q~ ~ C c~ ~ .
~ ol ~ ~ o ~
~1 O Z . r-l ~10 V 'C ~ ~J ~ J
O Z L ~,c~ 0,o , ' ,:

V~ ~1 1_ ~ C~ V O : , b o _ ~ E :~

_ , c~ ~ ~

O E E E E c) ~ ~7~ > ~ E
.; . C:~ O O O O O O C C ~
~ co ~O CO ~D CO ~G -- -- ~ _ . _ C~

E - E - E-- - c -- O V o :~ ~r ~ o ~ c - ~ - ~ 7 ; ' : L ~ ~ o 1~ V o ~ ~ L o ~ c V ,~
. o l E Q o l E Q o l E O , l I C ' ~ V C
Z ~
.` :
:, ' ~ . ~

- 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.

Claims (17)

THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:

1. A novel therapeutic composition comprising a pharmaceutically acceptable carrier containing a therapeutic-ally effective amount of at least one substance selected from the group consisting of an ethereal monosubstitution of a mono-saccharide derivative having the general formula S-O-Y, and organic acid and inorganic acid salts thereof, wherein S is the residue of a monosaccharide selected from the group consist-ing of pentoses, hexoses and heptoses which has been derivatized with at least one substance selected from the group consisting of (a) at least one aliphatic alcohol containing 1-18 carbon atoms to produce an acetal group at the site of at least one available hydroxyl residue, (b) at least one aldehyde contain-ing 1-18 carbon atoms to produce at least one acetal group at the site of at least one available hydroxyl residue, (c) at least one ketone containing 1-18 carbon atoms to produce at least one ketal group at the site of at least one available hydroxyl residue and (d) at least one organic acid residue con-taining 1-18 carbon atoms to produce an ester group at the site of at least one available hydroxyl residue, and Y is selected from the group consisting of cyclic monovalent nitrogen con-taining organic radicals and residua and monovalent organic radicals and residua having the general formula , wherein Rl is a divalent organic radical having a linear carbon chain length of about 1-7 carbon atoms and R2 and R3 are select-ed from the group consisting of -H, -OH, -SH, halogen and mono-valent organic radicals and residua having a linear carbon chain length of about 1-7 carbon atoms, but excluding a composition containing 3-O-3'-(N',N'-dimethylamino-n-propyl)-1,2-O-isopropyl-ideneglucofuranose, 3-O-4'-(N'-methylpiperidyl)-1,2-O-isopropylidene-glucofuranose, 3-O-2'-(N',N'-dimethylaminoethyl)-1,2-0-isopropyl-idenaglucofuranose, 3-O-3'-(2',N',N'-trimethylamino-n-propyl)-1,2-0-isopropylideneglucofuranose, 3-O-2' (N',N'-dimethylaminopropyl)-1,2-O-isopropyl-ideneglucofuranose, 6-O-3'-(N',N'-dimathylamino-n-propyl)-1,2-O-isopro-pylidenegalactopyranose, 6-O-2'-(N',N'-dimethylaminopropyl)-1,2-O-isopropyl-idenegalactopyranose, 3-O-3'-(N',N'-dimethylamino-n-propyl)-1,2:5,6-di-O-isopropylideneylucofuranose, 3-O-4'-(N'-methylpiperidyl)-1,2:5,6-di-O-isopropyl-ideneglucofuranose, 3-O-2'-(N',N'-dimethylaminoethyl)-1,2:5,6-di-O-isopropylideneglucofuranose, 3-O-3'-(2',N',N'-trimethylamino-n-propyl)-1,2:5,6-di-O-isopropylideneglucofuranose, 3-O-2'-(N',N'-dimethylaminopropyl)-1,2:5,6-di-O-isopropylideneglucofuranose, 6-O-3'-(N',N'-dimethylamino-n-propyl)-1,2:3,4-di-O-isopropylidenegalactopyranose, 6-O-2'- (N',N'-dimethylaminopropyl)-1,2:3,4-di-O
isopropylidenegalactopyranose, -N',N'-dimethylamino-isopropyl-2,3:5,6-di-O-iso-propylideneglucofuranoside, and pharmaceutically acceptable organic and inorganic acid salts thereof.

2. A novel therapeutic composition in accordance with Claim 1 wherein y is , Rl is a hydrocarbon radi-cal having a linear carbon chain length of 1-3 carbon atoms, and R2 and R3 are selected from the group consisting of hydrogen and hydrocarbon radicals having a linear carbon chain length of 1-3 carbon atoms.

3. A novel therapeutic composition in accordance with Claim 1 wherein Y is selected from the group consisting of -(n-propylamino), -(N',N'-dimethylamino-n-propyl), -(N',N'-dimethylaminoisopropyl), -(N',N'-methyl piperidyl), -(N',N'-dimethylaminoethyl), -(N',N'-diethylaminoethyl), and -(2',N',N'-trimethylamino-n-propyl),

4. A novel therapeutic composition in accordance with Claim 1 wherein Y is -(N',N'-dimethylamino-n-propyl).

5. A novel therapeutic composition in accordance with Claim 1 wherein the said ethereal monosubstitution of the monosaccharide derivative has a general formula selected from the group consisting of:

(a) (b) (c) wherein X and Z are selected from the group consisting of H, OH, and monovalent hydroxyalkyl, alkoxy, and alkoxyalkyl radicals containing up to 3 carbon atoms, W is selected from the group consisting of H and monovalent alkyl, alkenyl, cyclic alkane, cyclic aromatic, and acyl radicals containing 1-18 carbon atoms, and Y represents the same organic radicals and residua as set out in Claim 1, and one of the said OH groups, X or z in said formula is replaced by -OY.

6. A novel therapeutic composition in accordance with Claim 5 wherein Y is is a hydrocarbon radi-cal having a linear carbon chain length of 1-3 carbon atoms, and R2 and R3 are selected from the group consisting of hydrogen and hydrocarbon radicals having a linear carbon chain length of 1-3 carbon atoms.

7. A novel therapeutic composition in accordance with Claim 5 wherein Y is selected from the group consisting of -(n-propylamino), -(N',N'-dimethylamino-n-propyl), -(N',N'-dimethylaminoisopropyl), -(N-methyl piperidyl), -(N',N'-dimethylaminoethyl), -(N',N'-diethylaminoethyl), and -(2',N',N'-trimethylamino-n-propyl).

8. A novel therapeutic composition in accordance with Claim 5 wherein Y is -(N',N'-dimethylamino-n-propyl).

9. A novel therapeutic composition in accordance with Claim 5 wherein the monosaccharide is a hexose.

10. A novel therapeutic composition in accordance with Claim 9 wherein Y is is a hydrocarbon radi-cal having a linear carbon chain length of 1-3 carbon atoms, and R2 and R3 are selected from the group consisting of hydrogen and hydrocarbon radicals having a linear carbon chain length of 1-3 carbon atoms.

11. A novel therapeutic composition in accordance with Claim 9 wherein Y is selected from the group consisting of -(n-propylamino), -(N',N'-dimethylamino-n-propyl), -(N',N'-dimethylaminoisopropyl), -(N-methyl piperidyl), -(N',N'-dimethylaminoethyl), -(N',N'-diethylaminoethyl), and -(2',N',N'-trimethylamino-n-propyl).

12. A novel therapeutic composition in accordance with Claim 9 wherein Y is -(N',N'-dimethylamino-n-propyl).

13. A novel therapeutic composition in accordance with Claim 5 wherein the monosaccharide is selected from the group consisting of glucose and galactose.

14. A novel therapeutic composition in accordance with Claim 13 wherein the glucose is monosubstituted in the 1-O- or 3-O- position and the galactose is monosubstituted in the 6-O- position.

15. A novel therapeutic composition in accordance with Claim 14 wherein Y is is a hydrocarbon radi-cal having a linear carbon chain length of 1-3 carbon atoms, and R2 and R3 are selected from the group consisting of hydrogen and hydrocarbon radicals having a linear carbon chain length of 1-3 carbon atoms.

16. A novel therapeutic composition in accordance with Claim 14 wherein Y is selected from the group consisting of -(n-propylamino), -(N',N'-dimethylamino-n-propyl), -(N',N'-dimethylaminoisopropyl), -(N-methyl piperidyl), -(N',N'-dimethylaminoethyl), -(N'-N'-dimethylaminoethyl), and -(2',N',N'-trimethylamino-n-propyl).

17. A novel therapeutic composition in accordance with Claim 14 wherein Y is -(N',N'-dimethylamino-n-propyl).