Note: Descriptions are shown in the official language in which they were submitted.
86a~S
;. ......
The invention relates to medicaments based upon adenosine
`5'-thioethers having especially antiviral and antitumoral
actions.
The methylation of proteins, nucleic acids and numerous
metabolites is a well-known reaction, in the course of which
specific enzymes, called methylases or methyl-transferases,
eEfect the transference of a methyl group supplied by S-
adenosyl methionine at certain precise points of the sub-
stratum, especially nitrogenised bases or sugars intervening
in the constitution of the nucleic acids.
It has recently been shown that tumoral tissues contain
higher proportions than normal tissues of methylated nucleic
-bases. In particular it has been noted that there is a
relationship between the hypermethylation of the nucleic acids
and the transformation of normal cells into malignant cells.
.
In correlation with this phenomenon, it has recently become
known that the messenger RNA of some tumorigenous viruses,
contains specific methylases.
It is known that research workers are already inter-
ested in the study, especially in VitYo~ of the regulation
of the methylases by natural or synthetic inhibitors. One
; known natural inhibitor is constituted by 5'-S-adenosyl-L-
homocysteine 1', hereinafter designated by the abbreviation
SAH, which is produced on the reaction of transference of
the methyl group supplied by S-adenosyl methionine; SAH is
itself a powerful inhibitor of methyl-transferases, especi-
ally in vitro. These same research workers have already
had the idea of effecting the synthesis of a certain
number of analogues of S-adenosyl-homocysteine and of
testing their inhibiting action towards methyl-transferases
. .
. ,~'~ ' .
,
- ,. ;
:3 ~686;QS
-
: in vitro . All the synthetic inhibitors which they have
described have proved much less active th~n SAH used under
the same conditions, if not inactive .
The invention is based upon the dlscovery that at
least certain of these analogues of SAH were on the contrary
ef~ective and had a methylase-inhibitor li]ce activity in vivo
with respect to methylases,more particularly the specific or
non-specific methylases of the oncogenous viruses. This dis-
covery was the more unexpected as SAH studied under the same
conditions is substantially in~ctive or inactived, perhaps by
intracellular hydrolysis or degradation.
Thus the invention concerns more particularly the
application as inhibitors of the oncogenous transformation of
tissues,(that is the transformation which induces the formation
of tumors, -o~ viral i~fections and espPcially those caused
by oncogenous viruses, of cellular divisions induced
by mitogens, of substances responding to the following formula :
NH-X
N _
~ I -
:- - R - S - CH2 . N ~ N~5~ ~.
~ /
'' \l ~/ ''
~.
OH 1H
wherein X is hydrogen or a benzoyl group and R is a
R
21r R2
. . .
- . . . . . .
~06~0S
with either R1 or R2 being an atom of hydrogen, a hydrocarbon
group comprising from 1 to 7 atoms of carbon, an OH, SH, CH2SH,
COOH, COCH3, NH2, NHCONH3 or SCH3 group; it also being possible
for R1 and R2 to form a phenyl or pyridyl ring with the CH .
group on which they are fixed; n is equal to O or 1.
A particular class of the compounds as above defined
is that wherein X is hydrogen and R is a -(CH2) - CH ~ 1 group
with R1 being 1 atom of hydrogen, a hydrocarbon group
comprising 1 to 7 atoms of carbon, an CH, SH, CH20H, CH2SH, ~ ~ .
COOH, COCH3, NH2 or NHCONH3 group and R2 being an atom of ~
hydrogen or an OH, SH, CH20H, CH2SH, C30H, COCH3, NH2 or
NHCOCH3 group; it also being possible for R1 and R2 to form a
phenyl ring with the CH group on which they are fixed; n
is equal to O or 1 .
: Compounds of particular interest by way of agents ~ .
inhibiting oncogenous viruses present in tumorous tissues are ; :~
those in which the radical R is an alkyl group comprising from
1 to 5 atoms of carbon, and possibly too at least one of the
following functional groups : - -NH2, -SH~ - COOH
Compounds of this type which contain also a SCH3 group
are also of particular interest~ .
Standard.examples of these inhibiting agents are those .
in which R is a - CH2 - CH2 - NH2 ; ~ CH2 - CH2 - SH;
- CH2 - CH - NH2 group.
COOH
. . .
Particularly interesting products are constituted by ....
` the S-isobutyl adenosine derivatives of formula :
. - . . .
~ ~
.' ~ '.
'~ .
.1 . .
:` ~'.,
. . . . .. .. . . .
10~136~S
-
NH - X
~ CH~- C~2 _ 5 C}12
: O O
wherein isas defined aboveO
The product in which X is hydrogen (hereinafter
: designated under the abbreviation SIA) is characterised by
a particularly high inhibiting activity, while being of a
remarkable innocuousness.
Further examples of inhibitors are-also those in
which R has one of the following significances
CH3
CH2 ~ CH2 - CH2 - CH3
~CH2 CH2 - CH2 - CH2 - CH3
tCH2)6 ~ CH3
CH2 c6~
~` CH2 CH(NH2) - COOH
CH2 - CH2 - NH2
CH2 - CH2 - NHCOH3
CH2 - CH2 - CH(NHCOCH3) - COOH ;
'! . :
, CH2 - CH20H ' ~:
CH2 - CH2 - COOH
CH2 - CH(OH) ~ CH20H :
m - pyridyle
CH2 S CH3
; .
~, .
- : - . . , ~. . , .:
o~ :~
CH2 - CH(NH2) - COOH (isomer D)
CH(CH3) - CH2 - CH
CH2 - CH(OH) - CH20H
C~12 - CH = CH2
The inhibiting activity of the said substances is
evidenced by a prevention of the transformation of normal
cells into mali~nant cells or, when the latter have already
developed, by an interruption of their multiplication. Admitted-
ly it is observed that the a~ents according to the invention
can likewise induce a retardation of the multiplication of
normal cells. However it is noteworthy, as evidenced by the
results of the pharmacological tests which will be described ;
hereinafter, that this effect is reversible as regards the
development of the normal cells, but irreversible as regards
the malignant cells.
Thus these properties make the agents according to
the invention acti~e principles of great value, utilisable in ;
the constitution of medicaments ~or the prevention or treat-
ment of malignant tumours and viral inections .
The majority of the compounds responding to the
above-indicated general formula has already been described.
It will also be recalled that several processes have been
described for their preparation.
- It will be recalled, as reminder, that these products ~-
can be obtained especially :
-by displacement o~ the p-toluene sulphonate group at 5' of
! ~ .
2', 3'-0-isopropylidene 5'-0-p-toluene sulphonyl adenosine~
~` with the aid of a thio-alcoholate of formula RSH, in which
R has the significance indicated above, especially within
dimethyl formamide or liquid ammonia (known as Baddiley's
method )
;`, ..
'
.. . . . .
- . : .. : . ... .. .
.
. .
,. . . , . , .: . ' . ~ .: ,
:~Q16t3605
- -by direct halogenation of the C5' of adenosine ribose by
thionyl chloride in hexamethyl phosphorotriamide, the obtained
5'-deoxy-5'-chloro-adenosine then being placed in contact with
the corresponding thio-alcoholate RSH within an aqueous
solution of sodium hydroxide (Kikugawa's method). Such
methods were recalled and developed in the doctorate thesis by
Michel Legraverend, maintained the 10th February 1975 at the
Orsay Centre of the Paris-South University and entitled "study
of the enzymatic methylation of ribonucleic acids of in v~tro
transfer, inhibition of an N2 guanine methyl transferase and
synthesis of new inhibitors".
Some of the compounds falling within the scope of
the general formula, though not disclosed 'per se r in the
prior art, are homologues of the known compounds encompassed
by the general formula. For example, as illustrative of such
compounds there may be mentioned the case where R is
-CH2-S-CH3 ~'-S-deoxy(methylthiomethylthio)adenosin ~ which
is prepared from the 5'-deoxy-5'-chloro-adenosine according to
the procedure described previously and which melts at 98-99C,
R is -CH(CH3)-CH2-CH3 melting at 93-95C and the N(6)-benzoyl-
SIA. This latter compound can be prepared as follows:
the specific benzoylation of the nitrogen at the ~ -
6 - position on the nucleus of adenosine is carried out
.:
according to the method described by RANGANATHAN et al.
(J. Org. Chem, 39, (1974), 290).
10 mM of distilled benzoyl chloride are added drop-
wise to a 30 ml pyridine solution containing 3 mM of SIA atOC.
The solution is stirred for 2 hours at ambient temperature and
`' ' '
- 7 -
,~
- . ~ . : ... ~ : . . . . .
:
361~i
.
thereafter 40 ml of NaOH 2 N are added to said solution.
After standing for one hour the reaction mixture is acidified
at 0C by adding acetic acid thereto. The solvents are evapo-
rated and the product extracted with chloroform. 1'he organic
phase is scrubbed successively with a sodium bicarbonate
solution, with HCl l N, with water, then dried on Mg SO~ and
finally evaporated;.
A chromatographically pure compound is obtained :
Rf = 0.82 ( chloroform/methanol 8/2)
MP = 210 - 21~C.
:, /
/
.''` - / '~ .
~;
- / ' ~
~ / '
'
~'` /
`; ,
/
' / .,
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: , , ,: ~ ~ : -:
.
' . ~ ' ' ' ' ~ . , :, ,
`~
~068~0~i
Further characteristics of the invention will also appear in
the course of the following description of pharmacological tests
carried out with the above-defined substances. In the course of
this description reference will be made to Figures 1 to 5 of
the drawings, which illustrate diagrammatically the effects
produced by the said substances under the conditions which will
be explained later.
" lo Effect of SIA and SAH upon the growth of chicken embryo
fibroblasts, respectively healthy and infected by Rous's
Sarcoma virus.
~
a) Effects of SIA and SAH upon the growth of normal chicken
embryo fibroblasts.
This effect was studied under the following conditions.
Embryo fibroblast cells were seeded in HAM F-10 medium
(described in the article by R.G. HAM Exp. Cell. Res., 29,
515-526 (1963)), in Falcon boxes of 60 or 35 mm. diameter, at
the rate of 3 x 105. The cells adhere to the bottom of the -~
box. Each time a count must be effected, the cells are
detached with the aid of a 0.12% trypsin solution. The count
is effected with the microscope with the aid of a haemocytometer.
The inhibitor utilised is introduced at the beginning of ;
the experiments into the medium at the desired study concen-
trations. The medium is renewed on the second day and the
fourth day with the inhibitors. On the fifth day the inhibitors
are eliminated by repeated renewal of the medium.
The curves 1, 2 and 3 in Figure 1 respectively represent
the multiplication of the cells (expressed on the ordinate axis
by the number N of cells) as a function of time (expressed in
days on the abscissae axis) in control cultures (curve 1), in
the presence of SA~ at a concentration of 1 millimol (mM)
.' '
.
. ' :
.: . ,
~86Q5
(curve 2) and in the presence of SIA at a concentration of
1 mM (curve 3).
Figure 1 shows that SIA and SAH retard the growth of
normal cells. However this effect is reversible, even after
5 days. The growth in fact resumes rapidly/ as soon as the
inhibitor has been eliminated.
b) Inhibition of the viro-induced transformation of the cells
by the 5'-thioethers of adenosine.
; System studied: chicken embryo fibroblasts infected by
Rous's Sarcoma virus (RSV).
The culture medium is constituted by the HAM F-10 medium
containing 5% o~ calf serum.
Chicken embryo fibroblast cells are seeded into the above-
indicated medium, in Falcon boxes of 35 mm. diameter, at the
rate of 106 cells per box. These cultures are infected by a
:~ dilute virus suspension permitting the obtaining of 60 to 100
centres of transformed cells per box. The inhibitor is added
for a duration of 48 hours, either immediately after the
infection (series A), or 2 and 4 days after the infection
. 20 (series B and C). After 48 hours of contact the medium is
` changed to eliminate the inhibitor and the centres formed are
- counted 7 days later. During this time the medium is renewed
periodically. -
`~ The results appear in Table I below. In the left-hand
column the inhibitors and the utilisation concentrations of
these inhibitors are indicated. The numbers of centres and
the inhibition percentages observed for the series A, B and C
-~ are respectively given.
-- 10 _
,
~6~6QS
TABLE I
Effect of SIA and SAH upon the viro-induced (RSV) trans-
formation of chicken embryo fibroblasts
. .... ____
Number of centres ~ inhibition
Control 72
SA~I 1.0 mM 58 19.5 )
SIA 0.5 mM 23 68 ) series A
SIA 1.0 mM O 100
Control 110
SAH 1.0 mM 112 O )
SIA 0.5 mM 60 45 ) series B
SIA 1.0 mM 6 95 )
Control 72
SAH 1.0 mM 60 17 )
SIA O A 5 mM 2 97 ) series C
SIA 10 mM O 100 )
The examination of this table shows that SIA is a very
powerful inhibitor of the oncogenous transformation of chicken
embryo fibroblasts. At a concentration of 1 mM, SIA completely
` 20 inhibits the malignant transformation, in irreversible manner.
This inhibition remains irreversible at least for 7 - 8 days.
` SAH, a natural inhibitor of transmethylases, studied for ;
comparison, hardly inhibits the transformation, probably by
reason of its degradation (intracellular hydrolysis). The -
following table shows the results obtained with analogues of
SIA.
In the following Table II there appear the results obtained
with analogues responding to the general formula indicated
above. They are identified in the table by the significance
of their R group. Moreover this table indicates:-
the concentration of each inhibitor utilised,
the duration of contact of the inhibitor with the culture,
the percentage of inhibition of the oncogenous transformation
: of the chicken embryo fibroblast.
- 11 -
'`' '' ',:
~96~60S
TABLE II
Activities of various analogues upon the chicken embryo
fibroblasts infected by Rous's virus
Group R of Conc. ~ime of ~O lnhlbition
the analogue contact of the trans-
formatlon
-CH3 0.5 mM 24 hours 70
,. 0.5 mM 48 " 95
-(CH2)2 - CH3 0.5 mM 24 " 44
ll 0.5 mM 48 " 83
-(CH2)3 - CH3 0.5 mM 24 " 62
0.5 mM 48 " 86
CH2 - CH - NH2 1. mM 48 " 87
CH2 - CH2 SH 0.5 mM 24, " 26
0.5 mM 48 " 84
1. mM 48 " 96
COOH
-CH2 - CH - NH2 1. mM 48 " 76
-CH2 - CH - COOH 1. mM 48 " 58 . :
-(CH2)6-CH3 0.1 mM 48 " 33
-CH2 ~ C H 0.1 mM 48 " 36 ~ .
1. mM 48 " 98
0.5 mM 48 " 93
-CH2 - S - CH 0-5 mM 48 " 100
0.25 mM 48 " 95
-cH2-cH(NH2)-cooH 0.5 mM 48 " 70 ~: ;
(D-isomer) .
( 3) C2H5 0.5 mM 48 " 99
-CH2-CH(OH)-CH2OH 0.5 mM 48 " 70 ;~
-CH2-CH=CH 0.5 mM 24 " 99
-cH2-cH-(cH ) 0.5 mM immediate 99
. _ . .. __
- 12 -
' ' .'. . " ' :: :
, . ' ~ ,"'`""-:;'.
.'`' ~ ' : .
.~ S
X is hydrogen except the last one (~) (in all the compounds
listed in table II in which X is a benzoyl group).
This latter table shows the fact that all the tested
substances exert a significant inhibiting action upon the
oncogenous transformation of chicken embryo fibroblasts.
2. Effect of S-isobutyl - adenosine upon the oncogenous
transformation of mouse cells by the Sarcoma Murin virus
(M-MuSV)
. .
The experiments were e~ected ~pon mo~se ~ibroblast
cells (~Lg cells) seeded the previous evenin~ upon H~ medium
in Petri boxes (5.104 cells/35 mm. Petri box); these cells `~
are placed in contact for 2 hours with the M-MUSV virus,
then returned into HAM medium containing or not containing the
substance to be tested, in the present case SIA. The cells
remain in contact with the inhibitor for 24 or 48 hours. The
: : ~'
inhibitor is eliminated by changing of the medium, and the
centres formed are counted 3 days after the eli~ination of the
inhibitor. The results are indicated in the following Table ~-
III, in which there appear the doses of inhibitor utilised, the
contact time and the number of transformed centres.
~ '
,, .
.:
.'`~ .'' , ,~:
: ..
~ ` ' ' ,
- 12 bis - ~
. ' . ''' : . ' ' ':
~!~6~6~S
~`
:`
TABLE I I I
;
. .... __ __
SIA Contact time Number of centres
transformed
._ ....... _.__
0 24 hours Confluent centres
2 mM ,~ 148
_ ':
,~ 0 48 hours Confluent centres
0.5 mM ,l 218
mM __ 18 :~
One notes the proliferation of the malignant cells in
the control culture, whereas SIA very efficaciously inhibits
the malignant transformation of the mouse cells by the Sarcoma
Murin virus.
3. Influence of SIA upon intracellular macromolecular
synthesis.
.. .. _ _
; The foregoing test have permitted of demonstrating
the fact that SIA had the effect of arresting cellular
division, this effect being both particularly vigorous
and irreversible in relation to cells having suffered an
oncogenous transformation. This effect can likewise be
.. ~ ~ . .
demonstrated by the study of the action of the inhibitor ~ -
upon macromolecular synthesis. This action was demon-
strated upon cultures of normal cells and transformed
''''
.
.'~ ` .
.
`: - 13 -
..
-
~L~68~0S
. cells under the conditions set forth under 1.) above, on the
occasion of the study of the effect of adenosine 5'-thioethers
upon the growth of chicken embryo fibroblasts, by isotopic
!. marking of these cultures, by incorporation of [3H] leucine, of ;:
[3H] thymidine and of [3H~ uridine respectively i.n normal and
tran.sformed cells of different cultures, of control cells on the
one hand and of cells cultivated in the presence of SIA, for ~ ~ .
durations respectively of 24 and 48 hours, on the other.
The results of these tests are the subject of the graphic
representation in Figure 2, which shows the percentages of
incorporation in the cells of the different cultures~
of [3H] leucine (results grouped under bracket A),
of [3H] thymidine (results grouped under bracket B),
: of [3H~ uridine (results grouped under bracket C).
The results grouped under the brackets n concern normal cell
cultures, those grouped under bracket t cultures of transfo.rmed cell.
The vertical elongated zones in solid lines (the length of
which is proportional to the percentage of incorporation of the .
marker) designated by the reference numerals 1 concern the control .
.......... cultures, without inhibitor; the solid line zones designated by the .
reference numerals 2 concern the percentages of incorporation of
the marker in the cultivated cells, maintained in the presence of
1 mM of SIA for 24 hours, and the zones 3 are representative of
the percentages of incorporation of the marker in the cultivated
. cells, maintained in the presence of lmM of SIA for 48 hours. .
.. The dotted line zones are representative of the percentages
of incorporation of the marker when the latter has been introducedinto the cultures 24 hours after the elimination of the inhibitor,
at the concentration and after the contact durations which were
indicated above.
`~ '~' .
. .
. - 14 -
6~6~i
Figure 2 shows clearly that in the normal cells the incorpo-
rations of [3H] leucine, [3H] thymidine and [3H~ uridine are
inhibited substantially in the same manner in the presence of
SIA. However this effect is reversible. In fact a resumption
of the macromolecular syntheses is noted as soon as the inhibitor
is eliminated.
In the presence of the inhibitor, it is noted that the same
doses of SIA exercise a greater effect of inhibition of the
development of the transformed cells than that of the normal
cells, and above all that the maintenance of the inhibitor in
contact with the transformed cells for a sufficient time,
especially for 48 hours, involves an irreversible inhibition of
all the incorporations in the transformed cells, that is the
arresting of the macromolecular syntheses (especially of the
proteins, of the deoxyribonucleic acids and of the ribonucleic
- acids, to which there correspond respectively the three markers
utilised, namely [3H] leucine, [3H] thymidine and [3H] uridine)
and consequently arresting of the cellular division.
; ~. Demonstration of the antimitogenic properties of
adenosine 5'-thioethers.
The description of the following tests aims to demonstrate
the antimitogenic properties of adenosine 5'-thioethers, of which
SIA is representative. The antimitogenic effect of the tested
substance results from the capacity which it possesses for
inhibiting the mitogenic action of substances known as having
this effect in relation to lymphocyte cultures. The results
obtained in relation to mouse spleen lymphocytes, rabbit spleen
lymphocytes and human lymphocytes are the subject of the graphic
representations in ~igures 3, 4 and 5 respectively.
- 15 -
' :~
:
,
0~86~)S
` The lymphocyte cultures were effected in accordance with
the technique described in the article by C. Bona et Coll.
published in "The ~ournal of Immunology, vol. 112, No.6, June
1974, 2028" and in the presence of the mitogenic agents identified
below and of the substance to be tested, in the present case SIA,
at two different concentrations : 0.1 and 0.3 mM, for 48 hours as
regards the mouse spleen lymphocytes (Fig. 3), 72 hours as regards
; the rabbit spleen lymphocytes (Fig. 4) and 120 hours as regards
the human lymphocytes (Fig. 5). In each case cultures of these
same lymphocytes are effected in the presence of the mitogenic
agents alone (control cultures). The mitogenic or antimitogenic
effect according to cases of the mitogenic agents and of the SIA
is set forth by the radioactivity of the lymphocytes resulting
from the incorporation of tritiated thymidine therein.
The utilised mitogenic agents are the following:-
Con A : Concavaline A : T mitogenic in mouse and rabbit, T and B
~ in man.
;I PHA : Phytohaemagglutinine : T mitogenic in mouse and rabbit,
T and B in man.
ALS : Antilymphocytary serum : T mitogenic in man.
PPD : Tuberculin : specific mitogenic of T cells in the case where
the subject has previously been sensitised by mycobacteria.
NWSM : Hydrosoluble mitogenic agent extracted from Nocardia cells:
mitogenic of B cells in mouse and rabbit.
Anti-B4 : Serum directed against the genetic determinants of the
light immunoglobuline chain of rabbit, carried by the
: B cells of rabbit: thus it is a B mitogenic of rabbit.
` PG : Peptidoglycan of E.coli : B mitogenic for mouse and rabbit.
LPS : Lipopolysaccharide of Salmonella enteridis :
B mitogenic for the mouse.
Lip. A : Lipid A or mitogenic principle of LPS.
- 16
: ~'
0~l~36~5
The doses expressed in microgrammes (~), or in dilutions
as regards ALS or Anti-B4 serum, of the antimitogenic agents used
in each of the described tests are indicated in the following
table, respectively, in connection with the cultures of mouse
spleen lymphocytes, rabbit spleen lymphocytes and human
lymphocytes:
Mouse Rabbit Human
spleen spleen lymphocytes
lymphocytes lymphocytes
Con A 3 ~ 5 ~ 5 ~
PHA 0~2 ~ 1 ~ 1 y
ALS - - 1/100
PPD - - 10 r ~
NWSM 10 ~ 50 ~ 100
Anti-B4 - 1/50
PG 50 ~ 100 ~ - -
; LPS 10 ~ - -
Lip. A 10 ~
The results are grouped in Figs. 3 to 5 for each selected
mitogenic agent. The lengths of the elongated zones are pro-
` portional to the measured radioactivity R (strokes per minute
multiplied by 10 3). The zone simply defined by a solid line
corresponds to the results obtained in the presence of the -
mitogenic agent alone, the zone hatched with diagonal lines to
- the results obtained in the presence of the mitogenic agent and
SIA at a concentration of 0.1 mM and the zone hatched with
horizontal lines to the results obtained in the presence of the
mitogenic agent and SIA at a concentration of 0.3 mM.
The continuous horizontal line represented in each Figure
corresponds to the incorporation of the marker in control
cultures, in the absence both of the mitogenic agent and of SIA.
The incorporated radioactivities measured for cultures of lymphocytes
in the presence of SIA alone have also been represented. The
- 17 -
' ` -
36~)5
'.~ '::,
corresponding results are indicated above the abbreviation SIAwhich is to be found in each of Figs. 3 to 5.
The drawings show the remarkable antimitogenic properties
of SIA. A notable inhibition is observed of the mitogenic action
of the selected mitogenic agents, in most cases for an SIA
concentration of 0.1 mM. In all cases it is very great at a
concentration of 0.3 mM, from which it results that adenosine
5'-thioethers not only inhibit cellular division, but likewise
counteract effectively the action of mitogenic agents.
5. Toxicity tests
a) Acute toxicity.
Three batches of five mice, weighing 20 to 25 g., were the
object of these tests. SIA in suspension in physiological serum
was administered to them in one single intraperitoneal injection.
The doses administered to the mice of the first batch were lO0 mg.
per kilo, to those of the second batch 200 mg. per kilo and to
those of the third batch 500 mg. per kilo.
The mice were examined after 1 hour, 2 hours and 24 hours.
Mortality was zero, even after several weeks.
::
b) Chronic toxicity.
These tests were effected upon five control mice and
fifteen treated mice. -
The controls received physiological serum intraperitoneally,
at the rate of l ml. per lO g. of weight. The other mice received
.. .
l ml. of a suspension containing l mg. of SIA (in physiological ~
. .
serum). The injections were effected in the rhythm of 30
injections in two months.
After the 28th injection, one single dead mouse is found, ~
probably rather by reason of the trauma caused by the injection '
than by the content of the injected suspension.
- 18 -
.
~L~6~il6~5
Thus, these results display a remarkable innocuous-
ness of SIA in mice.
The adenosine 5'-thioethers constitute active prin-
ciples of value for the constitution of medicaments utilisable
for the prevention or treatment of malignant tumours and viral
infections. Their major antimitogenic properties also make
them valuable active principles for the constitution of me-
dicaments for the treatment or prevention of leukaemias and
myelomas.
The invention is thus concerned with pharmaceutical
compositions containing the compounds as defined hereabove,
particularly an effective dose thereof, in association with a
pharmaceutically or physiologically acceptable vehicle.
Particularly the invention is concerned with compo-
sitions in which the active compounds are associated with
solid or liquid physiologically acceptable carriers suitable
for oral administration, or in which they are associated with
carriers suitable for their administration by the rectal route.
~ According to a preferred embodiment of the invention,
said compositions are administrated by the parenteral route.
The invention is concerned in particular with liquid composi-
tions containing at least one of the compounds as above defi-
ned associated with an injectable, sterile liquid vehicle~ -
As is self-evident and as moreover already appears
from the foregoing, the invention is in no way limited to
those of its manners of application and realisation which have
been more especially envisaged; on the contrary it includes
all variants thereof .
- 19 _
