CA1178956A - Cephalosporin analogs - Google Patents

Abstract

A B S T R A C T
The present invention relates to 7-acylamino-1-carba-3-cephem-4-carboxylic acid and derivatives thereof represented by the following general formula [1]:

[I]

(wherein x represents an acyl group, R2 represents hydrogen atom or a protecting group of carboxylic acid and R2 represents hydrogen atom, a lower alkyl group or a lower acyloxy group). These compounds posses excellent anti-bacterial activities against Gram-positive and Gram-negative bacteria.

Description

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.

XHN R
9 ~ ~
o~N~. [I]
11 ~ CO2 .~,.

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 23 ~ ~ N C~2 24 H ~ ~ Ac : 0 COO~

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 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 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 .

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]
. ~.. ~ . . .

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 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 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 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}.

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~-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.

1~789S6 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.

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~

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.

~ ~ ~ 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~.

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

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 17 ~

1!1 712r7 ~ 0COCH~3 N3 ~ ~oCOCH3 o ireduct- O
COOH tion CO~H

22 ~ t-~

2~

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 1~7~956 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.

~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 _- ' ~.' ' 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~

~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.

~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 . ~ .. , 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 ~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

Microorganism MIC (~g/ml) IS
Vibrio percalans KY4174 2 ~rwinia aroides KY3241 Staphylococcus aureus KY4279 18 :~`
Escherichia coli KY4271 8 Bacillus subtilis KY4273 2 Proteus vulgaris KY4277 8 22 Shigella sonnei KY4281 8 Salmonella typhosa KY4278 Klebsiella penumoniae KY4275 4 2s 2~

31 ~.

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.

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).

3l ~,~

~. 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 -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).

~9 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 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 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.

.~

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 .

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 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 1~789S~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) 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:

zg 3 1 ~, .

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.

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:

24 ~ ~C'~i H ,CH3~ ClHCONH ~
~OCo F ~ ~ N ~-26 CO~u CO~H

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-: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.

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.

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.

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 ~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

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 -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.

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:

20 H H Tr~ S ~ -21~2N~ ~ CH3 ~ ~o~ H C~13 222~ ~-~u oJ~I~ .......

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:

4 , S ~S

6 IlCONEICl-I~ CC~ CH~
N0~
COltf3-1 O~H
a 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.

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~`~

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 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~.

~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) 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:

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) ~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.

I;

l7 'q4;
2l "~
~3 2~

~9 I ~ 54 - ~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 3l c<~

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 ~

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

~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 17 .
r . I .
Ig _ 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 ? ~

;' '`, :~, 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 l4 IS H 1~ Hl ~ CON~
l6 0 ~N-OCH~ ~ N ~
17 COOtBuCOOtBu 18 ` ~

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.~

~ 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) 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 31 ~ .

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 17 }3 ~ COl`Tg~_~
l8 O N ~ ~ ~ N ~
l9 COOtBu OH COOtBu 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) ~,.~ ,....

~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) 3l ~ .

~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 87 ~N~ ~ N ~ SCHaCON
28 o~N~ ~ N ~
COO~Bu COOtBu 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

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

~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

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 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

22 cootgU COO~I

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

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 23 ~xample 37 _ 24 Preparation of (~)-cis-7-cyanomethylthio-acetamido-2-carboxy-1-azabicyclo[4,2,0] oct-26 2-en-8-one 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

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 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 ~ ;-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 ~ -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

~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~.

16 IR(KBr)vmax : 3325, 1770, 1715, 1690, 1655, 1635 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 23 ~ N ~ H CO~ ~

26 ~ COOH

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 27If2N ~ ~ ~ CH -COI~H
23~ N ~ S03I-I
COOH
29 COO~

~ . .

~ -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.

- 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~ :

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 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 ~

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.

~.~78~5!~
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.

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.

~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 ~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 - ~

12 I ~
O~ ~- N ~
G00 ~ U

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.

1) Preparation of 2-[4-(3~butenyl)-3-azido-2-oxoazetidin-22 -~
l-yl]-2-diethylphosphonoacetate-t-butylester.

In this Example, 447 mg (1.78 mmole) o~ t-butyl-~-aminodiethylphosphonocacetate {the com~ound has the fol].owing 2s properties: an oily product; IR(neat)vCax 3400, 1735-1745, 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) 268(M )} is dissolved in 25 ml of anhydrous ether and 164 mg (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 ~

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) 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) 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~}:

29 ~ N

COO~

~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 ,~

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}: ~.

H H
12 N3~,~
l3 o~N~

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) 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 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) 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 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 stirring. The mixture is stirred at room temperature for one hour and 900 mg o~ Molecular Sieve 4A and 700 mg of magnesium su~fate are added. After stirriny for 1.5 hours, the reaction 22 -~
mixture i5 subjected to filtration under reduced pressure.

The resulting filtrate is concentrated to obtain a pale yellow - oily product. Anhydrous benzene (30 nl) is added to the product 2s and the resulting solution is again concentrated to obtain 2.82 g of an oily product. The presence of a Shiff's base in the product is confirmed by N~.R spectrum. The oily product is dissolved in 56 ml of dried cyclohexane and 56 ml of anhydrous 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) 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) 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) 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.

2~

-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 .

2s -~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 ~8 N ~ H~N ~

2tBu COl~u 2~

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.

1~7~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:

6 `t~-8 ~OO
lg ~P'D'.,~,ff.
21 The present compound is produced by the following 22 processes 1) and 2).

1) Preparation of the t-butyl ester of 2-[4--(2-methyl-3-butenyl)-3-azido-2-oxoazetidin-1-yl]-2-diethylphosphono-acetate ~ "^~,CH3 29 O ~ t .2 BU

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.

'"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).

-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 ~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) 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:

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.

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 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:

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) 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 27 1) Preparation of (~)-cis-7~-azido-4rl-ace~oxy-1-azabicyclo 28 [4, 2, 0] oct-2-en-8-on-2-carboxylic acid:

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 l7 21 Preparation of (~)-cis-7~-amino-4~-acetox~-1-lB azabicyclo [4, 2, 0] oct-2-en-8-on-2-carboxylic acid:

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

~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 ~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 16 H~N ~ HlN ~

17 CO,tBU ColH

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 ~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

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. ,~

~ 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 CF3 ~

C~H C02H
16 ~ -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

Claims (13)

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A process for producing cephalosporin analogs represented by the general formula [I]:

[I]

[wherein X1 represents a group selected from the follow-ing three groups:
1) a group represented by the formula:

(wherein B represents a phenyl group, a hydroxy phenyl group or a thienyl group, and A2 represents a hydrogen atom, an amino group, a hydroxyl group, a carboxyl group or a sulfo group);
2) a group represented by the formula:

(wherein Ba represents a phenyl group or a hydroxy phenyl group, and A8 represents a lower alkyl group having 1 to 4 carbon atoms); and 3) a group represented by the formula:

(wherein B1 represents an amino substituted thiazolyl group, a furyl group or a thienyl group, and A11 repre-sents a hydrogen atom or a lower alkyl group having 1 to 6 carbon atoms);
and R2 represents a hydrogen atom or a lower alkyl group having 1 to 4 carbon atoms];
and pharmaceutically acceptable salts thereof, which comprises acylating cephalosporin analogs represented by the general formula [II']:

[II']

(wherein R2 has the same significance as defined above) or a functionally equivalent compound with carboxylic acid represented by the general formula [III]:

X2COOH [III]

(wherein X2 has the same significance as X1 wherein the substituent is protected if necessary) or a reactive derivative thereof and, if necessary, the substituent in the group X2CO- being eliminated in a conventional manner and, if necessary, converting to pharmaceutically acceptable salts thereof in a conventional manner.

2. The process in claim 1, wherein X1 in the general formula [I] is X1a representing a benzyl group or a group represented by the formula:

(wherein B1a represents a 2-aminothiazol-4-yl group, a 2-furyl group or a 2-thienyl group, and A11 has the same significance as defined in claim 1), and X2 in the general formula [III] is X2a having the same significance as X1a (wherein the substituent is protected if necessary).

3. The process in claim 1 or 2, wherein the hydro-gens at the 6- and 7-positions in the formula [I] have "cis" configuration.

4. The process which comprises reacting (2-chloro-acetylamino-4-thiazolyl)-2-syn-methoxyiminoacetic acid chloride with a salt of (?)-cis-7.beta.-amino-4.alpha.-methyl-2-carboxy-1-azabicyclo[4,2,0]oct-2-en-8-on to form the (?)-cis-7.beta.-[2-(2-chloroacetylamino-4-thiazolyl)-2-syn-methoxyiminoacetamido]-4.alpha.-methyl-2-carboxy-1-azabicyclo-[4,2,0]oct-2-en-8-on and after removing the 2-chloro-acetyl group recovering the (?)-cis-7.beta.-[2-(2-amino-4-thiazolyl)-2-syn-methoxyiminoacetamido]-4.alpha.-methyl-1-azabicyclo[4,2,0]oct-2-en-8-on-2-carboxylic acid.

5. The method which comprises reacting (2-chloro-acetylamino-4-thiazolyl)-2-syn-methoxyiminoacetic acid chloride with a salt of (?)-cis-7-amino-2-carboxy-1-azabicyclo[4,2,0]oct-2-en-8-on to form the (?)-cis-7-[2-(2-chloroacetylamino-4-thiazolyl-2-syn-methoxyimino-acetamido]-1-azabicyclo[4,2,0]oct-2-en-8-on-2-carboxylic acid and after removing the 2-chloroacetyl group recovering the (?)-cis-7-[2-(2-amino-4-thiazolyl)-2-syn-methoxyimino-acetamido]-1-azabicyclol4,2,0]oct-2-en-8-on-2-carboxylic acid.

6. The method which comprises reacting (?)-cis-7-amino-1-azabicyclo[4,2,0]oct-2-en-8-on-carboxylic acid with 1N-phenylacetylchloride and recovering the (?)-cis-7-phenylacetamido--azabicyclo[4,2,0]oct-2-en-8-on-2-carboxylic acid.

7. The process which comprises reacting a salt of (?)-7.beta.-amino-4.alpha.-methyl-1-azabicyclo[4,2,0]oct-2-en-8-on-2-carboxylic acid with phenylacetylchloride and after hydro-lyzing the compound obtained recovering the (?)-cis-7.beta.-phenylacetamido-4.alpha.-methyl-1-azabicyclo[4,2,0]oct-2-en-8-on-2-carboxylic acid.

8. Cephalosporin analogs represented by the general formula [I]:
[I]
[wherein X1 represents a group selected from the following three groups:
1) a group represented by the formula:

(wherein B represents a phenyl group, a hydroxyphenyl group or a thienyl group, and A2 represents a hydrogen atom, an amino group, a hydroxyl group, a carboxyl group or a sulfo group);
2) a group represented by the formula:
(wherein Ba represents a phenyl group or a hydroxyphenyl group, and A8 represents a lower alkyl group having 1 to 4 carbon atoms); and 3) a group represented by the formula:
(wherein B1 represents an amino substituted thiazolyl group, a furyl group or a thienyl group, and A11 represents a hydrogen atom or a lower alkyl group having 1 to 6 carbon atoms);
and R2 represents a hydrogen atom or a lower alkyl group having 1 to 4 carbon atoms];
and pharmaceutically acceptable salts thereof, when prepared by the process defined in claim 1.

9. The compounds in claim 8, wherein X1 in the general formula [I] is X1a having the same significance as defined in claim 2, when prepared by the process defined in claim 2.

10. (?)-cis-7.beta.-[2-(2-amino-4-thiazolyl)-2-syn-methoxy-iminoacetamido]-4.alpha.-methyl-1-azabicyclo[4,2,0]oct-2-en-8-on-2-carboxylic acid, when prepared by the process defined in claim 4 or by an obvious chemical equivalent.

11. (?)-cis-7-[2-(2-amino-4-thiazolyl)-2-syn-methoxy-iminoacetamido]-1-azabicyclo[4,2,0]oct-2-en-8-on-2-carboxylic acid, when prepared by the process defined in claim 5 or by an obvious chemical equivalent.

12. (?)-cis-7-phenylacetamido-1-azabicyclo[4,2,0]oct-2-en-8-on-2-carboxylic acid, when prepared by the process defined in claim 6 or by an obvious chemical equivalent.

13. (?)-cis-7.beta.-phenylacetamido-4.alpha.-methyl-1-azabicyclo-[4,2,0]oct-2-en-8-on-2-carboxylic acid, when prepared by the process defined in claim 7 or by an obvious chemical equivalent.