Note: Descriptions are shown in the official language in which they were submitted.
1178956 ;.
1 Detailed Explanation of the Invention
2 The present invention relates to 7-acylamino-1-
3 carba-3-cephem-4-ca~boxylic acid represented by the following
general formula [I], the nomenclature of carbacephem referri;lg ;-
to Journal of the American Chemical Society Vol. 96, 7584,
6 1974, and derivatives thereof.
7 --
XHN R
9 ~ ~
o~N~. [I]
11 ~ CO2 .~,.
12
13
14 (wherein X represents an acyl group, R represents hydrogen
Is atom or a protectlng group of carboxylic acid and R2
16 represents hydrogen atom, a lower alkyl group or a lot~er
17 acyloxy group).
18 So far, as carbace~hem compounds, those ~hich have substituted
l9 methyl group at 3-position such as (+)-l-carbacephalotin
represented by the formula
21
22
23 ~ ~ N C~2
24 H ~ ~ Ac
: 0
COO~
26
27
28 are known and some are reported to have an antibacterial
29 activity [the same reference above and J. Med. Chem., 20,
551, (1977)~. As the result of various studies to obtain
3I novel analogs of cephalosporin with potents antibacterial
56
1 activity, the present inventors previously made an invention
2 relating to cephalosporin analogs, represented by the follow-
3 inq general formula [II~, that is, C-4 or C-5 substituted
4 carbacephems (numbering system shown in the structure ~II]
is used hereinafter) and salts thereof
, ~
Xl ~ ~2 [II]
N~ ~
Il .~OOR3
12
3 [wherein Xl represents amino group, azido group or phthalyl-
imino group, Rl represents hydrogen atom, a halo group,
~15 hydroxyl group, a lower alkoxy group, an aryloxy group, an
6 aralkyloxy group, an acyloxy group, a lower alkylsulfonyloxy
group, an arylsulfonyloxy group,
8 a lower alkylthio group, an arylthio group, an aralkylthio
19 group, a lower alkylsulfinyl group, an arylsulfinyl group,
an aralkylsulfinyl group, a sulfonium group represented by
21 the ~eneral formula -S R4R5 whérein R~ and R5 may be the
22 same or-different and repr~sent a lower alkyl, aryl or
2~ ~ralkyl group, a lower alkylsulfonyl group, an arylsulfonyl
24 group, an aralkylsulfonyl group, a quarternary ammonium -
2s group represented by the general formula N R6~7R~ wherein
2~ R6, R7 and R8 may be the same or different and represent
27 a lower alkyl, aryl or aralkyl group, an arylseleno group
28 or an arylseleninyl group, R2 may be the same significance
29 as Rl or represents a lower al~yl group, a lower alkyl group
substituted with halogens, azido group, nitrile yroup or an
31 amino group represented by the general formula N RgRlo
~ .
1~7~9S~
;~ . .. ~ ,
1 wherein R9 and Rlo may be the same or different and represent ---
2 hydrogen atom, a lower alkyl, aryl or aralkyl group, and R3
3 represents hydrogen atom, a substituted or unsubstituted ~
4 alkyl, aryl, aralkyl or silyl group3 and salts thereof. : -
S As the result of further studies, the present inventors
6 successfully attained to unexpectedly highly potent antibacterial
7 cephalosporin analogs by acylating a compound represented by :-
8 the general formula [II]
g - -
' ' .,
11 Xl ~ 2 -
12 ~ ~ [II]
OOR3 .
14
6 (whereln Xl is NH2, Rl is H, R2 is ~1, a lower alkyl group.or
17 a lower acyloxy group and R3 is Rl).
18 The compound of the present invention is represented
19 by the general formula [I]
. ~.. ~ . . .
21
22 ~ R ~.
z3 O ~ l [I]
24 I CO~R
2s
z6
Z7 {wherein X represents an acyl group represented by the general
28 formula XlCO wherein Xl represents the following five groups:
29 l) cyano methyl group or a group representèd by the general
formula (Al~-nB-CII-
31 A2 ~_
-
~ ~78956
I lwherein B represents an unsaturated six membered carbocycle
2 such as cyclohexenyl group,cyclohexadienyl group, phenyl
3 group, etc. or a five or six membered heterocycle such as
4 furyl group, thienyl group, pyrrolyl group, thiazolyl group,
oxazolyl group, isothiazolyl group, isoxazolyl group,
6 imidazolyl group, pyrazolyl group, triazolyl group, tetra-
? zolyl group, pyridinyl group, pyrimidinyl group, pyrazinyl
8 group, pyridazinyl group, triazinyl group, 5,6-dihydro-1,4-
g dithin-2-yl group, etc., Al represents substituent(s) which
0 is selected from hydrogen atom, hydroxyl group, a lower
Il alkoxy group having 1 to 4 carbon atoms, a halo group, nitro
12 group, amino group,aminomethyl group, methylsufonamido group,
l3 a lower acyloxy group having 1 to 4 carbon atoms, n is a
1~ number from 0 to 5, and A2 représents hydrogen atom, amino
group, hydroxyl group, carboxyl group or sulfoxyl group~,
16 2) a group represented by the general formula
17 (AltnB~CH~
18 ! NHCON ~
19 [wherein Al, B and n have the same significance as defined
above, A3 and A4 are the same or different and represent
21 hydrogen atom, a lower alkyl group having 1 to 4 carbon
22 atoms, a group represented by the general formula
23 -C-A5 (wherein A5 means a lower alkyl group having 1 to 4
24 carbon atoms) or a group represented by the general formula
OA
-P ~ ~ (wherein A6 and A7 are the same or different and
26 OA7
27 represent hydrogen atom, a lower alkyl group having 1 to 4
28 carbon atoms or an alkali metal) and -N / 3 also means
29 ~ A4
~ 8
a group represented by the general formula -N N-A8
31
-- 4
1~7895~
I (wherein A8 and A'8 are the same or different and mean
2 hydrogen atom or a lower alXyl group having 1 to 4 carbon
3 atoms) or a group represented by the general formula
O A'g
-N ~N-Ag (wherein Ag represents hydrogen atom, a lower
6 alkyl group having 1 to 4 carbon atoms, methylsulfonyl group
7 or furfurylideneimino group, and A'g represents hydrogen
8 atom or a lower alkyl group having 1 to 4 carbon atoms)],
9 3) a group represented by the general formula
~o (Al~n f
~ NHCOAlo
12 [wherein Al, B and n have the same significance as defined
13 above, and Alo represents a substituted aryl group or mono-,
1~ bi- or tricyclic heterocycle(s) such as a group represented
IS by the follgwing formulae:
, ~ , HN~N ~ YH ~ (Y is oxygen or sulfur atom),
~ ~ and naphthyridinyl group such as
2I which may have substituents such as those represented by A
22 group on ring(s)]
23 4) a group represented by the general formula
2~ (Alt--nB
NOAll
a6 Iwherein Al, B and n have the same significance as defined
27 above and All represents hydrogen atom, a lower alkyl group
28 having 1 to 6 carbon atoms, a lower alkenyl group having
~ 2 to 6 carbon atoms, a lower alkinyl group having 2 to 6
carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms
131 or an aryl group those groups being unsubstituted or
1~ ~ 5
~789S~
~ substituted with suitable substituent(s) such as carboxyl
2 group, cyano group, a halo group, carbamoyl group or lower
3 alkyloxycarbonyl group having 1 to 4 carbon atoms],
~ 5) a group represented by the general formula
~1-Z-CH2-
6 [wherein Bl represents trifluoromethyl group, cyanomethyl
7 group or a group represented by (Al~-nB- (Al, n and B have
8 the same significance as defined above) and Z represents
g oxygen atom or sulfur atom],
Rl represents hydrogen atom or an ester-protecting group con-
ll ventionally employed in the field of the chemistry of penicil-
12 lins and cephalosporins, that is, an alkyl group having 1 to 5
13 of carbon atoms such as methyl group, ethyl group, n-propyl
~ group, isopropyl group, n-butyl group, isobutyl group,
t-butyl group etc., a halogenated alkyl group having 1 to 5
16 of carbon atoms such as chloromethyl group, 2,2,2-trichloro-
~7 ethyl group, 2,2,2-trifluoroethyl group etc., an arylmethyl
18 group having 7 to 20 of carbon atoms such as benzyl group,
19 diphenylmethyl group, triphenylmethyl group, etc., an aryl-
methyl group having 7 to 20 of carbon atoms and having methoxy
2I group, nitro group, etc., on the phenyl ring, a substituted
2z silyl group such as trimethylsilyl group or triphenylsilyl
23 group or a group enzymatically or nonenzymatically readily
21 eliminable ln vivo, for example, a group represented by the
general formula -CHOCOR4 wherein R3 represents hydrogen
26
27 atom or a lower alkyl group having 1 to 6 of carbon atoms,
28 ~4 represents a lower alkyl group having 1 to 6 of carbon
29 atoms, a lower alkoxy group having 1 to 6 of carbon atoms
or phenyl group, etc. and ~2 has the same significance as
3~ defined above}.
9S6
1 In case that Rl in the general formula lI] is
2 hydrogen atom or that X is an acyl group having a free
3 carboxyl group or amino group, the compounds represented by
4 the general formula ~I] may be salts of pharmace~tically
acceptable, inorganic or organic bases or acids.
6 The compounds represented by the general formula
7 [I] include all stereoisomers at the 6-, 7- and 4-positions
8 and mixtures thereof. Among the stereoisomers, those which
g have cis configuration at the 6- and 7-positions have higher
antibacterial activities than trans-isomers, so the cis-
11 isomers are more useful as antibiotics. The --NOR group in
~2 the acyl group has the following two geometrical isomers,
~3 i.e. syn and anti.
~4 -C-CO- -C-C~-
11 11
Is N-OR RO-N
16 (syn) (anti)
Since the syn-isomer is superior to the anti-isomer
~8 in antibacterial activity, the syn-isomer is more useful as
19 antibiotics. In general, it is known that thiazolyl group
S~l
21 represented by H2N ~ ~ show reversible interconversion
22 with thiazolinyl group, as shown below, and both are usually
23 dealt with as identical. In the present specification, both
2~ isomers are represented by thiazolyl group.
H2N ~` ~ HN
26 \N
27 thiazolyl thiazolinyl
2t The compounds represented by the general formula -
29 [I] are produced by acylating the compounds [II'~ represented
by the general formula [II] wherein X is NH2, Rl is H, R2 is
3~ H, a lower alkyl group or a lower acyloxy group having l to 5
11~956 ^'' '' '
1 carbon atoms and R3 is Rl according to Flow Sheet I.
2 Flow Sheet I
pl 0~ 1
8 [II'~ [I]
9 ~"4,~
(wherein ~l, R2 and X have the same significance as defined
11 above).
12 The acylating reaction (condensation) is carried
13 out according to a conventional method of acylation employed
1~ in the field of the chemistry of penicillins and cephalosporins.
The compounds represented by the general formula [I]
6 are prepared by the condensation of the compound [II']
1I repre5ented by the general formula [II'], the salt thereof
18 or a compound functionally equivalent thereto (these are
19 referred to "7-amino compound" hereina~ter) and a carboxylic
acid represented by the general formula [III]
21 X2COOH [III]
22 or a reactive derivative thereof and~ if necessary, followed
23 by deprotection of protective group in the group X2- or -COOR
2~ in a conventional manner. In the formula [III], X CO
2s corresponds to XlCO, that is, X in the compound represented
26 by the general formula [I] and x2 represents group Xl defined
27 above or suitably protected Xl group in case that Xl has
28 group(s) which is susceptible to acylation such as hydroxyl
29 group, amino group, carboxyl group or mercapto group.
31 ,~
l:t7~gS6
1 x2 represents the following five groups
2 concretely:
3 1 ~ ) cyanomethyl group or a group represented by the general
formula:
(A l~-nB IH
6 A'2
7 (wherein B has the same significance as defined above, A'l
R represents substituent(s) which is selected from a group of
g hydrogen atom, hydroxyl group, a protected hydroxyl group,
lo a lower alkoxy group having l to 4 carbon atoms, a halo
Il group, ni~ro group, a protected amino group, a protected
12 aminoethyl group, methylsulfonamido group, a lower acyloxy
13 group having 1 to 4 carbon atoms, n is a number from 0 to 5,
1~ and A'2 represents hydrogen atom, a protected amino group,
hydroxyl group, a protected hydroxyl group, carb~xyl group,
a protected carboxyl group, sulfoxyl group or ~ protected
sulfoxyl group),
2') a group represented by the general formula:
(A'1--t~B~CH~
NHCON~
2 0 (wherein A'l~ A3, A4, B and n have the same significance as
21 defined above.)
2 2 3') a group represented by the general formula:
23 (All~B-fH-
NHCOAl o
24 (wherein A'l, Alo, B and n have the same significance as
2 5 defined above.)
2 6 4') a group represented by the general formula:
2 7 ( l-t-n 1l
NOAl 2
2 a (wherein A'l~ B and n have the same significance as defined
g
~:1789S6
I above and A12 represents hydrogen atom, a lower alkyl group
2 ha~ing 1 to 6 carbon atoms, a lower alkenyl group having 2 to
3 6 carbon atoms, a lower alkinyl group having 2 to 6 carbon
4 atoms, a cycloalkyl group having 3 to 6 carbon atoms or an
S aryl group, those groups being unsubstituted or substituted
6 with suitable substituents such as a protected carboxyl
7 group, cyano group, a halo group, carbamoyl group or a
8 lower alkyloxycarbonyl group having 1 to 4 carbon atoms)
9 5') a group represented by the general formula
B'l-Z-CH2-
11 (wherein B'l represents cyano group, trifluoromethyl group,
12 cyanomethyl group, a group represented by ~A'l~nB- ~A'l,
13 n and B have the same significance as defined above) and
14 Z represents oxygen atom or sulfur atom.
s As the protecting group~s) for amino group,
16 hydrox~l group, carboxyl group, etc., in the x2 group,
17 those which are employed in the chemistry of cephalosporins
18 or penicillins are used and those are shown below.
19 As the salt of Compound ~II'l, inorganic
2 o salts and organic salts such as hydrochloride, sulfate,
21 carbonate, phosphate, formate, trifluoroacetate,
2 2 malate, etc. are mentioned. Further, the sodium salt,
2 3 potassium salt, calcium salt, ammonium salt, organic amine
2 4 salt, etc. of the carboxylic acid represented by the general
2 5 formula [II'] wherein Rl is hydrogen are exemplified.
2 6 The salts are prepared in a conventional method.
2 7 As the compound functionally equivalent to Compound
2 8 lII'], 7-monosilyl or 7-disilyl amino derivatives of Compound
2 9 [II'] are exemplified.
-- 1 0
1~789S6
1 As the reactive derivatives of the carboxylic acid
2 represented by the general formula [III], X2COOH, l) an acid
3 halide, 2) an acid anhydride, 3) a mixed acid anhydride, 4)
4 an active ester, 5) an active thioester, 6) an acid azide,
etc. are exemplified.
6 Condensation (acylation) reactions using the above
7 derivatives are explained in the following.
8 l) Method using an acid halide
g A 7-amino compound and an acid halide are subjected
0 to condensation reaction in an inactive solvent, preferably
11 in the presence of a proton acceptor. The acid halide is
2 prepared in a conventional manner.
As the proton acceptor, inorganic and organic
bases, preferably sodium carbonate, potassium carbonate,
sodium hydroxide potassium hydroxide, triethylamine, N-
l6 methylmorphorine, pyridine, etc. are used.
~7 As the solvent, any inactive solvent which does
~8 not affect the reaction, preferably water, an ether such as
19 tetrahydrofuran, dimethoxyethane, etc., an ester such as
ethyl acetate, etc., an amide such as dimethylacetamide,
21 hexamethylphosphoric triamide, etc. or a sulfoxide such as
22 dimethylsulfoxide, etc. is used solely or in combination.
23 The reaction is carried out at a temperature of
24 -20 to 40C, preferably from 0C to room temperature.
2s 2) Method using an acid anhydride
26 A 7-amino compound and an acid anhydride are
27 subjected to condensation reaction in an inactive solvent.
28 The acid anhydride is prepared in a conventional
29 manner.
As the solvent, any solvent which does not affect
31 the reaction, preferably the same solvents as those in
~`~
1~7895
1 the above Method, is used. The range of temperature in
2 the reaction is the same as the above Method.
3 The typical method using carbodiimide such as
4 dicyclohexylcarbodiimide is exemplified as acid anhydride
method.
6 3) Method using a mixed acid anhydride
7 A 7-amino compound and a mixed acid anhydride
8 are subjected to condensation reaction in an inactive
g solvent.
lo The mixed acid anhydride is prepared in a
Il conventional manner, for example the method wherein a
12 corresponding carboxylic acid, X2COOH and a chloroformic
13 ester such as ethyl chloroformate, isobutyl chloroformate,
14 etc. are reacted in the presence of a base.
As the solvent, any inactive solvent which does
16 not affect the reaction, generally an unhydrous solvent
17 or a mixture of water and an unhydrous solvent, preferably
18 the same solvent as in the above Methods, are used.
19 The range of temperature in the reaction is the
same as in the above Methods.
21 4) Method using an active ester
22 A 7-amino compound and an active ester are
23 subjected to condensation reaction in an inactive solvent.
24 Solvents and reaction temperature are the same
as those of the above Methods l to 3.
26 ~S the active ester, a phenylester such as p-
27 nitrophenyl ester, trichlorophenyl ester, etc., a methyl
28 ester having an electronegative group such as cyanomethyl-
29 ester, etc., and N-oxydiacylimide ester such as N-hydroxy-
succinimide ester,etc. is exemplified.
31
l2 -
11789~6; ;'''~';"'
The active ester is prepared in such conventional ~ -
2 method that a corresponding carboxylic acid and a hydroxyl
3 compound are reacted in the presence of a dehydrating
4 condensation reagent such as dicyclohexylcarbodiimide, etc. -
s 5) Method using an active thiolester
6 This method is carried out in a similar method as
7 in Method 4~.
8 As the most preferable thiolester, p-nitrothio-
g phenyl ester which is prepared by a mixed acid anhydride
~o method or dicyclohexylcarbodiimide method is exemplified.
6) Method using an acid azide
12 A 7-amino compound and an acid azide compound are
l3 subjected to condensation reaction in an inactive solven-t in
a similar manner as in the above ~ethods l to 5.
The acid azide is prepared in such method that the
16 hydrazide of a corresponding carboxylic acid is reacted with
nitrous acid at a temperature of -20 to 0C.
8 In case that reactive derivatives of -the carboxylic
19 acid mentioned above have such a group as amino group,
hydroxyl group, carboxyl~ group or mercapto group, which is
21 susceptible to acylation, those groups are preferably protected
z2 with a suitable protecting group in a conventional manner prio~
23 to the conclensation reaction with amino compounds.
z~ As the suitable protecting group, those used ln
z5 the field of the synthetic chemistry of penicillins and
z6 cephalosporins are mentioned.
27 As the amino-protecting group, t-butyloxycarbonyl
28 group (Boc), benzyloxycarbonyl group (~bz), trichloroethyloxy-
29 carbonyl group, trityl group, formyl group, chloroacetyl
group, trialkylsilyl group, proton, ~-diketon, ~-ketoester,
31 etc. are preferably exemplified. As a compound protected
1 - l3 -
~.~7895~ ~ ~
I with a proton, the compound represented by the formula: -
3 ~ CHCOC~
4 NH3CQ
is exemplified. As a compound protected with a ~-ketoester,
6 the compound represented by the formula:
7 Ar-CH-COOH
8 ~ ~11
g ' '
~ ~0
11 ~ OEt
~2 is exemplified.
13 As the hydroxyl-protecting group, benzyl group,
14 benzyloxycarbonyl group, trityl group, tetrahydropyranyl
group, t-butyl group, etc. are exemplified. Since the
16 reactivity (nucleophilic activity) of most amino groups
17 .is higher than that of hydroxyl group, the protection
18 of the hydroxyl group is not necessarily essential depending
19 on the employed method of acylation.
As the carboxyl-protecting group, t-butyl group,
2I benzyl group, p-methoxybenzyl group, p-nitrobenzyl group,
22 benzhydryl group, etc. are mentioned.
23 As the mercapto-protecting group, benzyl group,
2~ trityl group, benzyloxycarbonyl group, p-nitrobenzyl group,
etc. are mentioned.
26 Deprotection of the above protecting groups are
27 carried out in a conventional method used in the field of
28 the synthetic chemistry of penicillins and cephalosporinS.
29 As the method of deprotecting amino-protecting
groups, the following are exemplified.
31
~ ~ ~ l4 -
:
li7~9S6
1 Boc group is deprotected by a method using an ;:-
2 acid such as formic acid, acetic acid, trifluoroacetic acid,
3 hydrochloric acid, etc.
4 Cbz group is deprotected by a catalytic reduction ;
5. or a method using hydrogen bromide-acetic acid, etc.
6 Trityl group is deprotected by a catalytic reduction
7 or a method using an acid such as trifluoroacetic acid, etc.
8 Formyl group is deprotected by hydrolysis using an
g acid or an alkali.
o Chloroacetyl group is deprotected by a method using
thiourea. ~ -
12 Trialkylsilyl group is deprotected by hydrolysis.
3 Proton is deprotected by neutralization.
4 The additive with ~-diketon or ~-ketoester is
Is removed by acid hydrolysis.
16 As the methods of deprotecting hydroxyl-protecting
17 groups, the following are exemplified.
18 Benzyl ~roup is deprotected by a catalytic reduction
19 or a method using hydrogen fluoride.
Cbz group is deprotected by a catalytic reduction or ~*~
21 a method using hydrogen bromidé-acetic acid.
22 - Trityl group is deprotected hy a catalytic reduction
23 or a method using trifluoroacetic acid, etc.
24 Tetrahydropyranyl group is deprotected by acid
hydrolysis. ;
26 t-Butyl group is deprotected by a method using an
27 acid such as trifluoroacetic acid, hydrogen bromide-acetic
28 acid, hydrogen chloride, etc.
29 As the methods of deprotecting carboxyl-protecting
groups, the following are exempliÆied.
15 _
~.~ 7~956
1 t-Butyl group is deprotected by a method using
2 an acid such as trifluoroacetic acid.
3 Benzyl or p-nitrobenzyl group is deprotected by
4 a catalytic reduction or method using ~ewis acid such as
AQCQ3, etc-
6 Benzhydryl or p-methoxybenzyl group is deprotected
7 by a catalytic reduction or a method using hydrogen bromide-
8 acetic acid, hydrogen chloride-methanol, trifluoroacetic
9 acid, etc.
As the method of deprotecting mercapto-protecting
11 groups, the following are exemplified.
12 Benzyl group is deprotected by a method using
13 hydrogen fluoride, etc.
14 Cbz group is deprotected by a method using hydrogen
bromide-acetic acid, trifluoroacetic acid, etc.
16 p-Nitrobenzyl group is deprotected by a catalytic
~7 réduction. The deprotection mentioned above may well be carried
18 out concomitantly with the conversion of Rl group to hydrogen
19 atom, that is deesterification.
If desired, the acylating reaction is preferably promoted
21 by silylating the starting compound [II] with a silylating
æ agent such as trimethylchlorosilane-base, hexamethyldisilazane,
23 N,O-bistrimethylsilylacetamide, etc~ to solubilize the start-
2~ ing material in organic solvents and to activate the amino
2s grou~.
- 16 -
r~ ~
~JI
1:~7895~ -
Reference Flow Sheet ~I'] . ^
4 R . . .;-
OHCCX~C~-C~-cH2 -~ ~ ~ N3C~2Cce `
6 (l' R=~C~ ) ~H~ P(OEt~2 ~ ( )2 --
8 CO~R CO2R
~2:R=H
9 ~2 :R=C~
2 ~l3~ ~ N~3 IO~-osoS~ N3~ I.o
3 ~ 1 0 N~.~'(OÉt)2
1~ C1R ~ C2
1s ~3:R=H ~ R=H
16 \3 :~=CH3 ~ ~' R=C~3)
R ~IlN ~ ~ R
~ duct-onO N~ ~ d~ ster
CO~R CO2,R CO
22 ~'-D~-C~ ) ~ (6 P~=C~3) r/ (7 R-~
2~ ~ ` ~ eductio~
2s O
2G ! C0~7H[
27 1 ~8~
~3':R=C'~3J
28
29
3 ~
1 ~ 17 -
~789S6
Reference Flow Sheet [II']
4 .
60 ~ R N3 ~
ClR , COjR
85:R=H g
- ~
2, 3 j~OCOCli3 H2N~/cCO"E~
reduct- O 1 deesteri-
13 0 COO~ tion COOR fication
1 0 . 1 1 --
\ 'deesteri-
\ fication
16
17 ~
1!1 712r7 ~ 0COCH~3 N3 ~ ~oCOCH3
o ireduct- O
COOH tion CO~H
21 12 1 13
22 ~ t-~
23
2~
26
27
28
29
31 ~.,~
1:1789S6
. ':
I In the Reference Flow Sheet [I'] and [II'],
2 compound 6, 7, 6', 7' 11 and 12 are examples of the starting
3 compounds represented by the general formula ~II'].
4 Preparation of the starting compounds is described in -
Reference Examples ~elow.
6 Compound [I'] represented by the general formula
7 [I] ~herein ~l is H can be produced by elimination of the
8 carboxyl-protecting group according to Flow Sheet [II].
g - Flow Sheet [II]
~ R ~ R2 ~-
l3CO~R CO2H
~4
l5[I ] [I']
l7 (wherein Rl is a protecting group of carboxylic acid and
18 R2 and X have the same significance as defined above). ~;
Is The deesterification reaction is carried out
according to a conventional method employed in the field
21 of the synthetic chemistry of penicillins and cephalosporins.
22 - As the reaction which converts -COORl group to
23 -COOH group, l~ catalytic reduction, 2) acidolysis, 3)
24 cleavage reaction using a Lewis acid, 4) hydrolysis, 5)
2s reduction other than catalytic reduction using reclucing
26 agents and 6) a method using an esterase are exemplified.
27 Rach method is explained in detail below.
28 l) Catalytic reduction
29 COOR group is converted to COOH group in the
presence of a catalyst in an atmosphere of hydrogen in an
31 inactive solvent. As the soivent, any solvent which does not
1 9
1~7~956
1 affect the reaction, preferably ethanol, water, tetrahydro-
2 furan, dioxane, ethyl acetate or acetic acid is used solely
3 or in combination. As the catalyst, palladium-carbon,
4 platinum oxide, palladium-calcium carbonate and Raney nickel
are exemplified. The reaction is carried out generally at a
6 pressure of 1 to 50 atoms and a temperature of G to 100C,
7 preferably at atmospheric pressure and room temperature.
8 This method is preferably employed in case that
g R1 -is benzyl group, p-nitrobenzyl group, diphenylmethyl
group, p-methoxybenzyl group, etc.
2) Acidolysis
COOR group is converted to COOH group with an
13 acid in an inactive solvent. As the acid, hydrogen chloride,
l4 p-toluenesulfonic acid, trifluoroacetic acid, etc. are used.
As the solvent, any solvent which does not affect the reac-
6 tion, preferably ethyl acetate, benzene, ethanol, acetic
acid, dioxane, methylene chloride or chloroform, etc. is
~8 used solely or in combination.
19 The reaction is carried out at a temperature o
-15 to 50C, preferably 0 to 25C, for 10 minutes to 5 hours,
2I preferably 30 minutes to 3 hours.
22 This method is preferably employed in case that
23 R is t-butyl group, trityl group and so on.
2~ 3) Cleavage reaction using a Lewis acid
2s COORl group is converted to COOH by cleavage
26 reaction in the presence of a Lewis acid in an inactive
27 solvent. As the solvent, any solvent which does not affect
2s the reaction, preferably a mixture of a nitroalkane such as
29 nitromethane and a haloalkane such as methylene chloride is
used. As the Lewis acid, aluminum chloride, boron trifluo-
3l ride, titanium tetrachloride, tin tetrachloride, etc. are
I ~ - 20 -
1~7~956
1 mentioned. The acid is used in an amount of l.0 to l.5 ~--
2 molar equivalents to Compound [I'~. The reaction is
3 preferably carried out in the presence of an aoent uptaking
4 carbonium cation, such as anisole. The reaction is carried
s out at a temperature of 0 to 50C, preferably at room
6 temperature for l to l0 hours.
7 - This method is preferably employed in case that
8 Rl is p-nitrobenzyl group and so on.
9 4) Hydrolysis - -
COORl group is converted to COOH group by hydrolysis
1 in the presence of an acid or alkali in an inactive solvent.
~2 As the acid, p-toluenesulfonic acid, hydrochloric acid,
13 acetic acid, etc. are mentioned. As the solvent, any solvent
14 which does not affect the reaction, preferably 2 % a~ueous
methanol, N,~-dimethylformamide or acetic acid-water-tetra-
16 hydrofuran, etc. is mentioned. The reaction is carried out
1l at a temperature of 0 to 50C, preferably 15 to 25C for l0
18 minutes to 2 hours. -
This method using acids is preferably employed in
case that Rl is t-butyldimethylsilyl group.
21 As the alkali, calcium carbonate is preferably
22 used in-an amount of l to 6 molar equivalents to Compound ;~.
23 [I"]. As the solvent, any solvent which does not affect
24 the reaction, preferably tetrahydrofuran-water, dioxane- -
water or acetone-water, is used. The reaction is carried
26 out generally at a temperature of 0 to 30C for 30 minutes
27 to 24 hours.
28 This method using alkalis is preferably employed
29 in case that Rl is methyl group, ethyl group and so on.
31
-- 2
~78956 --`
~ 5) Reduction using reducing agents (other than catalytic
2 reduction)
3 COOR group is converted to COOH by reduction in
4 an inactive solvent. As the reduction, a method using
zinc-acid is exemplified. As the solvent; acetone, water,
6 dioxane, tetrahydrofuran, ethanol, acetonitrile, N,N-
7 dimethylformamide and acetic acid are used solely or in
8 combination. As the acid, hydrochloric acid,form~c acid
9 and acetic acid are mentioned. The reaction is carried out
at a temperature of 0 to 100C, preferably 0 to 40C for l
Il to 10 hours. The amount of zinc used for the reaction is
12 usually 1 to 10 molar equivalents.
l3 If appropriate reagents are employed, the protect-
14 ing group in group X can be eliminated simultaneously with
the deesterifieation.
16 The present compounds represented by the general
]7 formula [I] wherein R is H, as mentioned in Example below,
have so exeellent antibacterial activities against Gram-
l9 positive and Gram-negative bacteria that they are expected
to be useful as antiinfectional agents or antibacterial
2l agents against these bacteria. The compounds represented by
22 the general formula [I] wherein Rl is other than H can also be
23 used as the starting material of the compounds represented
24 by the general formula [I] wherein Rl is H.
The invention includes within its scope pharma-
26 ceutical compositions comprising, as an active ingredient,
27 Compound [I] or a pharmaceutically aceeptable salt thereof
28 in association with a pharmaceutical carrier or diluent.
29 The compounds of this invention are administered by parenteral
(intramuscular, intraperitoneal, intravenous, or subcutaneous
31 injection routes) , or~l or rectal ~ute o- admilis~rat-.c); arcl
- 22 -
_- ' ~.' ' f-.; !
`,','`--'.,~
11789S~
1 can be formulated in dosage forms apnro~riate f~r each route of
2 administration.
3 Preparations according to this invention for
4 parenteral administration include sterile aqueous or non-
aqueous solutions, suspensions, or emulsions. Examples of
6 non-aqueous solvents or vehicles are propylene glycol,
7 polyethylene glycol, vegetable oils, such as olive oil, and
8 injectable organic esters such as ethyl oleate. Such dosage
g forms may also contain adjuvants such as preserving, wetting,
lo emulsifying, and dispersing agents. They may be sterilized by,
2I for example, filtration through a bacteria-retainin~ filter,
12 by incorporating sterilizing agents into the compositions,
13 by irradiating the compositions, or by heating the composi-
14 tions. They can also be manufactured in the ~orm of sterile
solid compositions which can be dissolved in sterile water,
16 or some other sterile injectable medium immediately before
~7 use.
18 Compositions for oral administration may be
19 presented in a form suitable for absorption by the gastro-
Z0 intestinal tract. Tablets and capsules for oral administration ~:~
21 may be in unit dose presentation form, and may contain conven-
22 tional excipients such as binding agents, for examples, syrup,
23 acacia, ~elatin, sorbitol, tragacanth, or polyvinylpyrrolidone;
z4 fillers, for examples, lactose, sugar, maize-starch, calcium
phosphate, sorbitol or ~lycine; lubricants, for exam21e,
26 ma~nesium stearate, talc, polyethylene ~lycol, silica; dis-
27 integrants, for example, potato starch or acceptable wettin~
28 agent such as sodium lauryl sulphate. The tablets may be
29 coated according to methods well known in the arts. Oral
liquid preparations may be in the form of aqueous or oily
31 suspension, solution, emulsions, syrups, etc. or may b~
-- 23 --
~17~956
1 presented as a dry product, for reconstitution with water or
2 other suitable vehicle before use. Such liquid preparations
3 may contain conventional additive such as suspending agents,
for example, sorbi.ol syrup, meth~l cellulose, ~lucose sugar-
syrup, gelatin, hydroxyethylcellulose, carboxymethylcellulose,
6 aluminum stearate gel, e~ulsifying agents, for example,lecithin
7 or sorbitan monooleate; non-aqueous vehicles, which may
8 include edible oils, for example, almond oil, coconut oil,
g propylene ~lycol, or ethyl alcoholi preservatives, for example, ~Y`
methyl or propyl p-hy~roxybenzoates or sorbic acid.
11 1Compositions for rectal administration are prefer-
12 ably suppositories which may contain, in addition to the
13 active substance, excipients such as cocoa butter or a
4 suppository wax.
The dosage of active ingredient in the compositions
6 of this invention may be varied; however, it is necessary
7 that the amount of the active ingredient shall be such that
18 a suitable dosage form is obtained. The selected dosage
19 depends upon the desired therapeutic effect, on the route of
administration, and on the duration of the treatment. ,~
21 ~enerally, dosage levels of between 5 to 350 m~/k~. of
22 body weight daily are adminis-tered to mammalian patients
23 to achieve an antibiotic effect.
24 As the pharmaceutically acceptable salt of compound
2s (I), inorganic salts and organic salts such as hydrochloride,
26 sulfate, carbonate, phosphate, formate, malate, etc. are
27 mentioned. Further, the sodium salt, potassium salt, calcium
2~ salt, ammonium salt, organic amine salt, etc. of the carbox-
29 ylic acid represented by the general formula (I) wherein Rl
is hydrogen are exemplified. The salts are prepared in a
31 conventional method.
- 24 -
~7~9S~ :
1 The following Examples show practical embodiments ~~
2 of preparing the present compounds,
3 :::
4 Example l -~`
Preparation of (~)-cis-2-carboxy-7-[2-(thiophene~
6 2-yl) acetylamino]-l-azabicyclo [4, 2, 0] oct-2-en-8-one
7 represented by the following formula: -
9 - ' j' ~ ,i
~S ~ CH~Co~
11 . ~ .. ,
00
13
In this Example, ~8 mg (0.489 mmole) of the amino
16 compound which is obtained in Reference Example 8 is dissolved
17 in 2 ml of deionized water and l ml of acetone and 84 mg of
18 aqueous sodium bicarbonate is added thereto. To the mixture,
1g 78 mg of 2-thienylacetylchloride dissolved in 0.5 ml of acetone
is added dropwise under ice cooling. Then the mixture is stirred
21 for 30 minutes and washed with ethyl acetate. The obtained
22 aqueous layer is adjusted to a pH of 2.0 with hydrochloric acid.
23 The resulting white suspension is extracted three times with 5 ml
2~ of ethyl acetate. The ethyl acetate layer is washed with satura-
2s ted sodium chloride solution, dried with anhydrous sodium sulfate
26 and concentrated under reduced pressure to obtain a yellow oily
?,7 product. The product is charged on a column packed-with 4.0 g
28 silica gel (Wako-gel ~ C-200, product of Wako Junyaku Co.,
29 Ltd., the same silica gel is used in the following Examples and
Reference Examples) and elution is carried out with chloroform.
31 The obtained syrup is treated with chloroform-ethanol to obtain
1 - 25 -
~17~956 ~` -
1 30 mg of crystals. The crystals are identified as the desired
2 compound based on the following properties. Yield 20.1 ~.
3 Melting point: 181- 183C ---
4 IR(KBr) vmax : 1775, 1690, 1650, 1615 -
S NMR(CD3OD)~(ppm): 7.16 -6.88(3H, m), 6.18(1H, t),
6 5.16(1H, d, J =5Hz), 3.80(1H, m), 3.37(2H, s),
7 2.5 -1.30(4H, m) :
9 Example 2
o Antibacterial activities of (i)-cis-2-carboxy-7-
11 [2-(thiophene-2-yl) acetylamino]-l-azabicyclo [4, 2, 0] oct- --
12 2-en-8-one obtained in Example 1 are shown below. Regular agar
13 dilution method is employed at pH 7Ø
14
Microorganism MIC (~g/ml)
IS
Vibrio percalans KY4174 2
16
~rwinia aroides KY3241
17
Staphylococcus aureus KY4279
18 :~`
Escherichia coli KY4271 8
19
Bacillus subtilis KY4273 2
Proteus vulgaris KY4277 8
21
22 Shigella sonnei KY4281 8
Salmonella typhosa KY4278
23
Klebsiella penumoniae KY4275 4
24
2s
26
27
2~
29
31 ~.
I - 26 -
1~78~S6
I Example 3.
2 Preparation of (+) -cis- 7-[2-(2-tritylamino-4- -
3 thiazolyl)-2-anti-metho~ minoacetamido]-2-t-
4 butyloxycarbonyl-1-azabicyclo[4,2,0]oct-2-
en-~-one:
8 HlN N ~CC~I
~ ~~~~~~ ~CO~
,~
I~ .
~2
~3 ,~lethod a)
14 In this Example,73 mg (0.307 mmole) of (+)-cis-
7-amino-2-t-butyloxycarbonyl-1-azabicyclo~4,2,0]oct-2-en-8-one
16 obtained as in Reference ~xample 9 and 135.9 mg (0.307 mmole)
17 of 2-(2-tritylamino-4-thiazolyl)-2-anti-metho~yimlnoacetic
18 acid are dissolved in 2 mQ of anhydrous methylene chloride,
l9 and 69.6 mg (0.307 mmole) of dicyclohexylcarbodiimide
dissolved in 1 mQ of anhydrous methylene chloride is added
2I thereto with stirring under ice cooling. The mix-ture i5
22 stirred for three hours and is allowed to react at a ' j,~
23 tem~erature of 10C overnight. The reaction mixture is
24 ~ashed ~lith 1~ aqueous nhosphate, saturatecl sodium
2s bicarbonate and saturated sodium chloride solu-tion. The
26 washing is dried with magnesium sulfate and concentrated
27 under reduced pressure to obtain 257 mg of a crucle product.
28 The product is purified by column chroma-togra~hy using 12 g
29 of silica c~el and a solvent of n-hexane ancl ethyl acetate
(1:1) to obtain 73 mg (35.9~) of the desired compound as
31 a pale yellow glass.
- 27 -
117895~ :;
I IX(KBr)vmax : 1780, 1725, 1695(sh), 1690, 1635
2 ~rlR(CDC~3) ~(ppm): 8.4~(d,1H,J=6.41lz), 7.25(s,15H),
3 6.30(t,1H,J=3.~), 5.35(t,1H,J=6.4IIz), 4.05
4 (s,3H), 2.5-1.6(m,4H), 1.52(s,9H) -
s ~lethod b)
6 ~ In this ~xample, 524.9 mg (1.18 mmole) of 2-(2-
7 tritylamino-4-thiazolyl)-2-anti-methoxyiminoacetic acid
8 is dissolved in 10 mQ of dried tetrahydrofuran. To the
g solution, 1.18 m~ (1.18 mmole) of lN-N-methylmorpholine-
o tetrahydrofuran and 1.18 mQ (1.18 mmole) of lN-isobutyl-
1l chloroformate-tetrahydrofuran are added at a temperature
2 of -30C and the mixture is stirred for ~0 minutes.
solution of 235 mg (0.987 mmole) of (+) -cis-7-amino-
l~ 2-t-butyloxycarbonyl-1-azabicyclo[4,2,0]oct-2-en-~-one
in~5 mQ of anhydrous methylene chloride is added dropwise
to the above mixture. The mixture is allowed to react for
30 minutes and further stirred at 0C for 2 hours. To the
18 reaction mixture is added 10 mQ of ethyl acetate and the
19 mixture is washed with water, saturated sodium bicarbonate
and saturated sodium chloride solution. The washing is
2I dried with anhydrous sodium sulfate and concentrated under
2z reduced pressure to obtain 865 mg of a crude product.
23 By silica gel chromatography using 40 g of silica gel
2~ according to ~ethod a), 580 mg (37.5~? of the desired
2s compound~is obtained. The ir and nmr spectra of the
26 product agree with those of compound prepared by Me-thod a).
27
28
29
3l ~,~
I - 28 -
~. 1 7895~
~ Exam~le 4.
2 Preparation of (+)-cis- 7-[2-(2-amino-4-
3 thiazolyl)-2-anti-methoxyimino-acetamido)~
4 1-azabicyclo[4,2,0]oct-2-en-8-on-2-carbo~ lic
acid:
6 .
H ~ 7~ H N ~ ~
8 CCOI~H N ~CONH
9 HJC ~ ~? HJCO-N ~OIH
13 In this Example, 500 mg (0.754 ~mole) of ~+)-cis-
1~ 7-[2-(2-tritylamino-4-thiazolyl)-2-anti-methoxyimino-
~5 acetami~o]-2-t-~utyloxycarbonyl-1-azabicyclo[4,2,0]oct-2
~6 en-8-one obtained in Example 3 is dissolved in
~7 a mixture of 5 m~ o~ trifluoroacetic acid, 2.5 mQ of
~B anhydrous methylene chloride and 2.5 mQ of anisole. The
19 solution is allo~7ed to stand at 0C for 3 hours and 40
minutes and then concentrated under reduced pressure.
21 To the concentrate, 5 mQ of 50~ aqeuous acetic acid is
22 added. The mixture is stirred at room temperature for
23 3 hours and is concentrated under reduced pressure. The
2~ concentrate is well triturated ~ith ether and filtered to
2s _ . obtain Z44 mg of a crude product. The product is purified
26 by column chromatography with 10 mQ of Diaion~HP-10 and a solvent
27 of methanol and water (2:5 by volume, the same shall apply
28 hereinafter~ to obtain 90 mg (32.7~) of a pale yellow powder.
?9 IR~Br)~mamx : 1760, 1670, 1630
NMR(CD3OD)~(ppm): 7.97(s,1H~, 6.40(m,1H), 5.51(d,1H,
3l J=5.011z), 4.05(s,3H), 4.3-3.7(m,1H), 2.6-l.l(m,4H)
1; - 29 ~
1178956 :-
I Example 5.
2 Preparation of (+) -cis- 7-[(R)-2-phenyl-2-t-
3 butyloxycarbonyl-aminoacetamido]-2-t-butyloxy-
4 carbonYl-l-azabicyclo[4, ,O]~ct-2-en-8-one:
s .
6 ~ (R)
7 H2N ~ ~ IHCONH
8 - ~0
9 - ' COl~U ~-~
.
2 ~lethod a)
3 In this Example, 81 mg (0.34 r~ole) of (+)-cis--
7-amino-2-t-hutyloxycarbonyl-1-azabicyclo[4,2,0]oct-2-en-
8-one and 94.0 ~y (0.34 mmole) of (R)-N-t-butyloxycarbonyl-
6 phenylglycine arè dissolved in 2 mQ of anhydrous methylene
~7 chloricle. ~ solution of 77 mg (0.3~ mmGle) of dicyclo-
8 hexylcarbodiimide in 1 mQ of anhydrous methylene chloride
19 is added to the solution under cooling with ice and sodium
chloride. The mixture is allowed to react under cooling
21 with ice for 2 hours and t~o drops of acetic acid is added
22 thereto. The mixture is stirred for 20 minutes and filtered
23 under reduced pressure. The cake is washed with 20 mQ of
2~ ethyl acetate. The filtrate and the washing are combined
2s and 20 mQ of ether is added thereto. The mixture is washed
2~ With 1% a~ueous phosphoric acid, saturated sodium blcarbonatc
27 and saturated sodium chloride solution. The washirlg is dried
28 with anhydrous sodium sulfate and concentrated under reduced
29 pressure to obtain 187 mg of a crude product. The product
is purified by silica gel chromatograPhy with 9 g of silica
3l gel and a solvent of n-hexane and ethyl acetate (1:1) to
I ~ 30 -
9S6
I obtain 104 mg (64.9%) of the desired compound as a colorless
2 glass.
3 IR(Ksr)vmax : 1770, 1750, 1720, 1630
4 N~IR(CDCQ3)~(p?m): 7.32(s,SH), 6.31(m,lH), 5.90(m,lH), ---
i 2.5~-1.70(m,4H), 1.50(s,9H), 1.40(s,9H)
6 ~lethod b)
7 In this Example, 297.3 mg (1.18 mmole) of (R)-N-
8 t-butyloxycarbonylphenylglycine is dissolved in 5 mQ of
9 anhydrous tetrahydrofuran, and 1.18 mQ (1.18 mmole) of
o lN-N-methylmorpholine-tetrahydrofuran and 1.18 mQ (1.18 mmole) of
~ J- i~u chloroformate-tetrahydrofuran are added at -30C.
12 The mixture is stirred for 30 minutes and 234 mg (0.983
13 mmole) of (+)-cis-7-amino-2-t-butyloxycarbonyl-1-azabi-
14 cyclo[4,2,0]oct-2-en-~-one dissolved in 5 mQ of anhydrous
IS m~thylene chloride is aclded -thereto. The mixture is
l6 allowed to react at a temperature oF -3~C for ~5 minutes
l7 and at 0C for 4 hours and lS minutes. The reaction
18 mixture is diluted with 15 mQ of methylene chloride and
l9 is washed successively with water, l~-HCQ, water and
saturated sodium chloride solution. The washing is dried
~, ,~,~,
21 with anhydrous sodium sulfate and concen-trated to obtain
22 5~8 mg of a crude acyl-compound. Purieication by silica
23 gel chromatoc~raphy with 28 g of silica gel is carried out
2~ accord.ing to ,~ethod a) to obtain 322 mg (69.4~) of the
2i desired compoundas a colorless glass. The ir and nmr
26 spectra of the product agree with those of the compound
27 prepared in !1ethod a).
28
~9
31
-- 3
li7.~S~
1 Examole 6.
2 PreparatiOn of ( 1 )-cis-
3 7-[(R)-2-phenyl-2-aminoacetamido~-1-azabicyclo-
14~2~o]o~t-2-en-8-on-2-carboxylic acid:
s
6 ~ fHCONH ~
8 NHBOC o"L~ t
g ~ CON~
NH~
CO,~
In this Example, 2~0 mg (0.59 mmole) of ('j-cis-
l3 7-[(~)-2-phenyl-2-t-butyloxycarbonylaminoacetamido~-2-t-
1~ butyloxycarbonyl-1-azabicyclol4,2,0]oct-2-en-8-one obtained
in Example 5 is dissolved in 2.5 mQ of anhydrous
J6 methylene chloride and 2.5 m~ of anisole, and 5.0 mQ of
1~ trifluoroacetic acid is added under ice cooling. The
~s mixture is allo~ed to stand for 4 hours an~ 50 minutes
19 under ice cooling and concentrated. To the concentrated
residue, 10 mQ of ether is added and the mixture is stirred
21 at room temperature for one ho~r to form a precipitate.
22 ~he precipitate is collected by fiitration to obtain 202 mg
23 (70.9%) of the desired compound as a pale yellow powder.
21 IR(KBr)vmax : 1765, 1680, 163~
2s 2IMRtD2O ~ith DSS as an internal standardj~(ppm~:
26 7.51(d,5H), 6.31(m,1H), 5.19(s,1H), 4.95(d,1H),
27 3.8-3.5(m,1H), 2.6-2.9(m,4H)
2~ Separation of the diastereoisomers of (~)-cis-
29 7~ )-2-phenyl-2-aminoacetamidol-1-azabicyclo
[4,2,0]oct-2-en-8-on-2-carboxylic acid:
3l The compound ( sn mg) obtained by the above method
I - 32 -
., "
~,
. . . ,~
89S6
I is dissolved in 150 m~ of water and the solution is subjected
2 to high speed liquid chromatography using Bondapak~C-18
3 (product of T~aters Co.) as a carrier and a sol~ent of 7%
4 methanol and 0.21~ potassium hydrogen phos~hate eight times.
~ The isolation of two fractions are monitored by a spectro-
6 scopic analysis at a wave length of ~54 nm. ~fter removing
7 methanol under reduced pressure, each fraction is lyophilized~
8 The dried matter i5 dissolved in water and adsorbed on a
g column packed with 20 mQ of Diaion~HP-10 (product of
~0 ~litsubishi Kasei Kogyo Co., Ltd.). The column is washed
Il with 200 mQ of water and elution-is carried out with 20%
12 ethanol. The fractions, positive to ninhydrin test are
collected and lyophilized to obtain 14.0 mg o. A-isomer
~J and 2~.6 mg of B-isomer as white powder of potassi um sal t .
A:' more polar ~raction
16 ~ ~22 (~ater, C=0 5); 7
17 IR(KBr)vmax : 1750, 1690, 1640
P~(D2O)~(p~m): 7.51(5H,s), ~.l5(lH,t,J=3.9Hz),
19 5.~0(1H,d,J=4.9Hz), 5.19(1H,s), 3.~8(11~, octet,
J=8.6, 3.7, 4.9Hz), 2.41-l.al(aH,m)
2~ B: less polar fraction
22 [~]22 (H2O, c=0.5): +57.2
--1
23 IR(KBr)vmax : 1760, 1690, 1640
2~ PPI~(D2~)~(p~m): 7.51(5H,s), 6.08(1H,t,J=4.2Hz),
2; 5.41(1H,d,J=4.9Hz), 3.83(1H,octet,J-8.6, 3.7,
26 4.9Hz), 2.2~-1.01(4H,m)
21 Taking Structure Activity Relationship of _
23 cephalosporins into consideration, the less polar ispmer
29 which has dextro []D value and stronger antimicrobial
activity than the more polar isomer as shown in table below
31 is assigned to have 6(~)7(~) absolute configuration.
- 33 -
.~
1-~7~9S6
1 ~xample 7
.
2 Preparation of (')-cis- 7-~(R)-2-phenyl-2-(4-
3 ethyl-2,3-dioxo-l-piperazinylcarbonyla~ino)-
4 acetamido]-2-t-~utyloxycarbonyl-l-azabicyclo
[4,2,0]oct-2-en-8-one:
7 ~ fHCONH
g ~a ~ f co~ ;
- Et
,~
13 J~eth~d a)
1~ In this Example, 68 mg (0.286 mmole) of (~)-cis-
7~amino-2-t-butyloxycarbonyl-l-azabicyclo[4,2,0~oct-2-en-
8-one and 100.4 m7 (0.2~6 mmole) of ~)-2-phenyl-2-(4-
ethyl-2,3-dio~o-l-piperazinylcarbonylamino) acetic acid
8 are dissolved in 2 mQ of anhydrous methylene chloride,
~9 and 70 m~ (0.315 mmole) of dicyclohexylcarbodiimide
dissolved in l m~ of anhydrous methylene chloride is added
21 under ice cooling. The mixture is stirred for 6 hours and
22 further stirred at a temperature of 10C overnight. The
23 reaction mixture is filtered and the cake is washed with
2~ methylene chloride. The filtrate and the washing are
2s combined and washed successively with 1% phosphoric acid,
26 saturated sodium bicarbonate and saturated sodium chloride
27 solution. The washing is dried with anhydrous sodium
28 sulfate and concentrated to obtain l89 mg of a crude acyl
2g compound. The product is purified by silica gel chromato-
graphy usins 9 g of silica gel and a solvent of n-hexane
31 and ethyl acetate (l:2) to obtain 43 mg of a more
1 ~ 34 ~
11~7~3956
polar isomer, 20 mg of a less polar isomer and 11.1 mg
z of a mixture of the two isomers. The total yield is
3 54.1%.
4 .he more polar isomer -
I~(CI~CQ3)vmax : 1780, 172~, 1695(sh), 1685
6 Nr~(CDCQ3)~(o~m): 7.73(d,1H,J=7.0E~z), 7.37(s,5H),
~ 6.25(m,lH), 5.7-5.0(m,2H), 4.3-3.0(m,7H),
8 2.6-0.7(m,4H), l.SO(s,~), 1.20(t,3H)
9 The -less polar isomer : ~.
o I~CHC~3)vmamx : 1780, 1695, 1685, 1620
Il NPIP~(CDCQ3)~(ppm): 7.77(d,1H,J=8.0Hz), 7.30(m,S~),
l2 6.21tm, lH), 5.67-5.33(m,2~), 4.5-3.2(m,7H),
13 3.5-l.O(m,4~), 1.50(s,9~1), 1.2~(t,3H)
l~ Method b)
IS ~ In this Example, 428.5 mg (1.13 mmole) of (R) 2-
phenyl-2-(4-ethyl-2,3-dioxo-1-piperazinylcarbonylamino)
11 acetic acid is dissolved in 10 mQ of dried tetrahydrofuran
l8 and 1.25 mQ (1.25 mmole) oL lN-N-methyLmorpholine-
t9 tetrahydrofuran and 1.25 m~ (1.25 mmole) of lN-isobutyl-
chloroformate-tetrahydrofuran are added at a temperature
21 of -30C. The mixture is stirred for 30 minutes and 235 mg
22 (1.13 mmole) of (+)-cis- 7-amino-2-t-butyloxycarbon
23 azabicyclo[4,?,0]oct-2-en-8-one dissolved in 5 m~ of
~ anhydrous methylene chloride is added. The mixture is
; allowed to react for one hour and stirred at a temperature
26 of 10C overnight. The reaction mixture is diluted with
27 20 mQ of ethyl acetate and washed successively with water,
28 0.1~-HCQ, saturated sodium bicarbonate and water. The
29 washing is dried with anhydrous sodium sulfate and concen-
trated to obtain 570 mg of a crude acyl com~ound. The
31 product is purified and fractionated according to ~lethod a)
I - 35 _
-
11789S~
l except that 27 g of silica gel is used to obtain 73 mg of
2 a more Dolar isomer anfl 61 m~ of a less polar isomeI (total
3 yield 65.4~). The ir and nmr spectra of the iso~ers
a~ree with those of the isomers o~tained by ~ethod a).
' -
6 Example 8.
~ Preparation of (~)-cis- 7-[(R)-2-phenyl-2-(4-
8 ethyl-2,3-dioxo-l-piperazinylcarbonylamino)
9 - acetamido]-1-azabicyclo[4,2,0]oct-2-en-8-on-2-
carboxylic acid:
Il
¦~ CO,~ CO ~ -
(N~ 0 ~N ~ 0
16~t .
J7
18 In this Example, 103 mg (0.2~8 mmole) of (_) cis-
19 7-[(~)-2-phenyl-2-(4-ethyl-2,3-dioxo-l-piperazinylcarbonylamino)
acetamido~-2-t-butyloxycarbonyl-l-azabicyclo[4,2,0]oct-2-en-
~t 8-one (the less polar isomer) obtained in Example 7 is
22 dissolved in a mixture of S mQ of trifluoroacetic acid,
23 5 m~ of methylene chloride and two drops of anisole. The
24 mixture is allowed to react at 0C for 2 hours and
2; concentrated under reduced pressure. To the concentrate,
26 dried benzene is added and the mixture is again concentrated
27 to obtain an oily product. To the product, ether is added
28 and the mixture is stirred at room tem~erature to form a
29 yellow precipitate. Crude product (104 mg) is collected
by filtration as a yellow powder. The crude product is
31 dissolved in ethyl acetate and extracted with 5 m~ of
- 36 -
i.
~ . j
117~9~j ` - ` `
I saturated sodium bicar~onate three times. The extracts - -
2 are wasled with ethyl acetate. ~he ~lashing is adjusted to
3 a pH of 2.5 ~ith 0.5N-HCQ under ice cooling and extracted
4 ~ith 5 mQ of ethyl acetate three times. The e~tract is
washed t~ith saturated sodium chloride solution, dried ~ith
6 magnesium sul ate, and concentrated under reduced pressure
7 to obtain 41 mg (46.0~) of a pale vello~ po~der.
8 IP(KBr)v~ax : 1775, 1720, 1685, 1620(sh)
g - N~R(CD3O~)~(ppm): 7.31(s,5H), 6.33(t,1H,J=4.0Hz),
5.40(m,~H), 4.30-3.1(m,7H), 2.40-0.7(m,4H),
Il 1.27(t,3H)
l2 From strong antimicrobial activity as shown in
13 t~ble belo~7, this compound is assigned to have 6(~)7(S)
l4 absolute configuration.
~xample 9.
_ _ .
16 Prep~ration of (~) -cis-7~-~(R)-2-phenyl-2-t-
~7 - butyloxycarbonylaminoacetamid]-4~-methyl-2-t-
18 butvloxvcarbonyl-1-azabicyclo[4,2,0]oct-2-en-
l9 8-one:
23 ~2N ~ ~ CE~ ~L~ J j J
24 0~ ~ CO~u
In this Example, 132 mg (0.53 mmole) of (i')--~J-t-
26 butyloxycarbonylpllenylglycine is dissolved in 5 mQ of
27 anhydrous tetrahydrofuran, and 0.53 mQ (0.53 mmole) of
28 lN-N-methylmorpholine and 0.53 mQ (0.53 mmole) of lN-
29 isobutyl chloroformate are added thereto at a te~perature
of 0C. The mixture is stirred for 15 minuteS and 0.07 mQ
3J (0.5 mmole) of triethylamine and 1.44 Mg (0.5 mmole) of
1- 37 -
1~789S~i ~
I the hydrochloride of (') -cis- 7B-amino-4~-methyl-2-t-
2 butyloxycarbonyl-1-azabicyclo[4,2,0]oct-2-en-8-one obtained in
3 Reerence Examole 12 are added thereto. The mixture is stirred
4 at a temoerature of 0C for one hour and then at a tempera-
ture of 5 to 10C overnight.
6 - The reactlon mixture is diluted with 10 mQ of
7 ethyl acetate and washed successivel~ with 10% citric acid,
8 saturated sodium bicarbonate, and saturated sodium chloride
g solution. The washing is dired with anhydrous sodium
sulfate and concentrated to obtain a crude acyl compound.
l The product is purified by column chromatography 30 g of
silica gel and a solvent of n-hexane and ethyl acetate
~3 (3:1, by volume). ~he desired co~pound [150 mg (61.7%)]
14 is obtained as a powder.
lS ~ IR(KBr)vmax : 3290, 1780, 1720, 1685, 1665
16 ~JIR(CDCQ3)~(Ppm): 7.34(5H,s), 6.60, 6.49(lH,d
17 . respectively,J=7Hz), 6.11, 6.04(1H,d respectively,
18 J-2Hz), 5.66, 5.60(1H,~, respectively,J=7Hz),
l9 5.18(2H,m), 3.~6(lH,m), 2.46(lH,m), 1.75(2H,br),
l.Sl(9H,s),1.42(9H,s), 1.15, 0.98(3H,d respec-
- . ~
21 tively, J=7.5Hz)
22
2 Exam~le 10.
24 Preparation of the trifluoroacetate of (+)~-cis-
7B-[(R)-2-phenyl-2-aminoacetamido]-4~-methyl-2
2G carhoxy-l-azabicyclo[4,2,01oct-2-en-8-one:
27
28
zg
3 1 ~, .
3 8 -
11789S~ - ~
2 ~ H H '
3 ~ CO~I ~, H C~~ ~HCOL~
4 ~ BOC
~ CF3CQ.2E~
C02~u ~ C02H
8 In this ~xample, 100 mg (0.21 mmole) of (+)-cis-
9 7~-[~R)-2-phenyl-2-t-butyloxycarbonyl aminoacetamido]-4
lo methyl-2-t-butyloxycarbonyl-l-azabicyclo[~2~o]oct-2-en
~1 8-one obtained in Example 9 is dissolved in 1 m~
of anhydrous methylene chloride and 1 mQ of trifluoroacetic
acid is added thereto under cooling on an ice bath. The
14 mixture is allowed to stand at a temperature o~ 0 to 5C
f~r 3.5 hours with occasional shaking. The reaction
16 mixture is concen~rated under reduced pressure. The con-
centrate is triturated Jith 5 m~ of anhydrous ethylether
18 and the ether layer is removed by decantation. The
19 treatmen~ is repeated three times and the resulting cake
is dried under reduced pressure to obtain 70 mg (75%) of
21 the desired compound as a po~7der.
22 IR(KBr)vmax : 3450(sh), 3230(sh), 3060, 2950-2~00,
23 1769, 1695(sh), 1681(sh), 1673
2~ ~1R(D~1SO-c16)~(ppm): 9.29(lH,t,J-~Mz), 7.49(5l1,s),
2s : 6.11, 6.04(lH,d respectively, J=2Hz), 5.30(lH,
26 m), 4.97(1H,d,J=411z), 3.~2(1H,br), 2.44(br,
27 partly overlapping wi-th the signal o~ DMSO-d6),
28 1.82(2H,br), 1.14, 0.91(3H,d, respectively,
29 J=7.5Hz)
Exampl _11. -
3I Preparation of (+) -cis-7~-[(R)-2-phenyl-2-t-
~ 39 -
~7~95~ ,
I butyloxycarbonyl-acetamido]-~-methyl-2-t-
2 butvloxycarbonyl-1-azabicyclol4,2,0]oct-2_en-8-
3 one:
H~N ' ' CH3 ~ (~ H ~I3
~ ' l~ ~c
6 ~o~tEU co2 ~
7 In .his Exam21e, the sa~e procedure as in
8 Example 9 is employed except that 144 mg of (+)-
9 ~ci~s- 7~-amino-4a-methyl-2-t-butyloxycarbonyl-l-azabicyclo-
l~4~2,03OCt-2-en-8-one obtained in Reference Example 11 is used
as a starting ~ompound. As a result, 140 mg (57.6%) of the
~2 desired compound is obtained.
--1
13 I~(XBr)vr~C~amx : 3330, 1792(sh), 17g2, 1730, 1692, 1675
14 N~(CDC~3)~(ppm): 7.34(5H,s), 6.73, 6.60(1H,d
~s respectively, J=7Hz), 6.28(1H,t,J=6I~z), 5.60(1H,
l6 m), 5.43-5.18(2H,m), 3.82(1H,m), 2.55(1H,m),
17 1.69(2H,m), 1.51(9H,s), 1.41(9~1,s), 1.09, 1.03
18 (3H,d respectively, J=7Hz)
19 ExarnDle 12.
Preparation of (+)-cis-
21 7~-[(R)~2-phenyl-2-aminoacetamido]-4a-methyl-2-
22 carboxy-l~azabicyclo[4,2,0~oct-2-en-8-one:
23
24 ~ ~C'~i H ,CH3~ ClHCONH ~
~OCo F ~ ~ N ~-
26 CO~u CO~H
27
28 In this Example, the same ~rocedure as in
29 Example 10 is employed except that ~0 mg of (+)-
cis- 7~-[(R)-2-phenyl-2-t-butyloxycarbonyl-aminoacetamido]-
31 4a-methyl-2-t-butyloxycarbonyl-1-azabicvclo[4,2,0]oct-2-en-
- 40 -
:1~ 7895~
~ 8-one obtained in Examplé 11 is used as a starting
2 compowld. A~ a result, 73 mg tlO0~) of the desired
3 compound is obtained.
4 ~ I~(KBr)vT,Iax : 3430, 3200, 3060, 2960-2650, 1780(sh),
1770, 1695(sh), 1680
6 N~l~(DMSO-do)~(ppm): 9.36(1H,d,J=8Hz), 7.47(5H,s),
7 6.28(1~,d,J=6Hz), 5.40(1H,m), 4.98(1H,m), 3.70
8 (lH,br), 2.45(br, partly overlappi~g ~ith the
g signal of DMSO-d6), 1.80(2H,m), 1.06, 0.95(3H,d
respectively, J=7.5Hz)
Il ~The above compound (30 mg) is dissolved in 100 ~1 of
l2 water and the solution is subjected to high speed liquid chro-
l3 matography using Bondapak C-18 (product of Waters Co.) as a
~4 carrier and a solvent of 7 % methanol and 0.2N potassium hydrogen
phosphate five times. The isolation of two fractions are monitored
16 by a spectroscopic analysis at a wave length of 254 nm. After
17 removing methanol under reduced pressure, each fraction is
l8 lyophilized. The dried matter is dissolved in water and adsorbed
19 on a column packed with 20 ml of Diaion HP-10. The column is
washed with 200 ml of water and elution is carried out with 20 %
~l ethanol. The eluate is taken in 20 ml fractions. The fractions
22 which are pOSitive to ninhydrin-test are collected and lyophilized
23 to obtain 8.9 Tng (yield 29 ~) of C-isomer which elutes earlier in
24 the high speed liquid chromatography and 11.4 mg (yield 38 %) of
zs D-isomer which elutes later. These are the potassium salts of the
26 desired compound. Specific rotations of the isomers are as
27 follows.
- 41 -
f~
1~78956
1 C-isomer ~a~D5 -38.4 (H20, C - 0.5)
2 D-isomer ~a~D5 ~4.23 (H20, C = 0.52)
3 D-isomer
4 IR(KBr) vmamx : 3420, 1760, 1695, 1633
PMR(D20)~(ppm): 7.51(5H, s), 6.10(1H, d, J ~ 5.1),
6 5.47(1H, d, J = 4.7), 5.19(1H, s), 3.89(1H, m),
7 2.45(1H, m), 1.44 - 1.04(2H, m), 1.00(3H, d,
8 J = 7.4).
g D-isomer which has stronger antimicrobial activity
o than diastereomixture ~(+) form] as shown in table below is
assigned to have 6(R)7(S) absolute configuration.
- 42 -
1~78g56 ;.~.,. i.-
1 Example 13. -~ -
_
2 Preparation of (+)~cis- 7~-[2-(2-tritylamino-4-
3 thiazolyl-2-methoxyiminoacetami;~c~]-T~-~ethyl-2-t
4 butyloxycarbonyl-1-azabicyclo~4,2,0]oct-2-en-8-
one:
7 H~N: ~ CH3 ~ ~ CCCNH
8 ~ CO ~ ~C~
9 "~ ' C0.~3u j~
~-~ethod A.
1l ~n this Example, 8~ mg (0.35 mmole) of (+)-cis-
12 7~-amino-4~-methyl-2-t-butyloxycarbonyl-1-azabicyclo
l3 [4,2,0]oct-2-en-8-one is dissolved in 1.~ mQ of anhydrous
14 methylene chloride and 155 mg (0.35 mmole) of 2-(2- --
IS tritylamino-4-thiazolyl)-2-anti-methoxyiminoacetic acid
16 is added. Further, 1.5 mQ of anhydrous dloxane is added
17 to the mixtu~c to make it more homogeneous. To the mixture,
18 80 my (0.39 mmole) of dicyclohexylcarbodiimide dissolved
19 in 1 mQ of dioxane is added and the resulting mixture is
stirred at a temperature of 5 to 10C overnight. The
21 resulting white precipitate is filtered off and 10 mQ of
22 ethyl acetate and 5 mQ of ether are added to the filtrate.
23 The mixture i5 washed successively with 5 mQ of l~ cold
2~ phosphor1c acid three times and saturated sodium bicarbona-te.
2s The washing i.s dried with anhydrous sodium sulfate and
26 concentrated under reduced pressure to obtain 2~0 mg of
27 the crude desired compound as a semlsolid. The crude
28 compound is charged on column packed with 27 g of silica
29 gel and elution is carried out with n-hexane and ethyl
acetate (2:1). The eluate is concentrated under reduced
3I pressure to obtain 170 mg (72~) o.f the desired compound. ~r~ri.
I -43 -
117~3~5~ ~
~ ~lelting point (recrystallized from n-hexane and
2 ethyl acetate): 214.5-Z15.5~C
IR(KBr)~max : 3240, 17~0(sh), 1762, 1730, 1665, 1636
~ N~(cDcQ3-cD3oD)~(p~m): 7-3!(153 H,s), 6.64(2/3H,s), -- -
s 6.34(1H,d,J=7Hz), 5.48(~/3~,d,J=5.5Hz),
6 5.43(1/3H,d,J=5.5Hz), ~.O~(lH,s),
? 4.01(2E~,s), 3.90(1H,m), 2.~6(1H,m), 1.75(2H,m),
8 - 1.5~,(3H,s), 1.53(6H,s), 1.1~(3H,d,J=7Hz)
g Method B. . F~-.
~o In this Example, 243.9 mg (0.05 mmole) of 2-(2--
1~ tritylamino-4-thiazolyl)-2-anti-methoxyiminoacetic acid is
l2 dissolved in 5 mQ of anhydrous tetrahydrofuran and 0.55 mQ
l3 (0.55 mmole) of l~-N-methylmorpholine is added. To the mixture,
l~ 0.55 mQ (0.55 mmole) of lM-ip,u chloroformate-tetrahydrofuran
is added dropwise at a ternperature of 0C with stirring
l~ and the mixture is further stirre~ for 15 minutes.
17 Triethylamine [0.11 mQ (0.5 ~mole)] is added to the mixture
18 ~ollo~/ed b~ addition of 1~4 m~ (0.5 ~ole) of the hydro-
ls chloride of (~) -cis-7~-amino-4~-rne-th~1-2-t-butyloxycarbonyl-
1-azabicyclo[4,2,0]oct-2-en-8-one. The mixture is stirred
21 at a temperature of 5 to 10C overnigh-t and concentrated
under reduced ~ressure. Ethyl ace-tate (10 mQ) i5 added
to the concentrate and the resultiny mixture is washed
successively with 5% hydrochloric acid, saturated sodium
chloride solution, saturated sodium bicarbonate and
q6 saturated sodium chloride solution. The washing is dried
with anhydrous sodium sulfate and concen-trated under_
reduced pressure to obtain a crude product. The product
~9 is charged on a column packed with 25 g of silica gel and
elution is carried out with n-hexane and ethyl ace-tate
31 (5:3). The eluate is concentrated under reduced pressure
I - 44 -
~1789S6
. . .
to obtain 250 m~ (74.0~) of the desired compound. Physical
2 properties of the compound agree with those of compound
3 pre~ared by ~lethod ~.
Exam~le 14.
6 Preparation of (+)-cis- 7~-[2-(2-amino-4-
7 thiazolyl)-2-anti-methoxyiminoacetamido]-4~-
8 methyl-2-carboxyl-l-azabicyclo[4,2,0]oct-2-en-
9 - 8-one: . - G~;
_~S
l2 H~CO-N ~"C~ ~CCOI`IH~ CHJ
,4 C2 ~U
-CO2H
1s
l6
l7 In this Example, 70 m~J (0 103 rnmole) oE (+) -cis-
l8 7~-~2-(2-tritylamino-4-thiazolyl)-2-anti-methoxyimino-
l9 acetamido]-4a-methyl-2-t-butyloxycarbonyl-1-azabicyclo
[4,2,0]oct-2-en-8-one obtained in Example 13
2l is dissolved in 0.5 mQ of anhydrous methylene chloride
22 and 0.1 mQ of anisole. The mixture is cooled to 0C
23 followed by addition of 0.5 mQ of trifluoroace-tic acid.
2~ The resulting mixture is allowed to stand on an ice bath
2s for 3.5 hours.
26 The reaction mixture is concentrated under
27 reduced pressure. The concentrate is -triturated with 5 mQ
28 of anhydrous ethylether and filtered to obtain a white
29 powder. The powder is dissolved in 2 mQ of 50% acetic
acid. The solution is allowed to stand at room temperature
3l for 2.5 hours and then at a temperature of 5 to lO~C
,~ - 45 -
1178956 -
I overni~ht. The soluti~n is further allo~^~ed to stand at
2 room temperature (25C) for 6 hours and concentrated under
3 reduced pressure to obtain a glassy product. ~he glassy
4 product is l~ell triturated with ether and filtered. The -
filtrate is dried to obtain 20 mg (5L%) of the desired
6 eompound.
--1
7 I~(KBr)vmax : 3480, 3300, 1770, 1680, 1635
8 Nr~(DMSO-d6)~(ppm): 9.17(lH,d,J-8Hz), 7.50(1H,s),
9 ~ 7.24(2H,m), 6.31(1H,d,J=5Hz), 5.52(1H,m), ~*
o 4.00(3H,s), 2.65(br, partly overlapping with
1I the sianal of D~SO-d6), l.7o(2H~m)~ 1.06(3H,d,
12 J=7.5Hz)
l3
l4 Exam~le 15.
Preparation of (~) -eis-7R-[2-(2-tritylamino-4-
thiazolyl)-2-methoxyiminoacetamic1O~-4R-methyl-2-
17 t-butyloxyearbonyl-1-azabicyelo[4,2,0]oet-2-en-
18 8-one:
19
20 H H Tr~ S ~ -
21~2N~ ~ CH3 ~ ~o~ H C~13
222~ ~-~u oJ~I~ .......
24
2s
26In this ~xample, the same procec1ure as in
27Example 13 Method B is employed except that 202 m~ _
2~ (0.7 mmole) of (+) -cls- 7~-amino-4~-methyl-2-t-butyloxy-
29 carbonyl-1-azabicyclo[4,2,0]oct-2-en-8-one is used as a
starting compound. As a result, 251 mg (53%) of the
3I desired eompound is obtained.
~ - 46 --
11789S6 ~
I ~ielting point: 2'01.~-202.0C
2 IR(KBr)vmax : 3225, 1780(sh), 176~, 1725, 1668, 1635
3 N~IR(CDC~3-CD30D)~(ppm): 7.35(1/2H,s), 7.30(15H,s),
4 6.55(1j2H,s), 6.14(1H,d,~=2Hz), 5.33(1H,d-d,
J=5Hz), 4.09(3/2H,s), 4.01(3/2H,s~, 3.96(lH,m),
6 2.45(1H,m), 2.05(2H,m), 1.53(9/2H,s), 1.52(9/2H,
7 s), 1.16(3H,d-d,J=7.5Hz) -
9 Example 16. '
0 Preparation of (+)-cis- 7~-[2-(2-amino-4-
11 thiazolyl?-2-methoxyiminoacetamido]-4,B-methyl-
12 2-carboxy-1-azabicylo[4,2,0]oct-2-en-8-one:
13
4 , S ~S
6 IlCONEICl-I~ CC~ CH~
N0~
COltf3-1 O~H
a
19
In this Example, the same procedure as in ~,~
... . . .
21 Example lA is employed except that 70 mg of (i)-
22 cis-7~-[2-(2-tritylamino-~-thiazolyl)-2-methoxyimino-
23 acetamido~-4~-methyl-~2-t-butyloxycarbonyl-1-azabicyclo
2~ [4,2,0]oct-2-en-~-one obtained inlExample 15
2s is used as a ~starting compound. As a result; 22 mg (56%)
26 of the desired compound is obtained.
2Y IR(KBr)~mamx : 3460, 3280, 1780(sh), 1770, 1670, 1630'
28 NrlR(D~ISO-d6)~(ppm): 9.24(1/2H,d,J=~Hz), 9.17(1/2H,d,
29 J=3Hz), 7.50(1/2H,s), 7.25(2H,m), 6.78(1/2H,s),
6.10(1}1,d,J=2Hz), 5.47(1H,m), 4.00(3/2M,s),
31 ' 3.85(3/2H,s), 2.60(br, nartly overlapping with
1~7~5~ ~
1 the signal of D~ISO-d6), 1.91(2H,m), 1.12(3~,
2 d-d,J=7.5Hz)
4 Example 17.
Preparation of (+) -cis- 7~-[2-(2-chloroacetyl-
6 amino-4-thiazolyl)-2-syn-metho~yiminoacetamido]-
7 4~-methyl-2-carboxy-1-azabicyclo[4,2,0]oct-2-en-
8 3-one:
g . , - ~,. .
O ~ C-~C~H >2C ~ ~ ~CONH l~ C~l~
OlH o N
13 CO~H
. .
6 In this`Example, 172 mg (0.62 mmole) of (2-
chloroacetylamino-~-thiazolyl)-2-syn~methoxyiminoacetlc
~8 acid is suspended in 3.6 mQ of anh~drous dichloromethane
19 and 68.9 rng (0.68 mmole) of triethylamine is added there-to
to make the solution homogeneous. Under cooling on an ice-
21 sodium chloride bath, 129 mg (0.62 mmole) of phos~horus
22 pentachloride is added to -the mixture with stirring and
23 the resultinq mi~ture is stirred ~or 1.5 hours. n-Hexane
2~ (13.3 m~) is added to the mixture and the supernatant is
removed by decantation. Anhydrous tetrahydrofuran (1.3 mQ)
26 is added to the residue to obtain an acid chloride solution.
27 On the other hand, 160 mq (0.52 mmole) of the
~8 trifluoroacetate of (~)-cis- 7~-amino-4~-methyl-2-carboxy-
~9 1-azabicyclo[4,2,0]oct-2-en-8-one obtained in Reference Example
13 is dissolved in one mQ of 50~ tetrahydrofuran-water, and
31 209 mg (~.06 ~mole) of triethylamine is added thereto.
48
117~9~
1 The mixture is added to the acid chloride solution under
2 ice cooling with stirring. After stirring at the same
3 temDerature for 1.5 hours, the mixture is adjusted to a
4 p~ of 4 to 5 with lN-hydrochloric acid and extracted with --
s lO m~ of ethyl acetate three times. The ethyl acetate
6 extracts are washed with sodium chloride solution, dried
7 with anhydroùs sodium sulfate and concentrated under
8 reduced pressure to obtain 147 mg (52.1%) of the desired
g compound. - P~`~
--1
lo IR(KBr)vmax : 1765, 1680
1~ P.~IR(DMS~-d6)~(ppm): 7.40(1H,s), 6.32(1H,d,J=5.2~z),
.~
12 5.53(1H,m), 4.35(2H,s), 3.90(3H,s), 2.50(1H,m),
3 1.9~-1.27(2H,m), 1.10(311,d,~J=7~5Hz)
IS ~xample 18.
____ .
16 Preparation of (+)-cis- 7~-[2-(2-amino-4-
l7 thiazolyl)-2-syn-methoxyiminoacetamido]-4u-
~ methyl-l-azabicyclo[4,2,~]oct-2-en-8-on~2-
19 carboxylic acid:
21 C~CH~CONH ~ ~ - H H H~N ~ ~
CCOI~H ~ ~ CH~ N CC~I H H CH3
22 ~ ~ ~ O~H
2s
26
27 In this Example, 147 mg (0.321 mmole) of the
73 chloroacetyl compound obtained in Example 17 is
29 dissolved in ~.5 mQ of dimethylsulfoxide and 2.5 mQ of
dimethylformamide, and 47 mg (0.6~ mmole) of thiourea is
31 added thereto at room temperature with stirrin~.
I -49 -
~1'7~gS~
I The mix~ure is stirre~ for 14 hours. ~fter addition of
2 ether, the supernatant is removed by decantation and the
3 residue is dissolved in a small amount of ~imethylsulfoxide.
4 The solu,ion is adsorbed on a column packed with 10 m~ of
s Diaion~HP-10. The column is ~ashed with 240 mQ of water
6 ana elution is carried out with a solvent of methanol and
7 water (1:10 to 1:2). The eluates are collected and
8 methanol is removed under reduced pressure. The residue
g is again adsorbed on a column packed with 10 mQ of Diaion~ i
~o HP-10 and the column is washed with 500 m~ of water.
Elution is carried out ~ith a solvent of methanol and water
~2 (1:1). The eluates are collected and concentrated under
3 reduced ~ressure to obtain 50.2 mg (41.1~) of the des1red
1~ compound.
IS ~ IR(KBr)vmax : 1760, 1680, 1655
16 PMR(DMSO-d6)~(ppm): 9.27(1H,d,J=9.OHz), 7.15(2H,br),
17 6.75(1H,s), 6.31(1H,d,J=a.2Hz), 5.5811H,br),
18 3.85(3H,s), 2.60(1H,m), 1.67(2H,br), 1.08(3H,d,
l9 J=8Hz)
21 Exam~le 19.
22 Preparation of (~)-cis-7 -[2-(2-chloroacetyl-
Z3 amino-4-thiazolyl)-2-syn-methoxyiminoacetamido]-
2~ 1-azabicyclo[4~2~0]oct-2-en-8-on--2-carbo~ylic acid:
2;
26 H H cecH~coNH~
27 ~I~N ~ ~ CC~ 7~?--
28 0 ~ 1~
29 CO~H C02H
3l
-- ! ~ 50 -
.
~ ~ 7f~9S6 ~
1 In this E~ample, 54.2 mg (0.195 mmole) of 2-
z chloroacetylamino-4-thiazolyl-2-Syn-methoxyiminoacetic acid
3 (syn ty~e) is suspended in 0.98 mQ of anhydrous methylene
4 chloride and 23.4~ m~ (0.195 mmole) of triethylamine is - -
added. Phosphorus pentachloride 40 8 mg (0.195 mmole)
6 is added to the reaction mixture under ice cooling.
7 ~fter stirring for 20 minutes, 3.92 mQ of n-hexane is added
8 to the mixture and the supernatant is removed by decantation.
g The residue is dissolved in 1.96 mQ of tetrahydrofuran to ~.
o obtain an acid chloride solution.
Il On the other hand, 45.g mg (0.155 mmole) of the
l2 trifluoroacetate of (+)-cis-7-amino-2-carboxy-1-azabicyclo-
[4,2,0]oct-2-en-g-one obtained in Reference Example ~ is
l4 dissolved in 2 mQ of 50% tetrahydrofuran-water and 47.4 mg (0.469
Is m~ole) of tri~thylamine is added thereto. To the solution,
l6 the above acicl chIoride solution is added under ice cooling
17 and the mi~ture ls stirred for 2 hours. The mixture is
l8 adjusted to a pl~ of 2.0 with 10~ hydrochloric acid and
19 extracted three times with ethyl acetate.- The extracts
are washed with saturated sodium chloride solu-tion. The
2I washing is dried ~ith anhydrous sodium sulfate and
22 concentrated to obtain 80 mg of the desired compound as
23 a pale yellow powder.
z4 IR(KBr)~max : 1760, 1710, 16fiO
2s p~lR(Drulso-d6)~(ppm~: 16.6~(1H,br), 9.39(1H,d,J=8.8Hz),
26 7.47(1H,s), 6.31(1H,t), 5.51(1H,d-d,J=5.5, 8.8Hz),
27 4.38(21-1,s), 3.89(3H,s), 2.54-0.~(4~1,m)
28
29
31 ~,~
Il- 51
1.~789~J
I Example 20.
2 Preparation of t+)-cis~ 7-[2-(2-amino-4-thiazolyl)-
3 2-syn-~ethoxyiminoacetamido]-1-azabicyclol4,2,0]
4 oct-~-en-8-on-2-carboxylic acid:
s ' '
6 C~CHlCO~ H2N ~ S~
~ CCOI~ H H CCO~H H H
g ~ ~OCH ~ ~ ~ NOCH~ ~
02H o.~H
~ .
I In this Exam~le, 80 mg of the com~ound obtained
13 in Exam?le 19 is dissolved in 0.96 m~ of di~ethyl-
l~ acetamide. To the solution, 27.5 mg of thiourea is added
ar room temperature with stirring and the mixture is stirred
~6 'or 14 hours. A'~er addition of ether, the supernatant is
17 removed by decantation to obtain a red oily residue. The
1~ residue is purified by chromatography using
l9 Diaion~HP-10 to obtain 19.2 mg of the desired compound.
zo I~(X~r~vmax : 1760, 1670, 1630
2l P~5~D~SO-d6)~: 9.26(1H,d,J=8.6Hz3, 7.11~2~,br),
22 6 75(1~,s), 6.30(1H,~), 5.47(1H,d-d,J=5.4, 8.8Hz),
~3 3.89(3H,s), 2.5-1.0~4H,~)
21 ~
. .
~, Example 21.
26 Preparation of (~)-cis-7~-(2-thienylacetamido)-4
27 acetoxy-1-azabicyclo[4,2,0]oct-2-en-8-on-2=
2~ carboxylic acid:
23
3~
! - 52 -
... .
1~7~5~ ~
~2~"0COCH~ 3~LC:{ICON OCOCH3
COOH OOH
8 In this Example, 126 mg of (+)-cis-7~-amino-4~-
g acetpxy-l-azabicyclo[4,2,0]oct-2-en-8-on-2-carboxylic acid
o obtained in Reference Example 14 is dissolved in 3.0 mQ of dioxane
Il and 4.0 mQ of water. The solution is cooled on an ice and
12 sodium chloride bath. To the solution are added 105 mg of
13 sodium bicarbonate, and 84 mg of 2-thienylacetylchloride
14 dissolved in 1 mQ of clioxane. The mixture is s-tirred for
o~e hour. ~hen, the reaction mixture is adjusted to pEI 2.0
16 ~1ith lN-hydrochloric acid and extracted three times with
l7 ethyl acetate. The extracts are combined and washed with
18 saturated sodium chloride solution. The washing is dried
IrJ with anhydrous sodium sulfate and is subjected -to
filtration. The filtrate is concentrated and the concen-
21 trate is charged on a column packed with 20 g of silica
22 ge]. Elution is carried out with a mix-ture of chloroform
23 and ethanol (20:1, by volume). ~he f~actions containing
2~ the desired compound are co~bined and concentrated to
dryness to obtain 89.1 mg of the desired compound as a
26 pale yellow powder. Yield 47~. Properties of the compound
27 are set forth below.
28 IR(KBr)vmax : 1780, 1745, 1660
29 N?~R(CDCQ3 -~ CD3OD)~(ppm): 7.27-6.93(3H,m), 6.39(1H,d,
J=5.4Hz), 5.43(1H,d,J=4.911z), 5.40(1H,m),
31 3.79(2~-1,s), 2.10-1.26(2H,m), 2.06(3H,s)
I - 53 -
~78g5~
I AntibaCterial activities of the Compounds
2 obtained in Examples 4, 6, 8, 10, 12, 14, 16,
3 13, 20 and 21 are deter~ined by Heart In,usio}~ ar Dilution
4 .ethod (p~; 7.2). ~he results are s;~own in ~he rollowin~ table. --
Cefazolin is used as a reference.
11
12
13
I;
16
l7
18
19
'q4;
2l
"~
~3
2~
26
27
28
~9
31
I ~ 54 -
Image
- 55 -
~i7~395~ ~;
: . :
I l : Staphylococcus aureus 209-p
2 2 : Staphylococcus aureus Smith
3 3 : Sta?hylococcus epidermidis
4 4 : Escherichia coli ~IHJC-2
5 : Escherichia coli Juhl
6 6 : Klebsiella pneumoniae 8045
7 7 : Klebsiella pneumoniae Y-60 - ..
8 8 : Serratia marcescens T-26
9 9 . Serratia marcescens T-55
lO : Proteus mirabilis 1287
ll ll : Proteus vulgaris 6897 .
12 12 : Proteus morsanii KY4298
13 13 : Proteus rettgeri KY4289
l4 14 : Pseudomonas aeruginosa 145
lS l.S : Pseudomonas putida F264
16 a : The compound obtained in Example 4
17 b : " 6
18 c : ~ isomer obtained in Example 6
l9 d : B isomer obtained in F.xample 6
e : The compound obtained in Example 8
2I f : " .10
22 g : " 12 .
23 h : " 14
2~ 16 ~ ~
2; j : 1~
26 k : " 20
27 l : " ' 21
~3 g': D-isomer obtained in ~xample 12
29
3l c<~
I - 56 -
1:1789S~ `;.`
~ . . .
I Example 22
2 Preparation of (+)-cis-7-phenylacetamido-1-azabicyclo
3 ~4, 2, 01 oct-2-en-8-on-2-carboxylic acld
s
6 ~ CH2CO ~
8 - COOH
g ~ ~
Il . .. .
2 In this Example, 50.1 mg (0.28 mmole) of (+)-cis-7-amino-
3 l-azabicyclo [4,2,0] oct-2-en-8-on-2-carboxylic acid obtained
l~ as in Reference Example 16 (or 8) is dissolved in 1 ml of water
and 2 ml of acetone, and 55.5 mg of sodium hydrogencarbonate is
16 added. To the mixture, 0.3 ml of lN-phenylacetylchloride-
17 methylenechloride solution is added at a temperature of -20C
18 and the mixture is stirred at the same temperature for 20 mlnutes.
19 After the temperature is raised to room temperature, the reaction
mixture is stirred for 2 hours. Then, the mixture is adjusted
21 to pH 1.5 with 10 % hydrochloric acid under ice cooling and
22 extracted three times with 5 ml of ethyl acetate. The extract
23 is washed with saturated sodium chloride solution, dried with
2~ anhydrous sodium sulfate and concentrated under reduced pressure.
2s The resultin~ crystals are recovered by filtration and washed
26 with a small amount of ethyl acetate to obtain 39.7 mg (47.8 %)
27 of the desired compound. Properties of the compound are set forth
28 below
Melting point: 192.0 - 193.0C
29 cm-1
IR(Ksr) vmax : 1770, 1690, 1650, 1615
NMR(CD30D) ~(ppm): 7.22(5H, s), 6.40(lH,t,J =4.0), 5.33(lH,
d, J= 5.0), 3.57(2H, s), 2.5 -1.5(9H, m) '
I!- 57 -
~17~9S$
1 Example 23
2 Preparation of (+)-cis-7~-phen~lacetamlde-4~-methyl-
3 l-azabicyclo [4, 2, O] oct-2-en-8-one-2-carboxylic acid
4 ;
6 ~_~
~ CH CONH
7 ~2 ~ 3 -:~
8 . O
COOH
11
~c
In this Example, 399 mg (1.28 mmole) of the trifluoro-
13 acetate of (+)-7~-amino-4~-methyl-1-azabicyclo [4, 2, 0] Oct-2-cn-
14 ~-on-2-carboxylic acid obtained as in ~eference Lxample 18 (or
13) is dissolved in 6.4 ml of water and 12.~ ml of acetone, and
6 432 mg (5.14 mmole) of sodium hydrogencarbonate is addecl.
The mixture i6 stirred under ice cooling. After adding 258.5 mg
s (5.14 mmole) of phenylacetyl chloride, the mixture is allowed to
lg react under ice cooling for 3 hours. The reaction mixture is
adjusted to pH 3 with 0.1N hydrochloric acid and extracted five
21 times with lO ml of ethyl acetate. The extract is washed with
22 saturated sodium chloride solution, dried with anhydrous sodium
23 sulfate and concentrated under reduced pressure. Ether is added
2~ to the concentrate and formed crys-tals are recovered by filtra-
2s tion. Thus, 242 7 mg of the desired compound is obtainecl.
26 The water layers obtained in the above ethyl acetate
27 extraction are again extracted five times with lO ml of ethyl
28 acetate. Crystallization process described above is repeated
29 to obtain 27.6 mg of the desired compound. To the residual water
layers is added saturated sodium chloride solution and the
31 obtained mixture is extracted five times with lO ml of ethyl
. . . ~ ., .
~ - 58 --
1~78gS~
I acetate. Crystallization process described above is repeated
2 to obtain 21,3 mg of the desired compound. Total amount of the
3 desired compound is 293.6 mg~ Yield 72.6 ~. Properties of ~
4 the compound are set forth below. -~ -
S IR(KBr) v ax : 1775, 1698, 1655
6 NMR(CD30D) ~(ppm): 7.31(5H, s), 6.48(lH, d, J= 5.0),
7 5.42 (lH, d, J = 5.2), 3.85(1H, m), 3.60(2H, s), -`
8 2.62(1H, m), 1.60(2H, m), 1.10(3H, d, J = 7.5)
. .
~ . , .
l2
l3
14
16
17 .
r . I .
Ig
21
22
23
24
26
27
28
29
31
_ 59 _
~. 1.7895~
I Example 24 ~-----2 Preparation of (~)-cis-7-cyanoacetamido-2-t-
3 butyloxycarboxyl-1-azabicyclo~4,2,0] oct-2-en- -~
4 8-one ----
7 H~N ~ NCCH2C0
8 O 1~ O
9 . COOtBu COOtBu
1 0 - _ .
11 .. -. ~
In this Example, 42.5 mg (0.5 mmole) of cyanoacetic
acid and 57.5 mg (0.5 mmole)of N-hydroxysuccinimide are dis-
l4 solved in 2.5 mQ of dried dioxane, and 103.3 mg (0.5 mmole)
of N,N-dicyclohexylcarbodiimide is added to the solution.
6 The mixture is stirred at room temperature for one hour.
The resulting crystals are removed by filtration and the
8 filtrate is concentrated under reduced pressure. To the -
, .- ,~, .
lg obtained residue is added 5 mQ of dried dichloromethane
and an insoluble material is removed by filtration to
. ., . ~,.
21 obtain a solution of succinimide ester of cyanoacetic acid.
22 On the other hand, 123 mg (0.45 mmole) of
23 hydrochloride of l~)-cis- 7-amino 2-t-butyloxycarbonyl-1-
24 azabicyclo[4,2,0] oct-2-en-8-one thereinafter referred to
2s as amino-compound) and 0.077 mQ of triethylamine are
26 dissolved in 5 mQ of dried dichloromethane. To the
27 solution is added the above solution of succinimide
28 ester of cyanoacetic acid. The mixture is stirred at
29 room temperature for 4 hours and then is concentrated
under reduced pressure. The thus obtained oily product
3l is subjected to silica gel column chromatography using
I - 60 -
? ~
;' '`, :~,
1 a: 7~39S!~
I a mixture of chloroform and methanol (15:1). The fractions
2 containing the desired compound are combined and concen-
3 trated under reduced pressure to obtain 112 mg of the
4 desired compound Yield: 73.4~
IR(NaCQ)vCamx : 3310, 2265, 1765, 1720, 1680,
6 N~1R(CDCQ3)~(ppm): 7.90(lH), 6.32(lH,m), 5.35(lH,m),
7 3.75(1H,m), 2.03(6H,m), 1.52(9H,s)
g ! Exarnple 25
lo Preparation of (+)-cis-7-[2-(2-furyl)-2-syn-
11 methoxyiminoacetamido]-2-t-butyloxycarhonyl-1-
12 azabicyclo[4,2,0] oct-2-en-8-one
13
l4
IS H 1~ Hl ~ CON~
l6 0 ~N-OCH~ ~ N ~
17 COOtBuCOOtBu
18 ` ~
19
2~ The same procedure as in Example;24 is repeated x~w
21 except that 158 mg (0.934 mmole) of 2-(2-furyl)-2-methoxy-
22 iminoacetic acid is used in place of cyanoacetic acid and
23 that 247 mg (0.9 mmole) of the amino-compound is used.
24 As a result, 313 mg of the desired compound is obtained. ;~
Yield: 89.3~.
26 IR(NaCQ)vmax : 3310j 1765, 1725, 1715, 1685, 1655
27 N~R(CDCQ3)~(ppm): 7.65(lH,m), 7.52(lH,m), 6.73(lH,m),
28 6.46(1H,m), 6.29(1H,m), 5.40(1H,m), 4.02(3H,s),
29 3.78(1H,m), 2.19(4H,m), 1.50(9H,s)
U.~
I - 61 -
~ 31 7~95~ - ~
I Example 26
2 Preparation of (+)-cis-7-cyanomethylthioacetamido-
3 2-t-butyloxycarbonyl-1-azabicyclo~4,2,0] oct-2-
4 en-8-one
7H~N ~ N~CH~SCH~CONH
8O N ~ o~N ~
COOtBu COOtBu
g ~",,c~
I I .
12 The same procedure as in Example 24 is repeated
13 except that 65.6 mg (0.5 mmole) of cyanomethylthioacetic
14 acid is used in place of cyanoacetic acid. As a result,
136.5 mg of the desired compound is obtained. Yield: 86.3%.
16 IR(NaCQ)vmax : 3315, 2250, 1760, 1725, 1715, 1675,
17 1655, 1650
18 NMR(CDCQ3)~(p~m): 7.90(1H,m), 6.32(1H,m), 5.38(1~,m),
19 3.7215H,m), 2 18(4H,m), 1.52(9H,s)
2D
2I Example 27
22 Preparation of (+)-cis-7-[2-(2-furyl)-2-hydroxy-
23 iminoacetamido]-2 t-butyloxycarbonyl-1-azabicyclo
2~ [4,2,0] oct-2-en-8-one
2s
26~I H ~ COl~H
27H~N ~ ~ ~ N~ ~
28o~N~ OH COOtBu
COOtBu
29
31 ~ .
62
i~l 7~95~; .. ~ `
..
I The same procedure as in Example 24 is repeated
2 except that 156 mg (1 mmole) of-2-(2-furyl)-2-hydroxyimino-
3 acétic acid is used in place of cyanoacetic acid and that
4 137 mg (0.5 mmole) of the amino-compound is used. As a
result, 115 mg of the desired compound is obtained.
6 Yield: 61.3%.
7 IR(NaCQ)vCamx : 3290, 1770, 1720, 1690, 1680, 1655
8 NMR(CDCQ3)~(ppm): 7.51(4H,m), 6.49(1H,m)~ 6.28(lH,m),
9 - 5.46(lH,m), 3.88(lH,m), 2.18(4H,m), 1.49(9~E,s)
Il Example 28
12 Preparation of (~)-cis-7-[2-(2-thienyl)-2-
13 hydroxyiminoacetamido]-2-t-butyloxycar~onyl-1-
14 azabicyclo[4,2,0] oct-2-en-8-one
16
17 }3 ~ COl`Tg~_~
l8 O N ~ ~ ~ N ~
l9 COOtBu OH COOtBu
21
æ The same procedure as in Example 24 is repeated
23 except that 85.6 my (0.5 mmole) of 2-(2-thienyl)-2-hydroxy-
24 iminoacetic acid is used in place of cyanoacetic acid and
2s that 137 mg (0.5 mmole) of -the amino-compound is used.
26 As a result, 103 mg of the desired compound is obtained.
27 Yield: 52.6%.
28 IR(KBr)vmax : 1760, 1720, 1675, 1655, 1650
29 NMR(CDCQ3)~(ppm): 8.02(1H,m), 7.22(4H,m), 6.18(lH,m),
5.37(1H,m), 3.78(1H,m), 2.09(4H,m), 1.46(9H,s)
31
~,.~ ,....
- 63 -
~78956
1 Example 29
Preparation of (+)-cis-7-(1-tetrazolyl)
3 acetamido-2-t-butyloxycarbonyl-1-azabicyclo
4 ~4,2,0] oct-2-en-8-One
H H ~ -CH~C0
8 21 ~ O 1~ 1
g - COO~Bu COOtBu
Il .
12 In this Example, 79 mg (0.55 mmole) of (l-tetra-
3 zolyl)acetic acid and 137 mg (0.5 mmole) of the amino-
14 compound are dissolved in 5 mQ of N,N-dimethylformamide
and 113 mg (0.55 mmole) of N,N-dicyclohexylcarbodiimide
6 is added to the solution. The mixture i5 stirred at room
~7 temperature for 2 hours and 50 mQ of lN sodium bicarbonate
18 is added thereto. The resulting crystals are removed by
ls filtration and the filtrate is extracted with 50 mQ of
ethyl acetate two times Ethylacetate layers are
21 combined, dried with magnesium sulfate, and then concen-
22 trated under reduced pressure. The thus obtained oily
23 product is purified by silica gel chromatography to obtain
24 157 mg of the desired compound. Yield: 90.1~
2s IR(NaCQ)vmax : 3290, 1765, 1725, 1715, 1685, 1680,
26 1670, 1~55
27 M~IR(CDCQ3)~(ppm): 8.94(1H,s), 8.24(1H,m), 6.29~1H,m),
2~ 5.61(3H,m), 3.77(1H,m), 2.16(4H,m), 1.49(1H,s)
29
3l ~ .
- 64 -
~7~
1 Example 30
2 Preparatlon of (+)-cis-7-(2-tetrazolyl)
3 acetamido-2-t-butyloxycarbonyl-1-azabicyclo
4 [4,2,0] oct-2-en-8-one
s
N-
7 H2N ~ ~~-CH2CON.'I
8 N ~ O
9 .COOtBu COOt~3u
I'
12 The same procedure as in Example 29 is repeated
13 exce~t that 79 mg (0.55 mmole) of (2-tetrazolyl)acetic acid
14 is used in place of (l-tetrazolyl)acetic acid. As a result,
142 mg of the desired compound is obtained. Yield: 81.4~.
`1
l6 IR(NaCQ)~max : 3300, 1760, 1725, 1705, 1690
l7 NMR(CDCQ3)~(ppm): S.60(lH,m), 8.52(lH,s), 6.29(lH,m),
18 5.47(2H,s), 5.35(1H,m), 3.72(1H,m), 2.15(4H,m),
Is 1.50(9H,s)
~, g~
2I Example 31
22 Preparation of (~)-cis-7-(4-pyridyl)thio-
23 acetamido-2-t-butyloxycarbonyl-1-azabicyclo
24 [4,2,0] oct-2-en-8-one
26 H H
87 ~N~ ~ N ~ SCHaCON
28 o~N~ ~ N ~
COO~Bu COOtBu
29
31
- 65 -
1~78gSf~
I The same ~rocedure as in Example 29 is repeated
z exceot that 181 mg (0.725 mmole) of hydrobromide Qf 4-
3 pyridylthioacetic acid is used in place of (l-tetrazolyl)
4 acetic acid. As a result, 148.6 mg of the desired
comoound is obtained. Yield: 76.3%.
6 I~(NaCQ)~max : 32~0, 1775, 1725, 1715, 1690
7 N~lR(CDCQ3)~(ppm): 8.38(2H,m), 8.24(lH,m), 7.25(2H,m),
8 6.25(1H,m), 5.35(1H,m), 3.78(2~1,s), 3.78(lH,m),
9 - 2.13(4H,m), 1.52(9H,s)
1I Example 32
12 Preparation of (+)-cis-7-(2-tetrazolyl)
13 acetamido-2-carboxy-1-azabicyclo[4,2,03 oct-
14 2-en-8-one
~S
16 21
17 L~ c~co~ L~ -C~IlCO~`IH
18 O ~ 0
COOtBu CO(:f~
2l
22 In this Examr~le, 3 mQ of cooled trifluoroacetic
23 acid is added to 150 mg (0.431 mmole) of (+)-cis- 7-(2-
24 tetrazolyl)acetamido-2-t-butyloxycarbonyl-1-azabicyclo
2s [4,2,0] oct-2-en-8-one (hereinafter referred to as the
26 ester-com~ound) obtained in Example 30. The solution is
27 stirred for 10 minutes and then is concentrated under
28 reduced pressure at room temperature. To the resulting
29 highly viscous substance is added 10 mQ of ether. The
mixture is stirred for 20 minutes and then is filtered to
31 obtain a solid ~roduct. The product is dissolved in 2 mQ
- 66 -
~178g5~ `'-'-`''
I of methanol and the mixture is allowed to stand on an ice
2 bath for one hour. The resulting crvstals are filtered
3 and are dried to obtain 111.5 mg of the desired compound.
4 Yield: 87.3%-
I~(KBr)vCm : 3340, 1765, 1720, 1685, 1635, 1555
7 Example 3~
8 Preparation of (+)-cis-7-(1-tetrazolyl)
9 -- acetamido-2-carboxy-1-azabicyclo[4,2,0] oct-
lo 2-en-8-one
Il
N =1
13 1 ~ C~C0'~-~ I CHlCO~
N ~----~N==)
14 ~1~ N ~
_ COOtBu O COCH
16
17
18 The same procedure as in Example 32 is repeated
19 except that 70 mg (0.201 mmole) of (~)-cis-7-(1-tetra-
zolyl)acetamido-2-t-butyloxycarbonyl-1-azabicyclo[4,2,0j
2~ oct-2-en-8-one obtained in Example 29 is used in place of
22 tlle ester-compound. As a result, 53.6 mg of the desired
23 com~ound is ob-tained. Yield: 91.2%.
2~ IR(KBr)~max : 3280, 1770, 1725, 1675, 1635, 1565
2s
26 Example 34
27 Preparation of (+)-cis-7-(4-pyridyl)thio-
2s acetamido-2-carboxy-1-azabicyclo[4,2,0] oct-
29 2-en-8-one
31
-67
1~l7~95Çj
3 N ~ SCHlCO~ N ~ S ~12CG~ ~
COOtB~ O COOH
8 The same procedure as in Example 32 is re2eated
g except that 105 mg (0.27 mmole) of (+)-cis-7~(4-~yridyl) ~J
o thioacetamido-2-t-butyloxycarbonyl-1-azahicyclo[4,2,0]
Il oct-2-en-8-one obtained in Example 31 is used in place of
l2 the ester-compound. As a result, 70 mg of the desired
compound is obtained. Yield: 78.0~.
I~ Br)~max : 1760, 1708, 1680, 1630
Example 35
17 Preparation of (+)-cis-7-cyanoacetamido-2-
18 carboxy-1-azabicyclo[4,2,0] oct-2-en-8-one
~9
NCCH~CONH I~CC~CONH
21
22 cootgU COO~I
23
24
2s
26 The same procedure as in Example 32 is repeated
27 except that 112 mg (0.367 mmole) of (-~)-cis-7-cyanoacetamido-
28 2-t-butyloxycarbonyl-1-azabicyclo[4,2,0] oct-2-en-8-one
29 obtained in Exam~le 24 is used in place of the ester-
compound. As a result, 81 mg of the desired compound is
3~ obtained, Yield: 88.6~.
~ 68
~?~
~i7~9Sf~
2 IR(KBr)v ax : 2270, 1755, 1703, 1690, 1655, 1630
3 Example 36
4 Preparation of (+)-cis-7-[2-( -furyl)-2-syn- --
metlloxyi~minoacetamido]-2-carboxy-l-azabicyclo
6 [4,2,0] oct-2-en-8-one
~ ~CONH ~ ~ CONH
lo ~{)c~ ~ .. N-OCli~
O COO Bu COOH
11
12
13
1~ The same procedure as in Exam~le 32 is repeated
IS except that 131 mg (0.336 mmole) of (~)-cis-7-[2-(2-
6 furyl)-2-methoxylminoacetamido]-2-t-butyloxycarbonyl-1-
azabicyclo[4,2,0] oct-2-en-8-one obtained in ~xample 25
18 i5 used in place of the ester-compound. ~s a result,
19 78 my of the desired compound is obtained.
Yield: 69.6~.
21 IR(KBr)vmax : 1760, 1690, 1660, 1640, 1620
22
23 ~xample 37
_
24 Preparation of (~)-cis-7-cyanomethylthio-
acetamido-2-carboxy-1-azabicyclo[4,2,0] oct-
26 2-en-8-one
27
28
29
- 69 -
1~ ~8g5~
3 ~ NCC.~2SC~CO.~H NCC~SC~ CO~
4 ~ --~D ~
O COO Bu O COOH
8 The same procedure as in Example 32 is repeated
9 except that 136 mg (0.387 mmole) of (+)-cis-7-cyanomethyl--
thioacetamido-2-t-butyloxycarbonyl-1-azabicyclo[4,2,0]
1l oct-2-en-8-one obtained in Example 26 is used in place of
2 the ester-compound. As a result, 100 mg of the desired
13 compound is obtained. Yield: 78.0~.
4 I~(KBr)vmax : 2250, 1755, 1730, 1665, 1635
6 _ample 38
l7 Preparation of (+)-cis-7-[2-(2-furyl)-2-hydroxyl-
18 iminoacetamido]-2-carboxy-1-azabicyclo[4,2,0] oct-
l9 2-en-8-one
2l ~ COI~H > ~ CON~I
~ ~ ~ N ~
23 OH C00tBU OH COOH
24
26
27 The same procedure as in Example 32 is repcated
28 except that 115 mg (0.3 mmole) of (+)-cis-7-[2-(2-furyl)-
29 2-hydroxyiminoacetamido]-2-t-butyloxycarbonyl-1-azabicyclo
[4,2,0] oct-2-en-8-one obtained in Example 27 is used in
3I place of the ester-compound. As a result, 92 mg of
i- 70 -
~ ~7~956
I the desired compound is obtained. Yield: 69%.
2 IR(KBr)~mmax : 1760, 1720, 1680, 1660, 1635
Example 39
Preparation of (+)-cis-7-12-(2-thienvl)-2-
6 hydroxyiminoacetamido]-2-carboxy-1-azabicyclo
7 [4,2,01 oct-2-en-8-one
C~ CONH > ~ ~CONH
OH O COOtBu OH COOH
l3
The same procedure as in Example 32 is repeated
16 except that 103 mg (0.263 mmole) of (+)-cis-7-12-(2-
~7 thienyl)-2-hydroxyiminoacetamido]-2-t-butyloxycarbonyl-1-
18 azabicyclol4,2,0] oct-2-en-B-one obtained in Example 28 is
l9 used in place of the ester-compound. As a result, 71.6 mg
of the desired compound is obtained. Yield: 81.2%.
21 IR(KBr)vCmx : 1760, 1690, 1665, 1630
22
23 Example 40
24 Preparation of (+)-cis-7-l2-~5,6-dihydro-1,4-
dithiinyl)acetamido]-2-carboxy-1-azabicyclo
26 [4,2,0] oct-2-en-8-one
27
28
29
31
~ ;-
1~7895$
' , . .
3 H2~ C~2-Co~
coor~ COOH
8 In this Example, 176 mg (1 mmole) of 2-(5,6-
9 dihydroxy-1,4-dithianyl)acetic acid and 115 mg (1 mmole)
0 of N-hydroxysuccinimide are dissolved in 6 mQ of dried
ll dioxane a~d 206 mg (1 mmole) of N,N-dicyclohexylcarbodi-
12 imide is added to the solution. The mixture is stirred
13 at room temperature for 2 hours. The resultin~ crystals
14 are removed by filtration and the filtrate is concentrated.
S The obtained residue is dissolved in 6 mQ of dried dichloro-
16 methane.
l7 On the other hand, 109 mg of hydrochloride of (~)-
l8 cis-7-amino-2-carboxy-1-azabicyclo[4,2,0] oct-2-en-8-one
l9 obtained by the same procedure an in Reference Example 16
and 0.21 mQ of triethylamine are dissolved in 6 mQ of dried
2I dichloromethane. To the solution is added the above solution
22 of dichloromethane and the mixture is stirred at room
23 temperature for 10 hours and then is concentrated under
24 reduced pressure. To the obtained residue is added 10 m~
of water and an insoluble material is removed by filtration.
26 The filtrate is adjusted to pH 2.5 with 2N hydrochloric acid.
27 After cooling, the solution is filtered and dried to-obtain
2s 120 mg of the desired compound. Yield: 70.5%.
29 Ip~(Ksr)vmmax : 3275, 1765, 1690, 1648, 1630
31
~ -72 -
1~7~195~
I Example 41
2 Preparation of (+)-cis-7-[(R)-2-phenyl-2-
3 hydroxyaeetamido]-2-carboxy-1-azabicy~lo
4 [ 4,2,0] oct-2-en-8-one
6 O ~ OH ~ ~
COOH COOH
9 -
~o
12 The same proeedure as in Example 40 is repeated
13 exce~t that 152 mg of D-mandelie aeid is used in place of
14 2-(5,6-dihydroxy-1,4-dithianyl)aeetic aeid. As a result,
142.5 mg of the desired compound is obtained. Yield: 90.1~.
--1
16 IR(KBr)vmax : 3325, 1770, 1715, 1690, 1655, 1635
17
18 Example 42
.
19 Preparation of (+)-eis-7-(2-phenyl-2-carboxy-
acetamido)-2-carboxy-1-azabicyclo[4,2,0] oet-
2l 2-en-8-one
22
23 ~ N ~ H CO~ ~
26 ~ COOH
27
28
29 In this Example, 90 mg (0.5 mmole) of phenylmalonic
aeid is dissolved in mixed solution of 5 m~ of dried ether
31 and 61 mg (0.51 mmole) of thionyl ehloride. To the solution
~- 73 -
- ; .
117~956 ~
1 is added a drop of N,N-dimethylformamide and the mixture
2 is heated under reflux for 2 hours. Then the mixture is
3 concentrated under reduced pressure. To the obtained
4 residue is added 5 mQ of dried benzene and the mixture is
again concentrated. The resulting residue is dissolved in
6 dried ether and to the solution is added a solution
7 wherein 103 mg (0.47 mmole) of hydrochloride of (+)-cis-
8 7-amino-2-carboxy-1-azabicyclo[4,2,0] oct-2-en-8-one
g obtained by the same procedure as in Reference Exam~le 16
is dissolved in 2 mQ of 0.5N sodium bicarbonate and 1 mQ
11 of ether under ice cooling. The mixture is stirred at
12 room tempe'rature for one hour and then adjusted to pH 2
l3 with lN hydrochloric acid. The mixture is extracted with
14 20 m~ of ethyl acetate five times. The extract is _
evaporated to dryness and the obtained solid is fully
IC suspended in 5 mQ of ether. The suspension is filtered
and dried to obtain 100 mg of the desired compound.
I~ Yield: 60.1~.
19 IR~KBr)vmmx : 1755, 1720, 1700, 1660, 1630
21 Example 43
22 Preparation of triethylamine salt of (-~)-cis-
23 7-(2-phenyl-2-sulfoacetamido)-2-carboxy-1-
24 azabicyclo[4,2,0] oct-2-en-8-one
26
27If2N ~ ~ ~ CH -COI~H
23~ N ~ S03I-I
COOH
29 COO~
31
~ . .
~ -74 -
1~78956 --
. . ,
I In this Example, 109 mg (O.S mmole) of hydrochloride
2 of (+) -cis- 7-amino-2-carboxy-1-azabicyclo[4,2,0~ oct-2-en-
3 8-one and 101 mg of triethylamine are dissolved in 6 mQ of
4 dried dichloromethane. To the solution is added 195 mg (0.5
mmole) of the ~-triethylammoniumsulfophenylacetic
6 ethylcarbonic acid anhydride under ice cooling. The
7 mixture is stirred for 1.5 hours and then the solvent is
8 removed by distillation. The resulting residue is dissolved
9 in 5 mQ of water and the solution is adjusted to pH 1.5 with
lN hydrochloric acid. Then the solution is extracted with
l 10 m~ of n-butanol two times. The n-butanol layers are
12 combined and concentrated under reduced pressure. The
l3 obtained solid is washed with 2 mQ of dichloromethane and
14 dried to obtain 196 mg of the desired compound. Yield:
81.3%-
16 IR(KBr)vmax : 3275, 1770, 1725, 1682, 1628.
17
18
19
21
22
23
24
26
27
28
29
- 75 _
_ "i ~ :
il7~5~
I Antibacterial activities of the Com~ounds
2 Examples 32, 33, 34, 36, 38, 41 and 42
3 are determined by Heart Infusion Agar Dilution Method
4 (pH 7.2). ~he results are shown in the folloT~ing table.
._ _ ~
~icro- ~5inimum- Inhibitory Concentration (u~/ml)
I organism ~
9 ~ . m n O ? ~ r s t
. . .. ~
1 ' 6.25 25 1.56 >100 SO 3.12 3.12 ~0.05
ll 2' 12.5256.25 >100 >100 25 >25 0.4
~2 3 ~25 5012.5 >100 >100 25 25 0.73
4 ~100 50 100 100 100 25 25 1.56
~ ' 5 1100 50 100 25 50 50 25 1.56
lS I 6 150 50 25 1.56 12.5 12.512., 0.4
16 7 1100 50>100 12.5 >100 50 >25 1.56
l7 ' 8 >100>100~100 >100 >100 >100 >25 >100
18 ' 9 1 >100>100>100100 >100 100 >25 50
Ig ,10 >100~100>100 12.5 100 100 >25 6.25
ZO I11 >100>100>100 1.56 >100 >100 >25 25 ~-~
21 112 , >100>100>10012.5>100 >100 >25 >100
22 13 1100 50>100 12.5 6.25 100 >25 25
23 ~1~ >100>100>100 >100 >100 >100 >25 >100
2.~ >100>100>100 >100 100 >100 >25 >100
2~ :
26
27
23
29
31
I - 76 --
1~l7~9S6
1 1 : Staphylococcus aureus 209-p
2 2 : Staphylococcus aureus Smith
3 3 : Staphylococcus epidermidis
4 4 : Escherichia coli NIHJC-2
5 : Escherichia coli Juhl
6 6 : Klebsiella pneumoniae 8045
7 7 : Klebsiella pneumoniae Y-60
8 8 o Serratia marcescens T-26
g - 9 : Serratia marcescens T-55
10 : Proteus mirabilis 1287
11 11 : Proteus vulgaris 6897
12 12 : Proteus morganii KY4298
13 13 : Proteus rettgeri KY4289
14 14 ; Pseudomonas aeruginosa 145
15 : Pseudomonas putida F264
16 m : The compound obtained in Example 32
l7 n : " 33
18 o : "
19 P : " 36
2I r : " 41
22 s : " 42
23 t : Cefazolin
2q
26
27
28
29
31 ~.
~.~78956
Example 44
Preparation of 7~- ~R)-2-para-hydroxyphenyl-2-
aminoacetamid ~-4~-methyl-1-azabicyclo ~,2, ~oct-2-en-8-on-
2-carboxylic acid:
HO ~ CHCON ~ CH3
NH2 N ~
C02H
In this example, the same procedure as in example
12 is employed except that 40 mg of (+)-cis-7~- ~R)-2-para-
hydroxyphenyl-2-t-butyloxycarbonyl-aminoacetamid ~-4~-methyl-
2-t-butyloxycarbonyl-1-azabicyclo ~,2, ~oct-2-e~-8-one, which
is obtained in the same procedure as in example 9, is used as
a starting compound. As a result, 29.3 mg (85%) of the less
polar isomer of the desired compound is obtained.
5 : +12;8 ~M Phosphate buffer (pH 7.0), c=0.
IR (KBr) vmamX : 3420, 3260, 1760, 1685
PMR (D2O)~(ppm): 7.35(2H, d, J = 8.0), 6.96(2H, d,
J = 8.0), 6.10(1H, d, J = 5.1), 5.45(1H, d,
J = 4.9), 5.11(1H, s), 3.92(1H, m), 2.45(1H, m),
1.50 - 1.08(2H, m), 1.01(3H, d, J = 7.1)
From the strong antimicrobial activity shown in
the table below, this compound is assigned to have 6(R)7(S)
absolute configuration.
- 78 -
~.~78~5!~
Example 45
Preparation of 7~-t(R~-2-phenyl-2-(4-ethyl-2.3-d~oxo-
l-piperazinylcarbonylamino)acetamido]-4-a-methyl-1-azablcyclo
C4,2~0~oct-2-en-8-on-2-carboxyllc acid:
<~ CON~c~3
Co C02~1
~N~O
O
Et
In this example, the same procedure as in Example 8
is employed except that 55 mg of (~)-cis-7~(R) 2~pheny~-2-
(4-ethyl-2,3-dioxo-1-piperaz~nylcarbonylamino~acetamido]~4-a-
methyl-2-t-butyloxycarbonyl-1-azabicycloC4,2,0]oct-2-en 8-one
(the less polar isomer) obtained in the same procedure as
in Example 7 is used as a starting material. As a result
35.7mg ~72 ~) of white powder is obtained.
la]D : -36.3 (MeOH, c = 1.0)
IR (KBr) ~ : 3450, 3300, 1775, 1720, 1683, 1630
PMR ~CD3OD)~(ppm): 9.90(1H, d, J = 6.4), 7.80(5H, br),
6.39~1H, d, J = 4.9), 5.46(2H, d, J = 4.9), 4.00
~2H, m), 3.85 - 3.40~6H, m), 2.48(1~, br), 1.20
(5H, m), 1.04~3~, d, J = 7.4)
From the strong antimicrobial activity shown in
2~ the table below, this compound is assigned to have 6(R)7(S)
absolute configuration.
- 79 -
1~789S6
Example 46
Preparation of 7~-t(R)-2-para-hydroxyPhenyl-2-
(4-ethyl-2,3-dioxo-1-piperazinylcarbonylamin~)acetamido~-4-a-
methyl-l-azabicyclo~4,2,0]oct-2-en-8-on-2-carb~xylic acid:
B~llCON~oll
1. o
Ct~ C02H
~N~O : i
~N~O
Et
ln this example, the same procedure as in Example 8
is employed except that 57 mg of ~')-cis-7~-[(R)~2-para-
hydroxyphenyl-2-(4-ethyl-2,3-dioxD-l-piperazinylcarbonylam~no)
acetamido]-4-~-methyl-2-t-butyloxycarbonyl-1-azabicyclo~4,2,0
oct-2-en-8-on obtained in the s~me procedure as in ~xample 7
is used as a starting material. As a result 40 mg (78X) of
white powder ls obtained.
IalD : -7.1 (MeOH, c = 1.0)
IR (KBr) ~Cmax : 3430~ 3300, 1773, 1721, 1685, 1620
PMR (CD30D)~(ppm)- 7.28(2H, d, J z 8.5), 6.78(2H, d,
J = 8.5), 6.00(1H, d, J = 5.1), 5.37(1H, s), 5.36
(lH, d, J = 4.4), 4.00(2H, m), 3.85 - 3.35(6H, m),
2.40(1~, br), 1.37 - 1.05(5H, m), 1.01(3H, d, J z 6.8)
From the strong antimicrobial activity shown in
the table below, this compound is assigned to have 6(R)7(S)
absolute configuration.
- 80 -
~78956
Example 47
Preparation of 7~-~(R) 2~para-hydroxyphenyl-2-
aminoacetamido~ azabicyclo[4,2,0~oct-2~en~8~on-2-carboxylic
acid:
H ~ HCON
~H2
co2H
In this example, the same procedure as in Example 6
is employed except that 78 mg of (+)-cis-7-[(R)-2-para-
hydroxyphenyl-2-t-butyloxycarbonyl-aminoacetamido]-1-aza-
bicyclo[4,2,0]oct-2-en-8-one obtained in the same procedure as
in Example 5 is used as a starting material. As a result
29.5 mg (80%) of an isomer of the desired compound is obtained.
~]15 : ~107 5
IR (KBr) vmamX : 3450, 3290, 3090, 1700(SH), 1685, 1640
PMR (D20)~(ppm): 7.36(2H, d, J 8.8), 6.95(2H, d,
J = 8.8), 6.06(1H, t, J = 3.9), 5.40(1H, d, J = 4.6),
5.12(1H, s), 3.84(1H, m), 2.22(2H, m), 1.62(1H, m),
1.12(1H, m)
From the strong antimicrobial activity shown in
the table below, this compound is assigned to have 6(R)7(S)
absolute configuration.
Antibacterial activities of the compounds in Examples 44,
45, 46 and 47 are determined by Heart Infusion Agar Dilution
Method (pH 7.2). The results are shown in the following
table.
~7895Çi
Micro-
organism
1 0.4 0.4 0.781.56
2 1.56 1.56 3.1212.5
3 0.78 0.78 3.126.25
4 6.25 6.25 0.4 12.5
6.25 1.56 0.4 12.5
6 3.12 ~0.05 ~0.053.12
7 100 6.25 12.5 12.5
8 50 12.5 12.5 >100
9 12.5 3.12 0.78 50
-~lo 25 3.12 3.12 50
11 100 ~0.05 ~0.05>100
12 100 1.56 3.12>100
13 >100 6.25 12.5 25
14 >100 50 25 >100
1.56 ' 3.12>100
- 82 -
~17~956
I The following Reference Examples show practical
2 embodiments of preparing the starting compounds used in the
3 Examples.
. .
Reference Example 1
6 Preparation of (+)-cis-2-t-butyloxycarbonyl-7-azido-
7 l-azabicyclo [4, 2, 0] oct-2-en-8-one {the cis-compound
8 represented by the following formula:}
9 - ~
11 H
12 I ~
O~ ~- N ~
G00 ~ U
14
16 The present compound is produced according to the
17 following Processes 1 and 2. Hereinafter, cis and trans refer to
l8 the stereochemistry at the 3- or 4-position of 2-azetidinone
19 ring or at the 6- or 7-position of l-azabicyclo [4, 2, 0] octane
2u ring.
21
1) Preparation of 2-[4-(3~butenyl)-3-azido-2-oxoazetidin-
22 -~
l-yl]-2-diethylphosphonoacetate-t-butylester.
23
In this Example, 447 mg (1.78 mmole) o~ t-butyl-~-
24
aminodiethylphosphonocacetate {the com~ound has the fol].owing
2s properties: an oily product; IR(neat)vCax 3400, 1735-1745,
26
27 1020 -1060, NMR(CDC13)~(ppm), 4.20(d-q, 4H), 3.83(d, lII, J =
20Hz), 1.76(br, 2H), 1.50(s, 9H), 1.35(t, 6H); Mass(m/e)
28
268(M )} is dissolved in 25 ml of anhydrous ether and 164 mg
29
(1.96 mmole) of 4-pentene-1-al is added to the solution.
The ~olution is stirred at room temperature for one hour and
31 ~
- 83 -
1~78956
1 200 mg of Molecular Sieve (4A) (the procluct of r,~ako Junyaku Co.,
2 Ltd., the same molecular sieve is used hereinafter) and 150 mg
3 of anhydrous magnesium sulfate are added to the solution. The
4 mixture is stirred for one hour.
The reaetion mixture is subjected to filtration under
6 reduced pressure and the filtrate is concentrated under reduced
7 pressure to obtaln a pale yellow oily product. .~nhydrous
8 benzene is added to the product and the mixture is concentrated
9 under redueed pressure to obtain a pale yellow oily produet. ~~
o The presenee of shiff's base in the procluet is confirmed by
nuelear magnetie resonance spectrum. The product is dissolved
12 in 12.5 ml of cyclohexane and 12.5 ml of anhydrous benzene,
13 and 0.369 ml (2.66 mmole) of triethylamine and 200 mg of
4 Moleeular Sieve 4A are added to the solution. Azidoaeetyl
chloride,[319 mg (2.66 mmole)] dissolved in 12.5 ml of cyclo-
6 hexane is added dropwise to the mixture with stirring at room
17 temperature in 1.5 hours. The reaetion mixture is furthcr
18 stirred ~or 30 minutes and diluted with 10 ml of benzene, The
l9 reaction solution is washed with 5 ~6 diluted hydrochloric aeid,
saturated sodium bicarbonate, deionized water and saturated ,~
2I sodium ehloride solution, dried wlth anhydrous sodium sulfate
22 and concentrated under reduced pressure to obtain a brown oily -~
23 product which is identified as a crude product of the desired
24 compound. The oily product is charged on a
column paeked with 45 g o~ silica gel. Elution is earried out
26 with a ~nixture of n-hexane and ethyl acetate (1 : 2) to obtain
27 two types of isomers. Properties of the isomers are-set forth
28 below and they are identified as the isomers at the 3- and
29 4-positions, i.e. 345 mg of cis-isomer and 58 mg of trans-
isomer. Total yield is 54.2 %,
3l
~ 84
- 1.178g~S~
I Cis-isomer
2 IR(CHcl3)~max 2120, 1775, 1770~sh), 1750,
3 1740(sh), 1645
~ NMR(CDC13)~(ppm): 6.13 - 6.33(1H, m), 4.93 - 5.17(2H, m),
4.50 -4.93(2H, m), 3.80 -4.40(5H, m), 1.93 - 2.17
6 (4H, m), 1.50(9H, s), 1.33(6H, t)
7 Trans-isomer
~ IR(CHcl3)vmax 2120, 1780, 1755, 1750(sh), 1650
g NMR(CDC13)~(ppm); 5.43 - 6.20(1H, m), 4.80 - 5.30(2H, m),
lo 3.75 -4.75t7H, m), 2.0 -2.50(4H, m), l.50(9H, d),
~1 1.17(6H, m)
12
13 2) Preparation of (~)-cis-2-t-butyloxycarbonyi-7-azido-
azabicyclo t4, 2, 0] oct-2-en-8-one.
In this Example, 298 mg (0.716 mmole) of cis-2-[4-
(3-butenyl)-3-azido-2-oxoazetidin-1-yll-2-diethylphosphono-
~1 acetate-t-butylester obtained above is dissolved in
18 8.5 ml of dioxane and 2.5 ml of deionized water, and 30 mg of
19 osmium tetroxide is added thereto. The solution is stirred
for 30 minutes. Powdered sodium periodate [496 mg t2.32 mmole)~
~l is added to the black reaction mixture in 20 minutes.
22 After stirring for 1.5 hours, the reaction solution is extracted
23 with 50 ml of ether three times. Ether extracts are combined
~ and washed with saturated sodium chloride solution. The resulting
solution is dried with anhydrous sodium sulfate and concentrated
2~ under reduced pressure to obtain a dark-brown oily product.
27 The product is charged on a column packea with 5 g of silica
28 gel and elution is carried out with a solvent of bqnzene ànd
ethyl acetate (1 : 2). Fractions which are positive to 2,4-
dinitrophenylhydrazine reaction are collected and concentrated
31 to obtain 235 mg of an oily product which is the cis-isomer of
i- 85 -
a~
~r; -
1~l78956 ;
1 the aldehyde compound. The oily product is dissolved in 15 ml
2 of anhydrous acetonitrile. Sodium hydride [50 ~, 27.1 mg (0.563
3 mmole)] is added to the solution in a stream of nitrogen with
4 stirring at room temperature.
s After stirring for 20 minutes, the reaction mixture is poured
6 in 20 ml of 2 % aqueous acetic acid and the solution is
7 extracted with 50 ml of ether four times. Ether extracts are
8 combined and washed with saturated sodium chloride solution.
g The-resulting solution is dried with anhydrous sodium sulfate ~*~-
and concentrated under redueed pressure to obtain 180 mg of an
11 oily product whieh is identified as a crude product of the
12 desired cis-compound.
The oily product is charged on a column packed with 5 g of
siliea gel and elution is earried out with a solvent of n-hexane
and ethyl acetate (3.5 : 1, hy volume). I1~hite crystals (91 mg)
l6 of the desired compound are obtained. Yield 51 ~.
Properties of the compound are as follows.
~8 Meltiny point: 64.5 -65.5C
l9 IR(ClIC13)vmax : 2130, 1790, 1730, 1640
N~IR(CDC13)~(ppm) 6.30(1H, t, J =4Hz), 4.93(1H, d, J =
21 5Hz), 3.80(lH, q), 1.6 -2.6(4H, m), 1.52(9H, s)
22
Reference Exam~le 2
23 _ .
24 ~ Preparation of (-~)-trans-2-t-butyloxycarbonyl--7-
2s azido-l-azabicyclo [4, 2, 0] oct-2-en-8-one {the trans-compound
26 represented by the following formul~}:
27
28 H H
N3
29 ~ N
COO~
3 1
- 86
~78g5~
I In this Example, 767 mg (1,84 mmole) of trans-2~[4-
2 (3-butenyl)~3~azido-2-oxoazetidin-1-yl]~2~diethylphosphonoacetate~
3 t~bu~ylester obtained as in Reference Example 1 -1) is dissolved
4 in 22 ml of dioxane and 6.5 ml of deionized water, and 100 mg
of osmium tetroxide is added thereto. The mixture is stirred
6 for 30 minutes. Powdered sodium periodate [1.5 g (7.04 mmole)]
? is added to the black reaction mixture in 30 minutes.
8 ~fter stirring for one hour, the reaction mixture is extracted
9 three times with 150 ml of ether. Ether extracts are combined
0 and dried with anhydrous sodium sulfate. The resulting solution
1l is concen~rated under reduced pressure to obtain an oily product.
2 The oily product is charged on a column packed with 20 g of
~3 silica gel and elution is carried out with a solvent of benzene
14 and ethyl acetate (1 : 2). Oily product (561 mg) is obtained
from the fractions which are positive to 2,4-dinitrophenyl-
l6 hydrazine reaction. The product is the trans-compound of the
17 aldehyde compound. The ~rocluct is dissolved in 6 ml of anhydrous
l8 acetonitrile and 61.~ mg (2.56 mmole) of 50 % sodium hydride is
l9 added thereto. The mixture is heated to a temperature of 50C
and allowed to react for ten minutes. The reaction mixture is
21 poured in 6 ml of 2 % aqueous acetic acid and extracted four times
22 with 50 ml of ether. ~ther extracts are combined and dried wi-th
23 anhydrous sodium sulfate.
24 - The resulting solution is concentrated under reduced pressure
to obtain an oily product. The oily product is charged on a
26 column packed with 20 g of silica gel and elution is carried
27 out with a solvent of n-hexane and ethyl acetate (3.5 : 1).
28 White crystals (218 mg) of the desired compound are obtained.
29 The compound is identified as trans-compound of the desired
compound. Properties of the compound are as follows.
31 ,~
- 87 -
1178gS~ ''
I Melting point: 80.5 - 81.5C ~ -
2 IR(cHcl3)vcm : 2110, 17gO, 1720, 1635
3 NMR(CDC13)~(ppm): 6.27 (lH, t), 4.28(lH, d, J =2Hz),
. .
4 3.53(1H, q), 2.0 -2.6(4H, m), 1.63(9H, s)
s
6 Reference Example 3
? Preparation of (+)-cis-2-carboxy-7-azido-1-azabicyclo
8 [4, 2, 0] oct-2-en-8-one {The cis-compound represented by the
9 following formula}: ~.
11
H H
12 N3~,~
l3 o~N~
14 OOH
16
17 In this Examp].e, 55 mg (0.224 mmole) of (+)-cis-2-t-
18 butyloxycarbonyl-7-azido-1-azabicyclo [4, 2, 0] oct-2-en-8-one
19 obtained in Reference Example 1 is dissolved in 2 ml of trifluoro-
acetic acid and the solution is allowed to stancl at room tempera-
21 ture for 10 minutes. The solution is concentrated under reduced
22 pressure. Benzene is added to the concentrate and the resulting
23 solution is concentrated under reduced pressure to obtain 51 mg
2~ ~ of a yellow semisolid. Properties of the semisolid are as
follows and the semisolid is identified as -the desired carboxylic
26 acid. Yield 100 ~.
27 IR(cHcl3)vmax 2120, 1770(sh), 1760, 171-5, 1635
2B Ni'lR(CD30D)~(ppm)~ 6.48(lH, t, J =4Hz), 5.10(lH, d,
29 J = 5Hz), 3.83(lH, q), 1.1 -2.5(4H, m)
31
- 88
117~9S6
I Reference Example 4
2 Preparation of (+)-cis-2-t-butyloxycarb~nyl-4-bromo-
3 7-azido-1-azabicyclo [4, 2, 0] oct-2-en-8-one {The cis-compound
4 represented by the following formula}:
N3 Br
9 ' ~coo~u
11
2 In this ~xample, 50 mg (0.203 mmole) of (+)-cis-2-t-
3 butyloxycarbonyl-7-azido-1-azabicyclo [~, 2, 0] oct-2-en-8-one
~4 obtained in Reference Example 1 is dissolved in 2 ml of anhydrous
chloroform, and 36.0 mg (0.202 mmole) of N-bromosuccinimide and
l6 a catalytic amount of azobisisobutyronitrile are added thereto.
17 The mixture is heated under reflux with stirxing for 30 minu-tes
l8 and is diluted with 5 ml of chloroform. The diluted solution
l9 is washed with 3 ml of water and 3 ml of saturated sodium
chloride solution, and dried with anhydrous sodium sulfate.
21 T~e resulting solution is concentrated under reduced pressure
22 to obtain 53 mg of an oily product.
23 The product is charged on a column packed with 4.0 g
24 of silica yel and elution is carried out with a solvent of
n-hexane and ethyl acetate (3.5 : 1). An oily product (23 mg)
26 iS obtained. The product is identified as the desired cis-
27 compound from the following properties. Yield 33 %.
28 IR(CHcl3)~max 2120, 1790, 1730, 1620
29 NMR(CDC13)~(ppm): 6.33(lH, d, J = 6Hz), 5.07(lH, d, J =
5Hz), 4.93(1H, m), 4.50 -3.90(1H, m), 2.50 -1.72
3l (2H, m), 1.52(9H, s)
- 89
11789S~
-:
1 Reference Example 5
2 Preparation of (+)-trans-2-t-butyloxycarbonyl 4-
3 bromo-7-azido-1-azabicyclo [4, 2, 0~ oct-2-en-8-one {The trans-
4 compound represented by the following formula}:
H H
7 N3.. ~ Br
O~L
- , COOtBU
11 .
In this Example, 100 mg (0.407 mmole) of (+)-trans-
13 2-t-butyloxycarbonyl-7-azido-1-azabicyclo [4, 2, 0] oct-2-en-
14 8-one obtained in Reference Example 2 is dissolved in 5 ml of
anhydrous carbon tetrachloride and 72.4 mg of N-bromosuccinlmide
6 is added thereto. The mixture is heated under reflux with
stirring for 30 minutes. Thereafter, 10 ml of methylerlechloride
8 is added to the reaction mixture and the resulting mixture is
l9 washed with 5 ml of deionized water and 5 ml of saturated sodium
chloride solution. The resulting solution is dried with
2l anhydrous sodium sulfate and concentrated under reduced pressure
22 to obtain 102 mg of an oily product. ~he oily product is charged
23 on a column packed with 5 g of silica gel and elution is carried
24 out with a solvent of n-hexane and ethyl acetate (3.5 1).
2s An oily product (24 my) is obtained. The product is
26 identified as the desired trans-compound from the following
27 properties. Yield 18.1 %.
28 IR(CHcl3)vmax 2130, 1790, 1730, 1620
2~ N~lR(CDC13)~(ppm): 6.23(lH, d, J =6Hz), 5.93(1H, m),
4.37(lH, d), 4.00(1H, m), 2.93 -1.93(2H, m),
3l 1.50(9H, s)
,
117~9S~
I Reference Example 6
2 PreparatiOn of (+)-cis-2-t-butyloxycarbonyl-4~-
3 acetoxy-7~-azido-1-azabicyclo [4, 2, 0] oct-2-en-8-one {the
4 cis-compound represented by the following formula}: -
H H
7 N3 ~ -~COCH3 --
O ~ N
COOtBU
o
2 In this Example, 75 mg (0.219 mmole) of (+)-cis-2-t-
butyloxycarbonyl-4-bromo-7-azido-1-azabicyclo [4, 2, 0] oct-2-
en-8-one (the cis-compound obtained as in Reference Example 4)
is dissolved in 2 ml of acetic acid.
6 Under protecting the reaction system from light, 39.4 mg (0.241
l7 mmole) of silver acetate is added to the solution and the mixture
18 is stirred for 2 hours and 20 minutes. The reaction mixture is
lg subjected to filtration and concentrated under reduced pressure
zO to obtain a crude acetoxy product of the desired compound.
2I The product is charged on a column packed with 3.5 g of silica
22 gel and elution is carried out with a solvent of n-hexane and
23 ethyl acetate (3.5 : 1) to obtain 51 mg of an oily product.
2~ The product is identified as the desired cis-compound from the
2s following properties. Yield 72.1 %.
26 IR(CHcl3)vmax 2130, 1790, 1750, 1730(sh), 1635
27 NMR(CDC13)~(ppm): 6.21(lH, d, J = 5Hz), 5.42(1H, m),
28 5.01(1H, d, J =5Hz), 3.95(1H, m), 2.02(3H, s),
29 2.6 -1.7(2H, m), 1.53(9H, s)
! - g~ -
1178gS6 - ~
Reference Example 7
2 Preparation of (+)-cis-2-t-butyloxycarbonyl-4-methyl-
3 7-azido-1-azabicyclo [4, 2, 0] oct-2-en-~-one (the cis-cornpound
4 represented by the following formula):
s
N3 ~ CH3
9 ' ~
COOtBU
l2 The present compound is produced according to the
13 following processes 1 and 2.
,1
1) Preparation of 2-[4-(2-methyl-3-butenyl)-3-azido-2-
oxoazetidin-l-yl]-2-diethylphosphonoacetate tert-butylester
16
In this ~xample, 2.13 g (8 ~ole) of t-butyl-~-amino-
l7
diethylphosphonoacetate is dissolved in 80 ml of anhydrous ether
ls
and 902 mg (9.2 mmole) of 3-me-thyl-4-pentenal is added with
19
stirring. The mixture is stirred at room temperature for one
hour and 900 mg o~ Molecular Sieve 4A and 700 mg of magnesium
21
su~fate are added. After stirriny for 1.5 hours, the reaction
22 -~
mixture i5 subjected to filtration under reduced pressure.
23
The resulting filtrate is concentrated to obtain a pale yellow
24
- oily product. Anhydrous benzene (30 nl) is added to the product
2s
and the resulting solution is again concentrated to obtain 2.82
26
g of an oily product. The presence of a Shiff's base in the
27
product is confirmed by N~.R spectrum. The oily product is
28
dissolved in 56 ml of dried cyclohexane and 56 ml of anhydrous
29
benzene and 900 mg of Molecular Sieve 4A and 1.67 ml (i2 mmole)
of triethylamine are added. To the mixture, 1.43 g (12 ~nole)
31
- 92 -
h~
1178gSt~i
I o~ azidoacetylchloride dissolved in 56 ml of dried cyclohexane
2 is added dropwise in 1.5 hours at room temperature with stirring.
3 The mixture is further stirred for 30 minutes and 30 ml of
4 benzene is added thereto. The mixture is transferred into a --
separatory funnel and washed with 30 ml each of 10 % citric
6 acid, saturated sodium chloride solution, saturated sodium
7 bicarbonate, and saturated sodium chloride solution in that
8 order. The resulting solution is dried with anhydrous sodium
g sul-fate and concentrated under reduced pressure to obtain 2.8
o of an oily product. The presence of a mixture of two major
Il isomers in the product is ascertained by a thin layer chromato-
l2 graphy [silica gel, n-hexane - ethyl acetate (1 : 1)].
3 The product is charged on a column packed with 300 g of silica
4 gel and elution is carried out with a mixed solvent of n-hexane
~5 and ethyl acetate (1 : 1) to obtain 380 mg (yield 11.0 %) o~
16 the less polar isomer of the desired compound, 570 mg (yield
17 16.7 %) of the more polar isomer of the desired compound, and
l8 201 mg (yield 5.8 ~) of a mixture of the two isomers.
l9 The properties of each isomer are described below.
From the data, the more polar isomer is identified as the cis-
21 isomer of the desired compound.
22 The less polar isomer
23 IR(CHC13)vmax 2110, 1770, 17~5
24 N~R(CDC13)~(ppm): 5.40 -6.10(lH, m), 5.27 -4.90(2.5H, m)
2s 4.68(0.5H, d) 4.23(6H, m), 2.60 -1.77(3H, m),
26 1.53(9H, s), 1.37(6H, t, J - 7.0Hz), 1.10(3H, d,
27 J = 6.OHz)
2 The more polar isomer (cis-compound)
8 -1
29 IR(CHC13)vmax ; 2110, 1765, 1745
N~IR(CDC13)~(ppm): 5.45 -6.13(lH, m), 4.83 -5.20(2.5H, m),
4.67(0.5H, d), 3.97--4.'15(6H, m), 1.77-2.55(3H, m),
1 50(9H, s), 1.33(6H, t), 1.08(3H, d)
- 93
1178g~6 ......
1 2) Preparation of (+)-cis-2-t-butyloxycarbonyl-4-methyl-
2 7-azido-l-azabicyclo [4, 2, 0] oct~2-en-8-one (the Ci5-
3 compound represented by the following formula):
~ H
6 N3 ~ ~H3
8 O N ~
COO BU
g .~
In this Example, 240 mg (0.56 mmole) of the tert-
2 butyl ester of (+)-cis-2-[4-(2-methyl-3-butenyl)-3-azido-2-
13 oxoazetidin-l-yl]-2-diethylphosphonoacetate obtained in Reference
14 Example 7 - l) is dissolved in 6.6 ml of dioxane and 2 ml of
deionized water. Osmium tetroxide (20 mg) is added and the
~6 mixture is stirred for l0 minutes. Powdered sodium periodate
17 [3g0 my (l.82 mmole)] is added in small portions to the black
18 reaetion solution in 30 minutes.
19 After stirring for 40 minutes, the reaction solution is subjected
to extraction three times with 30 ml of ether and the extraets
21 are combined. The combined extracts are washed with saturated
22 sodium chloride solution, dried with anhydrous sodium chloride,
23 and concentrated to obtain 230 mg of an oily product. The oily
2~ product is charged on a column packed with 6 g of silica gel
2s and elution is carried out with a solvent of benzene and ethyl
26 acetate (l : 2). Fractions which are positive to 2,~-dinitro-
27 phenylhydrazine reaction are combined and concentrated to
28 obtain 185 mg of an oily product which is the cis-compound of
29 the aldehyde compound. The product is immediately dissolve in
8 ml of anhydrous acetonitrile and 21.6 mg (0.45 mmole) of 50 %
31 sodium hydride is added to the solution in a streatm of nitrogen
' ~ 94
117~95~
I with stirring at room temperature. ~fter stirring for 30 --
2 minutes, the reaction solution is poured into 15 ml of 2 %
3 aqueous acetic acid and the mixed solution is subjected to
4 extraction twice with 20 ml of ether. Ether layers are washed -
s with saturated sodium chloride solution, dried with anhydrous
6 sodium chloride, and concentrated under reduced pressure to
7 obtain an oily product.
8 The product is identified as a crude product of the desired
g eis-compound. The oily product is eharged on a column packed
lo with 20 g of silica gel and elution is carried out with a solvent
of n-hexane and ethyl acetate (3.5 : 1, by volume). The desired
12 compound (70 mg) is obtained as a colorless oily product which
13 crystallizes on standing. Properties of the product are
l4 deseribed below. Yield 48.1 O.
IR(KBr)vmax : 2110, 1784, 1715, 1623
16 NMR(CDC13)~(ppm): 6.30(4/5H, d, J -5.1Hz), 6.10(1/5H, d,
l7 J =2.7Hz), 4.98(4/5H, d, J =5.0Hz), 4.89(1/5H, d,
18 J =5.0Hz), 3.60 -3.90(1H, m), 2.65(1H, rn), 1.70 -1.80
l9 (2H, m), 1.51(9H, s), 1.20(3/5H, d, J =8.0Hz),
1.13(12/5H, d, J = 8.0Hz)
21 The obtained crystals are identified as a mixture of
22 4~-methyl isomer and 4~-methyl isomer in a ratio of about 4 : 1
23 by the n m r data mentioned above.
24
2~
26
27
28
29
3 1
I - 95 -
-
li~7895~
1 Reference Example 8
2 Preparation of (+)-cis-2-carboxy-7-amino-l-azabicyclo
3 [4, 2, O] oct-2-en-8-one
s H .~N `
6 .~L N~
CO~H
8 . . . - . . . ..
g ~
11 ~In this Example, 9l mg of (+)-cis-2-carboxy-7-azido-
~2 l-azabicyclo [4, 2, O] oct-2-en-8-one obtained as in Reference
13 Example 3 is dissolved in 6.5 ml of ethanol and 26 mg of lO %
14 palladium-carbon is added thereto. The mixture is stirred at
1s room temperature and at atmosp1leric pressure in a stream of
16 hydrogen for 2 hours. The mixture is subjected to filtration
~7
18 .
19
21
22
23
24
2s
26
27
28
29
31
-- 9 6
-~78956
I to remove the catalyst and the filtrate is concentrated under
2 reduced pressure. The concentrate is again dissolved in 10 ml
3 of methanol and 26 mg of 10 % palladium on carbon is added
4 thereto.' The mixture is subjected to catalytic reduction at `I
room temperature and at atmospheric pressure for 3 hours and
6 50 minutes and is subjected to filtration using a filter'aid,
7 Hyflo Super Ce~. The filtrate is concentrated under reduced
pressure to obtain 88 mg (100 %) of a semisolid product; The
g product is identified as the desired amino-compound based on the
~0 following da,ta. 1
Il IR(KBr)vmax : 3450, 2950, 1770, 1650
~7
~3 ~eCerence Example 9
l4 Preparation of (+)-cis-7-amino-2-t-butyloxycarbonyl-
lS l-azabicyclo [4, 2, 0] oct-2-en-8-one;
~6
17 H H H H
~8 N ~ H~N ~
2tBu COl~u
2~
22
73 In this Example, 178 mg (0.67 mmole) of (+) cis-7-
7~ azido-2-t-butyloxycarbonyl-1-azabicyclo ~4, 2, 0] oct-2-en-8-
25 one obtained in Reference Example 1 is dissolved in 10 ml of
26 ethanol and 25 mg of 10 % palladium-carbon (catalyst) is added
77 thereto. The mixture is stirred at room temperature~in a stream
23 of hydrogen for 50 minutes. The reaction solution is subjected
29 to filtration to remove the catalyst and the filtrate is con-
30 centrated under reduced pressure to obtain the desired compound
31 as a yellow oil product.
1- 97
1~7~gS~ '
The amount of the product: 159.5 mg
2 Yield: 100 ~
3 IR(CHC13)vmmax : 1775, 1725, 1640
g NMR(CDC13)~(ppm) 6.27(m, lH), 4.50(m, lH), 4.2 -3.1
(m, 3H), 2.6 -1.7(m, 4H), 1.5(s, 9H)
6 In case that the catalytic reduction is carried out
in ethanol containing an equivalent of hydrochloric acid, the
8 hydrochloride of the desired compound is obtained.
lo Reference Example 10
Il ~Preparation of (+)-cis-2-t-butyloxycarbonyl-4-methyl-
12 7-azido-1-azabicyclo [4, 2, 0] oct-2-en-8-one represented by
3 the following formula:
14
6 `t~-
8 ~OO
lg
~P'D'.,~,ff.
21 The present compound is produced by the following
22 processes 1) and 2).
23
1) Preparation of the t-butyl ester of 2-[4--(2-methyl-
24
3-butenyl)-3-azido-2-oxoazetidin-1-yl]-2-diethylphosphono-
acetate
26
27
~ "^~,CH3
29 O ~ t
.2 BU
3 1
I I - 98
117~9~i6 ~
, .:
I In this Example, 2.13 g (8 mmole) of t-butyl-~-amino ~-
2 diethylphosphonoacetate is dissolved in 80 ml of absolute ether
3 and 902 mg (9.2 mmole) of 3-methyl-4-pentenal is added with
4 stirring. The mixture is stirred at room temperature for one
s hour and 900 mg of Molecular Sieve 4A and 700 mg of magnesium
6 sulfate are added thereto. The mixture is stirred for 1.5 hours
7 and is subjected to filtration under reduced pressure. The
8 filtrate is concentrated to obtain a pale yellow oily product.
g To the product, 30 ml of anhydrous benzene is added and the
mixture is again concentrated to obtain 2.82 g of an oily product.
l The presence of a Shiff's base is confirmed by nmr spectrum.
2 The oily product is dissolved in 56 ml of dried cyclohexane and
3 56 ml of anhydrous benzene and 900 mg of Molecular Sieve 4A and
1.67 ml (12 mmole) of triethylamine are added thereto.
Azidoacetylchloride [1.43 g (12 mmole)] dissolved in 56 ml of
16 dried cyclohexane is added dropwise to the mixture with stirring
l7 at room temperature in 1.5 hours and the mixture is stirred for
8 30 minutes. The reaction rnixture is transferred into a separa-
l9 tory funnel together with 30 ml of benzene. The benzene layer
is washed with 30 ml each o~ 10 % ci-tric acid, saturated sodium ~4
21 chloride solution, saturated sodium bicarbonate and saturated
22 sodium chlo~ide solution. The resulting solution is dried with
23 anhydrous sodium sulfate and concentrated under reduced pressure
2~ to obtain 2.8 g of an oily product. The presence of two isomers
2s in the product is detected by thin layer chromatography [silica
26 gel, n-hexane and ethyl acetate (1 :1)]. The product is charged
27 on a column packed with 300 g of silica gel and elution is
28 carried out with n-hexane and ethyl acetate (1 1). The less
polar isomer (380 mg, yield 11.0 %) of the desired compound,
570 mg (yield 16.7 %) of the more polar isomer, and 201 mg
(yield 5.8 %) of a mixture of the two isomers are obtained.
99
'"5'`
~ !l7~ 6 - `
Properties of each isomer are set forth below. The
2 higher polar-isomer is identified as the cis isomer of the
3 desired compound.
4 The less polar-isomer (trans isomer)
IR(CHc13) max 2110, 1770, 1745
6 NMR(CDC13)~(ppm): 5.40 -6.10(1H, m), 5.27-4.90(2.5H, m),
7 4.68(0.5H, d), 4.23(6H, m), 2.60 -1.77(3H, m),
8 1.53(9H, s), 1.37(6H, t, J = 7.0Hz), 1.10(3H, d,
g - J =6.0Hz)
o The more polar isomer (cis isomer)
IR(CHcl3)vmax 2110, 1765, 1745
l2 NMR(CDC13)~(ppm): 5.45 -6.13(lH, m), 4.83 -5.20(2.5H, m),
l3 4.67(0.5H, d), 3.97 -4.45(6H, m), 1.77 -2.55(3H, m),
1.50(9H, s), 1.33(6H, t), 1.08(3H, d)
2) Preparation of (+)-cis-7-azido-2-t-butyloxycarbonyl-
16 4-methyl-7-azido-1-azabicyclo [4, 2, 0] oct-2-en-8-one
In this Example, 2~0 my (0.56 mmole) of the t-
1 butylester of (~)-cis-2-[~-(2-me-thyl-3-butenyl)-3-azido-2-
19 oxoazetidine-1-yl]-2-diethylphosphonoacetic acid obtained in
Reference Example 10 -1) is dissolved in 6.6 ml of dioxane and t~:
2l 2 ml of deionized water, and 20 mg of osmium tetroxide is added
22 thereto. The mixture is stirred for 10 minutes and 390 mg ~`
23 (1. 82 mmole) of powdered sodium periodate is added in small
24 portions to the black reaction mixture in 30 minutes.
Af-ter stirriny for 40 minutes, the reaction solution is subjected
2G to extraction three times with 30 ml of ether. The extracted
27 ether layers are combined, washed with saturated sodium chloride
23 solution, dried with anhydrous sodium sulfate and concentrated
29 to obtain 230 mg of an oily product. The oily product is charged
on a column packed with 6 g of silica gel and elution is carried
3l out with a solvent of benzene and ethyl acetate (1 :2).
I -- 1 00 --
-r~
117~g5~
1 Fractions which are positive to 2,4-dinitrophenylhydrazine
2 reaction are combined and concentrated to obtain 185 mg of an
3 oily product, the aldehyde compound of the desired compound.
4 The product is immediately dissolved in 8 ml of anhydrous
acetonitrile and 21.6 mg (0.45 mmole) of 50 ~ sodium hydride
6 is added thereto at room temperature with stirring in a stream
7 of nitrogen. After stirring for 30 minutes, the reaction mixture
8 is poured into 15 ml of 2 ~ a~ueous acetic acid and the mixture
g is extracted twice with 20 ml of ether. The obtained ether
o layers are washed with saturated sodium chloride solution, dried
Il with anhydrous sodium su1fate and concentrated under reduced
12 pressure to obtain an oily product. The product is charged on
13 a column packed with 20 g of silica gel and elution is carried
l4 out with a solvent of n-hexane and ethyl acetate (3.5 :1, by
volume). The desired product (70 mg) is obtained as a color-
16 less oily product in a yield of 48.1 ~. The product crystallizes
on standing. Properties of the product are as follows.
~8 IR(KBr)Vmax . 2110, 1784, 1715, 1623
lg NMR(CDC13)~(ppm): 6.30(4/5H, d, J = 5.1Hz), 6.10(1/5H, a,
J =2.7Hz), 4.98(4/SH, d, J = 5.OHz), 4.89(1/5H, d,
21 J = 5.0Hz), 3.60 - 3.90(lH, m), 2.65(1H, m), 1.70-
22 - 1.80(2H, m), 1.51(9H, s), 1.20(3/5H, d, J =8.OHz),
23 1.13(12/5H, d, J = 8.OHz)
24 Based on the data of nmr, the obtained crystals are identified
2s as a mixture of 4~-methyl isomer and 4~3-methyl isomer in the
26 ratio of about 4 : 1. The compounds can be separated by silica
27 gel chromatography using a solvent of n-hexane and e~thyl acetate
28 (3 : 1). The more polar isomer corresponds to 4~-CH3 isomer,
9 i.e. (+)-cis-7~-azido-4~-methyl-2-t-butyloxycarbonyl-1-
azabicyclo [4, 2, O] oct-2-en-8-one and the less polar isomer
31 corresponds to 4~-CH3 isomer. They have the following properties
-- 1 01
~178gS6
Irespectively.
~ .
H H H H
4~7 ~ CH3 ~3 ~ CH~ ~
6~ C02~Bu CO~u
7~ ~ CH3 isomer ~ a CH3 isomer
94~-CH3 isomer
Melting point: $4.0 -86.5C
l1 IR(KBr)vmax : 2135, 1783, 1715, 1622
12 NMR(CDC13)~(ppm): 6.13(1H, d, J =2.7Hz), 4.90(1H, d,
13 J = 5.01-1z), 3.93 -3.73(lH, m), 2.53(lH, m),
1~ 2.16 -1.75(2H, m), 1.53(9H, s), 1.20(3H, d, J = 6.0Hz)
s 4~-CH3 isomer
~6 Melting point: 82.0 - 84.0C
17 IR(KBr)vmax : 2120, 1790, 1721, 1630
18 N~R(CDC13)~(ppm): 6.33(1H, d, J =5.011z), 5.00(lH, d,
Is J = 5.5Hz), 3.89 -3.68(1H, m), 2.66(lH, m),
1.82-1.57(2H, m), 1.53(9H, s), 1.12(3H, d, J = 7.0Hz)
21
22 Reference Example 11 -
23 Preparation of (~)-cis-7~-amino-4~-methyl-2-t-
2~ butyloxycarbonyl-l-aZabiCyClo [4, 2, 0] oct-2-en-8~one:
26
27 h ~ 3 1 ~ ~ ~
29 C2 ~ 2
3l
!¦ -- 102 --
1~789S6
1 In this Example, 255 mg (0.67 mmole) of (+)-cis-7~-
2 azido-4~-methyl-2-t-butyloxycarbonyl-l-azabicyclo [4, 2, 0~
3 oct-2-en-8-one obtained in Reference Example 10-2) as a less
4 polar isomer is dissolved in 10 ml of ethanol and 100 mg of
s 10 % palladium-carbon is added thereto. The mixture is sub-
6 jected to catalytic hydrogenation for 1.5 hours and to filtra-
7 tion to remove the catalyst. The catalyst is washed with
8 methanol. The filtrate and the washings are combined and
g concentrated under reduced pressure to obtain a pale yellow ~-
o oily product. The product is dissolved in 8 ml of ethyl-acetate
1l and the solution is subjected to extraction five times with
3 ml of 10 % citriC acid. The water layer is adjusted to a
3 pH of 6 to 7 with potassium carbonate to obtain a white suspen-
14 sion. The suspension is extracted two times with 5 ml of ethyl
IS acetate and washed with saturated sodium ehloride solution.
16 The washings are dried with anhydrous sodium sulfate to obtain
17 177 mg (76.6 ~) of an oily product.
18 IR(CHC13)vmax 3400, 1770, 1720, 1630
Ig NMR~CDC13)~(ppm): 6.23(lH, d, J = 5.OHz), 4.53(lH, d,
J = 5.8Hz), 3.93 -3.47(1H, m), 2.56(1H, m), 1.92
21 (2H, br), 1.80 -1.60(2H, m), 1.50(9H, s),
22 . 1.31(3H, d, J =7.0Hz)
23 In case that the eatalytic hydrogenation is earried
24 out with an equivalent of hydroehlorie acid, the hydrochloride
of the desired compound is obtained.
26
27
28
29
3 1
l 0 3. ,-
'~'i" ?~-
~1789S6
.;....
I Reference Example 12 --
2 Preparation of (+)-Cis-7~-amino-4~-methyl~2-t-
3 butyloxycarbonyl-l-azabicyclo [4, 2, 0] oct-2-en-8-one:
s
6 H H H H
8 N~ ~ CH~ H~N ~ CH3
O;! B~l C.2~
g . ~
In this Example, 655 mg (2.35 mmole) of (+)-cis-7~-
l2 azido-4~-methyl-2-t-butyloxycarbonyl-1-azabicyclo ~4, 2, 0]
13 oct-2-en-8-one obtained in Reference Example 10-2) as a more
l4 polar isomer is dissolved in 6 ml of ethanol and 0~79 ml
lS (2.37 mmole) of 3N~HCl is added thereto. The mixture is
16 subjected to hydrogenation with 200 mg of 10 ~ palladium-carbon
l7 for 70 minutes. Methanol is added to the resulting mixture to
l8 dissolve the deposited salt of the desired compound. The
19 catalyst is removed by filtration and the filtrate is concen-
trated to obtain a crude product. The product is well triturated
2I with ether and filtered. The filtrate is dried to obtain 512 mg
22 (75.4 ~) of the hydrochloride of the desired compound.
23 Melting point: 216 -221C (dec.)
2~ IR(KBr)vmax : 3430, 2590, 1780, 1762, 1712, 1630
26 Reference Example 13
27 Preparation of the trifluoroacetate of (-t)=cis-7~-
28 amino-4~-methyl-2-carboxy-1-azabicyclo [4, 2, 0] oct-2-en-8-one:
29
- 104 -
1~789S~; ~
3 H2N ~ C~3 H~N' ' C~
~ C~3CO~H
o~ ~ O~H
7 In this Example, 196 mg (0.78 mmole) of (+)-cis-7~-
8 amino-4rl-methyl-2-t-butyloxycarbonyl-1-azabicyclo [4, 2, 0]
9 oct.2-en-8-one obtained in Reference Example 11 is dissolved
in 4.2 ml of anhydrous dichloromethane and 1.8 ml of trifluoro-
Il acetic acid is added thereto at room temperature with stirring.
~2 After 1.5 hours, the mixture is concentrated under reduced
13 pressure. The concentrate is subjected to azeotropic distilla-
14 tion with anhydrous benzene to obtain an oily product. The
product is triturated with ether and filtered to obtain 167 mg
16 (69.3 6) of a powdered desired compound.
17 IR(KBr)vrnax : 3460, 2980 -2500, 1780, 1685, i630
18 PMR(D2O, with DSS as an internal standard)~(ppm):
19 6.77(1H, d, J =5.8Hz), 5.00(1H, d, J =5.6Hz),
4.10(1H, m), 2.83(1H, m), 1.86(2H, m), ].15(311,
2I d, J = 8.0Hz)
22
23 Reference Example 14
__
24 Preparation of (')-cis-7~,-amino-4~-acetoxy-1-
2s azabicyclo [4, 2, 0] oct-2-en-8-on-2-carboxylic acid
26
27 1) Preparation of (~)-cis-7~-azido-4rl-ace~oxy-1-azabicyclo
28 [4, 2, 0] oct-2-en-8-on-2-carboxylic acid:
29
31
105
1178g56
~ ~ OCOCh; N~ ~ .OCOCH3
C00 ~ U C00!~
7 In this Example, 179 mg of (+)-cis-7~-azido-4~-acetoxy-
8 2-t-butyloxycarbonyl-1-azabicyclo [4, 2, 0] oct-2-en-8-one
g obtained as in Reference Example 6 is dissolved in 3 ml of
methylene chloride and 3 ml of trifluoroacetic acid.
1~ The solution is allowed to stand at room temperature for two
2 hours and concentrated to obtain 145 m~ of the desired compound
13 as a yellow powder. Yield 100 ~. Properties of the compound
1~ are set forth below.
IR(CHC13)vmax : 2130, 1790, 1715, 1445
16
l7 21 Preparation of (~)-cis-7~-amino-4~-acetox~-1-
lB azabicyclo [4, 2, 0] oct-2-en-8-on-2-carboxylic acid:
19
H H
J~c_oc~3 H2M ~ rCC!)CH3
23 C00H C0~H
2~
26 In this Example, 145 mg of (~)-cis-7~-azido-4~-acetoxy-
27 l-azabicyclo [4, 2, 0] oct-2-en-8-on-2-carboxylic acid obtained
28 as in Reference Example 14-1) is dissolved in 14 ml of ethanol and
29 40 mg of 10 ~ palladium-carbon is added thereto The mixture
is subjected to catalytic hydrogenation at atmospheric pressure
31 with stirring for one hour. The reaction mixture is subjected
; - 106 -
~78gs6
1 to filtration and the filtrate is concentrated to obtain 126 mg
2 of the desired compound.
4 Reference Example l5 --
Preparation of (+)-cis-7~-azido-2-t-butoxycarbonyl-
6 4~-hydroxy-l-azabicyclo [4, 2, 0] oct-2-en-8-one (the cis
7 compound represented by the following formula)
g . ~
0 N3 ,H ~ OH
~ ~ 0 ~ N
CCO
13
14
~5 In this Example, 200 mg of (+)-cis-2-t-butoxycarbonyl-
16 7-azido-l-azabicyclo ~4, 2, 0] oct-2-en-~-one obtained as in
17 Reference ~xample l is dissolved in 8.8 ml of carbon tetrachlo-
18 ride and 134.9 mg of N-bromosuccinimide and catalytic amount of
19 ~ azobisisobutyronitrile are added, The mixutre is heated
under reflux for 30 minutes. After cooling, the reaction
21 mixture is diluted with 5 ml of chloroform and washed with 3 ml
22 each of water and saturated sodium chloride solution. The -
23 washing is dried with anhydrous sodium sulfate and filtered.
2~ The filtrate is concentrated to obtain an oily bromo compound
2s which is the same oily product in Example 4. The product is
26 immediately dissolved in lO ml of acetone and 50 mg of silver
27 carbonate and 50 ~l of water are added. The mixture is stirred
28 at room temperature for lO minutes. The reaction mixture is
29 filtered and concentrated to obtain a crude product. The
product is charged on a column packed with 20 g o. sllica gel
31 and elution is carried out wi-th a mixture of n-hexane and ethyl
1- 107 -
~6~
~78956
l acetate (2 :1). The eluates are concentrated to obtain 86.4 mg
2 of the desired compound as pale yellow crystals. Yield 40.7 %.
3 Properties of the crystals are set forth below.
4 M.P. 100.0 - 101.0C
s IR(CHC13)vCm : 2130, 1790, 1635, 1630
6 N~IR(CDC13)~(ppm): 6.30(lH, d, J = 5Hz), 5.03(lH, d, J =
7 5.2), 4.47(1H, m), 3.93(1H, m), 3.20(1H, br),
8 2.1 -1.8(2H, m), 1.55(9H, s)
~ g
lo Reference Example 16
11 Preparation of (+)-cis-7-amino-1-azabicyclo [4,2, 0]
oct-2-en-8-on-2-carboxylic acid
13
14
16 H~N ~ HlN ~
17 CO,tBU ColH
18
19
In this Example, 300 mg of (t)-cis-7-amino-2-t-
2I butyloxycarbonyl-l-azabicyclo [4, 2, 0] oct-2-en-8-one obtained
22 as in Reference Example 9 is dissolved in 3.0 ml of methylene
23 chloride and 3.0 ml of trifluoroacetic acid is added. The
24 mixture is allowed to stand at room temperature for one hour
2s and 20 minutes. The reaction mixture is concentrated and
26 benzene is added to the residue. The solution is again con-
1 27 centrated to obtain 250 mg of the trifluoroacetate of the
28 desired compound as a yellow powder. Properties of the product
29 are set forth below.
IR(KBr)vmax : 1780, 1680, 1630
- 108 -
~89S~
1 The above trifluoroacetate is dissolved in 2 ml of
2 water and adjusted to pH 7.0 with saturated sodium bicarbonate
3 to form crystals. Thus, 129 mg of the desired compound is
4 recovered by filtration. Properties of the product agree with
those of the product in Reference Example 8.
7 Reference E~ample 17
8 Preparation of (+)-cis-7~-azido-2-carboxy-4~-methyl-
g l-azabicyclo [4, 2, G] oct-2-en-8-one
Il :
3 ~' ~ ~"
14 CO.~'C91L 02X
16
l7 In this Example, to 238 mg (0.703 mmole) of (_)-cis-
l8 7~-azido-2-t-butyloxycarbonyl-~-methyl-1-azabicyclo ~4, 2, O]
l9 oct-2-en-8-one obtained as in Reference Example lO is added
4 ml of trifluoroacetic acid and the mixture is allowed to stand
21 at room temperature for 10 minutes. The reaction mixture is
22 concentrated at 25C under reduced pressure. The concentrate ~-
23 iS subjected to extraction with 5 ml of dried benzene twice.
24 Then, 255 mg of the obtained oily product is dissolved in 5 ml
of ethyl acetate. The solution is extracted with 2 ml of 10 %
26 potassium carbonate twice and the water layer ls adjusted to pH
27 about 3 with 0.5N HCl. The solution is ex-tracted with 5 ml of
28 ethyl acetate twice and dried over anhydrous sodium sulfate.
29 The solvent is distilled off under reduced pressure to obtain
166 mg of the desired compound as an oily product. Yield 83.8 %.
31 The propduct crystallizes on standing. ,~
-- 1 09 -
~ s~ ~
1 ~L7895~ -
1 Properties of the product are set forth below. ~~
2 M.P. : 121.5 - 123.0C
~l
3 IR(CHcl3)vmax 2110, 1769, 1750, 1716, 1630
4 NMR(CD30D)~(ppm): 6.47(lH, d, J = 5.6Hz), 5.22(lH, d,
J =5.0), 4.2 -3.7(1H, m), 2.3 -2.9(1H, br),
6 1.11(3H, d, J = 7.2)
8 _ ference Example 18
g ~- Preparation of the trifluoroacetate of (+)~cis-7
lo amino-4~-methyl-1-azabicyclo [4, 2, 0~ oct-2-en-8-on-2-
1l carboxylic acid
12
CF3 ~
C~H C02H
16 ~ -
18
l9 In this Example, 200 mg of (+)-cis-7~-azido-2-t-
butyloxycarbonyl-4~-methyl-1-azabicyclo [4, 2, 0] oct-2-en-8-one
21 is dissolved in 2 ml of water and 2 ml of ethanol followed by
22 addition of 75 mg of 10 % palladium-carbon. -~-
23 The mixture is stirred in a stream of hydrogen gas at atmospheric
2~ pressure. After 20 hours, the reaction mixture is filtered under
2s reduced pressure. To the cake is added 2 ml of trifluoroacetic
26 acid.
27 After removing the catalyst by filtration,~the filtrate
28 is concentrated under reduced pressure followed by addition of
29 10 ml of dried ether. The crystals formed are recovered by
filtration to obtain 120 mg of the desired compound. Yield 43 ~.
3I Properties of the compound agree with those in Reference Example
13. Applicant
Name: (102) Kyowa Hak~o Kogyo Co., ~d.
Shukuo Kinos~ita, Representativc Director
~ 1 n
