CA1106374A - 0-2-isocephem antibacterial agents - Google Patents

Abstract

USSN 567,323 0-2-Isocephem Ahtibacterial Agents Abstract of the Invention There is described the stereoselective total synthesis of novel .DELTA.2,3 1,4-morpholine-2-carboxylic acids possessing a fused .beta.-lactam ring in the 1, 6-position and carrying a substituent cis to carbon 5 in the 7-position of the fused ring system represented by the general formula I

Description

~ ~'6 3 ~ 4 The presen~ invention prov~des stereoselective total synthesis o eertain novel substi~uted ~ 2'3-1, 4-morpholine-2-carbo~ylic acicls possesslng a fused ~-lactam ring i~ he 196-posit~on and carrying a sub~tituent cis to ca~bon 5 in the 7-position of the ~used ring system represented by ~he general ~ormula X ~ 4 ~ CH Z

" ' ' ' whesein Z is halo, hydro~yl, etherified hyd~oxyl or hydroxyl esteriied with a carboxylic or a sulfonic ~cid residue and X is azido, a~ino o~ acylamino. When X is acyla~ino, these acids ~and thei~ pha~Ddceu~ically acceptable salts and physiologically hydrolyzPd esters) sr~ potent antibacterial agents.
Also included ,~L this invent~on are various novel intermediates~ useful i;~ preparing the active ~-lac~am de~ivatives clescribed above and various processes for ~he produc~ion o~ the intermediates and active compounds~
The compounds ha~ng ~he ab w e general formula sepresent a new family of ~-lac~am antibiotics. They c~ ~e consiclered nuclear analogs of ~ephslosp~rins in ~hich the sulphur atom of the d~hydro~hiaz~e r~ng i~

. f i3'7~L
' , , - ' ' . .

replaced by an oxygen atom ~nd shlfted fr~ posit~on 5 to position 4 of the ~-lactam ring syst~m as numbered the formula above. The nomenclature to be used could ~ ~e the following:

; 2 8 1 " ^~
3 3-oxa-6-aza-bicyclo-~4,2,0]octa~-7-one.
. ~ 6 ~ 4 , . . .. .

However, Sheehan bas used the term O-cephe~ fos ~e structure ~3 ; O-C:epham ':;
[J.G. Sheehan and ~. Dadic, J. ~e~erocyclic ChPm., 770 ~196B)] and we propose the use of the term 0-2-i~ocepham for the basic system having ~he formula .

7~o 2 O--N ~J 3 .

The numerical pref~x indicates the position of the hetero-atom.
.~ " ' ' ' ' . .

2 ~

~o lllustrate the ~b~ve syst~m, the intermediate siE the formula ~ ~ .
~ ' ~~o ', 20~ 3 C~ H2C6H5.

~y ~e named benzyl 7~-amino-3-acetoxy~ethyl-~3-0-2-i80cephem-4-carboxylate and the compound of the ~or~ula 2 ~ H H
C~2C0WH ~ `

: ~ N
- CO~N
~ ~ y be named 7~-(2^ami~o~ethylphenylacetamido)-3-acetoxy-- ;~ ~ethyl- A 3-0-Z-isocephem-4- arbo~ lic acid. ~
There is~hus provided by the present inventio~ the :~ nove~ 0-2-isocephem compounds ha~ing the for~ula ~0 (~ ~ CH2Z-~T ~ C02H
where~n R is an acyl group and Z ls halo, hydroxyl, hydroxyl est.ssi~ied with a car~oxylic acid or a sulfonic 8cid residue or ethe~ied hydro~yl~.and easily cle~s~le esters and pharmaceueic~lly acceptable s~lt~ o s~id acids . .

. .
. ~

7~
~, . .
.
and esters.
T,he acyl group R c~n be chosen from 3 wide v æ iQty of o~ganic acyl rad~cals which yield produc~s of oved ~roper~ies and i~ prefesably a~ acyl radi~
~ich is contained in a ~3turally oc~r~ing or bio-- 8~nthet~cally, semi-syntheticall~ or totally-syntheti- -cally produced pharmacologically active N-acyl desi~ati~e . of 6-aminopen~cilla~ic acid or 7-a~in~cephalosporanic . .~cid. Examples o~ suitable acyl groups are defined:in:
the following gener 1 formulae, but lt ~hould be ~oted th~t this i~ ~ot intended to be an exh ustive list of all the possible acyl groups whicb may be used.

~ i) RaCnH2~CO-~ where Ra is a~yl. (carbocyclic or heterocyclic), su~--.
s~i~uted asyl~ cyclo~lkyI, subs ituted cycloalkyl, i . , .
c~e~oalkenyl, subs~ituted cycloalkenyl or a nonaroma~ic . . , -or mesoionic he~erocyclic group, and n is an integer ~so~ 1-4. The pre~erred Ra substi uents are (a~ aryl selected from phenyl, 2-~h~enyl, 3-thienyl, ~urylg 4-~oxazolyl, p~ idyl.~ ~eerazolyl, sydnone-3 or -4 9 i~idazoly~, naphthoyl, quinoxal~nyl, tria201yl, iso thLazolyl, thiadiazolyl, thiazolyl, oxazolyl, oxadiazalyl, pyrazolyl, furazan9 pyrazinyl, pyrimidinyl, pyridazinyl ~r triazinyl; (b) substit~ted aryl in which the aryl groups m,e~tioned abave under (a) are subs~ituted by .. .
.~ .

_ 4 _ . . '' ,,, . ~ r i3~4 . .
o~ or more radicals s~lected ~rcm chloro9 brom~, iodo, fluo~o9 nitro~ amino, cyano, ~lower)alkanoyloxy~ (l~wPr3-~lkanoyl, (lower)alkoxya~nino, (lower)alkoxy, (lower)-~lky~, ~low~r)aLkylamino~ hydroxy1 guanid~no, ~lower3-al~ylthio, carboxy, phenyl~ halophenyl~ trifluorome~hyl, di(lower)alkylamino, sulfamyl~ ~lower~alkanoyla~ino, phenyl(lower)alkylamido, cycloalkylamino, allylamido ~o~pholinocarbonyl, pyrrolidinocarbonyl, piperidinocar~
bonyl, tetrahydropyridino, fur~uryla~ido or N-alkyl-N-an~lino; (c) C3-C12 cycloalkyl, (d~ substituted C3-Cl~

.
~ cycloalkyl where the substituents are one or ~ore radicals .
selected fro~ chloro; bromo, fluoro, iodo? ni~ro, trifluoro-methyl,~ Cl-C4 alkyl, Cl-C4 alkylamino, Cl-C2 alkoxy or amino; (e~ C3-C12 oycloalkenyl~ said cycloalke~yl group h3ving 1 or 2 dou~le bonds; and ~f) substituted C3-C12 cycloalkenyl, said cycloalkenyl group having l or 2 double bonds and being substituted by one or more ~adicals selecte~ from chloro, bromo, fluoro, iodo, ~itro, trifluoromethyl, Cl-C4 alkyl, Cl-C4 alkylamino, Cl-C2 alkoxy or amino. The most preferred Ra groups ~re phenyl; phenyl substitu~ed by one or ~ore radlcals 8elected from chloro, bromo, iodo, fluoro, nitro~ amino,-(lower)alkyl, guanidino, (lower)alkylthiog cya~o9 (loweo~-~lkoxy, sulfamyl, (lower)alkylamlno, hydroxy7 acetoxy or trifluoromethyl; 2-thienyl; 3-thienyl, tetrazolyl;
8yd~0ne -3; sydnone -4; furyl 9 isothiazolyl; thiadi~Ozolyl ) optionally substituted with phenyl; oxadia~olyl optionally ~u~stituted with phenyl; hiazolyl; imidazolyl; triazoly~;
oxaz~lyl; pyridyl; fura:zan optionally substituted at the

3-p~sition with methoxy3 4-isoxazolyl optionally sub~
8tituted at the 5-posit~on ~ith methyl and at the 3-posi-tion with phenyl or halophenyl; 1,4-cyclohexadienyl;
l.-cyclohexenyl alld l-amirlocyclohexyl.
The mcst preferret acyl gro~ps of this categor~
are those in.wh~ch n is 1. Examples of t~is category include phenylacotyl, halophenylacetyl~ nitrophenylacetyl, aminophenylacetyl, ~(o-amino~ethylphenyl~propionyl, (lower)al~anoylaxyp~enylacety~ (e.g. p-acetoxyphenylacetyl)) (lower)alkoxyphenylacetyl (e.g, methoxyphenylacetyl, ethoxyphenylacetyl), ~lower)alkylph~nylacetyl (e,g, methyl-phenylacetyl or ethylphenylacetyl), hydroxyphenylac~tyl ~e.g. o-hydroxyphenylacetyl), (lower)alkylaminophenyl acetyl ~e.g. o-~ m- or po aminomethylphenylacetyl), o-m- or p- gu~nidinoph nylace~yl, o-carboxyp~enylacetyl, ~,N-bis~(2-chloroethyl)aminoph~nylpropion~l, thien-2 ~d 3-y~Lacetyl, 2- or 3- fur~lacetyl, 1,2,5-thiadiazole-3-acety:L, isothiazolyl-4-acetyl, 4-isoxazolylace~yl~
l-cyclohexenylacetyl9 2-aminomethyl-1-cyclohexenyl-acetyl, l-aminocyclohexylacetyl, 1,4-cyclohexad;enyl-~cetyl, 2-aminomethyl-1,4-cyclohexadienylac~ yl~
pyridylacetyl, tetrazolylacetyi (o~her heterocyclic g~oups of thi~ type are disolo~ed in U.S. 3~819,~23 - G - ~ .

_ .. ... .

~d 3,5l6,997~ or a syd~loneace yl ~roup as disclosed .
~n U.S. 3,6819328, 39530,l23 and 3~563,g83~ Other ~roups o thi~ type in~lude 3-phenyl-S-chlorophenyl-S~ethylisoxa~ol-4-ylacetyl ant 3-(~,6-dichlorophenyl~-5-methylisoxazol-4-ylace~yl or a group in which isox-~zolyl is replaced by isothiazole as disclosed in U.S~
3jS519440. 5till other example~ are o-, m- and p-,~2'-aminoethoxy)phenylacetyl ~as disc~osed in U.S.
3,759,905), 4,5-dImethoxycarbonyl-~,2~3-~riazol-l-ylacetyl or 4-cyano-l,2~3-triazol-l-yl-acetyl Sas disclosed in U.S~ 3,821,206~ and imidazo~ acety~
~as disctosed in U.S. 3,632,810;

(ii) ~2n~1C~ ~ , , where n is an integer from 1-7~ The alkyl group may be stralght or branched and, i desired, may be inter-r~pted by an oxygen or sulphur atom or substituted by, e~g~, a cyano group. E~a~ples o. th;s group include cyanoacetyl, valeryl, hexanoyl, heptanoyl, ethoxy carbonyl, octanoyl and butylthioacetyl. ~ pr~ferred acyl group i~ cyanoacetyI;
(~ i) CnH2n_lCO- .
where n ~.s an integer from 2-7. The alkenyl group m~y be str-ight or branched and, if desired, may be ' . ' , .

.. . .

r 3r74 .

~terrupted by an o~yge~L or sulphur atomO An e~am~le of this group is allylthioacetyl;
~v3 ~b ~-O-~-CO- ' ~here Ra is as defîned ~nder ~i) and i~ addition ~ay ~e benzyl, ~l-C~ alkyl or ~lower~al~oxycarbonyl and $~ and Rc which may be the same or dif~erent each .
represe~t hydrogen, phenyL, benz~l, phenethyl or Cl C6 alkyl. ~he praerred Ra substituents in this category are benzyl, Cl-C6 alkyl3 (low~r)alkoxyc~rbonyl and those mentioned under ~i) as being preferred aryl, su~s~i~uted aryl, cycloalkyl. (and su~stituted cycioalkyl~ and cycloal~enyl tand subs~ituted cycloalkenyl~ groups.
T~e most preferred Ra gxnup is phenyl. Examples ~f . this group include phenoxyacetyl, 2-phenoxy-2-phenyl-:acetyl, 2-phenoxypropionyl~ 2-phenoxybutyryl, benzyloxy-ace~yl~ 2-methyl-2-phenox~propionyl, p cresoxyacetyl, ~ - p-methylthiophenoxyacetyl and ethoxycar~onylacetyl;
`. ' (~ Rb ''`` RaS C CO
RC
where ~ s as defined under ~i~ and in addi~ion m~y be benzyl or Cl-C~ alkyl and Rb and R~ ha~e the meanings defined under (i~). The preferred R8 subs~i~uen~s ~n .. . .

, .

.. ~

~ . .

r 1~6;~74 .

t~is category arP benzyl, Cl-C6 alkyl and those m~ntioned ~sder (i) as belng prefE~Yed aryl, substituted aryl, cycloa-lkyl ~and substituted ycloalkyl) and cyclo-alkenyl (and substituted cycloalkenyl) groups~ The ~st pre~erred ary~ groups of this type. ar~ those ~n wh~ch Rb and Rc are.hydrogen and Ra is phenyl; phenyl ~ubstituted w~th one or more radicals selected fro~
chloro, bromo, iodo, fl~oro, nitro, amino~ (lower)-Y1 D (lower)alkythio, cy2no~ (l~wer)~lkoxy, (low~r~-alkylamino, hydroxy, acetox~ or ~rifluoromethyl;
3-pyridyl; or 4-pyridyl;

(~i) RaX (C~I2)mCO-where R~ is as defined u~der~(i) a~d in addition may be benzyl, X i5 oxygen or su~phur and m is an in~eger from 2-5, The pre~erred Ra groups are benzyl and ~hose ~entioned under ti~ as being preferred aryl~ subs~itu~ed ~ I, cycloalkyl ~and subs~ituted cycloal~yl~ and cyeloal~e~yl (and substituted cyoloalkenyl) groups.
A~ exan~lle of ~his group is S-ben~ylthiopropionyl.
~ v~'i3 RaCO-whe~e R'l is as defined under (i~. The preferred Ra grou~s are those mentioned under ~i) as beang preferred aryl, substituted arylg cycloalkyl (and su~stituted cycloalkyl) and cycloalke~yl (and substituted cyclo-alkenyl~ groups. The most pr~ferred aryl groups o _ 9 _ this categ~ory are those in which Ra is phenyl; phenyl ~ubstituted with one or mure rad~ als selected from chloro, bromo, iodo~ fluoro, nit~o, amino, ~lo~e~)-a~kyl, (lower)alkylthio, cyano, (lower)alkoxy, ~lower)-~lkylamino, di(7.ower)~1~yla~ino, hydroxy, acetoxy or tri1uoromethyî, and most preferably phenyl su~tituted . , .
8t the 2-position by c~rboxy or phenyl or at the 2- and 6-positions by methoxy; 2-ethoxynaphthoyl; 3-phenyl~
5-methylisoxazol-4-yl; 3-o-chlorophe~yl-5-methylisoxazol-

4-yl; 3-(2,6-dichlorophPnyl)-5 methylisoxazol-4-yl and l-aminocyclohexyl. Examples of this group inclu e 2,6-dimethoxyben~oyl~ benzoyl, 2-biphenyicar~on~}~ 2-amin~
.: . ~
~ methylbenzoyl, 2-c~rboxybenzoyl-2-phenylbenzoyl, 2-.~ .
thie~ylcarbonyl~ 3-thienylcarbonyl and 2-chlorobenzoy~;

~viii:) : Ra IH-CO- or R~-IH-CH~CO-y where Ra ~s as deined unde~ ~i) a~d Y is hydrazino, guanidino, ureido, thioureido and subs~ituted thiour~ido ~as disclosed i~ U.SD 3,741,962), ~llophanamido (as described in U.S. 3~4837188~, 3-guanyl-1-ureido( as in U.S. 3,796,709), 3~ fur~yl)ureido, cyanamino (~s in U.S. 3,796,709), 3-~benzoyl)ureido, azido, amino, acylamino (e.g. carbobenzoxyamino), a group o~tained by reacting ~he amino group of the 7-side , - - I

~ ~ 6 3 7~

' . ' .

c~a~n with an aldehyde or ketone (e.g. acetone, form-~ldebyde, acetaldehyde, butyraldehyde, acetylacetone, - ~ethyl acetoacetate, benzaldehyde, salicylaldehyde, ~e~-hyl ethyl ke one or ~thyl acetoacetate~, hydroxy, etherified hydroxy, est~sified hydroxy~ carboxy, esterified car~oxy (as disclosed for example in U.S.
3~282,926, 3,819,601 and 3,635~961 and including especially -C- O ~ ), trLazolyl, . .. .
tetrazolyl~ cyano9 halogeno, acyloxy (e.g. formyloxy cr ~lower)alkanoyloxy~,~rsulfo, sulfoamino or esteri~ied sul o. The preferred Ra subseituents ~re those mentioned : under ~i) as being preferred aryl, ~ubstitu~ed aryll .: .
cycloalkyl (and subsei~uted c~cloalkyl) and cycloalkenyl ; ~and subs~ituted cycloalkenyl) groups. Prefersed Y

~ubstituenes are hydrazino; guanidino; ureido; substi-tuted thiour~ido o~ the formula .P
~Rq in wh~ch RP is hydrogen or Cl-C8 alkyl and Rq is hydrogen7 ~ Cl-C8 alkyl, C2-C8 alk~nyl~ phenyl~ benzoyl, C~-C8 : ~lkoxy- Cl~C8 alkyl, ~car~o Cl-C8 alkoxy) Cl-C~ alkyl;

.

1 1 -- !

6~3r~
.

' allophanamido; 3-guanyl-1-ureido; 3-(2-furoyl)ureido;
3-~benzoyl)ureido; azido,; am~no; a group obtained by react~ng the amino group Y with acetone, formaldehyde9 ~ce~aldehyde, butyraldehyde, acetylace~one, methyl ~cetoaceta~e, benzaldehyde, salicy~aldehyde, methyl ethyl kPtone or ethyl acetoacetate; hydroxy; etherified hydroxy including especially tlower)alkoxyJ carboxy;
esterified carboxy including espec~ally 5-indanyloxy-carbonyl; triazolyl; tetrazolyl; cyano; cyanamino;
halogeno; formyloxy; (l~wer)alkanoyloxy; sulfo; or 8ulfoamino. Examples of this group include a-amino-phenylacetyl; ~-carboxyphenylacetyl; 2,2-dimethyl-5-oxo-, 4-phenyl-1-imidazolyl; a-amino-p-hydroxyphe~ylacetyl;
hydro~yphenylacetyl; ~-~ormyloxyphenylacetyl and other yl roups of this type dlsclosed in U.S. 3~81~,116 and 3,821,017; a-amino-a-?- or 3-thienylacetyl; a-amino-~-(3~chloro-4-hydroxy)phenylace~yl; a-amino~ 4-cyclc-. .
h~xadienyl)acetyl; a-azidophenylacetyl; a-amino-a~
cyclohexenyl)ace~yl; 2-car~oxy-a-3-thienylace~yl; a-amino- -~-~3,5-~dichloro-4-hydroxyphenyl)ace~yl, ~-amino-a-3- or 4-or ~-isothiazolylacetyl (as in U.S~ 3,579,506) and othsr ~-am~no and G-hydroxy-he~erocyclylacetyl groups as ~-sclosed for example in U.S. 39821~207, -- .

.

gix~ R~
. . . , ~ . .

Rd~ ~ -CO-.
. where Rd~ Re and R~ which may be the same or diferent saay each rep~esent Cl-C6 .alkyl, phenyl or substituted phenyl., The preen:ed phleny~ substituents are one.or more radicals selected f;~om chloro, bromo, iodo, fluoro~
tr~fluoromethyl~ nitro, aminQ, cyanot ~lower)aïkanoyloxy, ~lower) alkanoyl, (l~e~) alkoxyamirlo, ~lower~ a lkoxy, (lower) a~ kyl~ (lower) alt~12mino3 ~ydro~r, (lower) alkyl-thio, carboxy; di~lower)alkylamino or sul~amyl. An @xample of this group is triphenylmethylcarbonylO
,., ~x) Ra_~oc_ :. . , 11 , .' ' X

where Ra is as defined un'der (i~ and in additio~ may be hydrogen, Cl-c~ alkyl, halogen su~stituted Cl-C6 alkylJ

phenethyl, phenoxymethyl; benzyl or R~ and X is oxygen ; . ~ O
o~ sulphur. An example o~ such a group is C:l(CH2)2NHCO;

(xi) /~ ~2\
~' ' - (C~

.
hese Y is as de~ined lmder (viii) and n is an integer of ~-4. A m~lst preferred Y subs ituent is aminQ., . An ~ . . .
example of this g~oup i~ aminocyclohexanecar~onyl.
: . .

.

:: o .l3 `

37g~

.

(xll) ~minoacyl, for example Rgc~NH2) - (CH2~nCO-where n is an integer oi 1-10, or ~ 2~~~ nH2nAF (CH 2)mCO-~ere m is zexu or a~ integer fro~ 1-109 and n is 0, l, or ~; Rg is hydrogen or an a~kyl~ asy~, aral~yl o~ carboxy group o~ a group as de~ined under Ra in ~i) abave, and Ar is an arylene group, e.g. p~phenylen~
or 1,4-naphthylene6 Prei-erred aryl groups oi. the above formulae are ~hose in which Rg is hydroge~, ~lower)-alkyl, phenyl, benzyl or carboxy ~nd Ar is p-phenylene or. 1~4-nap~thylene. Exa~ples of such groups are disclosed in U~K~ 1,054,806. Exampl~s o groups of this ~ype include p-aminophenylacety~ and J-aminoadipoyl der~ved from naturally occurring amin~ acids and deri~-~t~res thereof, e.g. ~-benzoyl ~ inoadipoyl;
~ xiii) Substituted g~yoxylyl groups of the formula Rh-Ca-co_ where Rh i8 an aliphatic~ araliphatic or aromatic group.
The preiesred Rh groups are ~-thie~y~; 3-~hienyl;
~-naphthyl; 2-phenanthryl or a mono-, di- or tri substituted phenyl group, the s~sti~uents being ~elected from oh~oro, bromo, iodo, fluoro, amin~
d~ wer)alkylamino, (lo~er)alkyl, ~lower)alkoxy, ~itro o-r ~lower)alkanoylamino~ -Examples o~ this categor~
are disclosed in U.S. 3,546,219 and 3~573,294. Included ;

.
, ~ ~

, ~ 6 37 4 .

i~ th;s group are also the a-carbonyl derivatives of the above subs~ituted glyoxylyl groups formed for e~ample with hydroxylamQne, semicar~azideg thio-~emicarba2ide, isoniazide or hydrazine;
(XiY~ Ra~lH_CO_ 3~

'' , X='~

El-N-C-Ri Il, X

where Ra has the meaning defined under (i), X is oxygen or sulphur, X is ~xygen or imino ~nd Ri repres2n~s ~lower)al~yl, cycloalkyl having 4,5, 6 or 7 carbon - atoms, monohalo(lower)alkyl, dichloromethyl, tri-chloromethyl, (lower)alkenyl of 2-5 carbon atoms, RJ ~o ~k ~ (CR2)n- R ~ C~

.

C~3 ~

R~ . R3 ' ~ . H3C~V~r~k . ~ H3 : ~ .
-- 1 5 -- . _ ~ . ...... I
. .

! ,r ~3C

E~ CH3 ;; - . . .
- ~ ~8 an ~ntegex from O ~o 3 inclus~ve and each o~ Rk - ~nd R3 is hydroge~, nitro, di(l~wer~alkylamino, ~lower) ~lkanoylamino, (lower)alkanoyloxy, Cl-C~ alky~, C~-C6 al~oxy, sulfamyl, chloro, bromo, iodo5 ~luoro or tri-,~luoromethyl. The preerred Ra substituents are those m~tioned under (i) a3 being preferred aryl, substituted : aryl, cycloalkyI ~and substi~uted cycloalkyl) and , cycloal~enyl ~and substituted cycloalkenyl) groups.
Preferred acy? groups o~ this type are those in wh~ch ~a is 2-thienyl, 3-thie~yl; phenyl; or phen~l sub-~tituted by one or more radicals se~ec~ed from nitr~
t~lowe~alkylamano, {~ower)alkanoy~amino, amino, ydro~, (lower)alkanoyloxy, Cl-C~ alkyl, Cl~C6 al~oxy, sulfamyl, chlo~o~ bromo, iodo, fluoro or ~rifluaro-~ethyl; X is oxygen; X' is oxygen or imino and Ri is : ~lower)alkyl9 phenyl, 2-thienyl, 3 thienyl, 2-furyl o~ 5-nitro-2-furyl. The most preferred groups are th~se of the above formul~ where ~a is pheny~, p-hydrox~-phenyI, 2-thienyl or 3-thienyl; X is oxygen; X' is oxygen, and Ri is phenyl or 2-furyl. Examples are d~sclosed in ~S. 3,6~7,94g and 3,646 7OZ~

;' ' , .
. `- ' .

,.. ..

-r 3~il .

a~) . ' Ct~_ , 11 ~ . .

.
C~2~

p,i~W or Ri-C-NH-CH~CO-where Ra has the meaning.defined ~n (i) and Ri has the ~eaning defined in (xiv). Th~ preerred Ra subs~ituents are those ment~oned under (i) as being pre~er~ed aryl, substituted aryl, cycloalkyl (and ~ub~tîtuted cycloalkyl) and cyclo3 lkenyl ~a~ substituted cycloalkenyl) ~roups~ Preferxed ~i ~b8tituents include (-ower)alkyl, dichIorome~hyl, C4-Cj cy~loalkyl, 2-thienyl, 3-thieny1, p~enyla ~e~2yl~ haloben-yl~

C ~ c~3 C ~

- " . , ' , ~6n5 ~ ~d ~ C6~5 E~amples of this gruup ar~ disclosed in U.SO 3~626,024 a~d 3,692~779;

R -C~-CQ-o whese Ra has the meaning defined in (i) and Rl ls ~lower)-alkyl, C3-C12 cycloalkyl, aryl (especially phenyl~, a monocyclic heterocyclic radical having 5 or 6 atoms çscclusive of hydrogen which are C, S, ~ or 0, no more than 2 atoms being other than G, or a substituted mono-cyclic heterocyclic radical as defined above ~av~ng sne or more substituents selec~ed from halo, (lowerj-alky?~ ~lower)alkoxy or phenylO Examples of ~his grcup are disclosed ~n U.S. 3,778,436~ Most p~e~erred groups a~e (lower3a~kyl, phenylg th~enyl or furyI~
A preferred class o~ acyl groups are those of the formula O
Ar '~c~-a-.., wherein Ar' is a radieal of the formula Rm Rm R ~ ~ or R
in which Rml Rn and R are alike or dif~esent and each is hydrogen, hydroxy, (low~r3alky}, cyano, (lDwer)alkoxy, ~hloro, bromo, iodo, 1uoro, tri~luoromethyl, nitroa amlno, (lower~alkylamino, di(lower)alkylamino, ~lo~er)-~lk~noyl, ~lower~alhalloyloxy su~h as p-acetoxy or phenyl ~nd Y is amino or a group obtained by reacting the amino group with acetaldehyde, form~ldehyde or acetoner fluoro, chloro, bromo, iodo, hydroxy, (lower)alkano~loxy, carboxy, guanidino, 3-guanyl-1-ureido, 3-(2-fuxoyl)ureido, 3-benzoyl-ureido, sulfo, sulfoamino, ureido9 thioureido; ~lower)-a~koxy~ cyano~ cyanamino or indanylox~carbonyl. Parti-~ularly prefe~red Ar radicals are phenyl9 p-hydro*yphenyl, 4 hydroxy-3 9 5-dichlorophenyl, 3-chloro-4-hydroxyphenyl~
o-, m- or p- aminethylphenyl, 2-thienyl, 3-thienyl, .

l-cy&lohexenyl and 1,4-cyclohexadienyl. Particularly preferred Y gro~ps are amino, hydroxy and carboxy.
Set ~orth belo~ are formulae of the most preferred acyl - groups of this class:

, ~ Cl -CO- ; ~o ~ CH-CO-: - 19 ...

~L ~LIJ 6;3 ~

1~1~ C~ H-CO i;~ ~CH-CO- ;

-~C~-CO-~S~ X~2 ; ~ ;
,~:H-CO-, ' ~.
~3_CU-CD~ CH-CO- ;

e3CI~-GQ- ;
I~H . .
.. .
.--~ C~
I~S~ 1H , (C3_C~-CO-OH

C~ -CO- ~-CO- ;

, <~ IH_CO~ ; ~COOH
COOH S
.

; 20 -- ~

``
;3'7~

~, .

CH-Co- ; ~CM-CO-COOEI , COO~

CO and ' ~Cil-CO-~0 Of most interes~ are the acyl groups of éhe above c1ass :~ wbere the acid ArCH~X)COOH is of the D-series.
Other particularly preferred acyl groups for the co~pound~ of for~nula I are ,,P ~O~I3 C-CH2CO- o ~ co - . 0~3 N ~ ~

5 - C ~SCH~CO-;

.~ U
~~ cCO-; . ' where lU and V are alike or different and each i.s hydrogen, chloro or f1uoro;
,' .

-- 21 ~
..

r 1~ ;374 . ' , ' , . .
; ~CH2CO- ~3CH2CO~

- ~H2CO- ~--CHzCO-f-CH2C~- ~ ~CH-CO-O _ C ~ C=
- ' C6H5 OCU2CO~ SC~2CO-~2NH2 ~82C- . ~l-C7 al~;yl-CO_ ~CO- ' ~CH2CO-I~Lc~2co ~CH2CO-r . ~ '631~4 ~ '' , . .
.- .

~1 ~ 7 alkox~-Co~
~f ` N.H2 .
.~ . .
~C%2C-~-C~2CO-, ~CHzN~CO-~) OH
O--C--I--H ~H2C-' ~CO~ CO-.
2~2 e~SCH2CO-~_<CH2NH2 C ~2~H2 <~CH2CO_ and 6~:H2CO

"
,~ " .

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

,r ' ll~l~i379L

.
Substltuent Z :Ln ~ormulae I and II above m~y be h~lo (chloro, bromo, fluoro or iodo) " hydroxyl, hydroxyl esteri~ed with a car~oxyli~ acid or a 8ulfonic acid residue or etheriied hydroxyl. Estes-~fied hydroxyl groups include radicals of the formula wherein Rl is an acyl group, a ~lower)alkylsulfonyl group, an arylsulfonyl group or an aralkylsulfonyl group and are pre~erably those of the formula -OCOR2 or ~here~n R2 is hydrogen, amino, ~lower)alkyl te.g. methyl, ethyl, propyl, isopropyl9 butyl, isobu~yl, ~-butyl, 2myl9 hexyl) 9 C3-C7 cycloalkyl,(e.g. cyclobutyl, cyclopentyl, cyclohexy1, cyclopropyl or cycloheptyl), C3~C7 cy~ioalkyl-(l~wer)alkyl (e.g. cyclobu~ylmethyl, cyclobutylethyl, O
cyclopentylmethyl, cyelohexylmethyl, cyclopentylpropyl, etc.j, aryl ~e.g. phenyl or nap~thyl), aralkyl (e.g.
be~zyl, tetsazolylacetyl, 2~ naphthyl)ethyl or phenethyl), or 3ryloxyalkylene, e~g., phenoxymethyl, and R2 is ~lower)-~lkyl, aryl or aralkyl. The R2 and R2 groups above may be Qptionally substituted as by one or more (lower)alkoxy, ~lo~er)alkylthio, halogen, tlower)alkyl, nitro, hydroxy, ~cyloxy~ car~oxy~ amino, (lower)alkylamino or acylamino radicals. The most preferred R2 groups are hydrogen, tlower)alkyl ~especially me~hyl) and amino, The mos~

- ~4 -, r 7~iL

.
preferred R2 groups a~e (lower)alkyl (especially methyl),t~fluoromethyl and p-tolyl.
Etherified hydroxyl groups include radicals o~ the formula w~ere R3 is (l~wer)al~yl, ~lower)çycloalkyl, (lower)cyclo-~lkylo(lower)alkyl, aryl, arslkyl or a heterocyclic group, ~ny ~f said R3 groups bring optionally substituted by one or more ~lower)alkoxy, (lower~alkoxy(l~wPr)alkyl, (lower3-alkylthio, halogen~ (lower)alkyl~ (lower~ yc loa lkyl, (lower)alkenyl, nitro~ hydroxy, acyloxy, car~oxy, amino, di(lower)aLkylami~o, (lower)alkylamino, ~rifluorome~hyl~
~ryl, aralkyl or acylamino radicals.

Preferred R3 groups ~nclude benzyl and 5- or 6-membered heterocyclic radicals con~aining 1-~ atom~
~elected from N, O and S, said heterocyclic radicals being optionally substituted by one or more substituents .
selected from halogen, amino9 nitro, Cl-C4 alkyl"
: C3-C4 cycloalkyl, Cl-C4 alkoxy, C2-C4 ~lkenyl, tri1uoromethyl phenyl, benzyl, Cl-C4 alkylthio, Cl-C4 alkylamino, di Cl-C4 alkylamino or alkoxyalkyl of u~ to 4 car~on atoms.
Examples of preferred heterocyclic R3 groups include furyl, thienyl, pyrazolyl, imidazolyl, isoimidazolyl, ~riazolyl, tetrazolyl, thlazolyl, thiadiazolyl, thia~ria-zolyl, oxazolyl ~ oxadiazolyl, isothiazolyl, isoxazolyl, pyridyl, pyridazinyl, pyrazinyl" pyr~midinyl and triaz~nyl, s~d radicals being optionally substitu~ed by one or two of the above-men~ioned substitue~ts, ~st prefer-ably Cl-C4 alkyl9 Cl-C4 a:Lkoxy or trifluoromethyl groups. Particularly preXerred heterocyclic R3 group~
~nclude 1,2,3-~siazolyl, 1,2,4-triazolyl, 1,2,4-thi~d1azolyl, lj3,4~thiadiiazolyl, 1~3,4-oxadiazolyl, 1,2,3,4-te~razolyl, 2-methyl-1,3~4-thiadiaz~l~S-yl, 2-methyl-1,3,4-~xadiazol-5-yl a~d l-N-met~yl~etr3zolyl.
Pre~erred Z substituents in formula I and II are ~hose of the ~ormulae -halo, i~e. chloro~ bromo, .iodo~ fluoro; -OH; -OCHO; -OCOCH3; -OSO2C~3; -OSQ~CF3;
-OS02C6H4CH3 (para); -OCH2C6H5; and -OCONH~

When Z is hydroxyl, compounds o~ formulae I and II may also exist as the ~actones which are formed by internal esterification with the carboxyl groups.
The term "(lower~alkyl~' as used herein means both s~raight and branched chain aliphatic hydrocarbon radicals having from one to ten carbon atoms such as methyl, ethyl, propyl, isopropyl9 butyl, isobutyl, ~-bu~yl, amy~, hexyl, 2-e~hylhexyl" heptyl, decyl, etc., S~ailarly, where the ~erm "(lower) " is used as part o~ the description of another group" e.g. "(lo~er)-~lkoxy", it refers to the alkyl portion of such group w~ich is therPose descrlbed above in connection with "(lawer) a~ky~ " .

-- 26 -- , 637~4 .

: ' ' . ', ` ' .
The pharmaceutically acceptable salts re~er~ed to ~bove inc~ude the nontoxic carboxyl~c acid salts, e~g. nontoxic metallic salts such as sodium, potassiu~, c~um ~nd ~lUm~um9 the ammon~um salt and s~lts wl ~ontox~c amines9 e.g. tri,alkylam~nes, procaine, dibenzylamine, N-be~yl-3-phenethyla~ine~ l-ephenamine, N,N'-dibenzylethyl~nediamine~ N-alkylp~peridine and other amines which have been used to or~ salts o~
pen~c~llins and ceph~losporins. When a basic group .is prese~t, as w~2n it occurs in the 7-acyl group9 the present invention also includes the pharmaceutically acceptable acid ~ddi~ion salts, e.g. salts with mine~al ~cids such as hydrochlori~ hydrobromic~ hydroiodic, pho~phoric, sulfuric and salts with.organic aci~s such .
as maleic, acetic, citric, oxalic, succinic, benzoic, tar~aric 9 fumaric, mandelic, ascorbic and malic. The term "pharmac~utically acceptable salts" is also m~ant to include nontoxic ac;d addition sal~s of the easily cleavable esters refe~red to above. ~e ~ompounds which contain a basic group in radical R may al.so be present i~ ~he form of an in ernal ~altj i.e. in the form of She zT7itterion.

. , ' . , , , ' .
, - 2 7 - .
, r r ;3~74 .
~ . ' ' Th~ easily cleavable esters referred tc~ above include ester groups which are removable by methodsD
e.g. chemical o~ enzymatic hydrolysis3 treatmellt- ~Ith chemical reduci~ag agents ur~er mild conditions, isradi~tion with ultravioet light or catalytic:
hydrogenat~on, which do not result in any appreciable destruction o:E the remainin~ portion of the molecule~, Examples o~ suita~e esters ~nclude thos~ disclosed ~n U.S. Pat rlts 3,284~451 and 3,249~622 and U.K.
Pa'cents 1,.229,453 and 1,073,5300 Esters which hatJe been used pr~viously ~n penicilli~ and cephalosporin che~stry include for example benzhydryl, p-n~trobenzyl, betlzyl, trichloroethylg silyl such as trimethylsilyl, phenacyl, p-methoxybenzy~ acetnyi, p~thalidyl, indanyl ~nd ~lower)alkyl such as methyl, ethyl ar~d t-butyl.
Part~cularly preferred easily clea~rable esters ar~
those w~ich are hydrolyzed u~der physiological condi-t ons such as pivaloyloxymethyl, ace~oxyme~hyl; phtha~idyl, ln~an5rl and metho~yrne~yl., ~s ~he 0-2-isocephem campolsnds of t~e present ~vention may possess one or m~re asy=e~cric carbon , r ~ 63 1;~4 at~ms a the invention includes all of the po~sible e~ant~omesic and diastereom.ric forms of the compounds o~ the general formula II sh~wn above. Resulting ~ixtures of lsomers can be separated into ~he ~ndividual isomers ~ccording to methods which are knswn ~ se, e.g. fractiona~ crystallization, adsorp~
~ion chrom3tography or other ~uitable separation processes. Resulting racemates can be .separated into ; ~he an~ipodes in the usual manner a~ter introductionof suitable sal~-formlng grouplngs, eOg. by fbrmi~g mixture of diastereoisomeric salts with optically active salt-forming agents, sepasating the mixture ; lnto diastereoisomeric salts1 and converting the ~eparated salts into the ~ree rompounds 9 or by frac-tional crystallization from optically active solvents.
I~ will be appreciated that cPrtain of . he compounds of this ~nvention exist in various states of solvation and the anhydrous as well as solvated forms are within the scope of the invention.
. The ~ee acid compounds of the above general formula ~I whese R is acyl and physiologically hydrolyzed esters thereof tog~ther with the pharm~ceutically acceptable s~lts of such free acids and esters are useful as anti-bacterial agents. Certain of the 7-acylated comPounds of formula IIg e.g. t~ose in which Z is halo, ~ydroxyl, -OCHO, -OCH2C6H5 or sulfonyloxy (especially -OS02CH3, -OSO~CF3 or : -- ~9 ._ - . ~

1 3t~ 374L

~OC6~4CH3) are use~ul not 8nly as active antibacterial ~gents per se but as intermediates in preparation of other active 7-acylated deri.vatives within the scope o formula II~ The remaining compounds of the above general fosmula $I in~luding salts and esters thereo~
~ ~re valuable inte~mediates which can be converted lnto the above-mentioned pharmacologically active compounds a simple manner for example, as described belvw.
P~eferred embodiments of the p~esent in~ention are the c~mpounds of the formula H
R-~H ~ O

~ ~CH2R4 ' ' C0 IIa wherein R is an acyl group and R4 is halo, hydroxyl, -OC~O, -OCON~2, -OCOCH3, -O~H2C6H5 or -0S02CH3, and easily cleavable ~sters and pharmaceu~ically acceptable salts ~hereof.
Pre~erred compounds of f~rmula IIa are those in which : R is an acyl group selected from ~he acyl groups de~ined 8bove under (i) to ~xvi), Use of the acyl groups mentioned above as being preferred within categories ~i) to ~xv~) re~ults in active end-prcducts having the most advantageous pharmacological properties. The most preferred compounds of Formula IIa are those having the same preferred R-sidechain as listed abo~e in connection with Formula II.

r ~ 3'74 The present invention further provides various ; . novel intermediates useful in the synthesls of the 7~acylamido 0-2-isocephem compounds of ~ormula II
d~sc~ibed above.
Preferred embodiments of the present ~nventio~
are the ~ovel intermediatles having the formula H ~H
~3 ~ ~ O

,~ ~ ~ C~2Z

' ~V ," . C02R"

: wherein Z is halo, hydroxyl, hydroxyl esterified with carboxylic ac-id or a sulfonic acid residue or etheri-fied hydroxyl and R" is hydrogen or an easily cleavable ~ster carboxyl~protecting group, and salts thereof. The : preferred Z substituents are as defined above in connec-tion with the compounds of formula II.
The ~ost preferred intermediates of formula IV are those of the form~la H
~3~ ~0 C~2R4 C~3,2R
I~a .'~ , .

: ' . 31 -.

..r 3~L

~herein R4 is h~lo, hydro~yl, -OCHO, ~OCH2C6H53 -OCO~H2, -OCOC~3 or -OSO2C~30 l~termediate~ of foT~ula IVa in whlch R4 ls ~ -OS~2C~3 or -OCHO are of particular importance 8S

: ~tarting m~terials ~n preparing other intermedi~tes within ~he scope of form~lla IV~
Other preferred intermediates are the compounds ~:~ having the formula H
.. ~

: ~ ~2Z
CO R'~
~II 2 wherein Z is halo, hydro~yl, hydroxyl esterified with a carboxylir acid or a sulfonic acid residue or e~her~ d . . hydroxyl and R" is hydrogen or an easily cleavable ester carboxyl-protecting group, and salts thereof. The preerred Z substituents are those mentioned above as ~being preferred in connection with the compounds of fo:rmula II~
The most preferred intermediates of formu1a III
~e tho:;e o the formula ' }I ~1 O ~"
~Ia wherein R4 ls halop hydrcxyl, -OGHO, -OCH;,CfiH~ OCOC:~.i3"
., , . - 32 -i3 ~4 -OCONH2 or -OSO2CH3 and R" is hydrogen or an easily cleavable ester carboxyl~protecting group.
Preferred intermediates of formula IIIa are those in which R4 is -OSO2CH3 or -OCOR2, wherein R2 is hydrogen, amino or (lower~alkyl.
Most preferred compounds of formula IIIa are those in which R4 is ~OCONH2 or -OCHO and R" is hydrogen, and those in which R4 is -OCOCH3 and R" is hydrogen or one of the preferred carboxyl-protecting groups mentioned below.
The intermediates of formula III and IV may be in the form of the free carboxylic acid or a salt thereof or in the form where the carboxyl group is protected in a conventional mann~r such as preferably by esterification. The prote~ting group is selected so that it may be removed by methods which do not result in any appreciable destruction of the remaining portion of the molecule. Preferred carboxyl protecting groups are the easily cleavable esters as defined above including in particular benzhydryl, p-nitrobenzyl, trichloroethyl, silyl including especially trimethylsilyl, phenacyl, p-methoxybenzyl, acetonyl, (lower)alkyl such as methyl, t-butyl or ethyl, benzyl, triphenylmethyl, methoxymethyl, acetoxymethyl, phthalidyl, indanyl and pivaloyloxymethyl. Most preferred carboxyl protect-ing groups include pivaloyloxymethyl, methoxymethyl, indanyl, phthalidyl and acetoxymethyl.
The novel 7-acylamido compounds of formula II may be prepared by N-acylating a 7-amino intermediate of the formula H H
H N
1~
~ ~ N ~ CH2Z
III
C2R"

, ,~L .

wherein Z i9 hydroxyl~ hyclroxyl esterif~ed with a c~rboxylic acid or a sulfonic acid residue or ethesi-fied hydroxyl and Rl' is hyd~ogen or an easily cleavable ~ster carboxyl-~rote~ting group~ or a salt thereof3 with .an acylating acid of the formula , ~-COOH

wherein R is an acyl group, or wit~ its ~u~ctional equi-valent as an ac~lating agent for a primary amine and, if desired, converting the so-produced product to the corres-ponding 7-acylated product having a s~bstituent Z
d~fferent ~rom that in starting material III and9 if desired, ta) when R" is a carboxyl-pro~ecting group, converting the 7-acylate~ ester to the free acid compound or a physiologically hydrolyzed ester or a pharmaceut-ically aceeptable salt of said acid or ester, or (b) when R" is hydrogen, converting the 7-acylated carboxylic acid to a physiolo$i~cally hydrolyzed ester or a phar~aceutically 8cceptable salt of said acid or es~er and, if desired, resolving a resulting isomer mixture into its component ~somers, The 7~amino starting m~erials of general ormula III are of use primarily as intermediates in preparing thè pharmacoligically active N-acyl der~vatives of formula II. The free acids, ~hysiologically hydrolyzed esters and pharmaceutically acceptable salts of sald .
sclds ~nd esters of foxmula III, however, do possess ~ome antib~cterial activlty per se against various pathogenic microorganis~s.
. The 70acylamido 0~2-~socephem ~ompounds of formula II are prepared by N-acylation according to k~wn methods of the 7-amlno group of inte~nediat2 III with 8~ acylating 3cid of the formula R-COOH
wherein R is an acyl group, or wi~h its functional e~ui-valent as an acylating agent for a prim~ry amino group.
The.acylating agen~s for preparing the products of formula II are known, readily preparable by known methods or described herein.
Intermediate III may be acylated elther in the form of the free carboxylic acid (or salt thereo~) or as an easily cleavable ester ~or acid addi~ion salt thereof).
Pre~erred esters include benzhydryl, benzyl, p-ni~robenzyl, trichloroethylr silyl ~especially trimethylsilyl), phenacyl, p-methoxybenzyl5 acetonyl, (lower~alkyl including partic-~larly methyl, ethyl and t-butyl, triphenylmethyl, methoxy~
methyl,. acetoxymethyl~ pivaloyloxy~ethyl, phthalidyl and .indanyl. The procedures for preparing esters of car~oxylic acids are disclosed in the literature and are well-known to those ski:Lled ln the art of penicillin and cephal-osporin chem;stry.

1 1~6 37 ~
Prlor to the acylation reaction, any reactive ~ubst~tuents on the acyl~ting seid or derivative thereof, e.g. hydroxy, carboxyl or mercapto9 may be pro~ected by use of suitable protecting or blocking groups which s~e well-~nown to those skilled in the art o~ B-lactam chemistry, e.g. ~s by acylation or silylation. When the acylat~ng agent contains an amino functional group i~ the acy~ moiety9 the amino group is protected by a conventional amino-blocking group which may be readily ~emoved at the conclusion of the reaction. Examples o~ suitable amino.-protecting or blocking groups include t-butoxycarbonyl, carbobenzyloxy, 2-hydroxy-1-naphth-carbonyl, trichloroethoxycarbonyl J 2-e~hoxycarbonyl-l-methylvinyl and 2-methoxycarbonyl-1-methylv~nyl.
p~tic~larly valuable amino-bl~cking group is a p~oton, ~s ~n the acyl~tlng agent of the fo~mula ~ . ' ' ~ ~ CH-C-Cl N~2 HCl .
;' ~ . - 36 -63~
r Preferred amino-protecting groups. are t-butoxycar~nnyl, c~rbobenzyloxy, the proton and a ~-diketone or ~-ketoester as ln UoK~ Patent 1,123,333 o~ U~S. Patents 3,32S,479 and 3,316,247, e.g. methyl acetoace~ate, or . . a ~-ketoamide as in Japan 71/24714. When the t-butoxy-car~onyl, carbobenzyloxy, ~-ketaester, ~-diketone or ~ ~-ketoa~ide protecting groups are employed, it is ,' pref~rred to convert the,~cyla~$Rg acid containing ~he blocked amino group to a rnix~d anhydride~ e.g.
with ethyl or ~sobutyl chloroformate, before reaction with compound III or a salt thereof. After the acylation coupling reaction, the amino-protecting group and any other functional;protecting groups used may be removed by methods known per se to form the desired product of formula II~ th respect to amino protecting groups, :' the t-butoxycarbonyl group may be removed by use of formic acid, the carbobenzyloxy group by catalyt~c ~ hydrogenation, the 2-hydroxy-1-naphshcarbonyl group by acid hydrolys~s, the trichloroethoxycarbonyl group by treatment with zinc dust in glacial acetic acid, the ~ proton by neutralization, etc.
',`~ .Acylatian of a free amino group of a cephalosporin or penicillin nucleus is a well-known reaction, and any of the function21 equivalents of the carboxylic acid . . .
` RCOOH commonly used in penicillin or ~ephslosporin .. . .
chemist,ry as acylating agents for prim~ry amino groups nay be employed in acylating incermediate III. Examples o~ su~table acylating derivati~es of the fsee acid include the corresponding acid anhydrides, mixed anhydrides ~e.g. alkoxyformic anhydrides), ~cid halides, sc~d azides~
~t1ve esters and active thioestersO

A preferred acylating agent for preparing 7-acylamido c:ompounds containing arl a-~mino substituerit" e.g~
aminobenzyl, a~amlno-~-thienylmethyl, e~c. is the N-carboxy anhydride (Leuch's anhydride). I~ this 6txucture the group which activates the oar~oxyl group also serves to protect the amino $roup. Another p~eferred acylating agent for introducing a side chain containing an u-~mino functio~al group is the ~cid chloride hydrochlo~ide, of the ~ormula Ar-CI H-COC l NH2-~Cl which also serves a dual ~unction o~ carboxyl activatian and ~mino protection. Mention was made above o~ the u6e of enzymes to couple the free aeid with compound III. Included in the scope of such processes are the use of an ester, e.g. the methyl ester, of that fsee acid with enzymes provided by v~rious micro-organisms, e.g. those described by T. Takahashi et al., A.C.S., ~ . 4035-40~7 ~Ig72~ and by T. ~ara et ~ J. Antibiotics (Japan~ , 321-323 (1971) and ~n U.S. 39682,777. A p2rticularly preferred coupling ~g~nt for coupling~he acylating ~cid with compound III
~or a salt or ester thereof) is ~-e~hoxycarbonyl-2-ethoxy-1,2-d:ihydroquinoline ~EEDQ) as described in J.A.C.S., 90, 823-824 and 1652-1653 (1968) and U.S.
Patent 3,455,929.

The part:iclllar process conditions, e.g~ temper-ature, solv~-t, reaction time, etc. selected for the coupling reaction ~re determ~ned by the nature of the re8ctants a~d acylation method used and are known to those skilled in the art.

. ' The acyla~ing agents which may be used to form the N-acyl c~mpounds o~ formula II are kn~wn in the ~iterature along wi~h methods for ~heir synthesis o~ -~re disclosed in the exa~ples w~ich ollo~. In ~hose cases where the acylating agent contains one or more a8ymme~ric carbon atoms and ~hus exists in optically ~ctive forms, the compounds obtained using such an ; ~ .
Dcylatin~ agen~ are ordinarily ob~ained in racemic ~orm. ~hen the separate optical isomers are desired, the acylating agent can be resolved in a conventional ~ manner such as by reacting the free acid with cinchonine, ;: . . strychnine, brucine or t~e like, fractionally cryst~l-lizing to separate the diastereoisomeric salts and separately acidifying the solid phase and the liquid : phase to liberate the optical isomers.
: The 7-acylamido compounds of the present invention m~y be isolated ~n any of the way~ customarily employed .
: for the isolation of corresponding cephalosporin com-; pouhds. Form~tion of a desired pharmaceutically ~ccep~able carboxylic acid ar acid addition salt is .

~ 39 r . .

carried out by known methods, e.g. reaction of the ~cid of compound II (or ester ~n the case of acid addit~on 8~1t8~ w~th a~ appropriate base or acid.
At the co~clusion of the acylation reaction, ~he product obtained may be converted ~be~ore or after re~oval of any protecting groups) by methods known per se to ano~her desired product of formula II.
A compound of the formula . , H H
~--NH~-- --.. ~ 01 .

. ' ~ ~H2 ~ ' . .
II' ~02Rr ,~" .
: ~n which 2 is hydroxyl a~ R' is an.easily cleavable c~rboxyl-protecting group may be con~erted by ~cy~ation to the corresponding 7-acylamido c~mpound in which Z
is hydroxyl es~erlfied with a carboxylic acid ox a ~ulfonic acid residue. Acylation of the 3-hydroxymethyl group to produce carboxylic esters is preferably carried ou~ using an acid anhydride, e.g. acetic anhydride, ~n the pre~sence of an organic base such as pyridine~ Other convent:Lonal acylating agents m~y be used including acid ~ hslides (preferably acid chlorides), mixed anhydrides ;: or free acids ln the presence of condensing agen~s.
~ethods for e~terifying the 3-hydroxymethyl group o~ a cephalosporin sre known in the liter~ture, eO~. ~ee .~ , . .
- ~ o - , .
., ```\ :
r ~ ~qG63 ~L

U.S. 3,7~8,342, 3,532,694 ~nd U.K. 1,365,954, ~nd ~uch methods n~y be used with the novel 3-hydroxy-~ethyl ~-lactam deri~at~ves of the present in~ention~
The sulfonic acid esters m~y be formed by reaction of ~he 3-hydroxymethyl compot~nd with a suit~ble sulfonic acid derivative~.most preiEerably with a s~lfonyl halide such as methanesulfonyl chloride, p-toluene-~ul~onyl chloride or trifluoromethanesulfonyl chloride ~n the presence of an organic b~se~ The acyla~ing or ester~fying agent used is preferably one which will . .
~esult in format~on of the pre~erred 3-esteriied hydroxyl compounds mentioned above.
A compound of ~ormula II' in which Z is etheri ied hydsoxyl reduc~ble by hy~rogenolysis, e.g. benzyloxy, may-be converted to the correspondi~g 7-acy~a~ed 3.-hydroxymethyl compound by catalytic hydrogenation ~cording to methods known ~ se. Suitable hydrogena-t~on catalys~s ~nclude noble met~ls 9 most prefer~bly pallad~um or platinum a~d their oxides ~nd hydroxides3 end Raney nickel, said catalysts being optionally supported on a conven~ional carrier such as carbon, d~'comac~eous earth, etc. An especiall~ preferred catalys~: ~s 20Z Pd t0H);2. Preferred solvents f~r the ~, h~drogeIwlysi~ reaction are no~-reducible inert solvents 8uch 8g n~e~hanol, ethanol or ethyl acetate. The reactlon i8 preferably conducted at ~tmospher~o or ~lightly r ~ 4 .

ated pressure at room temperature~ When car~oxyl-proteoting group prese~ in compound II' are reducible ~y hydrogenolysls3 e.gO benzyl, p-nitrobenzyl, benzhydryl, etc.~ compound II' may be simultaneously de-blocked and reduced to the desired 3-hydroxymethyl fre~2 acid.
A compound of ormula Il' in which Z ls hydroxyl ~y ~e converted to the corresponding 7-acylated 3-etherified hydroxyl product by reacting the compound with an etheriying sgent by procedures used for cephalosporin compounds, e.ga those described in U-SO 3,665,003.
A compound of ~ormula II' in which Z ~s hydroxyl m~y be con~erted to the corresponding 3-heterocyclic ether compound by first forming a sulfonic acid ester ~t the 3-position~ e.g. ~y reacting the 3-hydroxymethyl compound with a sulfonyl chloride in ~he presence of ~n organic base, and then displacing the sulfonate residue with a heterocyclic alcohol in the presence of an organic base. The 3-he~erocyclic ethers m~y 31so be prepared by nucleophilic displacement of the corresponding 3-halomethyl compound, the prepar~t~on of ~hich is described below.
A compound of formula II' in whic~i Z is acetoxy ~y be converted to the corresponding 3~hydroxymethyl product by enzymJtic hydrolysis, e.g. by use of citrus ~cetyl es~erase.

.

' A compound of ormula Il' in which Z is hydroxyl y be converted to the corresponding 3-halomethyl eompo~nd by reaction with a suitable halogenatlng agent, eOg~ a phosphorus halide ~;uch as phorphoru~ trichlorlde~
pho~phorus tribromide~ phosphorus pentschl~ride~
phosphorus pen~a~romide, phosphorus oxychloxide, o~
phosphorus oxybromide. The 3-iodomethyl compounds m~y also be formed b~ treating ~he 3-bromomethyl or 3-chloro-methyl compound with ~n alkali metal iodide.
Compounds of formula II' in which Z is hydroxyl msy be converted to the corresponding compound~ having 7. - -OCONH2 by reaction in an inert organic solvent, e.g. benzene, with a source of cyanate ions, e.g., from an alkali metal cyanate, followed by ~reatment with tr~fluoroacetic acid. The cyanate ion ~nd trifluo~o-~cetic scid are preferably each employed in ~ lar ratio of about 2:1 with respect to the hydroxymethyl starting ma teria 1.
A compound o formula II in the form of the free scid or a salt thereof may be converted to a pharma-ceutically accep~able salt ~hereof or to a physiologic-~lly hydrolyzed es~er or pharm~ceutlcally acceptable ~alt thereof~ Similarly, the product of formula Il"
ln the ~orm of an easily cleavable ester or salt thereof ~ay be converted to the free ~cid product or a pharma-ceutic~lly scceptable salt thereof by removal of the ~sterifying group to form the free scid~ 2.g. by acid c o~ ~lkaline hydrolysi~, by enzym~tic hydrolysis (as ~it~ human or anim~l serum~, by hydrogenolysis or by treatment with chemi~al reagents known to remove par~ic-ul~r blocking groups, e.g. sodium thiophenoxide as in U.SO 39284,451, and subsequen~ treatment of the free acid with an acid or base to form a pharm~ceutically acceptable salt.
: The e~sily cleavable esters of the compounds of formul3 II ~re useful as intermediates in the produc-t~on of the free acid product. The pivaloyloxymethyl, ~cetoxymethyl, ph~halidy~, indanyl and methoxymethyl e~ters are also useful as active antibacterial agen~s since on oral administra~ion they are rapidly hydrolyze~
to the active metabolite. These esters a~e of partic-ular interest because they provide on oral administration dif~erent rates and amount~ of absorption and give dif-~ering concentrations of the active antibacterial agent in blood and tissues.

.

: ' :

;' ' .' ' , "'.

: The 7-~mino i~termediates o~ general formula.III
~y be prepared by selectLvely reducing ~ 7-~zido inter-- ~ediate of the formula ~3 ~ ~ O
, ~ ~C~2Z

IV' C2R

- .wherein Z is halo, hydroxyl, etherified hydroxyl or hydroxyl esterified with a carboxylic acid or sulfonic ~c~d residue and R' is an easily cleavable ester carboxyl-protecting group. The carboxyl-protected compound n~ay, i~ desired, be cleaved to produce the free-acid ~n~er~
medlate III which can be,converted to a salt by methods known ~ se.
Pre~erred reducing agen~s for use in preparing the intermediates of ~ormula III include chemical reduoi~g agents such as zinc and ammonium chloride, ~luminum amalgam 3nd hydrogen sulfide in the presence ~:of a base, e.g. triethylam~ne or ammonia. Catalytic hydrogenation ~y also be employed with such oatalysts .as noblle metals, pre~erably pla~inum or palladium ~ncluding derivatives thereof such as oxides, hydroxides and halides, or Raney nickel, said catalysts belng option-ally supported on 2 conventional carrier such as oarbon or diatom~ceous earth~ Catalyt~o hydrogenation is performed .

. _ 4g _ , .
.

w~th ~ non-reducible- inert solvent~ e.g. methanol, ethanol or ethyl acet~te~ and preferably at atm~spher~c or slightly elevated pressure at room ~mperature.

Compound III in the car~oxyl-prote~ted form or ~ salt thereof may be used directly as a st~rting material in the N-acyla~ion process discussed aboYe. Alternatively, the protected intermed~ate may be de-bloc~ed to ~or~ the free carboxylic ~cid which m~y then be op~ionally converted to ~ salt or to another carboxyl-protected ~orm, e.g. a phys~ologically hydrolyzed ester or salt thereof. By proper selection o~ reduction conditions ~nd protect~ng groups, azido intermediate IV' may be converred either simultaneously or in stepwise fash~on to ~he 7-amino free acid III. Thus, if mild hydro-genation condi~ions are used~ e.g. catalyt~c hydro-genation with 10% Pd-on~charcoal or a.mild chemieal reducing agent such as H2S in the presence of a base such as triethylamine or ammonia~ the azido group may be ~educed without concomitant removal of es~ers resistant to guch conditions, e.g. benzyl or p-n~trobenzyl. If ~trQngex reducing conditions are used such as 30%
Pd-on-diatomaceous earth, both the azido group and m~st reduc~ble esters will be slmul~aneously reduced.
When it is de~ired to prepare an intermediate III
where Z i9 etherifled hydroxyl, it m~y be nece~sary to select sufficiently mild reducing conditlon~, e.g.

.

~ - 46 -.......
- ' ~ 1~ 6 ~374 , H2S with base, so as not to afect a reduc~ble ether ~iety.
A preferred embod~ment of the present ~nvention ~he process eomprising the consecu~i~e steps of 1) selecti~ely reducing ~ 7-azido intermedi~te of the ~ormula IV' where Z is halo, hydroxyl, hydroxyl esteri~ied with a carboxylic acid or sulfonic acid residue or etheri~ed hydroxyl and R' is an easily cle3vable ester carboxyl-protecting group tc produce 8 c~rboxyl protected 7-amino intermediate of formNla III
~nd, if desired~ removing the carboxyl-protecting group to produce the corresponding free acid intermediate of ormula III or optio~ally a sal~ thereof; and 2~ N-acylating i~termedia~e III or a salt thereof with an acylat~ng acid of the formNla R-COOH where R is aD acyl group, or wi h i~s functional equiYalent as an aeylating agent for a prim~ry ~mine and, if desired, converting the so-produced product to the corresponding 7-aeylated pr~duct having a substi~uent Z as deff ned above di~ferent from that in compound III and, if desired9 (a) wheln R'7 is a.carboxyl-protectLng group, conver~ing the 7-acylated es er to the ~ree ac;d compound or a p~ysiologically hydrolyzed ester or a pharm~ceutically acceptable salt of said acid or ester, or (b) when R" ~s hydrogen, convexting the 7-acylated o~rboxylic acid to ~ physiologically hydrolyzed ester or a ph~rm~ceutically , ~ 36 3~
'" ' ' ''', ' ' .' .

~cceptable salt of sa~d ~cld or ester and~ i~ de~ired, r~solving a resulting isonner mixture into its component i~omers.
The 7~-~zido intermed~ates I~' may be prepared by ~arious methods depending on the nature o~ substltuent Z. When Z is a group -OCt)R2 derived from an acid ~2COOH
having a PKa Of between 3.5 to 5.5 and wherein R2 i9 hyd~ogen, amino, ~lower)alkyl, C3-C7 cycloalkyl, C3-C7 c~cloalkyl-(lower)alkyl, aryl, aralkyl or aryloxyalkylene, 8aid R2 groups being optionally substitutèd by one or more radicals sclec~ed from (lower)alkoxy, (lower)alkyl-thio, h~logeng tlower)alkyl, nitro, hydroxy~ acyloxy, carboxy, amino, ~lower)alkylamino or acylamino, inter-~cdiates o~ formula IVI may be prepared by cyclizing in an inerP organic solvent a compound of ~he formula ~, ~
`~3 ~
C (X) ~CH2X ' ) ": . . CO
.. '. V .

. . . .

' ' ' ' . .

: - 48 -.

r ~herein X and Xl whioh m~y be the same or dif~eren~
e~ch represen~ 8 halogen atom, preferably bromine, or iodine and most prefer~bly iodlne~ Rl is an easily cle~vable ester carboxyl-protecting gro~p and Y' ~epresents 3 suitable leaving group, preferably a group such as halo or sulfonyloxy, e.g. alkyl~ or subs~ituted alkylsul~onyloxy or asyl- or su~stitu~ed 2rylsulfonyloxy and most pxeferably ~ group selected from halo~ -OS02-(l~wer~alkyl including especially -OSO2C~39 OSO2cF3 and os~2c6H4~H3~ i an anioll of the onnula ~2COO derived from a carboxylic ~c~d ha~ ing a PKa f between about 305 and 5.5 and in which R2 is as deffned above.
The dihalide starting material V may be used in either of lts isomeric forms Y~
~ B
N3 ~

N ~ CH * ' . CO~R
Va or y~
H H l N3~) C~2X
~N ~, CO~R~
~b _ 49 _ ~6 , or a~ a mixture o isomers. ~or~ula V ~bove is intended t~ represent either of thle ~ndividu~l isomers or the ~xture. Any dihslide including a m~xed dihalide, e.g.
X~ , X' ~ Br, may be used but the most preferred c~pound is the diiodide. Co~pound V is rescted in an l~ert organic solvent, pr~ferably a polar organic salvent such as dimethyLfor~mide, with an excess o~ the base.
A~y base derived from a car~oxylic acid R2COOH
8ati8fying the pK~ conditions abo~e asld w~erein R2 is 88 defined above m~y be ~mployed in the cycllzation ~eac~ion.. The preferred bases ~re the anioas of the formula R2COO ln ~hich ~2 ~s hydrogen, ~lower)alkyl ant espec~ally methyl, phenoxymethyl and tetrazolyl-methyl. The most preferred bases are~the formate ~nd ~cee~te anions, e.g. ~rom an alkali ~etal, ammonium or substituted ammonium formate or acetate.
~ , The leaving group Y~ should be one which is e~ficiently ~isplaced under the conditions of the base cyclization reacticnO Sui~ab~e leaving groups include hal~ andl sulfonyloxy groups, i.e., alkyl- or substituted ~tlkylsulfonyloxy or axyl- or substituted arylsulfonyloxy.
most preferred Y' leaving group is t~e mesyla~e group.
Cyclization of compou~d V is con~Teniently carried ou~ ~t room temperature or below.

.

. r r . I

' St9sting materials oiE ~ormula ~ used ~n th2 above process may ~e prep~red as described below in the 8ection entitled l1Preparation of Star~ing Materials."
Brie~ly summarized, the reaction scheme ~s as shown ~n Flow Sheet I:

HON C~rbo~yl ~oteçtion as y_~cetallzatlon_ ~ .~ C~COCH3 j~
C02R ~

HON O~\ ' nitroso ) reduction ~ t ~
GO2R' CH3 N O'~ Schiff base ~ fi h tion as ~>
C02R' CH3 cinn~m~ldehyde .

. . .. . . . .

51 ~ ~.r `

.

1~ ~ol~ctam formation s with ~? ~zidoacetyl halide >
N~

C02R ' H
N3~ ~ ozonolysis~

O
~02R ' ~3 _ CHO ~elective aldehyde ; ~ O~~> ~ reduction _ _ ~>
; ~ ~ N~
; C02R ' ~ ~ CH OH conversion of N3 ~ 2 hydroxyl group to a ~ ~ more activated .: ~ ~ lea~ing group as by ~ ~_-N ~ halogenation or o~ ~ C ~ esterlfication wi~h C02R' ~ sulfonic c~d deriveti~e ' .
.

~637 H H 1:)50~CH3 ~I~J de-ketalization >
~o ~32R 3 H ~ OS02CH3 hydroxyl activation N ~ ~ ~ ~8 with triflic 3~ anhydride >
~N

CO ~' N H H 1 2 3~llene formation :: 3~ ~ with ba s e _ >
1 0~"2CF3

6 ~ 3 ., N _ ¦
~: . 3~ h~logenation 02~' .

Flow Sheet 1 ~

~ 3~ ~

Intermediates of formula IV' h~Ying Z either -OCHO
i - or -OCOR2 in which R2 is ~lower)~lkyl m~y be con~erted to the corresponding ~ntermediates of formula IV' wherein Z ~ hydroxyl by sub~ecting such compounds to acid hydrolysis, e.g. by treat~ment ~ith a miner~l acid ln an ~queous ace~one sol~ent system~
Intermediates of formula IV' in which Z i5 hydroxyl esteri~ied wlth a carboxylic acid or sulfonic acid residue m~y be prepared by esterifying a 3-hydroxymethyl inter-mediate of the formula : - .
H H
. ~3~ ~ O

N ~ H20H
C~)2R ' ,~ .
~ith a caxboxylic acid or sulfonic acid agent cap~ble of introducing the desired acyloxy or sulfonyloxy ~cyl ~o~ety at the 3-position. Esterification in this embodi-men~ of he presen~ invention may be performed in the sa~e m~nner as descr~bed previously in connection wi~h the modi-f~c~tiol~ of ~he ~ subst~ tuents in compounds of formula IIo A prefexred embodiment comprises reacting ~n in~ermedi~te IV' in which Z is hydroxyl with aceti anh~dr~de in the presence of an organic base to prepare the corresponding carboxyl-protected 3-acetoxymethyl 7-azido intermedla e.

:

" ' '' ~ . ' .
.~ ~

- ; /
iLg~.~6~

Another preferred embodiment i~v~lves esteriying - -intermediate IV' in which Z is hydroxyl with an ester-~fying der~vative of methanesul~onlc ac~d, trifluoro-methanesulfonic acid or p-toluenesulfon~c acid, e.g.
m~thanesulfonyl chloride, p-toluenesulfonyl chloride or triflic anhydride, in the presence ~f an organic base, e.g. triethylamine, and in an inert organic 801vent, e.g~ methylene ch~oride, to produce an inte~-mediate of formula TV' in which Z is methanesulfonyloxy, p-toluenesulfonyloxy or trifluoromethylsuIfonyloxy.
InSermediates of formula IV' in whLch Z is hydroxyl ~y be con~erted to the corresponding 3-halomethyl compounds by resction with a halogenating agent, e.g.
phosphorus halide such as phosphorus trichloride, phosphorus tribromide, phosphorus pentachloride, phosphorus~pentabromide9 phosphorus oxychloride o~
phosphorus oxybromide. The 3-iodome~hyl compounds may ~lso be formed by treating the 3-bromomethyl or 3-chloro-methyl compound with an alkali metal iodide.
Inl:ermediates of formula IVI in which Z ~s 3-halo-~ethyl or 3-sulfonyloxy m2y be con~erted t:o ~he corres-.ponding 3-heterocyclic ether intermediates by nucleo-ph~lic displacement with a heterocyclic alcohol in the presence o~ an organlc base.

h t~
' '' ' ' ' ' .
., .
, . . ' ' , .
C~rbamate interMediates o~ the formul~

~ ~ .

N ~ ~H OCONH
CO R' may be prepared by reacting the corresponding carboxyl-protected 7-azido 3-hydroxymethyl compound in an inert organlc so~vent~ e.g. benzene, with a sourc.s of eyanate ions9 e.g. an alkali metal cyanatel ~ollowed by tre~tment w~th trifluoroacetic acid. The cyanate ion and ~rifluoro-acet~c acid are preferably each employed in approxima~ely a ~:1 ~olar r~tio with respect to the azido star~ing ~terial. ~

An al~ernate method provi~ed by the present inv~ntion for preparation of intermediates of formula IV9 where 7. is hydroxyl comprises cyclizing with base ~n an inert organic s~lvent an intermediate o~ the formula H
_ J
3 ~
N ~ HO
~2R~

' wherein R' is an e~slly cleav ble ester carboxyl protecting group and Y' represerlts a suitable le~ving grollp, preferably a h~lo or a sulfonyloxy g~oup, eOg. alkyl- or substituted alkylsulfonyloxy or aryl~
o~ substituted arylsulfonyloxy and most preferably group selected from halo" -VS02-(lower)alkyl g p y S 2 3 ~ S02CF3 and OS02C6H4CH3 .
The base used in the cyciization of VI may be selected from a 6~ide variety of bases including especially bases of the following categories:

~) anions derived from carboxylic ~cids having 2 PKa ~ beeween 3.5 and 5.5;

(b) tertiary organic amines sllch as a 'crialkylamine (e.g., triethylamine) ~
pylidine, N-methylpiperidine~ N-methyL-:. morpholine , etc .;

~c) ~ cali metal hydrides~ e.g.~9 sodium . or potassium hydride; and ,....... . .

'' (d) organolithium compounds ~ncludingespecially lithium alkyls, e.g~, methy~
. llthium or butyl llthium.

~ ost preferred cycliz~tion bases are the ~cetate and ~ormate a~ions, e~g., from alka~i metal, ammonium or substituted ammon~um formates or ac~tates. The most preferred base is ~he acetaSe anion. The base is preferably used in a molar excess relative to compound ~I, and the reaction is conducted in an lnert organic solvent9 preferably a polar organic ~olvent such as dimethylsulfoxide or dimethylformamide.
The temperature: for the cyclization is ~ot critical, and room temperature may ~e used fF convenience, .

.
.
' . 53 ~

, r - Compou¢ld V~ used in the above proc ss may be p~epared by ~he procPdure~ described below in the 8ect~0n entitled "Preparation o Start~ng ~teri~ls".
A ~ummary of the reaction scheme is shown in Flow ~;heet 2: 4 BuI,i/ICA
(EtO) 2CHCO~1 :t CH3C2CH2~

(Eto)2c~coc~I2co2R1 nitrosation ketone reduction (E~O) C~lCOCC02Rt _ as_with a ~, 2 ~ borohydride - -- hydroxy~ protection ~EtO) C~ICHOHCC02R ' a s with 2 . Il dihydropyran NOH + H~

QT~P r ~EtO)2CHCHCCO2R' nitroso reduction ~, NOH as wi~h Al(Hg) OTHP Schiff base ~ormation ~E~0)2CHCHcHco2~' cinnam~ldehyde ?
N~2 OTHP ~-lactam formation (EtO)2C~IC~CHC02R' az~doacetyl halidé ~>

, _ 5~ _ -' :

~ ~ 6 ~7 ~

N3 ~ ozonolysis OTHP ----' . `' N ~ OEt ~:92R

~ - ~ select;ve aldehyde OTHP reductlon as with a borohydride OEt C02R ' H ~ conversion of 3 ~ fcH~oH hydroxyl group to O.THP a more activated ~I ~ 1 , eaving group as b "~ ,OEt halogenation or I ~ OEt esterification with a . CO2R' sulfonic acid d~rivative H H
3 _ - / CX20SO2CH3 remo~s 1 of OTHP HP a~ by _~
OEt~cid hydrolysis C~2R ~

-- ~o --H H

N _ CH20S02CH3 hydroxy.aldehyde formation as with ~
OH ~5% trifluoroacetic acid 7 Et C2R ' H H - .............. _ ................ _.
N ~ / CH20SO2CH3 Et: = C H
t ICA = isopropylcyclohexylamine CHO T~.~ = tetrahydropyran C2R ' 0 -- C6H5- , I
Flow Sheet 2 The 7~-azido intermediates of formula IV' where Z is etherified hydroxyl may be prepared by cyclizing with base in an inert organic solvent an enol intermediate of the formula H H Y' _ - I
3 ~ OH

o ~CH2 Z
VII C2R' wherein Z is an etherified hydroxyl group, R' is an easily cleavable ester carboxyl-protecting group and Y' represents a suitable leaving group, preferably a halo or sulfonyloxy, e.g. alkyl- or substituted alkylsulfonyloxy or aryl- or sub-stituted arylsulfonyloxy and most preferably a group selected from halo, -OSO2-(lower) alkyl including especially -OSO2CH3, -OSO2CF and -OSO C H CH .

, . .

j The base used to cyc:lize enol VII may be any of the bases mentioned abo~e in connection with the cyclization of intermediate VI and is p~eferably a ~cetate or forma~e ~rllon and most preferably the ~cetate anion~ e.g., from an alkali metal, an~non~
or 6ubstituted ammoni~ acetate. Cyclixation is carr~ ed out in an inert organic sol~7erlt, e . g ., methylene chloride, dimethylforTnamide or dimethyl-8Ulioxide, wlth an excess of baseD and may conven-~ently be done either at room ~emperature or under re~lux~ -Compound VI used in the above process m~y be preparedby the general method described below in the section entltled "Preparation of~Starting Materials". A sun~nary o~ ~he reaction scheme is shown in Flow Sheet 3:
' , .
3~CH20Na ClCH 2C02H - > ~CH2 H2 2 > ~CH20CH~CO~E~
O

Li~ H2C2R ' ~SH OCH ~
> I 2 2 n~r ~-- t i-~n >

C2R ' .

. . . ~ ~2 _ ~ 6 ~

, O
HON ~ ~elective carbonyl CH2OCH2~ reduction as with >
~:02R ' ~ borohydride OH
HON ~ CH20CH2~ hydr~yl proeectlon ~O2R~ di~ dropyran t ~THP
~ON ~ ~ OC~ sele~tive nitroso CB2 20 ~ reduct~on as with ~O2R~ Al(Hg) ' aT~P
H ~ l r Schiff base : ~ ~H2OCH2~ formation as_with cinnam~ldehyde C02R ' .
. ~
lactam formation OTHP as with I azidoacetyl halide N~ CH20CH2~1 ' .

.~ .
,' `' ' ' " .

H H

N - _ ~ ,o,z,o,n:o,I,y,s,is, , , ~ O~HP ~~ ------------C2R ' H H CHO
N - ~ / selective aldehyde 3~ - ~ OTHP reduction as with I a borohy~rride `''~~~~
o~ , ~ \~CH2 0CH20 C2R ' H H CH OH conversion of N _ _ ~ 2 hydroxyl group to 3 ~ ~ O~HP a more activated I leaving group as by N CH OCH 0 halogenation or ~ ~ 2 2 esterification with a v I sulfonic acid derivative C2R' ' H H
= = CH OSO CH b) oxidation as N _ _ / 2 2 3 with CrO3 ~ ~ Ch20C~20 a~ 10~ HCl hydrolysis ) C2R ' N - ~ CH20So2CH3 ~ OH
~N ~ CH20CH20 C2R ' Flow Sheet 3 :
:, . . .

6~3r~
r The pharmaceu~ically actlve conpounds of the p~esen~ invention are potent antibacterial agents use~ul ~n the tr2atment o~ infectious diseases in poul~ry and animals, includi~lg man~ caused by m~ny Gram;positive and Gram-nega~ive bacteria. The active compounds are a~so o~ ~alue as nutritio~al supplements ~ animal feeds and as agents for the txea~me~t of ; m~titîs in cattle.
- Illustrative examples of the preparation of : starting materials and compounds of the present ~nve~tion follow. These examples are given in illustration of, but not ~n limitation of, the present invent~on. All temperatures are in degrees : Centi~rade. DMF rep~esents dimethylformamide, THF
s~ands for tetrahydrofuran and EEDQ is the amide bo~d formi~g reagent having the structure .

; ~ OEt The 7-acylamido compounds prepared in th~
examples which follow all have the hydrogen atoms at , . . .
carbons 6 and 7 cis with respect to each other and, ~nless ind;cated~ the products are racemic mixtures ln the sense that they are composed o~ equal parts o~ the two isomers having ~he following s~ructures:
.' .
H ~ H H
~S~ ~ <

`' C~/O ~0-r ~ 6 , - Preparation Of Start~
.

lo Preparation o~ intPrmediates o~ formulae H ~ 53~02CH3 H H OS02CH3 ~J Br and ~J ~H2Br ~N~CH2B~ ~ ~Br C2R ' C2R

8e,nzyl_Oximino-A~etoacetate . .
~ H3~ ~ C02CH2~
d NOH
1.1 -, The procedure was essentially the ~ame as that used t~ make the corresponding ethyl es~er by H. Adkins ~nd J. Reevea JACS 60, 1328 (1938).
In a three necked one liter flask fi'cted with a thermometer9 a dropp~ng funnel and a m~gnetic stirrer we~e placed 173 g. (0.9 mole) of benzyl acetoacetate [The benzyl acetoaceta~e was prepare~ as described by Bzker et al., 30 Org. Chem. 17~ 91 ~1952)] and 130 ml.
o glaci,al acetic acid. The co~ten~ were coolçd in a~ ice bath and ~ solution of 69 ~. (l mole) of .
80dium ~itrite in 130 ml. of water was added over a period of half an hour; the ~emperature was kept at O to 10 C4 After the react~on mixture was ~tirred ~or one hour at room temperature9 400 ml. of ~ater ~as ~dded and the s~irring was continued for an ~dditional ~wo hours. The reaction mix~ure was extracted t~ree times w~th 200 ml~ portions of diethyl etherO The d~ethyl ether extracts were c~mbined~ washed once with waeer, three times with saturated sodium bicar-bon~te solution and once with brine. After drying over anhydrous sodium sulfate, the die~hyl ether solu-tion was evaporated leaving tl.l] as a clP~r oil whic~
solidi~ied upon trituration with petroleum ether (30 - 60~ to give 186.5 g. (93 . 2%) of white solid.
Its NMR spectrum was consistent wit~ the assigned structure. Generally the~product was used as such ~n subsequent reaction but it can be recrystallized ~rom toluene, m.p. 81 - 82 C.

Benzyl Qximino-Acetoacetate Ethylene Ketal CH3~ CO2CH2~ ~ ~ CH3 ~ C~- ICl-CO2CH
NOH . ~ NOH

1.1 ;~ ~

.
I~ fl two lite~ flask fitted with a ~ean Stark wster separator and a condenser were placed 186.5 g.
.

.

~0.85 mole) of benzyl oximino-acetoaceta~e ~1~13~ 62 g.
(1 mole) of ethylene glycol, 800 ml. of ben~ene (reagent grade) and 2 g. (10.5 mmole) o~ p-toluenesul~onic acid nohydra~e. The reaction mixture was boiled at re~lux until 15 ml. of water was removed (3 hours). The benzene solution was washed once with saturated sodium bicar~
~onate solution and once with brine. After drying over anhydrous sodium sulfa~e, the benzene solu~ion was eY~por~ted, leav~ng 212 g. (94Z yield) of benzyl oximino- :

acetoacetate ethylene ketal ~2~1] as a light yellow oil' Its NMR spectrum was consistent with the assigned structure. ~enerally, the product ~as used as such in su~sequent reactions b~t one of the isomers can be crystallized2 from toluene-petroleum.either (b.p. 30-60CC.); m.p. 52C.
Anal. Calc'd. for C13H15N05: C,-58.86; H, 5.70; ~, 5.28.
Found : C, 58.97; H, 5.68; N, 5.12 1~ A mixture of the ~ and anti isomers.
2. Only 35% of the oil could be crystallized.

- 6~ -.

Slelective C~ 0~ reduction >~ CH- C02CH20' : O OH ~ NH2 2.1 3.1 ~reshly prepared aluminum amalgaml ~from 27 g. of : ~luminum foil) in a three-necked one li~er ~lask was covered with 500 ml. of diethyl ether. The flask was ~- fitted with a mecha~i~al stirrer~ a condenser, and a dropping funnel. A solution of benzyl oximino-aceto-acetate ethylene ketal ~2~1~ (132.5 g.; 0.5 mole) in .
3~0 mlO o~ wet diethyle~her' was added dropwise at su~h ., ~ .
ra~e as to m~intain boiling at reflux. After stirring for four hours, the reaction mixture was filtered through a Buchner funnel. The filtrate was evapor-ted leaving 110 g. of yellowish oilO The oil was d~ssolved in 800 ml. of dry diethylether and dry hydrogen chloride gas was bubbled into the solution until no further precipi~ation occurred. The white precipita~e was filtered off and washed once with diethyl~ether and then dried Ln vacuo. . This provided 108 g. of benzy~ am~noacetoacetate ethylene ketal hydrochloride3 ~3vl]; m~pO 157-158C.
.

.

, . ;

, , .

Anal.Calc'd. for C13H17N04.HCl: C, 54.26; H~ 6.31, ~, 4.87.
Fo~nd: C, 53.96; H~ 6.19;
N, 4.60.
. To obtain the free base, the hydrochloride salt ; was suspended in 500 ml. of diethylether and concen-trated ammonium hydroxide was added with shaking un~il . the solid went into solution. The diethylether layer was separated and washed twice with brine. Aftsr drying over anhydrous sodium sulfate, the solvent was evapor-ated leaving 90 g. (71% yield) of colorless oil.
', 14 The aluminum amalgam was prepared essentially as . described in A.I. Vogel ("Practical Organic .
: Chemistry"j 3rd, Edn.~ Longemans Green & Co., London, (1957), p. 1983 except for the following modific~tion:
~) 5% NaOH was used.
(b) The second washing with ethanol was omitted.

.

r 1~ ;3 ~c~ Dry die~hylether was used for washing and most of the water must be drained.
2. The diethylether was saturated with water by sh~king wth water ~n a separatQry funnel. - ~ -3. The product can be stored as the hydrochlorlde sal~.

Schi~ Base Formation And ~-Lactam Formation ; ~ ,0/--\0 ~ ' ~0~~1 - ~ ~ formation ~ ~ ~
- C~ ~ - CH:- C02C~20 ~ 03 3.1 ~ 4.1 H H
N

5,~1 .
..~
In ~ one liter flask fitted with a Dean Star~ water 8epara~0r and a condenser were placed 70.3 g. (0.28 mole) benzyl aminoacetoaceta~e ethylene ketal [3.l]~ 37 g. ~0~28 mole) cinnamaldehyde, and 750 mlO of methylene chloride . , ~ , . .

~ 63~ 7fl (reagent grade~. The mixture was boiled at reflux fur 30 minutes and then 400 ml. of methylene chloride were distilled o~f. The concentrated solution was then dried over anhydrous ~sodium sulfate and ~hen eva-porated to dryness in vaouol. The residual oll w~s checked by NMR to ensure that Schiff base forma~on was complete be~ore continuing on to the next step.
The freshly prepared Schif~ base ~4.1~ was diluted with 600 ml. of methylene chloride2 and cooled ~o 0C.
~ice-salt bath3. Triethylamine (31.1 g.; 0.308 mole) was added and then a solution of 36.2 g. ~0.308 mole) of azidoacetyl chloride3 in 362 ml. of methylene chlor~de2 was added aropwise at 0C. over a period of one hour. The reaction mi~ture was stirred for ~n additional hour at room temperature4 and then-evapor-~ted on a rotary evapora~or at reduced pressure while being heated on a 35C. water bath . The residue was diluted with 500 ml. of diethylether and filtered.
The filtrate was wa~hed twice with brine and dried.
over anhydrous sodium sulfate. Evaporation of t~is ~olution yielded ll7.5 g. (94% yield) of styryl ~-lactam ~5.1]. Its NMR and IR spectra are consistent with the assigned structure and indicate the presence o~ a mixture of isomers, diasteriomeric at the ear~on a to the car~onyl of ~he benzyl ester.
1~ Th~s evaporation must be done to ensure comple~e . .

.

` ~ ~
6~

:~

Sshiff base ~orma~ioslO
20 All the methylene chloride used ~n ~he cyclo-add~tion reaction w3s reagent grade which was . first dried over molecular sieve (Type 4A) and ; th~n over anhydrous c:alcium chloride~ It was stored thereafter over molecular sieve (Type .', ~A).
3. 3O H. Boyer and J. Horner, J. Amer. Chem. Soc.

~ (1955), 77, 951~

; 4O The reaction mixture can be kept overnite at ; 0 i~ necessary.

5. This operation is necessary to ensure complete ~-laetam forma~ion.
.~

: ,, :
~-~ H H ~

, ~ ~ CH 2 ( ) ~ ~
.; ~2CH2~ CH3 ~:02CH2)~ -5.1 6.1 Styryl ~-lactam ~5.1~ tll7.5 g.; 0.262 mole) was dissolved in one liter of methylene chloride (reagent .grsdc), cooled to -50 to ~60~C. in a dry ice-acetone ~ath, ~lld ozonized until a faint blue-green color appeared. The solution was then flushed with nitrogen until the oolor faded. Methylsulfide (100 ml.) was 3~4 ,~ . .

~dded to the ^50C~ solut:ion, wh~ch was ~hen allowed ~o slowly reach 25~ as ~hle cooling ~ath gradually ~lted. It was kept over~ite at ro~m temperature u~der nitrogen and then it was washed twice with . lZ sod~um bicar~onate solution, twice with brine, dried over anhydrous sodium sulfate, and evapora~ed to dryness. The resulting oil triturated four .
times with 100 ml. portions of petroleum ether (b.p.
3~-60C.~ to remove benzaldehyde. The oil was then triturated carefully with diethylether whereupon ~t solidified. The solid was fil~ered off and dr~ed to provide 75 g. t7~-5%) of aldehyde [6.1] as a ~ixture of isomers diasteriomeric at the carbon a d to the carbonyl of the benzyl es~er. Recrystall~za-tion of 16.1] ~rom ether gave white crystals, m.p.

101-102C. (corrected).

Anal. Calc'd. for C17Hl~N406 : C, 54.54; H, 4.84;

N, 14096.

Found : C, 54.75; H, 4~7;

N~ 14.89.

' .
CHO N ~ ~
3 ~/ /~~ Na~H4 ~ ~ ~ C2~1d20' 8 a L

The ~ldehyde [6.1] (116.3 gO; 0.31 mole) was tis-~olved in 600 ml. of THF (reagent grade) and the solution was then cooled to -10C. (ice-methanol bath). Sodium ~orohydride (5 . 88 g .; 0 . 155 mole3 was added and the reaction mixture was stirred 1 hourO lOZ aqueous ~iydro-chloric acid was a~ded until the mixture was slightly ~cidic~ then 600 ml. brine was added. The THF layer w~s separated and the aqueous phase was extracted twice ~ith 250 mlO portions o~ d;ethylethe~O The combined ; . organic phases were washed twice with 400 ml. portions o~ brine, dried over ~nhydrous sodius;l sulfate, and evaporated in vacuo to yield 117.3 g. of crude alcohol . . .

.;

. .

\

;6~'7 , 17~ n orange oil~ This oil w~ used ~s ~uch i~
the next reac~ion.
A ~olu~ion of methanesulfonyl chloride (37.8 g.;
0.34 mole) in lO0 ml~ of methylene chloridel was added d~opw~se at 00C. (ice-w~ter bath) to a stirring 801u-tion of alcohol ~7Ola tlO5~6 g.; 0.28 mole, triethyl-~mine (56.6 g.~ 0.34 mole) and one liter of me~hylene chloridel. Afterw~rds~ ~he reaction was stirred or 30 hours at 25C. It was then washed twice with brine (500 ml, port~ons), dried over anhydrous ~odium sulfate, and evaporated ~n vacuo. The resulting oil was dissolved in methylene chloride, treated with nori~e, and ~hen filtered over approximately 200 g, of activity I silica gel. The silica gel was hen washed with approxim~ely 2 l~ters of me~hylene chlorideO The filtrate W3S eva-por~ted to dryness and the resulting oil (116 g.) was covered with diethyletherO It crystallized on ~tanding ~o give 87.2 g. (8~% from [601~ 0~ mesyla~e ~8.l3 as off-white solid, m.p..97-99C ~corrected).
l. The methylene chloride used was reagent grade wh~ch had been further purified by passing ~ver column of c~lcium chloride and then storing over molecular sieve ~Type 4A).

i - 76 qSO2CH3 . H ~ OS02C~
~5 _ J de-~etal- N~
za~

: C02~I20 ~:2CH20 . 8.1 ~9.1 N~ J 3 SO2~:F3 OJ ~~ 5 ~2CH2 .
10'~
`~ ~ mixture o~ mesylate ~}ol~l ~3~19 g~; 6.43 mmole) ~nd 30 ml. of 95% trlfluoroacetic ac~d was stirsed ~t 25 fo~ 2 hours~ The mixture was diluted with 300 ml~
o br~ne ~nd extracted three times with methylen~
chloride (lOO ml~ portions3. The c~nbined extr~cts were w~shed three 'cime~ w~th w~ter (SO ml. portions, ~ntil neutr~ dried (anhydrous sl~dium sulfate~ ant e~spor~ted to drysle~ ~n vacuo le~vlng 3.17 g. o~ a ,br~ oilO ~ spectsa of thifi oil ~ndic~te the presence of ~907. enol [9.1~, . .
", .

.

.

Crude enol ~9,1~ (48.0 8-; 0.117 le) ~nd ~riflic a~hydride (33.0 g.; 0.11~ mole) were dissolved in 500 ~1. of methylene chlor~de ~nd the sulution W~9 then cooled to 0C. ~ice-water bath)J A solution of tri-ethyl~mine (11.8 g.; 0.117 mole) in.80 ml O of methylene chlor~de2 was added dropwise over a period of 40 mlnutes Wh~n the addition was complete, the ice-water b~th W~8 removed and the mixture was stirred at 25~ for 45 m~-nutes. The mixturP was then poured into 300 ml. of ice w~t~r and washed with c~ld water l~ntil the pH of the w~shings was approxim~tely 60 The extract W~8 : dr~ed (anhyd~ous sodium ~ulf2te) and ev~porated in v~cuo to glve 54.0 g. of crude trifla~e.~10.1] 88 a d~rk red : oil; This oil was disso~ved in 400 ml~ of benzene (USi) and passed through a ~ 1/2" p~d of ~ctivity III silica ~el. The pad w~ washed with 1 lo Of benzene. Ev~por-: a~ion of the benzene gave 38.3 g. of a yellaw oil. This oil wa~ carefully trltursted with 5Q ml. of absolute eth~nol and t~en cooled flt 0C. for 2 hours. The resulting white solid wa~ f~ltered off ~nd dried in vacuo to give 19.5 g. of tr$fla~e ~10.1~ a3 one isomer, m.p. 57-59C. (corrected).
Anal. C~lc~d. for Cl7~17F3N404S2: C, 37.67; H, 3.14;
~, 10.,~3; S, 11.82.
; Found : C, 37.40; H~ 3.12;
M, 10.43; S, 11~73,

- 7~ -1. Trifl~c ~alhydr~de W~3B prepared ~8 follow~:
170 gO ~100 ml.) CF3S03}1 ("Fluorochem~c ac~d"
3~I Comp~ny3 and 135 g. P20~; w~re mix d c~refully, . 8h~ken well, ~rld ~tored 18 hour~, pro~e~et_fr~m m~isture. The product was distilled ~rom the resulPlng solid mass using ~ fl~me, the fr~ction ~oiling 8~-90C. was collected. Re-distillation of this frac lon yielded ll9t45 g. (74%~ o~
~sr~flic anhydride boiling 82-84Co 20 The methylene chlor~de used was reagent gr~de which h~d been fur'cher purified by passing over column of calcium chloride and then stored ovsr molecul~r si~ve ~Type 4A)o ~ .
H OS02C~3 . H H OSo2cH3 J org~nic ~ N3 - ~ ~
~ OS02CF3 base 7 ~ ~C~2 d-- ~ CH3 o~ ~
o I c0 H lI J sO2cH3 Br ~ 3~ J ~l~r ~r ~Br C02t:H2~ !G02CH20' 2cl 13.

*Trade uark ~ 79 , Triethyl~mine ~1 g.; 0.1:~1 mole) W~18 ~dded to 9 ~tirxed ~olution of trlfl~lte [10.1] (5.42 g.; 0.01 ~le~
1~ 55 ml. of methylene chlor~de tA.R.~ at room temper~-tU~C~lo After s"cirring for five minute~ (at wh~ch po~rlt ~ ho~?~ complete fo~tl.on of ~llene ~ solutlon of bromine (10 ml~ of l~I ~olution ln Ct:14; 0.01 mol~) w~a ~dded dropwi~e~ A~ter ~ddition o the brom~ne, the mixture ~a~ concentrated, ~bsorbed onto Activity I

8~11ca gel ~nd dry column chrom~togrsph~d o~ Activity I
811ic~ gel by elut~ng with methylesle chlorid2 (USP?~
This yielded one fr~ction ~uniformly one spot by TL~) ~8hing 2 ~ 5 g . (457O) . Its IR, UVg ~nd N~ 8pectr3 ~ere consi~tent with the ~xpected d~romlde stru~ure ~J,3~1] . O
A~al~ Calc 'd. for C16H12Br2N406S: C, 34.8û; H, 2.92, Folmd: C, 35.25; H9 2.97;

~, 10 . 02 .

.~ ' ' , ' .

.

Prepar~tion of intermedl~te~ of ~r~u1ae OSO~CH
H . 3 23 ~ ~ .
3~ I mld - ~N ~C:H
23 1, l (aSC02CH20' .
.
OSO CH

N - I
3 ~
~ CH I
- - ~ N~
O CH
23.1~b) 2 2 . .... _ ---- .
A solution of triethylamine (101 mg.g 1.00 mmole) ~ 1,.4 ml, of methylene ~hloride was added wlth ~tirring 1:o ~1 801ution 0~ triflate ~10.11 (542 mg." 1.00 mmole) 5~4 ~ol. of m~thylene chloride at 0CO After al~owing ~he solution to w~r~ to 24 over 15 minute~, a ss:lu~ion oi~ iodine (254 mg., l.OO mrnole) in 7.5 ml. of methyle~e ~hloride was ~dded ~ith stlrring over 30 minu~Ps, then wJshed with water, dried, decolorized, filtered ssld the ~olvent evaporated in vacuo to give the di~odide ~23.1]
(588 mg.; 91% yield? $n gse~ter ^chan 95X purity. The IR ~nd NMR spestra were consisten~ for the pr~posed 8truc~ures.
~nal. Cslc'd. f~r C16H16N406~2S
N9 8.67; I, 39.28; S9 4.g6,.
Found: C~ 29.~6; H, 2"47;
N, 8.61; 3;y 39~37; S" 5.18.
.

'7 2~ Prep~rat-lon of intermedi~tes of formul~

~ ~ S2~H3 ~ 0~
- ~ ~ 'CHO
CO2~

BuLi¦ICA
C~3~2CH2~ 2 2 2 1.2 , 2.2 Butyl lithium (1.1 molesg 2~4 M in hexane) was ~dded under an atmosphere of dry nitrogen to 1 llter of dry . ~etrahydrofuran (freshly distllled from Lithium aluminum .
hydride) at -60C. I~ was ollowed by the dr~pwi~e ~ddition of isopropylcyclohexylamine (dried over pota~
~ium hydroxide p~iLIets~ ~200 ml., 1.1 moles~ and then benzyl acetate ~165 g.,l.l moles~. A~ter ~tirring for 3/~ hour at -60C., it was tre~ted fairly r~pidly w~h diethoxy ethyl acetate ~1~2~ (178 g., loOl mole~)~

The mixture w8~ ~llowed to come slowly to 20~. It w~8 ~hen cooled to 0~CO and acidified with 10% hydrochloric scid. E1:her wa~ ~dded and the ether extr~ct wa8 wa8hed w~th w~ter (twice) and then br~neO It was dried over ~nhydrous ~odium sulfate and evaporsted to gi~e 283 g~
(~00%)of ~-~eto e~ter l2.2l of ~uit~ble pur~ty to be used.directly~

' ' , ' 6j3~

, 1. P:repared by the method of Rr Moffett ~Org~nic 8yn~hesis, Coll~, Vol. XV5 po 42~) or more conven~er~ly by ~he m~thod o~E E. Bis~gni e'c ~1.
(Bull. Soc~ t:h~m" Fr. 1968, 637)., (~tO3 ~CHCOC~2C02CX20' ~ > (EtO) 2cHco~co2cH20 202 30~
A solu~iorl of the ~-keto e~ter 12.2J (283 g., 1.01 les) in gl~cial ~cetie acid (300 ml.) was cooled to 0C. and treated dropwise below 5C. with ~ solution of ~odium nitr~te tlOO g." 1.45 moles) in 250 ml. of wa~cer~
By-the end o the addition, the ~olid product has begun to ~onD.. After comin~ torroom temperature over 1 1/2 hours~ it was tre~'ced slowly with 400 ml. of water.
The sol~d was ollec~ed by filkra~ion and washed with old wa~er .
This solid was then dissolved in 1 liter o ether and washed w~th water and brln~. It wa~ dried over anhydrous ~odium ~ulfate and evaporated to give 288 g"
(g3%) of the ox~me [3.2] which was used without further pur~ fica tion .
For analysis, a ~ample was crystallized ~wice from ethcr/petroleum ether (b.p. 30-60) giving white needles w~th mOp. 95~97C"

.

q ~
Anal. Calctd. for C15HlgN06: C, 58.24; H, 6.~9; N~ 4.53.
Fouuld: C, 58.28; H7 6.37; ~, 4.510 8elective carbonyl (~0)2Cac~c~2cH2~ ~.eductin ~ (EtO~2C~CHOHCC~
~ith NaBH~ ~
~ H ' ~OH

3.2 4.2 A ~olution of the keto-ox~me r3.2] (288 g.~ 0~94 moles) in 1 liter of tetrahydrofuran wa~ cooled to O~C.
and tre~ted portionwl~e wi~h sodium borohydride (1708 g., 0.47 moles). After ~tirring ~or 3/4 hours st 0C., it w~ allowed to come to room tempersture over 1 1/2 hours.
It was then recooled ~o 0C.~ acidified c~efully with dilute hydrochloric,acid~ and extracted w~th e her~
The ether extract was washed with water ~nd brineO It wa~ dried over ~nhydrous sodium sulfate and evspor~ted to g~ve 241.5 g. (83%) of the alcahol [4.2~ whieh W~8 used without purlf~cation.

~rotection I HP
(E ~)~CHCHOH~COzc~2~ as with ~ (Eto)2cHcH~
bx ¢~
4.2 . 5~2 A ~olu~ion of ~he ~lcohol [4~2] (241.5 g.~ 0078 mol~s) ln 1 liter of ether W~8 tre~ted with dihyd~opyr~n . . ~ .

.

3'74L

.
, ' ~130 gO; 1~5 les~ ~nd then with 4.0 gO o~ p-t~luer~e-~ul~o~ic ~cid monohydrate. Some cooling w~ requlred to keep the in~tial react:ion at gentle reflux. When the ~nitial re~ct~on had ce~sed, ~t w~ stirred rooD~ temper~ture ~or 2 hours.
It was then poured OIltO 500 ml. o~ s~tur~ted sodium bicasbonate solution and the ether extract W~18 was~ed with w~ter and brine. It w~ dried over aTIhydrous sodi~ sulfate and ev~por~ted to give 356 g.
of the THP protected ~lcohol ~5.2] ~ It Wi~8 used a8 such in the ~ollowing reaction .
1. The crude m~xture contains [5.,2] as well a3 some di-T}lP compound ~6 .2 3 .

OTHP
(EtO) 2CHCHhC02CH20' OTHP
. 6.2 - .

OTHP Selec~cive ~TXP
~Et~)) 2C~CH~ 2CH2~ aSdWiti n ~ (EtO) 2 ::HCH~HcO
bH AI(Hg) N~2 5.~ ~.2 - A Isolution of the oxime [5.2] (64 g.; 0.143 moles) in tetrahydrofuran (200 ml.~ W~9 ~dded to ~ slurry of _ ~5 _ .. . .
.

;37gL

', ~lum~num ~malg~m (prep~red from 0072 mole~ of aluminum foil ~ccording to the m~thod o Vogel, "Pr~ctlcal Org~nic Chemistry", p~ 3-98) at such 8 rate ~8 to m~intain a vigorous re~c~on. A~ter stlrring for 3-4 hours, ~ fresh ~mount.of ~luminum am~lgam ~0.36 moles3 wa~ added at such a r~te ~8 to m~intain a vigorou~
react~on. After tirring for 16 hours, lt w~ filtered through celite w~sh~ng well with ether.
The ~iltrste wa evapo~ated ~nd the residue W~8 d~ssolved in ether ~nd tre~ted with a solution of ox~lic ., , acid (12.9 g., ~ol43 moles) in'ether. The mixture was the~ extr~cted with cold water twice and the c~mbined . ~queous extrac~ were acidified with concentrated ~mmonium hydroxide ~n the cold. It was then extracted with ether and the extract dried over anhydrou~ sodium .~ sul~a~e. ~vapor~tion gave 22.8 g. (4270) of the Rmine 7.2~

O~XP ~ I HP
~EtO)2CHCH~HC02CH2~ CH0 > (EtO)~CHCHI C02CH~
~2 7 a 2 ~ 8 ~ 2 pJ

N H
3~,, _ ~ , HP

. ~ X ~ OEt ', C02C~

9..2 .
~6 ... . .

63~74 The amine [7,2] (28.9 gO; 0.076 mole3) Wl~8 tre~'ced ~ith trans-clnn~m~ldehyde tlO.O g.; 0.076 mole~) in methylene chloride (200 ml.~ and refluxed ~or 2 hou~s.
It was then drled over anhydroug sodium s~ e for 8~veral m:Lnutes and ev~por~ted. The residue was dis~olved in a further 200 mi. of methylen~ chloride and then evapor~ted agsin. This procedure wa~ repeated t:wice more giving ~ residue with N~ showing almo~
complete Schiff base 18.2] formation.
The Schiff base ~8,2] was immedi~tely dlgsolved in ~ethylene chloride (200 ml.~dried over molecular ~eve~
~nd cooled to 0C. It was treated wlth triethyl~m~ne ~dried over potassium hydroxide pellets) (7.7 g.; 0.û76 moles) and then dropwi~e ~t O to 5C~ with a ~olution of azidoscetyl chlorlde (9.l g.; 0.076 moles) in dry methylene chloride (lO0 ml.) under.an atmosphere of ~ry nitrogen. When the ~ddition was complete, it was all~wed to come ~o room ~empera~ure for 16 hours and the~ xefluxed for 3J4 hour.
~ n cooling, the solution was w2shed with water, 170 hydrochloric acid, and brine. It ~as dried over ~nhydrous sodium sulfate ~n~ evapora~ced to g~ve 43.0 g.
(100%) of crude styryl compound ~9.2].

. . . - !

6 ~!74 .

.
,~ .

. ~ N3 - _ ~ 3 - C~IO
~ Ol~HP . 3 ~, ` ~ OTHP

; ~ N~ Et C ~J~OEt - CO~H;~0 C02C~
9.2 1002 The crude 8t:yryl compound ~9.2l ~14.4 g.; 0.025 moIes) was diL~solved in methylene chlorid~ (80 ml.) ~nd cooled to -60~C. ~n ~ dry ice: isoprop~nol bath~
Ozone was then bubbled through th~ 301ution until the appearance of a faint blue color persi~ted. It W~8 then flushed wlth vxyger. to remove excess ozon~ and ;; treated wich methyl sulfide (10~ to decompose the ozonide. It W3~ allowed to come to room te~persture over 4 hours ~nd then wa~hed wi~h w~er, 5% sodium biearbonate" water, ~nd brine. It W1~8 dried oves anhydrous sodium ulfate znd ev~por~ted. The re~ldue W~18 evaporated under high vacuum at 40-50C. for 18 hour~
to remove most of the benzaldehydeO The residue (12.1 g.) hsd ~ showing ~bout~ 50-607~ of fre aldehyde ~10.2] by ~ntegratiLon. It was used as such ir~ the next .~tep.

~: - 08.

.

6 ~ 7~

N H H selective N _ H
~; ~ ~ldehyde 3- - ,C~2OH
1 OTHP E reduceion ,r~ Q~HP
with NaBH4 ~ N~ ~ OEt C02C~120 ~t)2CH2d

10.2 11.2 The crude ~ldehyde ~10.2] (12.1 g.; 0.024 moles) was dissolved ~n tetrahydro~uran (100 ml.) and cooled to C. It was treated portionwise wlth powdered sodium borohydride ~0.46 g~; 0.012 moles) over a short period of time. After 1¦2 hour stirring at 0C., it was poured o~to ice-cold 10% ~cetic acid and the acidified mixture W2~ extracted with ether.
The ether extract was ~ashed with water ~twice), 10% sodi~n bicarbonate, and brine. It was dried over anhydrous sodium sul~ate and evapora~ed to give 10.9 g~, of crude alrohol ~11.2l~

.
H H hydroxyl ~ H
N3- -- CH2OH activation as N3- -~ CH2OSO2CH3 ~ith CH3SO2Cl > ~ Et ~:02CH2~ C02CH20

11~2 12.2 The crude alcohol ~ (12.8 g.) was di~solved .
,-g g 6;37~L

methylene chloride ~100 ml.) (dried over molec~lar ~eves~ and cooled to 0C. It was treated wlth dry ts~ethylamine t2.23 g.; 0,.022 moles) and then drop~ise w~th a solution o methanesulfonyl chloride ~2.51 g.;
a.O22 moles) in dry methy.lene chloride (30 ml.).
It w~s allowed to cosne to room temperature over 3 hours and then washed with water, 1% hydrochloric ~cld, and brine. It was dried over anhydrous sodium 6ulf~te and evapo~ated to give 11.8 g. of crude mesylated ~lcohol [12.2].

H H . H H
~ ~T'HP removal N3- - CH20s~2c~3 . ~ ~~~~ OTHP as wi~h ~ ~_~ff .: . 1 OEt l870 HCl in _ > ~ ~ OEt ~ _ N ~ ~ Et THF O' ~ OEt : bo CH ~ ` CO~CH2~
1~!o2 13~2 The crude THP-protected alcohol [12.2] (11.8 g.~ was : .dissolved in tetrahydrofuran (100 ml.) and treated wi~h coollng (keeping the temperature below 15C.) with 18%
hydrochloric acid (50 ml.)~ A~ter stirring fox an addi-. t~on~l hour in the cold~ thin layer chromatography showed ; los~ of the ~tarting m~terial.
It ~as diluted with water and extracted with ether~
. The ether extract was washed with water and brine. It wa~ dr~ed over anhydrous sodium sulfate and evaporated.
.

. .

~ 6~

The crude residue was pu~ified by dry-column chrom~tography on silica gel ~Activi~y III) us~ng ether as the solvent. This gave 3~5 g. of the acet~l ~13.2~. The y~eld is 28X overall from the amine [7.2¦.

H H H H
20502CH3 hydrolysis N CH20so2cH3 / OHas wi~h 3 ~
OEt 95% TF~ _ I OH
N ~ OEt ~ N ~ CHO

CO:!CH2~ ~02C~
1302 14.2 : The acetal ~13.21 (1.5 g.; 3 mmoles) was s~irred ~or 45 minutes with 5 ml. o 95% trifluoroacetic acid - an~ then tre2ted with methylene chlor~de and water.
The methylene chloride extract was washed with water, 10% sodlum bicarbonate, and brine. It was dried over : . snhydrous sodium sulfate and evaporated to give 1.0 g.
(78%) of crude hydroxy aldehyde ~14.2].

Preparation of intermediates of f~rmula H H Oso2cH3 . N3 ~ .

: ~ N ~ CH20CH2c6H5 CO~R' ~,. ..

~CH OH
2c2H . _ 2 ~ ~ H20CH CO H
~CH2~Na 2 2 1.3 2.3 Benzyl alcohol (6500 ml.) is added to a 12 liter 3-necked flask e~uipped with an efficient stirrer, condenser and a heating m~ntle. The alcohol is heated above the melting point of sodlum (^-100-110C.~ and sodium metal (600 g., 26 moles) is added in small pieces wlth vigorous stirring at a rate sufficlent to maintain a gentle re~lux and to prevent the accumulation of large ~mounts o~ sodium in the reaction flask (~ 3-4 hours).
The solution is then cooled to about 80C. and chloro-acetic ~cid ~1.33 (1120 g.; 12 moles) dissolved in ~he minimum amount of benzyl alcohol is added dropwise fairly rapLdly. The mixt~re is heated under reflux for 4 hours, cooled to room temperature ~nd diluted with 10 liters of ether. The precipitated sodium salt o benzyloxyacetic acid is collected by ~iltration~ washed with ether, dissolved in cold water and acidified to pH 3 with concentrated HCl. Extraction with CH2C12, drying and concentration gives 1952 g. (98% yield) of ~enzyloxyacetic acid ~2.3] as a crude oil which is esterified with~ut further purification. If d~sired, the ac~d can be purified by distillation: b.p. 130- .
132C.l0.1 mm.

; '' ' .
.. . .

1 1~63 Ethanol ,~( H20~H2CO~H ~ ' ' > IZfCH20CH2C02Et 2.3 3~3 Ethsnol (7 liters) in a 12 li~er l-neck flask ~qu~pped with condenser and drying tube is saturated w~th dry HCl. Benzyloxyacetic acid ~2.3~ (1952 g., 11.8 mole) is added ln one portion and the solution is heated under refIux for 5 hours. Most of the ethanol ~5 removed by distillation at normal pressure. The residue is cooled to about 5C. and diluted with about 4 I. of cold water~ Ex~raction with ether, washing of ~he ether with cold waterS saturated Na~CO3 solution and ngain with cold water, drying over Na~S04 and concen- -tration yields an oil which is purified by distillation to glve 1425 g. (62% yield) o ~3.3]; b.p. 138-140C./
8 mmO The NMR spectrum indicated a benzyl -CH2:
singlet at 4.57~ ~ aroma~ic H at 7~3 ~ , O-CH2-CO- at 4.07 ~ and an ethyl quartet and triple~ cen~ered at 4.18 and 1.25 0~ .

p'CX20CH2Ct)2Et -- ' >0~CH2OCH2~ C~2CO2C~2 ' O
3~3 4.3 All operations ~re carried out under an stmosphere o~ dried N2. A reaction flask (12 1. 3-neck flask with -- 9 3 . --stirrer, N~ inlet, 500 ml. addition funnel and cold temperature thermometer~ is charg,ed with 3 1. of drled tetrahydrofuran a~d cooled to -78C. in a bath of dry ~ce-2~c~0n~. Three bottles of ~,uli (2~4M in hexane, l.mole~bottle~ are added via a ~lexible adaptor under N2. N,N- isopropyl cyclohexy~amine (423,,8 g., 3 mole) added via rhe addition ~unnel over a pe~iod of . . .
about 15 minutes snd stirring is continued or 15 minutes.

Benzyl acetate (450.5 g., 3 moIe) is then added dropwise :-: over 2 30 m~nute period and the solution is stirred ~or 30 minutes. E~hyl benzyloxyacetate ~3.3] (555.0 g., ~: 2.85 mole~ is added dropwise over about 30 minutes and the cooling bath is removed at ,,he end o~ the addition.
.. . . . .
': ~ The reaction mixture is stirred and~allowed to come to . room temperature in abou~ 2 hours. When the internal temperature reaches 0-5, a voluminous solid separates.

A~hydrous ether (about 3 1.) is then added to complete ~: the precipitation. The solid is collected by flltra~ion . ~ ~nd washed with ether on the filter.

The solid is added to a vigorously stirred mixture ~:.
- of 250 ml. of concentrated HCl, 500 ml. of water, 500 ml.
of ice and 3 1. of ether. When s~lution is complete . the pbases are decanted and the aqueous phase is extracted ., ~ once more with ether. The combined e~her layers are washed with brine,, dried over N~2S04 and concentrated to g q, ' ' ' ' ' ' ` . , 1~6 ~

leave 659 g~ (78%) o oily benzyl ~-benzyloxyaceto-3cet~te~ [4.3]: TIC: (silica gel~ ether-pet. ether 2,.1~: 1 spo~, Rf. 0.5, ~ ~ indicates (aromatlc H a~
7,23 ~; benæyl ester C~2~ singlet a~ 5.05 ~ ; benzyl ether CH2, singlet at 4.42 ~ ; 0-CH2C0, singlet ~t 4.0~ ; and COCH2C029 singlet at 3.48 ~.
This ~il is used without purification in the next 8tep. Distill~tion of large amounts proceeds with ~xtensive decompos~tion but a small amount ~5 ml.) can be purified by distillation if necessary: b.p. 180-182/
0.05. The TLC and NMR of the crude product and of a distilled sample are identical. New spots appear on TLC if this oil is s-ored for some time.

.- .
HNO
2 ~ 0CtI2CH2~ C )2CH2~( O O NOH
4.3 5.3 A solution o~ sodium nitrite (165 g., 2.4 mole) in 660 ml. o water is added to the cooled ~-~10C.) solution of benzyl ~-benzyloxyace~ate from the step ~mmediat:ely above in 1100 ml~ of acetic acid. Th-e rate o~ addit:ion is regulated so that the temperature of the re9ction mixture does not exceed 25C. A white solid crystallizes out w~en about one-hal~ of the nitrite 801ution ~s added. A~ter the end of the addition the ~63'7~

'~ ' ; ' mixture ls stirred ~or an additional 30 minutes at lO'' ~nd diluted slowly with 2 1. of water. The solid oxim~
i~ collected by ~iltra~cion washed with cold water until the washings are neutral, and dried in vacuo (16 hours) t~ give 856 g. of oxime as a white solid. The o~ime is dissolved i~ methylene chloride~ the water removed by decantation, the org~nic phase washed with brine~
: dried over Na2S04 and concentrated until crystals st~rt appearing. Crystallization is completed by dilution with petroleum ether (30 - 60)~ Filtrati4n, washing : with petroleum ether and drying affords 585 g. t81%~
o~ oxime ~5.3~; m.p. 92-95, NMR indicates arom~tic H
at 7.27 and 7030 ~, benzyl ester CH2 at S.27~ an~ ether . CH2 at 4.5~ . This material is used in the nex~ seep.
: An analytical sample prepared in benzene-petroleum ether had the following properties: m;p. 96-g7.
Calcld for C17H18N05: C, 66.05; H, 5.23; N, 4.280 Found: C, 66.06; H, 5~25; N~ 4.23.
, -CH2~ co2cH2~ H2~GH2-~H C-C02CH20 NOH OH NOH
5.3 6.3 The keto-oxime ~5.3] (400 g., 1.2~ mole) dissolved ~n 1000 ml. dioxane is cooled to 5, stirred vigor.ously .
~nd t~eated port~onwise with finely powdered sodium ' . .

borohydride (23.4 g., 0.62 mole) over a 3~ minute period. The temp~rature remains at 10~ for a while ~nd then a vigorous reaction sets in which must be controlled with an ice-~alt bath (temperature goes up to 50-fiO). After 1 hour TLC (sllica gel, ether-petroleum e~her) shows disappearance of the starting material. The mixture is stirred an additional hour, poured lnto ice-cold dilute hydrochloric ac~d 3nd extracted with ether (2 x 1 1.). The combined extr2cts .are w~shed.with w~ter (5 x 1 1.) and with brine. Drying and concentration leaves a tan-colored solid which is recrystallized in benzene-petroleum ether to give 308 g.
(77% yield) o~ the alcohol-oxime [6.33 as a white solid, : m~p. 83-85. NMR indicates aromatic H at 7.23~ , ester : CH2 at 5.18~ , benzyl ether at 4.48~ 9 (triplet); H of - CHOH at 4.67~ , CH2 ether at 3.65~ and exchanageable H at 3.42 and 10005~ .
Calc'd f~r C12HlgN05 C, 65-64; H, 5.81; N, 4.250 ~ound: C, 65.62; H, 5.91; N, 4026.
. , : - 9 7 -... ..

, ,.

o~ ~ ~
~CH20CH20' --~ I R~H~ P "

02CH2~ , -I CH20C:H25~

6.3 ~ 7~3 H ~ I .

To a welI-stirred suspension of hydroxy-oxime E6.3~ (298 g., o.gl mole~ in dihydropyran (650 ml., 7.1 mole~ are added 15 to 20 drops of concentrated HCl and the mixture is stirred at room temperature.
After l/2 hour a clear solution is obtained and after 4 hours TLC indicates disappearance of the starting hydroxy-oxime. . The solution is diluted with technical ether (1.5 l.) and poured into a 10% NaHC03 solution~
The ether ph~se is washed with brine, dried and concen-trated to leave 402 g. of a yellow oil which consists of ~ mlxture of mono and bis tetrahydropyranyl derivatives.
The above oil (394 g.) dissolved in ether (technical 600 ml.) is added dropwise to freshly prepared alumlnum ~malgam covered wlth technic~l ether (about 1000 ml.) (the amalgam is prepared using 120 g. aluminum foil 3S
described in Vogel~ Practical Organlc Chemistry) at a .. . .
_ 9~,_ , - , , . , :

0~ 37~

r~te su~icient to maintain a vigorous reflux. The mixture is stirred 2 hours after the end of the ~ddition~ After removal of the insoluble mater~al~
by fil~ration o~er Celi~e, the filtrates are dried over Na2S04 and ~reated w:~th anhydrous oxalic acld ~72 g., 0.8 mole) dissolved in the minimum volume o~ e~hanol. After two hours at 0~5 the solld oxal~te salt is collected by filtration, washed wlth ether ~nd dried to give 215 g. of wbite solid, m.p. 115-119.
This crude oxalate is used for regeneration of the ~ree base.
An snalytical sample of the oxalate, prepared in methanol-ether gives: m~p. 136-137.
23H29N05.C2H204: C 61.34; H, 6-38; N 2 86 Found: C~ 61.2S; H, 6.50; N~ 2~76.
The free base [8.3~ is regenerated by adding the solid oxalate to a well s~irred mixture o~ an excess of ice-cold concentrated ammonîum hydroxide and ether:
152 g. (42%3, yellowish heavy oil. NMR indicates 2rom~tics:at 7.30 ~, benzyl ester CH2 at 5.1 ~, benzyl ether CH2 at 4.47~ and NH2 at 1.82 ~.

, _ 99 _ , ~7 ~

OTHP J I
~2 ¦ J~--~HO ~ OT~P
~f CH20CH2~ ~ N~H20C:H2~( C02C:H2~ O~!CH20 8 3 C,oG3/ 9 3 ~/

~OTHP
.~ od~N~ CH20CH2.
CO~ CH
10.3 A mixture of the amine ~8.33 (117.2 g.; 0.294 mole~
~nd cinnamaldehyde ~3808 g~; 0.294 mole) in 5.00 ml. of reagent grade CH2C12 was refluxed for one hour. The solvent was removed on ~he rotary evaporator and the residue redissolved in 500 ml. of fresh CH2C12. Part of ~he solvent was distilled at atmospheric pressure ~350 ml.~ snd the rem~inder was taken to dryness on the rotary evaporator to give an oil consisting of crude f g . 3 ~ ~
The 3bove oil was dlssolved in 400 ml. of CH2Cl~, -trie~hylamine (45 ml., 0.32 mole) was added and the 601ution was cooled to 0G. A solu~ion of azidoacetyi chloride (38.4 g~; 0.32 mole) in 200 ml. of me~hylene ~hloride waq added to the above cooled and ~tirred -- 100 - ' i solution in 1.5 hour. The! mixture was allowed to come ~1QW1Y to room tempe~ature and was stirrsd for 16 hoursO
It was then heated under reflux fPr 1 hour, cooled, ~sshed ~ith ice-cold water, with ice-cold satura~ed ~odium bicarbonate solution and with ice-cold brine 801ution, dried over Na2S04 and concentrated on the rotary evaporato~ to leave 171. 2 g . (98%) o~ an oil whose IR and NMR spectra are consistent with struc~ure ~10~3J.
I needed, puriflcation can be e~fected through chromatography over a column of silica gel. The crude product is normally used in the next step..

~/a~HP ~3;~C80 H2,0CH2~ N`~`CH~20CH2~' 02CH20 co2c~
10~3 11 . 3 A solution of the crude styryl compound l1003J
(117.6 g., 0.198 mole~ in 600 mll o~ CH2C12 was cooled -to -60C,. and ozonized until a faint blue color appeared.
Dimethyl sulfide ~75 ml.) was added and the solution was allowed l:o come slowly to room temperatureO After lB

hours~ the solution was washed sever~l times with iee-cold water, dried over Na2504 and concentr~ted on the ' ' .

,, . - 1 o 1 ~ .

~ 6 ~
;' .

.. . . .
: rot~ry evaporator to leave 104 g. of an oil. The pure ~ldehyde rll.3] (34.6 g~) was obtained ln 3270 yield ~fter chrom~tography on 12~0 g. of ac~ivity 2 8ilic~ gel using the dry column technique and ; eluting with ether-petroleum ether.
; ' M3 H H Selective H H
HP as with NaBH > N3 - - H
~CH20ca21~ ~2C1~2Ç~

1~.3 12~3 .
; The pure aldehyde ~11.3] (35.5 g., 0.068 mole) was : dissolved in 250 ml. of reagent grade THF, cooled to -5C., and treated porti~onwise under good stirring : w~th sodium borohydride (1.29 g., ~.034 mole). After one-half hour, the mixture was acid;fied with ice-cold ; 10% acetic acid and was extracted with ether. The combined ether extracts were washed with ice-cold water and ice-cold 1% NaHC03 solution, dr,.ed and concent:rated to leave 33.4 g. (94%) o crude alcohol . ~s ~n o:~l whose NMR spectrum is consiste~t with s~ruc~ure

12.31.
Thl~ alcohol was purified by column chromatography o~er si:Lica gel.

~.' ' , ' .
.' ' , ' '' ''' "'.

7~

N3 - ~ OH Hydroxyl act~vatlon H H
OTHP s by treatment with ~ 3 ~ SO2CH3 N ~ H20CH2~ meth nesulfonating ~ N ~ C 20CH

~2.3 1303 The purified alcohol [12.3] (15 g., 0.0286 mole) was d~ssolved in 100 ml. of CH2C12, cooled to 0C., treated with triethylamine t4.2 ml., 0.0~ mole~, and ~reated dropwise with methanesulfonyl chloride (3.44 g., 0.03 mole) dissolved in 50 ml. of CH2C12. A~ter 2 hours, the mixture W3S washed with ice-cold water and ice-col~
brine solut~on, dried and concentrated on the rotary evaporator to g~ve 15.7,g. (9270) of a thick syrup.whose NMR spectrum is consistent with structure [i3.3].

.
H H H H
N3 - _ SO CH Removal of protecting pTHP acid hydrolysis ~ ~ SO2CH3 , ` ~ H2CH20' ~ ~CH 20CH20 CO~C~23~ C02CH20

13~3 14.3 The mesylate [13.3] (15.5 g., 0.0257 mole) was dissolved ~n 1~0 ml. of THF and treated with 50 ml.
o~ 10% hydrochloric ~cid~ After 3 ~our hour stirring period, the solution w~s extracted with ether and the ~ 6 ~7 4 .

combined ether extracts were w~shed with ~a~e~ ~nd ~rine solution~ dried and concentrated on the ro~ary ~vaporator to leave 13;5 g. of an oil whose NMR spectrum - i8 consistent with structlure ~14.3~.

~f desired, the alcolhol [14.3~ can be purified by . column chromatogr~phy.

H H . H H

3~So2cH3 N3 _ ^
Oxidation as w~th ~> ~~ 2 ~N~I~cH OCH ~0 CrO3 ~ H~OCH20

14.3 15.3 The crude alcohol ~14.33 (17.4 g., 0.033 mole) dissolved in 170 ml. of r~agent grade acetone was treated, .~ under vigorous stirring, with a total of 12.3 ml. of Jones Reagent (CrO3-H2SO4-H2O, 0.033 mole) at such a rate that the Reagent was consumed before the next drop was added (orange to green). The mixture was diluted w~th water and extracted with ether. The combined ether layers were extracted with 140 ml. of ice-cold ~ . lZ NaOH solution, the basic ex~rac~s acidified immedi-asely wit:h ice-c~ld 1~% hydroohloric acid and extracted "` with e~her. Concentration of the neutral ether extracts gave 10,8 g. of an oil consisting ma~nly in unreacted 114.31. These were reoxidized as above. The combined .

637'~
.
. .

ether extracts containing the acldic compound were dried snd concentrated to give ~ totsl o~ 4. 6 ~S . of crude enol [15.3] as an o:il whose IR and N~ are co~sistent wi~h structure ~15.33.
: , '.

:; , . .

6 ~4 . , _-carboxylate ~from dibromide intermedi~
- ' , H H
N ~

~CH20COCH3 The mixture of dibromide starting materials rl2.1 and 13.1~ ~1.7 g.; 3 mmole) in 50 ml. of dimethylformamide SDMF) was stirred with potassium acetate ~1.2 g.; 3.5 equiv.) for 16 hours at room temperature. The mixture was then dilu~ed with 100 ml.. of diethylether ~nd 100 ml.
of~10% hydrochloric acid. The~ether layer was washed w~th 10% sodium bicarbonate solution (once; 50 ml~) an~
with brine (once; 75 ml.). The extract was then dried and evaporated in vacuo to leav~ 1.0 g. of a dark brown residueO This product was purified by passing through æhort column of activity III silica gel with methylene chloride elution. This gave a fraction contain~ng 700 mg.
~63~ yield) of a colorless oil whose spectral data confirm that.it has the tructure of the title product.
Crystallization from 2:1 benzene:petroleum e~her g~ve a white solld; m. p . 94-97~C O

- 1~6 -.

' Anal~ Calc'd for C17H16N406 Found: C~ 55~19; H, 4O47; N, 14.8g.

1. The potassium acetate used was analytical grade ~hich had been flnely ground before use. ~ater Wa8 probably absorbed by the acetate during this process.

~ .
Example 2 en ~ 0-2 carboxYLate (from diiodide intermediate) Diiodide intermediate ~23.1] (600 mg., 0O93 mmole) prepared according to the method described above w~s added to a solution of potassium acetate (380 mg., 3O9 mmole~ in 5 ml. of di~ethylformamide and stirred at 24C. ~or 20 hours. The solution was mixed with ether (25 ml.) and washed with water (5 x 25 mlO).
The ~ther mixture was dried (sodium sulfa~e) and evaporated in v cuo to give crude benzyl 7~-azido-3-acetoxymethyl- ~3-n-2-isocephem-4-carboxylate. NMR
i~d~cated approxLm~tely 35~ yield. NMR and TLC
est~blished th~t the product is identical with that produced in Example. lo ., ~ 6 3 7~
~ . . . .

Ex~mple 3 4-carboxylate H H

~ ~ CH20CHO

Sodium formate (2.86 g~, 42 mmole) was added to a 801ution of the dibromide mixture [12.1 and 13.1~ (2~86 g., 5.05 mmole) in DMF (10~ ml.) containing water (O.I ml.3.
The solution was stirred at room temperature for 72 hours, diluted with wa~er (100 ml.) and extrac~ed in ether ~5 x 75 ml.). The combin~d organic phases were washed with brine, dried (Na2S04~ and concentrated to leave 2.52 g. of an oil. The oil was purified by column chrom~-tography on silica gel (100 g.) eluting with 3:1 ether:
petroleum ether. The main fraction contained 0~70 g.
o~ pure benzyl 7~-azido-3-~ormyloxymethyl-~3-0-2-isocephem-4-carboxylate as a colorless oil. The NMR
spectrum of the product was in agreemen~ with the structure of the title product.

.

.

H

~H20Cl~CI::20C jj~5 Dibromide mixture [12.1 and 13.~3 (~05 g.~ 170~
~mole) was dissolved ~n 100 mi. o DMF and treated with .
p.otassium phenoxyace~ate (8.2 g., 43.0 m~ole). The reaction mixt F e was st~rred at 50-60C. for 18 hours.
On cooling, ~t was poured onto 500 ml. of w~ter and extracted with dichloromethsne, The extracts were washed sever~l t~mes with water and ~inally with ~rine.
Drying over anhydrous sod~um sulfate and evaporatlon ga~e crude product which was puxified by "dry-colun~"
chrom~ography on Activity III silica gel elu~ing ~th ether. The product was ob~ained as a middle cut ~3.1 g~9 38% yield) which was about 60% pure by ' ; ' ' ' , . .

''~ ' `'` "

., , '` -~

~ 3~
. , ' ' , .

~,5 meth~l- ~ -0-2-is DCG
4-carboxyla~e .. . .
H H

~; 3 , . '~7l~CH2H

. The crude hydroxy aldehyde starting material ~14.2 (50 g., 0.117 mmole) prepared above and powdered potassium aceta~e (50 g., 0.51 mmole) were s~irred in 350 ml. o~ dimethylformamide for 18 hours. The mixture .. W9S then diluted with water and extracted with methylene .~ . chloride. The extract was washed wi~h water sever31 times and dried over anhydrous sodium sulfa~e.
:- Evaporation gave 40 g. of a residue which was .
:~ pur~fied by dry-column chromatography on silica gel (Act~vity III) using ether as the solvent. This gave 7t0 g. (19% yield) of pure title product as a pale-yellow solid.

This compound was ~ound to be identical (IR, NMR~

. mixed m.p.), to the compound prepared in Example 8.

: For analysis, a s~mple was crystallized from ethyl .
~cetate/petroleum ether (30-60) giving white crystals with m.pO~88-90C.

Anal. Calc'd. for Cl$Hl4N405: C, 54.54, H~ 4,27; N, 16.96.

Found: C, 54.49; H, 4~23; N, 16.97~

, . - - .

~U~

.

~ 6 Benzyl 7~-Azido-3-benzyloxymethyl-~3-0-2-isocephem-4-carboxylate -H H
N - "~
. ~ O

C02CH2~

The crude enol starting m~terial ~1S.3] (4.6 g., 0.089 mmole) was dissolved in 75 ml, ~f CH2Cl2, treated with triethylamine ~1.55 ml., 0.11 msle) and heated under reflux for 3 hours. After cooling, the mixture ~as washed with 1070 hydrochloric acid, water~ 3%
NaHC03 solution and brine~solution. Drying (~a2S04) and coneentration left 3.56 g. of cr~de material which was purified by chroma~ography over silica gel to give 1.05 g. (28% yield) of pure ben~yl 7~-azido~3-benzyloxy-methyl-~3-0-2-isocephem-4-carboxylate~ IR and NMR of th~s compound were in agreement with the strueture of ~he title product.

' . Example 7 8e~zyl 7~-Azido-3-acetoxymethyl- A3-0-2-isocephem-4-carboxylate (acetylation of benzyl 7~-azido-3-hydroxy-meth~l-~3-0-2-isocephem-4-carboxylate Ben~yl 7~-Azido-3-hydroxymethyl-~3 0-2-i~ocephem-~ 37~

, 4-carboxylate (0.2 g.~ 0.6 mmole) was treated with 8 ml. of pyridine and them acetic anhydride (0.2 g., 2.0 mmole). A~ter stirring at room temperature ~or 3 hours, the re~ction mixture was diluted with wa~er ~nd extracted with ether. The extract was washed with cold 5% hydrochloric acid9 water, 1% sodium bicarbonate, and brine.. It was then dried over anhydrous sodium 8ulfate and evaporated to give 0.18 g. (83% yield) o~
the acetylated title product.
This compound was found to be identical (IR, NMR, mixed m~p.) to the compound prepared in Example lo Example 8 Benz~l 7~-Azido-3-hydroxymethyl-~3-0-2-isocephem-4-carboxylate (aci_ hydrolysis method) A) A solution of benzyl 7~-azido-3-formyloxymethyl-Q3 0 2-isocephem-4-carboxylate (2.0 g.) in acetone (15 ml.), water ~15 ml~) and concentrated hydrochloric acid (2.0 ml.) was stirred at room temperature for 2.5 hours. l'he solution wa~ extracted with CH~C12 (3 x 25 ml.) and the c:ombined organ~c extracts were washed with-brine, dried (NazSO~) and concentrated to give 1.77 g. o~ an oil whlch w~s purlfied by coiumn chromatography on silica gel (80 g.) to give 1.0 g. (52% yield) of pure title product identical (IR~ MMR, m~pO) with a sample prepared ~18~

, ~n Example 3 by cyclizatlon.
B3 The procedure of Part A wa~ repeated excep~
that ~he starting material was benzyl 7~-azido-3-~ acetoxymethyl-~3-0-2-lsocephem-4~casboxylate ant ; the resction time was 48 hours. The t~tle prod-~ct w~s produced in 20% ~leld.

I~
' ' ' , .~'' ' , ' , ,, ' .
~ . . .

: . ' ' E~9 te H

N ~ L CH20S02CH3 CO~CH2 ~P~:2 A solution of methanesulfonyl chloride (0050 ml., 6.5 mmole) ~n 10 ml. of methylene chloride was added dropwise with stirring to a solution of p-nit~obenzyI
7~-azido~3-hydroxymethyl~3-0-2-isocephem-4-oarboxylate 2.41 g., 6.43 mmole), triethyIamlne (0.97 ml., 7.0 mmole) and 75 ml. o~ methylene chloride at -10. After 1/2 hour at -10 and 1 hour at 24, the solution was washed with 5% hydrochloric acid, 2% sodium bicarbonate, ~nd ~ater ~85 mlO each), then the solvent was evaporated in vacuo to give the mesylate t~tle product~ 2,86 g.
(9870 yield), as a yellow foam. The NMR of the product ~s ~n agreemen~ with the proposed structure~
The! p-nltro~enzyl 7~-azido-3-hydroxymethyl-~3-0-2-$socephem-4-carboxyla~e starting material used above ' '' . . .

~6 3 ~4 may be prepared as ollows:
lo The dliodide ~ntlermediate of the ~ormul~

qS2CH3 ~3 ~
I

~N ~L--~H I
C~2CH2~ 2 w~8 prepared rom p-nitrobe~zyl ~cetoacetste acco~ding ~o the procedures o~ Preparat~ons 1 and lA (S.tarting M~terials) described above.
2. The dilodide intermed~ate (6.6 g.~ 9.6 ~mole~
was cyclized with potasslum form~te (2.54 g., 3V mmole) in a solution o~ 100 ml. DMF and 0.1 ml. water at 0.
After stirring for 5 hours with the cooling bath re.moved, the mixture was p~ured into 100 ml. of cold water and extracted w~th methylene chloride. After washing with w~tex conta~ning a little N~Cl, drying and evaporation in vacuo~ p-altrobenzyl 7~-azido-3-formyloxymethyl-~3-0-2-isocephem-4-carboxylate was recovered (5.3 g.) as brown o~l~
3. To a solution of 5~3 g. o~ the 3-formyloxymethyl ~ntermed~ate in 53 mlO o~ acetone w~s added 26 ml. o w~ter and 3~2 ml. of 12M HCl. The mixture was s~irred ~t 24 ~or 7 hours, then poured into 100 ml. w~ter and extractecl wlth methylene chloride. The combined extracts :: .

. .

~ 6 3 7 ~

were washed with w~ter containing a little sodium chloride, dr~ed and evaporated i~ vacuo to give 3.6 g. of a brown oil. The oil was absorbed from methylene chloride onto LB g. of sil~c~ gel and placed on a 72 g. silica gel column (grade 3, 5%
ether). The ~olumn was eluted with 200 ml. o ether3 ~hen with ether/ethyl acetate 3:1. The m~jor component (Rf 0.20) gave, on evaporation of the salvent in vacuo, yellow solid which was recrystallized f~om acetone-ether to give the 3-hydroxymethyl start~ng material of this example, 950 mg. ~17~5~Jo yield from the diiodide). m.p~ 147-148.

nalO Calc'd, for C15H13N~07: C, 48.00; H~ 3-49;
N, 18.66.
Found: C, 48.11; H, 3.61;
~; 18. 810 63~fi~

' ' ;. . .
~10 yl ?~-Azido-3-hydroxYm~thyl-~3-H

N
' ~0 An a~ueous solution of 0.05N sodium carbonate ~20 ml., 1.0 mmole) was added dropwise with st~ring to ~ solution of p-nitrobenzyl 7B-azido~3-hydroxy~
methyl-~3-0-2-isocephem-4-carboxylate taS prepared in Example 9) (0.38 g., 1.0 rlmlole) ln 50 ml, tetra-~- hydrofuran. The addition~ took 15 minutes and the solution was stirred for an additional S minute~, then d~luted with 20 ml~ brine and 2~ mlO ether. Th~ phases : ~ere separated and the organic phase was washed wi~h ; water and brine,: dried (Na25O4) and evapora~ed in vacuo to glve 0.2~ g. o~ a solid which was chroma~ographed ~dry~col~mn me~hod) on 15 g. ~ilica gel (activ~ty III) eluting with ether:ethyl acetate 3:1. The lactone (0.16 g., 72Z~ was obtained as 8 white crystalllne ~ol~dO M.p. 178-179d (ethyl acetate-ether~.
Ansl. Calc'dO for C8H6N4O4: C9 43.25; H~ 2.72; N~ 25~22O

Found: C, 43.10; H, 2~75; N~ 25.450 ' ~ ~.17 _ r ~L ~L¢63t74 .

.

carbamoyloxyme~y~ 3oo-2 .
~3~

: N ~ CH20CONH2 Gl2C~o2 p-Nitrobenzyl 7~-Azido-3-hydroxymethyl-~3-0-2-~socephem-4-carboxylate ~375 mg., loO mmoles) was dissolved in 15 ml~ o~ benz~ne and treated with sod~um cyanate (130 mg., 2.0 mmoles) followed by trifluoro~
~cetic acid ~0.16 ml., 2~ mmoles). After stirring or 2 hours, the reaction mixture was treated with 25 ml.
- o water and 15 mlO of ethyl acetate. The aqueous layer W8S extr~cted once wlth 15 ml. o~ ethyl acetate ~nd the combined organic extracts were washed with water and brine. Drying was over anhydrous sodium 8ulfate and evaporatlon in vacuo gave 400 mg. (9~%) of crude c2rbam~te product which showed only a minor impurity by thin-layer chrom~tography (silica gel, 10%
ethyl 8cetate in ether).

, ~ ' .

. . .

~ .
p-Nitrobenz 1 7B-Azido-3~ methvl H H

~Lc~2-o~

~ 02CH2~102 p-Nitrobe~zyl 7~-azido-3-hydroxymethyl-~3-0-2-~socephem-4-carboxyiate (0.70 g. 9 1~ 86 mmole) was dissolved in methylene chloride (S0 ml.) and triethyl-sm~ne (0.24 ml~ 8S mmole) was added. While stirring the solutio~, there ~as added dropwlse (i~ 5 minutes) solution of methanesulfonyl chloride (0D145 ml., 1 D 86 mmole) in methylene chloride (10 ml.). The resulting ~olution of p-nitrobenzyl 7~-azldo-3-methyl~ulfonyl-oxymethyl-~3-0-2-isocephem-4-carboxylate was s-ir~ed at room temperature ~or 1 hour, (pro.ected ~rom moist~.e by ~ calciu~ chloride drying tube). At ~he end of this time, an additional quantity of trie.hyla~ine (0.24 ml., 1.86 mmole) was added followed by a solu~ion of l-methyl-te~rszol-5-ol (0~186 g., 1~86 mmole) in methylene chloride (2~ ml.). The solution was stirred at room temperature ~or 5 days and the:~ the methylene chloride was ,emoved by evapor~t:ion in vacuo and replaced by chloroform (100 ml.). The solution was refluxed 20 hours, then cooled, wa~;hed successively with 10% HCl, w~ter and -- llg --brine, dried (Na~S04~ and e~aporated to dryness leaving 8 d~rk semi~solid tO~80 g.). This was chromatograph~d (dry~column) on silica gel III t25 g9) J eluting wi~h ether:ethyl acetate 3:1. The pure compound was obtained 88 ~ whlte solid, m~ p. 174-176d ~ethyl acetate). N~
8howed the compound to be the t~tle product.
~nal. Calc 'd. for C17H15N907: C, 44.64; H, 3O30; N, 27~56"
E'ound: C, 44.87; H, 3.48; N, 27~,59.

Exa~ le 13 BenzYl 7~-Amino-3-acetoxymethyl~ 0-2-isocephem-4-carboxylate ~. .
H R

2 ~ C1~0COCX3 ;; ~H20 S gas was slowly introduced into a stirring solution o~ benzyl 7~-azido-3-acetoxymethyl-~3-0~2-isocephem-4-carboxyla,te (793 mg.~ 2.1 mmole), triethyl-amine (210 mg., 2.1 mmole~ and 15 ~i. of methylene chloride for lO m~nu.es. The flask was then flushed with nltrogen to remove excess H2S and then evapora~ed to dryness in vacuo. The residue was partitioned between diethylether and 10/~ hydrochloric acid three ~imes. The combined acid extracts were backwashed twice with diethylether and then carefully alkalized with solid sodium bicarbona~e, saturated with sodium chloride ~ 63~

and extracted with methylene chloride (three times, 75 ml. portions). The ct~nbined extracts were washed wlth brine (once, 75 ml. portion), dr~ed tanhydrous ~od~um sulfate) and evaporated in vacuo to glve 415 mg.
(60% yield) of title product as a colorless oil.
Spectr~l data were consistent wi~h the assigned structure~
Anal. CalcId. for C17H18N206: C, 58.95, H, 5.24; N, 8.09~
Found: C, 58.39; H, 5.32; N, 7.950 . . .
Exam~le 1 ` ~ ' BenzYl 7B -Amino-3-benzy : 4-carboxylate H H
2~ ~ r 2C~2~
` ~ C2GH2~
.` Benæyl 7~-azido-3-benzyloxymethyl- ~3-0-2-isocephem- -- 4-carboxylate ~1.45 g., 3.5 mmole) was dissolved in 150 :- ml. o~ dry CHzCl~ and cooled to 0C. Triethylamine . (1 ml.~ 7 mmole) was added and, while stirring and cooling, H2S gas was passed through the solution until it was saturated. The solution was then allowed to come to room ternperature in 2 hours and concentrated on the rotsry evaporal:or. The residue was partLtioned between 10%

.

- !

, ;;37~

hydrochloric acid ~nd e~her. The mixt~lre o solld and ~queous phase was sep~rat~d and combined with the ~ubsequent aqueous w~shings of the ether phase. The . solids and combined aqueous phases were made basic with solld NaHC03 and extracted with CH2C12. The extracts were dried tN~2SC)4),and concentrated to leave 0.80 g. of crude title product. The NMR o~
the sample was in agreement with the structure proposed~

Example 15 Benzyl 7~-Amino-3-hydroxymethyl-~3-0 2-isoce~hem-4 carboxylate H H
2 ~ 0 . . ~ ~CH20H

; Benzyl 7~-azido-3-hydroxymethyl~3-0-2-isocephem-4-csrboxylate (1.0 g., 3 mmole) was dissolved in 60 ml.
~ ; of dry CH2C12 and cooled to 0C. Triethylamine (0075 ml., : 6 mmole) was added and, while stirring and cooling, H2S
g2S was passed through the solution until it was satur-~ted~ The solution was stirred at room temperature for 30 minutes and concentrated. Fresh CH2C12 ( ~30 ml~) was added and the solution was a8ain concentrated. This .

~ 6 3 7 ~
.

operatlon was repeated ~ second time. The residue w~s 8 yellow oil consisting oi. cr~de t~.tle product.

~ 16 .

7~-Amino-3-acetoxymethyL- ~ 3-0-2~isoce~hem 4-carboxylic scid H H
H21J~

N ~ H20COCH3 Benzyl 7~-azido 3-acetoxymethyl-~3-0-2-isocephem-4-car~oxylate ~S.0 g., 13.4 mmole) was dissolved ~n 300 ml. of absolute ethanol and treated with palladium chloride (102 g~9 7.1 mmoie)O It was then hydrogenated on ~ Parr Hydrogenator for one hour at roo~ temperature ~nd an initlal pressure of 50 p~s. i.g. It was then ~iltered~ washing with warm ethanol (150 ml~). The ~iltrate was evaporated in vacuo to yield 4.2 g. o~ a residue in the form of a solidified foam. The NMR was consistent with the hydrochloride salt of the ti~le product.
The ~ree amino acid title product is obtained by dissolving the hydrochloride salt in the mln~mum amount o~ w~ter and adjusting the pH to 3.5 with cold concen-trated aTmmonium hydroxide. The product is collec~ed by filtration ~nd wsshed with cold water and acetone.

- !

7~
The ~. .o acid ti~le product has ~p. 275~ (deco~-po~l~lon) and I~ and NM~ consisten~ with the deslred ~tructure.
A s~mple of the above compound (called BC~L66)was ~ound to inhibi~ D._pneumoniae A9585 at a concentration o~ 8 mcg./ml~ 9604 a~ 8 mcg./ml. and S. ~rc ~ a~ 32 mc~./ml.

~.Z

E~ nzyl 7~-amino-3~ methvltetrazol-5-yloxv_ met~l) - ,~3-0-2 -lsocePhem-4-carbox~,la te p-Ni~robenzyl 7~-azido-3~ methyltetrazoI-5-yls~xymethyl) -~3-0-2-isocephem-4-carbox~late (0. 26 gO, 0.57 mmole) was dissolve~ in methylene chloride (50 ml.) and nitrogen was bubbled throu~h the solution fox S
minutes. Triethylamine (0.145 mlO ~ 1.14 mmole) was added and the solution was stirred while hydrogen sulLide was gently bubbled into it for abou~ 1 1/2 minutes. The resulting solution was stirred at room temperature for 1 1/2 hours by which time there was no m~re gas evolutionO
The excess hydrogen sulfide was removed by a stream D~
~itrogen bubbled t~rough the solution which was ~n turn extracted with 10% HCl (10 ml.) and water (2 x 10 ml~?.
The combined aqueous extracts were carefully alkalized with 5% ~odiwm bicarbonate solu~ion~ and then extracted *ith methylene chloride (3 x 25 ml.). The organic extracts were washed with brine, dried (Na2S04), and evaporated on the aspirator to give the ~itle product as ~ white amorphous sol~ d ~0 ~,15 g . ~ O

11~6;~>^~;~

, ~ 8 Benz,yl 7@-phenoxyacetamido ,~ , .
H H
PC~2CN~,~O
~. N~CH20COCH3 i; C02,CH20 A mlxture of benzyl 7~-amino-3-acetox~methyl- ~3-0-2-isocephem-4-carboxylate (415 mg., 1.2 mmole) phenoxyacetic acid (183 mg., 1.2 mmole) EEDQ ~325 mg., 103 mmole) and 15 ml. of methylene chloride was : stirred at room temperature for 1.5 hours, The mixture W8S then washed with 10% H~l (once9 20 ml.), brine (once, 20 ml.), 10% NaHC03 solution (once, 30 ml.~, brine (once, 30 ml.3, and finally dried (anhydrous Na2S04) and ev~porated to dryness leaving a colorless oil. This oil was triturated with diethylether to give 410 mg. of crude amide title product in 71% yield.
This material was dry column chromatographed over activity III silica gel by eluting with diethylether to give 400 : . mg. of title product as a white crystalline solid; m.p.
~ 146C. (corr.).
25 24 28 C, 62~49; H7 5.04; N, 5.83.
Found: C, 62.58t H, 5.07; ~, 5.83.
. .

-- lq5 --1 The methylene chloride used in this experiment was reagent grade which had bleen further purified by passing - over a c,alcium chloride column.
: ' , ' ~19 7 ~PhenoxYacetamido-3-acetoxYmethyl- ~3-0-2-isocephem-4-carboxYLic acid i , ~IOCI~2CONH~

N~7LCHzococH3 : C02N
A mixture of benzyl 7~-phenoxyacetamido-3-acetoxy-` methyl-~3-0 2-isocephem-4,-carboxylate (100 mg., 0,20 mmole), 100 mg~ o~ 10% Pd-C and 50 ml. of THF was . hydrogenated for 15 minutes in a Parr hydrogenator with an ini~ial pressure of 40 p.s.i.g. The catalyst was ~iltered off and the filtrate evaporated to dryness leaving 50 mg. of white crystalline solid title produc~;
m.p. 160-170C. (corrO) ~decomposition) (recrystallized from ethanol.
AnaL Calcld. for C18H18N208: C, 55.38; H, 4.65; N, 7.18.
Found: C, 55.32; H, 4.88; N, 7,.13.
` A sample o the title product (called BC-L53) after ~olution in water and d~lution with Nutrient Broth was '~ , . .

., .

1~'6~7~
''' ~

was found to exhibit the :~ollowing Minimum Inhibitory Concentrations (M.I.C.~ in mcg,/ml, versus the ~ndicated microorganisms as determined by overnight incubation ~t 37C. by tube dilution. Cephalexin was included as a comparison compound.

, Tab M I.C. in mcF~./ml.
.~ ~
. Or~anism BC-L53 ~_-lexin ~D. pneumoniae A9585 .06 .13 . ~5% serum*
Str. pyogenes A9604 .13 /13 ~5% serum*
S. aureus Smith ~ A9537 l .25 S. aureus Smith ~ A9537 4 ,5 +50% serum S. ~ure~s BX1633-2 A9606 at 10 dil'n ~: S. ~ure~s BX1633-2 ~9606 125 4 ~t 10- dil'n S. aureus me~h.- A15097 16 32 resist.; a~ 10-3 dil 'n Sal. enteritidis ~ A95al 2 2 . E. coli Juhl ~ Al511gI6 4 : . ~. coli ~ A9675'125 8 K. pneumoniae ~ A997716 2.
.~ . .
Ko pneumoniae A15130>125 8 Pr. mir~bilis ~ A9900 4 4 Pr. morganli ~ Al5153>125 >125 ~s. ~eruginosa ~ A9843A>125 >1~5 serO sn~rcescen~ $ A20019>125 >125 Ent. cloacae A9656>125 >125 : Ent. cloacae A965763 2 Ent. cloacae A9659~125 >125 .
* 50Z Nutrient Broth - 45% Antibiotic Assay Broth ~ at lO 4 dilut~on.

., .

- 1 2 ~, -.

~ 3~

.

; ~2 0 ; ~ -Phenoxyacetamido-3-hYdrox~methyl- ~3-0-2-isoce ~ -4-c~h~xvli~ ~cid , ~ ~CH2CO~

~LCH20H

A. Acylation to prepare benzyl 7~-phenoxYacetamido-3-benzyloxyrnethYl- ~3-0-2-isocephem-4-carboxylate :: To benzyl 7~-amino-3-benzyloxymethyl-~3-0-2-isocephem-4-carboxylate (~.4 gt~ 1~01 mmole) dissolved in 100 ml. o~ CH2C12 was added EEDQ (0~25 g., I.01 :~ ~mole) and phenoxyacetic acid (0.1S4 g~, l.01 mmole).
`;~ The solution was stirred at room temperature for 2 hours, w~shed s~ccessively with a 1% NaHC03 solution, 10%
. hydrochloric acid and brine solution, dried over Na2S04 : ~nd conc~ntrated on the rotary evaporator to give 0.53 g.
o~ crude~ benzyl 7~-phenoxyacetamido-3-benzyloxymethyl-~3-0-2-isoc:ephem-4-carboxylate. Purification was achieved : ~hrough chrom~tography over silica gel and gave 0.35 g.
(66%) of pure benzyl ester.

-- 1~9 --.

~ 3 7 ~

, " ' ' ' . ' , ' .

8D ~Simultaneous de-bloc~:in~ at 3- and 4-_positions The benzyl ester intermediate of Step A (0.35 g~, 0c66 mmole) was hydrogenated a~ atmospheric pressure in a total o~ 55 ml. o~ ethyl acetate in the presence of 20% Pd(OH)2 (0.35 g.). When hydrogenation was complete (18 hours), the catalyst wa~ removed by filtration over Celite and the filtrates were concen~
tr~ted on thè rotary evaporator to give ~.18 g. of ~emi-solid which crystallized after trituration with eth2r. Recrystallization from acetone~ether gave - . 0.101 g~ of white solid; m.p. 157-1.59C. Spectral data, IR and NMR, were fulIy consis~ent with the struc ure of the tltle p~oduct.
This compound was found to be identical (}R, M~, .
mixed m.p.) to the compound prepared in Example 21-Anal- ~al~'d- for C16~16N27: C, 55.17; H, 4063; N, 8.04.
Found: CJ 55.06; H~ 4.59; N~ 7.99.
A sample of the ~itle produc~ (called BC-~43) aster ~olution in water and dilution with Nutrient Broth was found to exhibit the following Minimum Inhibîtory Concentrations (M.I.C.) in mcg./mlu versus the indicated microorganisms as determ~ned by overnight incubation a~
37C. ~y tube dilution.

*Trade Mark - 130:-63q9~

Table 2 M.I,C, in mcg./ml.
5~ BC-~43 Cepha-lexln D. pneumoniaeA9585 <~25 .13 ~5% serum*
5tr. pyogenesA9604 , ~.25 .13 ~5% serum*
S. aureus Smith$ A9537 <.25 .25 S. aureus Smith ~ A9537 2 t507~ serum S. aure~ls BX1633-2 A9606 4 at 10~3 dil 'n S. aureus BX1633-2 A9606 63 4 ~t 10-2 dil 'n S . aureus meth . - A 1509732 63 res~ stO; at 10-3 dil tn Sal. enteritidis ~ A9531 16 2 E. coli Juhl ~ A15119~125 8 E. coli A9675>125 32 K. pneumoniae ~ A9977 63 4 K, pneumoniae ~ A15130~125 ~ 6 Pr. m~rabilis ~ A99û0 63 4 Pr, morganii ~ A15153~125 ~125 Ps. aeruginosa ~ A9843A>125 >125 Ser. marcescens ~ A20019>125 >125 Ent. cloacae A9656>125 >1~5 Ent. cloacae A9657>125 4 Ent. cloacae A9659>125 ~125 ' * 50% Nutrien~ Broth - 45% Antibiotic Assay Broth 10 4 dllution.

, , .~ .

., .

ti379L

`
~PhenoxYacetam~do-3-hydroxYmet~yl~/\300-2-isoce,~hem-4-~rboxylic acid A. To the crude 7~-amino intermediate from Example 15 dissolved in 60 ml. of CH2C12 was added EEDQ (00741 g., 3 mmole) and phenoxyacetic acid (0.456 g.~ 3 mmole).
The solution was stirred at room temperature for 2 1/2 hours, washed successively with a 5% hydrochloric acid solution, water, a 1% sodium bicarbonate solution and water, dried (Na2S04) and concentrated to give 1.48 g.
of crude benzyl 7~-phenoxyacetamido-3-hydroxymethyl-~3-0-2~isocephem-4-carboxylate. Crystallization in ether con~aining a trace of ethyl acetate afforded 1.07 g.
of the pure benzyl ester as a white solid.

B. The benzyl ester from Part A (0.25 g.~ was hydro-genated in a Parr-shaker in ethyl acetate (50 ml.) with ~OZ Pd~OH)2 as the catalyst at an initial pressure of 60 psi of hydrogen. After 30 minutes, the catalyst was removed by filtration over Celite and the filtrates were concentrated to give the title product (0.20 g., 98% yield) as white crystals, m.p. 156-158C.
This compound was found to be identical (IR, NM~g mixed m.p.) o the compound prepared in Example 20.

~?~

:e A ~olution o~ methanesulfonyl chloride ~0.04 ml., 005 mmole) in methylene chloride (~0 ml.) was added dropwise to a cooled (S) and stirred solution of benzyl 7~-phenoxy~cetamido-3 hydroxymethyl~ 0-20 ~ socephem-4-car~oxylate (0 . 219 g ., 0 . 5 nmlole~ arld triethylamine (0.01 ml., 0.5 ~mole) in methylene.
chloride (10 ml.). The cooling bath was removed ~nd the solution was stirred for 2 hours a~t room emper-sture~ Aftsr cooling, thë solution was washed successively with 10% hydrochloric acid, diluted ~mmonla solution, water snd brine. Drying and concsntration left 0.26 g. of title product as ~
~emi-solid. NMR analysis veri~ied the identity of thc product.

- 133 - .., .

:

7~-(2-Thienylacetamido~-J~ 3-o-2-~id .

~CH2CONH~

' ~--CH20COCH3 .- O2H
' :
The crude amino acid hydrochloride prepared ~n Example 16 (4,2 g. J 13.4 mmole) was treated with 150 ml~
- of water, cooled to 0C., and ~reated slowly with sodium bicarbonate ~7.5 g., 90 mmole). A solution o~
2-thienylacetic acid chloride (4.5 g., 29 mmole) in ., : 50 ml~ of acetone was added slowly and the mixture ~tirred for an additional hour at 0C.
It was then extracted with ether (2 x 100 ml.) and the aqueous phase was 3cidified with dilute hydrochloric ; acid. The product was extracted into methylene chloride and the extract was dried over anhydrous sodium sulfateO
Evaporation in vacuo gave a crude residue which solidi-~ied on seeding and trituration with ether. The yield o~ this crude materifll ls 2.6 g. (51% yield from the ~zldo ester).

.

.

~ 6 ~

The product was purified by crystallization ~rom absolute ethanol giving 1.8 g. (35% yield from the azido ester3 o~ title product with m.p. 182 (decomposition~.
Anal. Calc'd. for C~6H16N207S: C, 50.52; H, 4~24; N, 7.37;
S, 8.43.
Found: C, 50.34; H, 4.41; N, 7.47;
S, 8.45.

U.V. ~ EtO~ 272, ~_ 8654.

The NMR and IR were consistent with the proposed structure. M.I.C. data for the above product (called BC-L58) $s shown in the following table.

~

1~6374 .

Table 3 . /ml .
2E~ BC-I,58 Cepha~
lexin lothin D . pneumon~ae A9585 . 03 . 25 , 06 ~5% serum* . 016 , 5 , 03 Str. pyogenes A9604 , .03 ,25 .06 ~5% serum* . 016 .13 . û3 S. aureus Smith ~ A9S37 ~5 1 .13 . . .25 ,~ .~)6 S. aureus Smith ~ A9537 l 1 .13 ~50~ serum 1 2 . 5 S. aureus BX1633-2 A9606 1 1 .13 at 10 3 dil'n .. 5 1 ,,13 S. aure~s BX1633-2 A9606 2 4 25 ~t 10- dil 'n 1 4 25 i ~ S . ~ureus meth~ - A15097 2 16 . 5 resist .; at 10-3 l 4 . 5 dil 'n Sal. enteritidis ~ A9531 . 5 2 ,25 .13 4 .25 E. coli Juhl ~ A15~19 8 ~ 16 ` 8 8 16 E:. coli i A9675 63 . 16 63 : 32 16 32 K.pneumoniae ~ A9977 1 2 ~ 5 ~ .5 4 ;~ . K.pneumoniae ~ A1513063 . 16 16 ` ~ Pr.mirabilis ~ A9900 1 4 .5 .. ~ 4 ~ . Pr. morganii ~ Al5153125 ~125 ~125 . >1~5 >125 >1~5 Ps. aeruginosa ~ A9843A~125 >125 >125 >1~5 >125 >125 Ser. marces~ens ~ A20019~L25 >125 >125 ~125 >125 ~125 ' . ....

T~ble 3 (cont 'd) M. I . C, in mc~ !ml Or~snlsm BC-L58 ~.- ~-lexin lothin Ent. clo*cae A9656~125 >125 ~125, - . ~125 >12$ >125 Ent. cloacae A965716 4 ;~ Ent. cloacae A9659>125 ~125 >125 ~125 >125 >125 ., .
* 5070 Nutrient Broth - 4570 Antibiotic Assay Broth $ ~t 10 4 dilution.

. .

Compound BC-L58 was resoïved irlto its ind~vldual ~omers designated BC-L68 (+ isomer) and BC-L69 t~ isomer) . Mo I.C~ d~ta for the isomers is provided bel~w.

- 13~ -i3'79 ;
, .
Table 4 ,, ~
M~, I . C ~, in mcg . /ml .
Or~anism BC-L68BC-L69 Ce~ha- Cepha-lexin lo~hin D. pneumoniae A9585 .016.,5 .5 ~03 ~57O ser~n*
S~r, pyogenes A9604 . 016. 5 .13 , 03 ~5% serum*
S. aureus Smith ~ AgS37 ,.06 4 .5 .06 S . aureus Smi~h ~ A9537 .25 32 .2 O 5 ~50% serum S . aure3s BX1633-2 A9606 .13 8 1 " 13 ~t 10- dil ~n S. aure~s BX1633-2 A9606 1 16 4 r25 at 10~ dil ~n .
S. ~ureus meth.- A15097 .25 8 4 O5 resi~t.; at 10-3 dilln S~l. enteritidis ~ A9531 .03 8 4 .25 E. col~ Juhl ~ A15119 , 4 ~125 8 16 E. coli ~ A9675 32 ~125 16 32 K. pneumoniae ~ A9977 .1332 4 K. pneumoniae ~ A15130 16 ~125 8 16 Pr. mirabilis ~ A9900 .1316 4 Pr. morganii ~ A15153 .125 >125 >125 >125 Ps. aeruginosa $ A9843A >125 ~125 >125 >125 Ser. marcescens ~ A20019~125 >~25 >125 >125 Ent. cluacae A965Ç >125 >125 >125 ~125 Ent. cloacae A9657 16 >125 4 4 Ent. cloacae A9659 >125 >125 >125 ~1~5 .

* 50% Nutrient Broth - 45/~ Antibiotic Assay Broth at 10 4 dil.ution.

~ v 3~
.

Benzyl 7~-phenoxyacetamido-3-acetoxymethyl-~3-0-2-~so~ce~he~r4--- acylaei~r f c orr~ n d ~n~
3-hydroxymethyl com~ound) ~ m~xture of benzyl 7~-phenoxyacetamido-3-hydroxy-methyl-~3-0-2-isocephem-4-carboxylate (1.07 g., Z.44 mmole), acetic anhydride (1 g., 9.8 mmole) and pyridine (15 mlO) was stirred at room temperature for 1.5 hours.
The reaction mixture was diluted with water and extracted with CH2C12. The organic phase was washed with water, 10% HCl and water, drled and concentrated to le~ve a solid. Recrystallization of the solid In ethyl acetate-ether gave l.0 g. of title product as white crystals, m.pO 1~4-146. This com~ound was found to be identical (mixed m~pi, IR and NMR) to the product of Example 18.
The benzyl ester may be converted to the corres-pondlng free acid compound by the procedure of Example 19.

- 1~0 --i3 ~.5 Separat~on of Diastereomers of ~CH-CONH~f b ~N ~'I`CH~OCOCH3 ' H H D- ~CHCO H
H2N ~ _ 1 2 ~CHCONH H H

~;~ co2CH2~l5 1 ~c `CH20COCH3 ~H2~ ~CH20COCH3 02CH2~ 02CH

~ .
A solution of ben2yl 7~-amino-3-acetoxymethyl-~3-0-2-isocephem-4-carboxylate (1.86 g., 5.4 mmole), : D-carbobenzoxyphenylglycine ~1.54 g., 5.4 mmole~ and EEDQ (1046 g., 6.0 mmole) in 50 ml. of methylene chloride was stir.red at 24 for 16 hours. The resulting solution ; was washled with 5% sodium bicarbonate, 10% hydrochloric ~cid and dilu~e sodium chloride solutions, then dried (sodium sulfate) and evaporated in vacuo to give a yellow tar, 3.27 g. The tar was dissolved in methylene chloride and absorbed on~o 16.5 g. of silica gel ~grade ITI) .

, . ~

~¢63~

.
whlch was placed on a chromatographic column of 165 g.
of silica gel. Elu~on with ether gave (ater some front running impurities) the major component as a partially separa~ed mixture of two diastere~mers. The 13s fraction (2.0 2-) ~shown by NMR to be a 34:66 : .
r8tio of isomers) was re-~bsorbed onto 10 g. of sillca gel and chromatographed on 100 g. of silica gel.
Elution with ether:hexane 9:1 changing to ether gave further separation of the isomers. The last fractio~
(371 mg.) was found to be a 15;85 mixture of two isomers.
The mixt-lre had ~ 5 - +26.5 (CHCl ) , ' .
. B.

H H H2/Pd > ~CHCO~H _ \~ t HCl ~H2 \~

C~O _~ N /~ ~ICl ~N ~1 OCH2; ~02CH2~ ~ 2 ~3 .
.~ .
To a solution of 325 mg. ~0.53 mmole) of the mixture of isomers from Part A in 8 ml. of ethyl acetate was added 8 ml. of ethanol~ 0.53 ml. of 1.00 M hydrochloric `, acid and 320 mg. of 30% palladium-on-diatomaceous-earth.
The mixture was hydrogenated at 24 and 1 a~mosphere until uptake o~ hydrogen ceased. The ca~alyst was 3 ~

removed by filtration and the solvent evaporated in vacuo to give the amino c3cid hydrochloride salt as an amorphous solid, 210 mg. N~R showed the product to be

15:85 ratio mixtuxe of isomers (the 85% isomer i5 ~rbitrarily des~gnated isomer B) and to contain about 1/3 mole o~ ethanol and l to 2 mole of water. The chemical purlty of the product w~s estimated from U.V. spectrum to be about 70%. Th~ product had ~25 _ ~72.0 (DMS0) and U.V. ~EatH 270 mm. t 6600).

IR and NMR spectra were in agreement with the proposed structure.
A sample of:the product (called BC-L65) was found to exhibit the following Minimum Inhibitory ConcentratLons.

.

. ~ 37'~

. .
T-lble S
M.I.C. in mc~./ml.
BC-L65 Cepha lexin D, pneumoniae A9585 ~06 o25 t5% serum*
Str. pyogenes A9604 , ,06 .25 ~5% serum*
S. aureus Smith ~ A9537 S. aureus Smith ~ A9537 4 2 ~SO% serum S. aureus BX1633-2 A9606 2 4 at 10-3 dil'n S~ aureus BX1633-2 A9606 8 4 at 10-~ dil'n S, ~ureus me~h.- A15097 4 8 resist.; at 10-3 dil'n Sal. enteritidis ~ A9531 . 5 4 E~, coli Juhl ~ A151L9 E. ~oli ~ A9675 4 16 R. pneumoniae ~ A9g77 1 4 K. pneumoniae ~ A15130 8 16 P~. mirabilis ~ A9900 ,5 8 Pr, morganii ~ A1515332 >125 Ps. aeruginosa t A9343A>1~5 >125 Ser. marcescens ~ A20019125 >125 Ent. cloacae A9656125 ~125 Ent, cloacae A9657 2 4 Ent, cloacae A9659 32 >125 * 50% Nul;rient Broth - 45% An~ibiotic Assay Broth ~ ~t 10 4 dilution.
'-.~' ' , .

: .

-- 1~4 --. .

3';7~

.
~3~E~ _6 -(2-Thieny1ace~am1do~-3~ methY1tetrazo~ ~10xy~
methY1~-~3-0-2-isoce~hem-4-c~rbox~71ic acid S H2CO~I. ~ 3 N
~CH2-0 A mixture conslsting of p-nitrobenzyl 7~-amino-3~ me hyltetrazol-5-yloxymethyl)- A3 0_2-isocephem_ 4-carboxylate tOo15 g.~ Q.35 mmole), EEDQ (0O086 g., 0.35 mmo1e~ and thienylacetic acid t0.05 g., 0.35 ~nole) ~n methylene chloride (SO ml. ) was kept at room temper-ature ~protected from mois~ture by a calcium ch10ride drying tube) for 16 hours. It ~as then washed succes-vely with 10% XCl~ water~ 5% sodium bicarbonate andbrine, dried (Na2S04) and evaporated to dryness to give 0.1~ g. of an amorphous solid which was ldentified ~y ` ~ as p-n~trobenzyl 7~-(2-Thienylace~amido3-3-~1-methyltetrazol-5-yloxymethyl)-f~3-0-2-isocephem-4-c3rboxylate~
: The p-nitrobenzyl ester (0.18 g., 0.32 mmole) was dissolved ln ethylacetate ~75 ml.) and n-butanol (10 ml~) ~n a 500 ~1. Par* bot~le. To this was added 20%
p~ll3dium hydroxide on Celite (0.27 gO) and O~lN hydro.
chloric ac~d ~3.2 ml., OJ32 mmole). The mixture w~s then sha~en under hydrogen ~inltial pressure 60 psi.) r ~ ~'J 6 3 IY4 , ' ' , for 2 1/2 hours. The solid was f~ltered off on Cel~te pad and the filtrat:e evaporated in vacuo down to fl vO~ oi~ about 2 ml., when a yellow solid ~eparated. Ether ~50 ~1~,) and 1% sodi~n bicarbon3te ~15 ml.) were sdded and, ~fter thorough shaking, the ~ueous phase was isolated and care~ully acidified with 10% hydrochloric acid~ On cool~ng, white crystals separated and were collected by suction filtra~ion, ws~hed wi~h cold water and dried to give the title product as a whi'ce solid (0.04 g~, m.p~ 190-192d.
AnalO Calc~d. for C16H16N606: C, 45.71 H, 3.84; N, 19.99., Found: C,. 45963; Hr 3.91; N, 20.11.

U.V. ~mUX - 7,900 ~THF) .
. . ' .

.

r ~

T~ble 6 2E~ BC-L72 lothin D. pneumoniae A9585.06 .~6 t5% sexum*
strO pyogenes A9604.,06 .,06 ~5% ser~m*
S . aureus Smith ~ A9537. 25 .13 S 1~ ~ureus Smith ~ A9537 1 . 5 ~50% serum S. sureus BX1633-2 A9606 8 o25 8~ 3 dil ' n S " aure~s BX1633-2 A9606125 0 5 at 10 dil 'n S~ aureus meth~- A150978 re~ist.; at 10~3 ~dil 'n Sal. enteritidls ~ A9531 .5 o25 E. coli Juhl ~ A1511932 16 E. coli $ A9675 ' 125 63 K. pneumoniae ~ A9977 2 . 2 K. pneumoniae ~ A1513063 16 Pr. mirabilis ~ A9900 2 Pr . rganii ~ A15153125 >125 Ps~ aeruginosa ~ A9343A>125 >125 Ser. marcescens ~ A20019>125 >125 Ent. cloacae A9656>125 >125 Ent. cloacae A9657. 16 4 Ent. cloacae A9659>125 >125 * 50k Nutrient Broth - 45% Antibiotic Assay Broth at lû 4 dilution.

.

.

6~74 ~3~ 7 arbamoylox~methyl-~3-0-2-, ~LCII2CONI~

: N ~ CH~OCONH2 p-NitrobenzyI 7~-az~do-3-carbamoyloxy~ethyl-~3-0-2-~ iQocephem-4~carboxylate (400 mg~g 0~96 mmoles) and ~ triethylamine (0.25 ml., 1.8 mmoles) were disso~ved in 30 ml. of methylene chloride~ Hydrogen sulfide was gently bubbled into the solution for 1 minute with stirring. After stirring,for 30 minutes, nitrogen w~s bubbLed through to remove most o~ the excess H2S
and it was then evaporated in vacuo.
The residue, p-nitrobenzyl 7~amino-3-carbamoyl-oxymethyl-~3-0-2-isocephem-4-carboxylate, was dissolved in 50 ~. of methylene chloride and treated with thienyl;
scetic acid (150 mg., 1.05 mmoles) and EEDQ (240 mg., ~0 mmoles~. The mlx~ure was stirred at room temperature for 18 hours and then washed with watera l% sodium bic~rbonate~ water, lZ hydrochloric acid, water and brine. It wa~ then dried over anhydrous sodium sulfate ~nd evaporated in vacuo to give crude product which W2S

148 ~
' ' .
~63~4 .
purified by dry-column chrom~ogr~phy on silic~ gel ~Activ~ty III) eluting with 25V/o ethyl acetate in ether. The yield of p-nitrobenæyl 7~ (2-thienyl-~cetamido~-3-carbamoyloxyme!thyl- ~3-0-2-isocephem-4-carboxylate was 150 mg. (29~o)o NMR and IR of the product ester were consisten~with the proposed structure.
To a solution of 310 mg. (0.80 mmoles) o~ the ester in 150 ml. of ethyl acetate and 50 ml. of n~butanol was ~dded 8 ml. of O.lN hydrochloric acid ~0.80 mmoles) and 410 mg. of 2070 palladium hydroxide on carbon. The mixture was hydrogenated at room temper~-ture on a Pa~r apparatus at 60 ?sig for 3 hours.
The catalyst was removed by filtration and the solvent was evaporated ~n vacuo, The residue was slurried with ether and extracted with l~/o sodium blcar-bonate. The bica~bonate extract was acidified with dilute hydrochloric acid and extracted with ethyl acetate~
It was dried over anhydrous sodium sulfa~e and evapora~ed in vacuo to glve a semi-solid residue, The residue W3S
, . . .
triturated with ether and filtered to give 25 mg. of a pale-yellow solid which was about 70-~0% pure by ~pectral da~a and analysis.
U. V. ~ THF _ 270 mm (~ = 10~351?o NMR and IR 3nalyses conf~rmed that the compound produced wag the title product.

.

3~
.

.
- T~ble 7 ., .
Il.~.C. in mc 1ml.
- ~ BC-~74 Cepha-lexi=n lo~hln 1)~ pneumoniae.A9585 16 1 ~13 . t5% ~erum*
SSr. pyogenes A9604 8 .25 .0 57~ ser~*
., .
~: S. aureus Smith A9537 .5 1 .13 S. a~reus Smith ~ A9537 ~63 2 .5 ~50% serum S. ~ureus BX1633-2 A9606 1 4 o25 : ~t 10-3 dil ~n S. aureus BX1633-2 A9606~125 8 .5 3t: 10 2 dil'n S O aureus meth. - A1509732 16 ~ ~ 1 resist.; a~ 10-3 d~l~n :' ~ .L ' Sal., enteritidis ~ A9531~125 4 025 E~ coli Juhl ~ A151~,9>125 8 16 E. coli ~ A9675~125 16 63.
K. pneumoniae i~ A9977~125 8 2 K~ pneumoniae ~r A15130>125 16 32 Pr. mirab~l~ s ~t A9900>125 8 Pr. mo1:ganii ~ A15153~125 >125 >125 P~. aeruginosa ~ A9343A~125 .>125 ~125 Ser. marcescens ~ A20019~125 >12; ~125 Ent. cloacae A9656>}25 ~125 >125 Erlt. cloacae A9657>125 4 8 Ent ., cloacae A9659>125 >1~5 ~}25 * 50% Nutr~ent Broth - 45% Antibiotic Assay Broth t ~t 10 4 dilution.

. .
.. - ' . .
- 150:-. .
, . . .

Example 28Preparation of 7~-[ a- (2-Aminomethyl-1,4-cyclohexadlenyl)-acetamido]-3-methyl- ~ 3-0-2-isocephem-4-carboxylic acid ~CH2CoNH~/
N~ ~ CH2OCOCH4 A. a- (2-Amin m thyl-1,4-cyclohexadi_nyl)acetic acid A solution of 16.5 g. (0.1 mole) of o-aminomethyl=
phenylacetic acid in 1.5 1 of liquid ammonia (which had been trea-ted with 50 mg. of Li to remove a trace of moisture) was slowly diluted with 500 ml. of dry t-BuOH.
To the solution was added in small portions 3.4 g. (0.5 atom) of Li over a period of 4 hours and the mixture was stirred for

16 hours at room temperature removing the l;quid ammonia in a hood and finally evaporated to dryness below 40 C. The residue was dissolved in 500 ml. of water and the solution was chroma-tographed on a column of IR-120 (H~, 700 ml.) resin ~ 20 and eluted with 1% NH~OH solution. Ninhydrin positive fractions of the eluate were combined and evaporated to ~ - 151 -, 3~4 `~-`

dryness. The residue was washed with four 50 ml. portions of hot acetone and recrystallized from 500 ml. of ethanol-water (1:1) to give 11.2 g. (67%~ of a-(2-aminomethyl-1,4-cyclohexadienyl)acetic acid as colorless needles. M.p. 183C.

' IR: maUx 1630, 1520, 1380t 1356 cm 1 ~ D2O -~ K2C3 2.72 (4H~ s~ H2 (2H, s, CH2CO), 3.20 ~2H, s, CH2--N~, 5.78 (2H, s, H ~ C=).

Anal. Calcd. for CgH13NO2: C, 64-65; H~ 7-84;
N, 8.38.
Found~ C, 64.77; H, 8.06; N, 8~44.
; B. a-[2-(t-Butoxyc'arbonylaminomethy'1'~'-1',4'-cyclohexa'dienyl]-ace'tic acid To a stirred solution of 8.0 g. (0.048 mole~ of a-(2-aminomethyl-1,4-cyclohexadienyl~acetic acid and 3.8 g. (0.096 mole~ of NaOH in 150 mlO of water was added a solution of 10.3 g. (0.Q72 molel of t-butoxycarbonylazide in 80 ml. of THF and the mixture was stirred for 18 hours at room temperature. The THF was removed under reduced pressure and the -residual solution was washed with ether (2 x 100 ml.), acidified with 6 _ HCl and extracted w-ith ether (3 x 100 ml.). The combined extracts were washed with water ~2 x lQ0 ml.) and a saturated NaCl solution (100 ml.), dried with Na2~O4 and evaporated to dryness.
The oily residue was triturated with n-hexane to give 10.5 g. (82%~ of colorless powder melting at 113C.

-:

~ - 152 -.

~R:~ PaUJ 3~70, 1715; 16~0, 15~0, 1280, 1160 cm 1.
N~ pp~ 1.45 t9~, s, ~-~u-H) J 2-73 (~ s~
C _ )~ 3~16 ~2~, s, c~co3; ~.76 (2H, d, 6~z~ C~
.90 (~Ig m, 1~, 5.66 ~H~ s, E_~C=)~ 10~6 ~ br s, t:OO~
An~lO Calcd. ~or C14H21N04: C~ 62.90; H, 7.92;

NJ 5.24.
~ound: C~ 63.i3;-E, 8.21, ~J~ 5.26.

.
C~ ~ } ~ bon~rlaminomethyl-l,4-cYclohexadien~
et-nidol-3--cetoxvmethyl-~3-0-2-isoce~hem-4-c~
To a stirred solution o equimolar amounts of ~-t2-(t-butoxycarbonylaminomethyl)-1,4-cyclohexadienyl)acetic acid and 294-dinitrophenol in eth~l acetate is added an equimolar amount 2f N~N~odicyclohexylcarbodiimide. The reaction mixture is stirred at room temperature or 3 hours. The separated d~cyclohexylurea is filtered off. The filtrate is evapora~ed to dryness to give the activated ester which is dissolved in tetrahydrouran. To th~s solu~ion is added a solution o~
7~-amino-3-acet:oxymeth~1- ~3-0-2-isoceDhem-4-carboxylic acid .and triethylamine in approxim~tely a 1-2 molar Drooortion, re~pectively,relative to the -~2-(t-butoxycarbonylaminomethyl)-1,4-cyclohexadi.enyl]acetic ac~d. The mixture is stirred at room tP~perature for several ~ours 3nd concentrated in vacuo.
The concentrate is washed ~ith ether~ ~cidified with dilute m~neral ac~d and extracted with ethyl aee~a~e. Th~ extracts - lS3 ~

637~4 .

~r~ washed with water and s~turated NaCl solu~ion and dried to give the title produc~0 D. 7~ 2-Aminometh21-lL4-c~clohexa~ienyl~acetamido~-3-scetoxymethyl-~3-0-2-isocephem-4-car~oxylic acid A so7u~ion of 7~ ra-(2-t-butoxycar~onylaminomethyi-1,4-cyclohexadienyl)acetamido~-3-acetoxymethyl- ~ 3-o-2-isocephem-4 carboxyiic ac~d in tri~luoroacetic acid is stirre~ zt 0C.
for one hour. To the so~ution is added dry ether un~il a precipitate ~orms. ~e precipitate is collected by fi~tra~ion3 s~spended in water and adJusted to pH6 ~o give the t~tle product.

Example ?~8 methyl- A3-0-2-isocephem-4-carboxylic acid , ~CH2NH2 ~C~

~2 A. ~ ox acetic acid A 801ution o ~-12-tt-butoxycarbonylaminomethyl~-1,4 cyclohexadienyl~-aceti~ acid ~1~33 ~., 5 mmoles) in 3%
~mmonium hydroxlde (10 ml.~ was hyd~ogenated at 40 ps;.

.

6i3r~3~

wîth pall~dium on charcoal (10%, 0.2 g.~. A ~heoretical ~mount o hydrogen was taken up in 3 hours. The catalyst was removed and the ~i~trate wa~ acidified to pH 2 with dil.
HCl and e~tracted with ethyl acletate ~2 x 50 ml.). The combined ex~ac~s were washed with wa~er (20 ml.), dried with Na2S04 and evapoEated under reduced pressure to a~ford ~n oil. ~1.34 g.) which solidi~ied on standing for several days. Recrys'callization from n-hexane - ethyl acetate gave 1.2 g. title product as colorless pris~ns mel~ing at ~18-119C, :I:R: YD~a~l 3450" 1730, 1660, 1510 cm~l.

S ppC13 1.58 (9H, s, ~-butyl-H), 1~.50 - 1.90 ~H, m, ~ 1. 90 - ~ .20 (4X, m, a llylic methylen~
3.18 ~2H, s, ~I2-C0~, 3~78 ~2H, d, 6 Xz, C~I2-P~), 5.OQ
(lH, br-s., NH), 809& (lX, br-s~r COC~I3.
.4nal. Cal~d- ror~ C14H23N0~ ~ 62042; H; 8-61;
~, 5.20.
~ou~d: C, 6~.12; II, 8 .77; ~J, 5 .37 0 B0 7B - r~- ~2-t-ButoxYcarbonylaminomethyl-l-cyclohexenyl~acetamidoJ-3-~cetoxymethyl- Q 3^0-2-isocephem-4-car~oxylic acid ; To a stirred solution of equimolar amounts of 12-(N-~-butoxycarbonylansinomethyl)-l-cyclohexen-l-yl~acetic acid and 2,4-d~nitrophenol ~ ethyl acetate is added an equimolar amount o~ N,N~-dicyclohexylcarbodiimide. The reaction mixture is . .
~irred for 1 hour ~t room te~perature and the precipi~ated dicyclohexylurea is filtered o~. The fi~trate is cooled ~o ?~3'^~
5~C. ~nd poured in~o a cold solution o 7~-amino-3-ace~oxy-methyl-/~3^0-2-isocephem-4-car~oxylic acid and excess tri-ethylamine in 50% ~queous T~. The mixture is stirred over-nlght at room temperature and washed wlth ether. The ~queous layer is acidiied with dilute HCl to precipita~e the ~itle product.

C. 7~-fa~Aminomethyi-l-cyclohexenyl)acetamido~-3-acetoxy-methyl-~3-0-2-isocephem-4-carbox~lic acid A solution of 7~-~a-(2-t^butoxycarbonylaminomethyl-1-cyclohexenyl)acetamido~-3-acetoxymethyl-~3-0-2-isocephem 4-c~rboxylic acid i~ trifluoroace~ic acid is stirred at 0C.
fQr 1.5 hoursO The mixture is diluted with ether to sep~rate the trifluoroacetate salt which is dissolved in water and neutralized to give the title product.

~;' ' ' .

Claims (25)

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

1. A process for preparing a compound of the formula III

wherein Z is halo, hydroxyl, esterified hydroxyl of the formula -OCOR2 wherein R2 is hydrogen, amino, (lower)alkyl, C3-C7 cycloalkyl, C3-C7 cycloalkyl(lower)alkyl, aryl, aralkyl or aryloxyalkylene, said alkyl, cycloalkyl, aryl or alkylene groups being optionally substituted by one or more (lower)-alkoxy, (lower)alkylthio, halogen, (lower)alkyl, nitro, hydroxy, acyloxy, carboxy, amino, (lower)alkylamino or acyl-amino radicals; or Z is esterified hydroxyl of the formula -OSO2R2' wherein R2' is (lower)alkyl, aryl, or aralkyl, said alkyl or aryl groups being optionally substituted by one or more (lower)alkoxy, (lower)alkylthio, halogen, (lower)alkyl, nitro, hydroxy, acyloxy, carboxy, amino, (lower)alkylamino or acylamino radicals; or Z is etherified hydroxyl of the formula -OR3 wherein R3 is (lower)alkyl, (lower)cycloalkyl, (lower)-cycloalkyl-(lower)alkyl, aryl, aralkyl or a heterocyclic group, said R3 groups being optionally substituted by one or more (lower)alkoxy, (lower)alkoxy-(lower)alkyl, (lower)alkylthio, halogen, (lower)alkyl, (lower)cycloalkyl, (lower)alkenyl, nitro, hydroxy, acyloxy, carboxy, amino, di(lower)alkylamino, (lower)alkylamino, trifluoromethyl, aryl, aralkyl or acylamino radicals; and R" is hydrogen or an easily cleavable ester carboxyl-protecting group selected from the group consisting of benzhydryl, benzyl, p-nitrobenzyl, p-methoxy-benzyl, trichloroethyl, trimethylsilyl, phenacyl, acetonyl, (lower)alkyl, triphenylmethyl, methoxymethyl, indanyl, phthalidyl, pivaloyloxymethyl ancl acetoxymethyl, or a pharmaceutically acceptable salt thereof; which process comprises selectively reducing a 7-azido intermediate of the formula IV' wherein Z is as defined above and R' is an easily cleavable ester carboxyl-protecting group and, if desired, removing protecting group R' to produce the corresponding free acid of formula III and, if desired, converting the free acid form of formula III to the above easily cleavable ester or salt thereof.

2. The process of Claim 1 wherein the 7-azido intermediate is reduced by catalytic hydrogenation with a catalyst selected from palladium, platinum or Raney nickel, said catalyst being optionally supported on a carrier, or by chemical reduction with a reducing agent selected from zinc and ammonium chloride, aluminum analgam or hydrogen sulfide and a base.

3. A process as in claim 1 wherein Z is esterified hydroxyl of the formula wherein R2 is hydrogen, amino or (lower)alkyl.

4. A process as in claim 1 wherein R" is hydrogen.

5. A process as in claim 1 wherein R" is benzhydryl, benzyl, p-nitrobenzyl, p-methoxybenzyl, trichloroethyl, tri-methylsilyl, phenacyl, acetonyl, (lower)alkyl, triphenylmethyl, methoxymethyl, indanyl, phthalidyl, pivaloyloxymethyl or acetoxymethyl.

6. A process as in claim 3 wherein R2 is hydrogen.

7. A process as in claim 3 wherein R2 is amino.

8. A process as in claim 3 wherein R" is hydrogen and R2 is hydrogen.

9. A process as in Claim 3 wherein R" is hydrogen and R2 is amino.

10. A process as in claim 1 wherein Z is -OCOCH3.

11. A process as in Claim 10 wherein R" is hydrogen.

12. A process as in claim 10 wherein R" is benz-hydryl, benzyl, p-nitrobenzyl, p-methoxybenzyl, trichloroethyl, trimethylsilyl, phenacyl, acetonyl, (lower) alkyl, triphenyl-methyl, methoxymethyl, indanyl, phthalidyl, pivaloyloxymethyl or acetoxymethyl.

13. A process as in claim 10 wherein R" is pivaloyl-oxymethyl, methoxymethyl, indanyl, phthalidyl or acetoxymethyl.

14. A compound having the formula wherein Z is halo, hydroxyl, esterified hydroxyl of the formula -OCOR2 wherein R2 is hydrogen, amino, (lower)alkyl, C3-C7 cycloalkyl, C3-C7 cycloalkyl(lowex)alkyl, aryl, aralkyl or aryloxyalkylene, said alkyl, cycloalkyl, aryl or alkylene groups being optionally substituted by one or more (lower)-alkoxy, (lower)alkylthio, halogen, (lower)alkyl, nitro, hydroxy, acyloxy, carboxy, amino, (lower)alkylamino or acylamino radicals; or Z is esterified hydroxyl of the formula -OSO2R2' wherein R2' is (lower)alkyl, aryl or aralkyl, said alkyl or aryl groups being optionally substituted by one or more (lower)alkoxy, (lower)alkylthio, halogen, (lower)alkyl, nitro, hydroxy, acyloxy, carboxy, amino, (lower)alkylamino or acylamino radicals; or Z is etherified hydroxyl of the formula -OR3 wherein R3 is (lower)alkyl, (lower)cycloalkyl, (lower)-cycloalkyl-(lower)alkyl, aryl, aralkyl or a heterocyclic group, said R3 groups being optionally substituted by one or more (lower)alkoxy, (lower)alkoxy-(lower)alkyl, (lower)alkylthio, halogen, (lower)alkyl, (lowex)cycloalkyl, (lower)alkenyl, nitro, hydroxy, acyloxy, carboxy, amino, di(lower)alkylamino, (lower)alkylamino, trifluoromethyl, aryl, aralkyl or acylamino radicals; and R" is hydrogen or an easily cleavable ester carboxyl-protecting group selected from the group consisting of benzhydryl, benzyl, p-nitrobenzyl, p-methoxybenzyl, trichloro-ethyl, trimethylsilyl, phenacyl, acetonyl, (lower)alkyl, triphenylmethyl, methoxymethyl, indanyl, phthalidyl, pivaloyloxymethyl and acetoxymethyl, and pharmaceutically acceptable salts thereof, whenever prepared by the process of claim 1 or by an obvious chemical equivalent thereof.

15. A compound as in claim 14 wherein Z is esterified hydroxyl of the formula -OCOR2 wherein R2 is hydrogen, amino or (lower)alkyl, whenever prepared by the process of claim 3 or an obvious chemical equivalent thereof.

16. A compound as in claim 14 wherein R" is hydrogen, whenever prepared by the process of claim 4-or an obvious chemical equivalent thereof.

17. A compound as in claim 14 wherein R" is benzhydryl, benzyl, p-nitrobenzyl, p-methoxybenzyl, trichloro-ethyl, trimethylsilyl, phenacyl, acetonyl, (lower)alkyl, triphenylmethyl, methoxymethyl, indanyl, phthalidyl, pivaloyloxymethyl or acetoxymethyl, whenever prepared by the process of claim 5 or an obvious chemical equivalent thereof.

18. A compound as in claim 14 wherein Z is esterified hydroxyl of the formula -OCOR2 wherein R2 is hydrogen, whenever prepared by the process of claim 6 or an obvious chemical equivalent thereof.

19. A compound as in claim 14 wherein Z is esterified hydroxyl of the formula -OCOR2 wherein R2 is amino, whenever prepared by the process of claim 7 or an obvious chemical equivalent thereof.

20. A compound as in claim 14 wherein Z is esterified hydroxyl of the formula -OCOR2 wherein R2 is hydrogen, and R" is hydrogen, whenever prepared by the process of claim 8 or an obvious chemical equivalent thereof.

21. A compound as in claim 14 wherein Z is esterified hydroxyl of the formula -OCOR2 wherein R2 is amino, and R" is hydrogen, whenever prepared by the process of claim 9 or an obvious chemical equivalent thereof.

22. A compound of the formula wherein R" is hydrogen or an easily cleavable ester carboxyl-protecting group selected from the group consisting of benzhydryl, benzyl, p-nitrobenzyl, p-methoxybenzyl, trichloroethyl, trimethylsilyl, phenacyl, acetonyl, (lower)-alkyl, triphenylmethyl, methoxymethyl, indanyl, phthalidyl, pivaloyloxymethyl and acetoxymethyl, and pharmaceutically acceptable salts thereof, whenever prepared by the process of claim 10 or an obvious chemical equivalent thereof.

23. A compound as in claim 22 wherein R" is hydrogen, whenever prepared by the process of claim 11 or an obvious chemical equivalent thereof.

24. A compound as in claim 22 wherein R" is benz-hydryl, benzyl, p-nitrobenzyl, p-methoxybenzyl, trichloroethyl, trimethylsilyl, phenacyl, acetonyl, (lower)alkyl, triphenyl-methyl, methoxymethyl, indanyl, phthalidyl, pivaloyloxymethyl or acetoxymethyl, whenever prepared by the process of claim 12 or an obvious chemical equivalent thereof.

25. A compound as in claim 22 wherein R" is pivaloyloxymethyl, methoxymethyl, indanyl, phthalidyl or acetoxymethyl, whenever prepared by the process of claim 13 or an obvious chemical equivalent thereof.