Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
2~931~
The pre~ent invention relates to the use of known 7-oxo-
7H-pyrido~1,2,3-de]-~1,4]benzoxazine-6-carboxylic acids
and e~ters a~ antivirally active medicaments.
The compounds to be used according to the invention are
known from ~P-A-253,235 either as anti-bacterial active
compound~ or as intermediates for ~heir preparation.
It has now been found, surprisingly, tha~ the compounds
li~ted below
9,10-difluoro-3-methyl-7-oxo-7H-pyrido~1,2,3-de]-
[1,4]benzoxazine-6-carboxylic acid
9-fluoro-3-methyl~10-(4-methyl-1-piperazinyl)-7-oxo-7H-
pyrido~l,2,3-de]-~1,4~benzoxazine-6-carboxylic acid
9-fluoro-3-methyl-10-~1-piperazinyl)-7-oxo-7H-pyrido-
[1,2~3-de3-[1,4]benzoxazine-6-carboxylic acid
9-fluoro-3-methyl-10-(3~methyl-1-piperazinyl)-7-oxo-7H-
pyrido[1,2,3-de]-~1,4]benzoxazine-6-carboxylic acid
9-fluoro-10-t4-methyl-1-piperazinyl)-7-oxo-7H pyrido-
~l~2~3-de~-tl~4]benzoxazine-6-carboxylic acid
ethyl 9,10-difluoro-2-methyl-7-oxo-7H-pyrido~1,2,3-de]-
[1,4]benzoxazine-6-carboxylate
9,10-difluvro-~-methyl 7-oxo-7H-pyrido~1,2,3-de]-[1,4~-
benzoxazine-6-carboxylic acid
9-fluoro-2-methyl-10-(4-methyl-1-piperazinyl) 7-oxo-7~-
pyrido[l,2,3-de]-[lr4]benzoxazine 6-carboxylic acid
ethyl ~,10-difluoro-7-oxo-2-phenyl~7H-pyrido[1,2,~-de]-
: .
.:: ~ - 1 -
~ ' ' ' .,
' t
~vg~o~
r 1, 4]benzoxazine-6-carboxylate
9,10-difluoro-7-oxo-2-phenyl-7H-pyridotl,2,3-de]-
[1,4~benzoxazine-6-carboxylic acid
ethyl 6~7-difluoro-9-oxo-9H-cyclohexa[1,2-b]pyrido-
~1',2',3,-de]-[1,4]benzoxazine-10-carboxylate
pos~e~s strong antiviral activity and are thus suitable
for use in combating viral diseases, in particular
hepatitis ~.
Preferably used in combating viral diseases, in part-
icular hepatitis B, are
9-fluoro-3-methyl 10-(4-methyl-1-piperazinyl)-7-oxo-7H-
pyrido[l,2,3-de]-tl,4]benzoxazine-6-carboxylic acid
9-fluoro-3-methyl-10-(1-piperazinyl)-7-oxo-7H-pyrido-
[1,2,3-de]-[1,4]benzoxazine-6-carboxylic acid
9-fluoro-3-methyl~10-(3-methyl-1-piperazinyl)-7-oxo-7H-
pyrido[1,2,3-de]-~1,4]benzoxazine-6-carboxylic acid
9-fluoro-10-(4-me~hyl-1-piperazinyl)-7-oxo-7H-pyrido-
~1,2,3-de]-~1,4~benzoxazine-6-carboxylic acid
9-fluoro-2-methyl-10-(4-me~hyl-l-piperazinyl~-7-oxo-7H-
pyrido~1,2,3-de]-[1~4]benzoxazine-6-carboxylic acid
ethyl 9,10-difluoro-2-methyl-7-oxo-7H-pyrido~1,2,3-de]-
[1,4]benzoxazine-6-carboxyla~e : ~
ethyl 9,10-difluoro-7-oxo-2-phenyl-7H-pyrido~1,2,3-de]-
: tl,4~benzoxazine-6-carbQxylate
: 25 9,10-difluoro-7-oxo-2-phenyl-7H-pyrido[1,2,3-de~-
[1,4~benzoxazine-6-carboxylic acid.
Particularly preferably u~ed in combating viral di~eases,
e A 28 862 - 2 -
20~3~6
in particular hepatitis B, are
9-fluoro-3-methyl-10-(4-methyl-1-piperazinyl)-7-oxo-7H-
pyrido~l~2~3-de~-[l~4]benzoxaæine-6-carboxylic acid
9-fluoro-3-methyl-10-(1-piperazinyl)-7-oxo-7H-pyrido-
~1,2,3-de3-~1,4]benzoxazine-6~carboxy1ic acid
9-fluoro-3-methyl-10-(3-methyl-1-piperazinyl)-7-oxo-7H-
pyrido~l,2,3-de]-[1,4]benzoxazine-6-carboxylic acid
ethyl 9,10-difluoro-2-methyl-7-oxo-7H-pyrido[1,2,3-de]-
[1,4]benzoxa~ine-6-carboxylate.
The compounds according to the invention which are listed
above can be prepared according to the processes which
are published in EP-A-253,235.
Demonstration of activity was carried out in hepatoma
cells (HEPG 2.2.15~ which were transfected with hepa-
titis B virus DNA. The results from the examples listed
below were obtained using the HBV test system described
in the following literature reference [Sells, M.A.;
Chen, ~.L., Acs, G., Proc. Natl. Acad. Sci. USA
pp. 1005 1009, vol. 84 (1987)]:
Transfected hepatoma cell (HEP G2.2.15) were incubated
with different concentrations of the particular compound.
By treatment of the transfected hepatoma cell line HEP
G2.2.15 with the compounds accordiny to ~he invention the
appe~rance of virus-specific HBV DNA in the supernatant
could be demonstrated as could a reduction in the le~el
of NBsAg. The content o:E ~W DNA ln the supernatant of
:~'
ke A 28 862 _ 3 _
2~931~6
the cell cultures was determined following precipitation
with PEG. Thi~ was effected with the use of a sample of
HBV genomic DNA which was not radioactively labelled
[Pauly, P., 1982, Ph.D. thesis, University o~ Gottingen,
F.R.G.; Xochel et al., 1990, EM~L, Accession No.
X 51790]. A reduction of extracellular HBV-DNA as well
as a decrease in the amount of HBV specific replicative
intermediates was observed. At the same time
demonstration of the effect on HBsAg formation took place
using a commercially available Elisa test. The indicated
IC50 values refer to the substrate concentrations which,
under the abovementioned test conditions, produce a 50 %
inhibition of the HBV DNA concentration in the
supernatant.
Table A:
~_
'`~ ~ 10 5~
3 0.1 > 100
4 0.1 50
6 0.5 - 100
~ At the same time, determination of~any cytotoxic effect
: of the compounds according to the invention ~hat might be
present was carried out by staining with crystal violet
?0 or by incorporation of radioaceively labelled thymidine
into the cellular DNAo
The effect of the compounds according to the in~-ention on
Le A 28 862 - 4 -
2~9~106
other cells was tested by incubation with HEL cells and
MEF cells. No effect on the vitality of these cells was
observed up to a concentration of 250 ~M.
As examples of areas of indication for the compounds
which can be used according to the invention, there can
be mentioned:
The treatment of acute and chronic viral infections which
can lead to a hepatitis, for example infections with
hepatitis viruses; additionally viral infections with
herpes viruses types 1 and 2, cytomegaloviruses,
varicella-zoster viruses and HIV.
Particularly preferred is the treatment of chronic
hepatitis B viral infections and the treatment of acute
hepatitis B viral infection.
Those pharmaceutical preparations which, as well as non-
toxic, inert, pharmaceutically appropriate excipients,
contain one or more compounds according to the invention,
or are composed of one or more ac~ive compounds ~ccording
; to the invention, are included in the present invention,
20 as are processes for producing these prPparations.
~he active compounds according to the invention should
: preferably be present in the abovementioned phanmaceuti-
cal preparations at a concentration of about 0.1 to 99.5,
preferably from sbout 0.5 to 95% by weight of the total
mixture.
:
:
~ ~e_A 28 B62 - s -
2~931~6
23189-7490
Besides the compound~ according to the invention, the
abovementioned pharmaceutical preparations can also
contain further pharmaceutical active compounds.
Production of the abovementioned pharmaceutical prepar-
ations i~ effected in a customary manner according to
known methods/ e.g. by mixing the active compound(s) with
the excipien~( 8 ) .
In order to achieve the desired results, it has in
general been found to be advantageou~, both in human and
veterinary medicine, to administer the active compound(s)
according to the invention in total quantities of about
0.5 to about 500, preferably 1 to 100 mg/kg of body
weight every 24 hours, optionally in the form of several
individual doses. A single dose contains the active
compound(s) preferably in quantities of about 1 to about
80, in particular 1 to 30 mg/kg of body weight~ It may,
however, be necessary to deviate from the indicated
dosages, specifically in dependence on the nature and
body weight of the sub~ect to be treated, the nature and
severity of the disease, the type of preparation and the
; administration of the medicament, as well as the time
period or interval within which administration take
place. ~ ~
The invention also extends to a commercial
package containing, as active pharmaceutical ingredient,
a compound selected from the active compounds listed
above, together with instructions for its use for combat-
ing viral diseases, particularly diseases caused by the
hepatitis B vlr~us.
Le A 28 862 ~ 6 -
, . .
2093~ ~
Starting com?ounds
Example I
Ethyl 3-(1,1-dimethoxy-2-propylamino)-2-~2,3,4,5-tetra-
~luorobenzoyl)acrylate
~ C-COOC2H5
F ~ F ~H
F HN`cH-cH(ocH3)2
CH3
6.4 g of ethyl 3-ethoxy-2-(2,3,4,5-tetrafluorobenzoyl)-
~ acryl~te are introduced into 8 ml of ethanol. While
:: cooling in ice, a solution of 2.6 g of 1-amino-
propionaldehyde dimethyl acetal in 15 ml of ethanol is
added dropwise. The mixture is allowed to come to room
~ temperature, is stirred for two hours and then is
: concentrated in vacuo.
~ ; Yield: 8 g (oil).
,:: ExamplQ II
Ethyl l-(1-1-dLmethoxy-2-propyl)-~-ox~-6,7,8-trifluoro-
4H-quinoline-3-carboxylate ~:
`:
:
' ~
Le A 28 862 - 7 -
209310S
F ~ COOC2Hs
F
CH-CH3
CH(OCH3)2
64 g of crude product from Example I and 2~ g of K2CO3 are
heated at 140C ln 370 ml of DMF for four hours. The
mixture is subsequently poured on to ice. The precipitæte
i8 taken up in CH2Cl2. The organic phase is washed and
~: 5 dried over Na2SO4~ After concentration in vacuo 50 g of
an oily solid are obtained, which is purified by washing
with ether.
Yield: 4~ g
M.p.: 125-126C
Example III
4-Oxo~ oxo-~-propyl)~6,7,8-trifluoro-4H-quinoline-
3-carbo~ylic acid : : : :
.
:
Le A 28 862 : : - 8 - :
, `
:
,
~93~ 0~
F ~ COOH
F ~ N
F CH-CH3
CHO
A.4 g of product from Example IX are heated at 140C for
our hours together with 15 ml of acetic acid, 13 ml of
water and 1.3 ml of sulphuric acid. After cooling to room
temperature, water is added and the precipitated solid is
isolated.
Yields 3.0 g
M.p.: 188-190C ~decomp.)
Exc~mple IV
Ethyl 2-(2~,3,4,5-tetrafluorobenzoyl)-3-(2,2-dimethoxy-
~ ethylamino)acrylate
:.`
O
`C-COOC2Hs~ :
F ~ F ~H
: F N~-c~H2cH(ocH3)z~
: In an analogous manner ~o Ex~mple II:the title compound
(oil) i~ obtained by~` reaction~ of ethyl 3-ethoxy-
~:: ~ : , :
-
Le A 28 862 - 9 -
. .
.
:::
. ,
- 20931 0~
2-(2,3,4~5 tetrafluorobenzoyl) acrylate with aminoacet-
aldehyde dimethyl acetal.
Example V
Ethyl 1-(2,2-dimethoxyethyl)-4-oxo-6,7,8-tri~luoro-4H-
quinoline-3-carboxylate
o
F ~ COOC2H5
CH2-CH(Oc~3)2
Cycli~ation of the product from Example IV is effected in
an analogous manner to the reaction in Example II and
: yields the title compound.
M.p.: 104-106C
Example VI
1,4-Dihydro-4-oxo-1-(2-oxo-ethyl~-6,7,8-trifluoro-4H-
: quinoline-3-carboxylic acid
O
F ~ COOH
F CH2-CHO ~
:
Le A 28 862 - 10 ~
20931~6
Hydrolysis of the product from Example V in analogous
manner to Example III yields the title compound.
Melting point 220-222C.
Preparation Examples
Exa_ple 1
9,10-Difluoro-3-methyl-7-oxo-7H-pyrido~1,2,3-de~-[1,4]-
benzoxazine-6-carboxylic acid
O
F ~ COOH
CH3
- 2.2 g of product from Example III are boiled for four
hours with 0.6 g of NaOH in a mixture of 20 ml of water
and 20 ml of ethanol. After cooling ~o room temperature,
the mixture is acidified with dilute hydrochloric acid
and the resultant solid i~ isolated.
Yield: 1~8 g
M.p.: 248-250~C (decomp.)
;~ 15 Example 2
9-Fluoro-3methyl-10-(4-methyl-1-piperazInyl)~7 oxo-7H-
pyrldo[l~2~3-de]-l1,4lbenzoxaziAe-6-carboxylic acId
~: :
.
:~
.
Le A 28 862
, :
2093~ ~
.
o
F ~ COOH
~ N ~ N
H3C-N J ~ CH3
1.6 g o~ product ~rom ~xample 1 and 2.9 g of N-methyl-
piperazin~ are heated in 20 ml of DMSO at 140C for
2.5 hour~. After cooling the mixture i9 concentrated
under high vacuum. The re~idue is mixed with water. The
precipitate i~ isolated and dried in vacuo at 50C.
Yield: 0O4 g
M.p.: 262-263C (decomp.)
, ~
The compounds listed in Table 1 are prepared in analogous
; manner to~Example 2:
,~':
:
;
:: ~
:
~: : : :
Le A 28 862 ~ ~- 12 - : :
~:
: :
-`
' ::
,
., : ' .
2~31~6
Table 1:
F ~ CO2H
~ CH3
Ex. ~. R1 ~.p.-C
3 HN N - 270-272 ~decomp.)
~ /
4 HN y N - 190-194
H3C
S Example 5
9-Fluoro-10-(4-methyl-1-piperazinyl)-7-oxo-7H-pyrido- -
[1,2,3-de]- L 1 J 4]benzoxazine-5-carboxylic acid
,~
:~ : o
F ~COOII
1--N ~ N
H3C NJ o~J
~: :
Le A 28 862 - 13 -
.
2~93~
2.85 g of 4-oxo~1-(2-oxoethyl)-6,7,8-trifluoro-4H-
~uinoline-3-carboxylic acid and 5.0 g of N-methyl-
piperazine are heated at 140C for 2.5 hours in 30 ml of
DMSO. Sub6equently all volatile material is removed under
high vacuum. The residue is mixed with acetonitrile with
~tirr~ng. The solid is isolated and dried.
Yield: 2.6 g
N.p.: > 300C
Example 6
:
Ethyl 9,10-difluoro-2-methyl-7-oxo-7H-pyrido~1,2,3-de]-
~1,4]benzoxazino-6-carboxylate
F~ ~ COOEt
F N
0~
CH3
5 g of ethyl 6,7,8-trifluoro-4-hydro~y-3-quinoline-
carboxylate, 3.5 g of chloroacetone and 6.& g of K2CO3 are
heated in 40 ml of DMF at 90C for ten hours. After
cooling, all volatile material is stripped off in vacuo.
The residue is ~tirred with water. The remaining solid
is isolated, dried and recrystallised from DMF.
Yield t 3.4 g
M.p.: 229-230~C
Le A 28 862 - 14 -
2093106
: Ex~mple 7
9,10-Di.fluoro-2-methyl-7-oxo-7H-pyrido[1,2,3-de]-
~1,43benzoxazine-6-carboxylic acid
O
F ~ ~OOH
0~
CH3
~: The title compound i~ obtained in an analogou~ manner to
the method of Example 6.
Yield: 2.4 g
M.p.: > 300C
; Example 8
:.'
9-Fluoxo-2-methyl-10-t4-methyl-1-[piperazinyl)-7-oxo-7H-
: 10 pyrido r 1,2,3-de]-[1,4]benzoxazine-6-carboxylic acid
~, ~
F ~ COOH
H3CN J ~ O ~
2.4~g of:product from~Example~7 and 4.3 g of ~-methyl-
piperazine are heated in 25 ml of :DMSO ~at~140C for
L~ A 28 862 -:15 -
:
.
.
.
~,
2093~
2.5 hours. Subsequently all volatile material is stripped
off in vacuo. The residue is stirred with water. The
remaining solid i~ isolated and dried.
Yield: 1.3 g
M.p.: 255-256C (decomp.)
Example 9
Ethyl 9,10-difluoro-7-oxo-2-phenyl-7H-pyridol1,2,3-de]-
[1,4]benzoxazine-6-carboxylate
O
F ~ ~ COOEt
F ~ N
0~
C6H5
5 g of ethyl 6,7,8-trifluoro-4-hydroxy-3-quinoline-
carboxylate, 5.9 g of ~-chloroacetophenone and 6.6 g of
R2CO3 are heated in 40 ml of DMF at 90C for 10 hours.
The mixture is subsequently concentrated in vacuo. The
residue is stirred with water and the solid is isolated
and recrystallised from D~F.
Yield: 2.4 g
M.p.: 251-252C
~ 9,10-Difluoro~7-oxo-2-phenyl-7H-pyrido~1~2,3-de]-[1,4]-
:: ; :
:
::
Le A 28 862 - 16 -
~'
2~931~
: benzoxazine-S-carboxylic acid
o
i F ~ COOH
F ~ N
': 0~
C6H5
1.9 g of product from Example 9, 6 ml of acetic acid,
~ : 4 ml of water and 0.5 ml of ~ulphuric acid are boiled for
`: ; 5 4 houxs. ~fter that the mixture is diluted with water and
the ~olid i8 separated off.
Yisld: 1.7 g
M.p.: ~ 300C
Example 11
Ethyl ~ 6,7-difluoro-9-oxo-9N-cyclohexa[1,2-b]pyrido-
l'~,2',3'-de]-[l,~benzoxazine-10-carboxylate:
~ : I
F ~ ~ C~C2Hs:
F ~ N
5~g ~of ethyl 6,7,8-trifluoro-~4-hydroxy-3-qulnoline-
Le ~ 2~862. : - 17 -~
2093~6
carboxylate, S g of 2-chlorocyclohexanone and 6.6 g of
K2CO3 are heated in 40 ml of DMF ~t 90C for 10 hours.
After that the mixture i~ concentrated in vacuo. The
residue is taken up in water and extracted with CH2Cl2.
The organic phase i~ dried and concentrated by
evaporation. The residue is separated with the aid of a
column (silica gel, eluent: toluenefethyl acetate 1:1).
Yield 0.6 g
melting point 198-208C.
L2 A 28 862 - 18 -
