Note: Descriptions are shown in the official language in which they were submitted.
~L2~
-- 2 --
BACK('7ROUND OF THE INVEI`ITI01`1
The present invention is directed to a new process for
the preparation of carbapenem derivatives having a 2-substituent
of the formula
-S-A-R
wherein A represents cyclopentylene, cyclohexylene or C2-C6
alkylene o~tionally substituted by one or more C1~C4 alkyl groups
and R represents a quaternized nitrogen-containing aromatic or
non-aromatic heterocycle attached to A through a quaternary
nitrogen atom.
`~he carbapenem derivatives prepared by the process of
the present invention are disclosed and claimed by my colleagues
Choung U. Kim and Peter F. Misco, Jr., in U.S. Patent Applica-
tion Se.rial No. 471,379 filed March 8, 1983, now U.S.
Patent No. 4,552,696. That patent, which corresponds to
West Ge.rman Published Patent Applic~tion 3,312,533,
discloses preparation of carbapenem antibiotics of the
formula
8 H
R
I ~ ~ S - A - N
RlF~ ''
COOR
IA
wherein R8 is hydrogen and Rl is selected from the yroup consistin~
of hydrogen; substituted and unsubstituted: alkyl, alkenyl and
alkynyl, having from l-10 carbon atoms; cycloalkyl and cycloalkyl-
alkyl, having 3-6 carbon atoms in the cycloalkyl ring and 1-6
carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl-;
and aralkynyl wherein th~ aryl moiety is Phenyl and the aliphatic
portion has 1-6 carbon atoms; heteroaryl, heteroaralkyl, hetero-
cyclyl and heterocyclylalkyl wherein the hereto atom or atoms in
the above named heterocyclic moieties are selected from the qroup
3~
-- 3
consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl
moieties associated with said heterocyclic moieties have 1-6
carbon atoms; wherein the substituent or substituents relative
to ,he above-named radicals are selected from the group consisting
of
Cl-C6 alkyl optionally substituted by
amino, halo, hydroxy or carboxyl
halo
O
-OCNR R
-CNR3R4
3 4
-NR R
~R
~ ~3R4
-502~R3R4
Il 3 4
-~CN~ R
O
R3C~ -
--Co2R3
O
-oCR3
~3
--S R.
Il g
-~~R
'
-- 4
-N3
-oSo3R3
-oSo2R3
-NR3So2R4
-NR3~=NR4
~3
-NR3Co2R4
-N02
wherein, relative to the above-named substituents, the
groups R3 and R4 are independently selected from
hydrogen; alkyl, alkenyl and alkynyl, having from 1-10
carbon atoms; cycloalkyl, cycloalkylalkyl and
alkylcycloalkyl, having 3-6 carbon atoms in the
cycloalkyl ring and 1-6 carbon atoms in the alkyl
moieties; phenyl; aralkyl, aralkenyl and aralkynyl
wherein the aryl moiety is phenyl and the aliphatic
portion has 1-6 carbon atoms; and heteroaryl, -
heteroaralkyl, heterocyclyl and heterocyclylalkyl wherein
the hetero atom or atoms in the above-named heterocyclic
moieties are selected from the group consisting of 1-4
oxygen, nitrogen or sulfur atoms and the alkyl moieties
associated with said heterocyclic moieties have 1-6
carbon atoms, or R3 and R4 taken together with the
nitrogen to which at least one is attached may form a 5-
or 6-membered nitrogen-containing heterocyclic ring; R~
is as defined for R3 except that it may not be hydrogen;
or wherein Rl and ~8 taken together represent C2-C10
alkylidene or C2-C10 alkylidene substituted by hydroxy;
A is cyclopentylene, cyclohexylene or C2-C6 alkylene
optionally substituted by one or more C1-C4 alkyl groups;
R2 is hydrogen, an anionic charge or a conventional
X~ ; .
'~
.
-
:
. ' ~ . ' : .
~x~
readily removable c~rboxyl protecting group, providing '.ha- when
R is hydrogen or a protecting group, there is also present a
counter anion; and
-N
-
represen,s a substituted or unsubstituted mono-, bi- or polycyclic
aromatic heterocyclic radical con~aining at least one nitrogen ~n
the ring and attached to A through a ring nitrogen, thereby
~orming a oua~ernary a~monium group; and pharmaceutically accept-
able salts thereof by the pr~cess shown m the following reaction
scheme:
R8 ~
Rl ~ diphenyl chlorophosphate
N ~ COoR2 -~>
III -
R H
Rl - ~ P(oc6H5)2 HS-A-OH
2'
COOR A = aikylene or
CS-C6 cycloallcylene
~:V
R8 H
.R I ~ S-A-OH . methanesulfonyl .chloride
~ --jCOC~R2 .' ~ ~3,,
V
R8 ~ .
R~ S-A-oso2cH3 ~)
~ ~--C
VI
Rl~ ~SOOR-2 ! AS~
II (~) = counter anion)
R8 H
R~
I 'A
Rl~ S-A-rC)
IA
U.S. Pat~nt No. 4,536,335 of Choung U. Kim and
Peter F. Micso, Jr., issued August 20, 1985, discloses
preparation o carbapenem antibiotics of the formula
R8 R R5
l ~ - A -N
O ~oOR2
. IB
; wherein R8 is hydrogen and Rl is selected from the group consisting
of hydrogen; substituted and unsubstituted: alkyl, alkenyl and
t alkynyl, having from 1-10 carbon a~oms; cycloalkyl and cycloalkyl-
alkyl, having 3-6 carbon atoms in the cycloalkyl ring and 1-6
carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and
aralkynyl wherein the aryl moiety is phenyl and the aliphatic
portion has 1-6 carbon atoms; heteroaryl, heteroaralkyl, hetero-
cyclyl and heterocyclylalXyl wherein the hetero atcm or atoms in
~ the above-named heterocyclic moieties are selected from the group
consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl
moieties associated with said heterocyclic moieties have 1-6
carbon atoms; wherein the substituent or substituents relative to
the above-named radicals are independently selected from the group
consisting of
r~
~2~
Cl-C6 alkyl optionally substituted by
amino, halo, hydroxy or carboxyl
halo
-oR3
O
oCNR3R4
-CNR3R4
- NR3 R4-
NR3
,~/
\NR3 R
-So2~R3R4
-NHC~NR3R4
O
R3 CNR4
-Co2R3
~O
- ocR3
-S~3
- SR9
Il 9
S R
-CN
N3 3
-OSO3R
-OSO2R
-NR3So2R4
.
: ~ '
~` ' .
', ~,
-NR3C-NR4
-NR3Co2R4
-N02
wherein, relative to the above-named substituents, the
groups R3 and R4 are independently selected from
hydrogen; alkyl, alkenyl and alkynyl, having from 1-10
carbon atoms; cycloalkyl, cycloalkyalkyl and
alkycycloalkyl, having 3-6 carbon atoms in the cycloalkyl
ring and 1-6 carbon atoms in the alkyl moieties; phenyl;
arlakyl, a:ralkenyl and aralkynyl wherein the aryl moiety
is phenyl and the aliphatic portion has 1-6 carbon atoms;
and heteroaryl, heteroaralkyl, heterocyclyl and
heterocyclyalkyl wherein the hetero atom or atoms in the
above-named heterocyclic moieties are selected from the
group consisting of 1-4 oxygen, nitrogen or sulfur atoms
and the alkyl moieties associated with said heterocyclic
moieties have 1-6 carbon atoms, or R3 and R4 taken
together with the nitrogen to which at least one is
attached may form a 5- or 6-membered nitrogen-containing
heterocyclic ring; R9 is as defined for R3 except that it
may not be hydrogen; or wherein Rl and R8 taken together
represent C2-C10 alkylidene or C2-C10 alkylidene
substituted by hydroxy; R5 is selected from the group
consisting of substituted and unsubstituted: alkyl,
alkenyl and alkynyl, having from 1-10 carbon atoms;
cycloalkyl and cycloalkylalkyl having 3-6 carbon atoms in
the cycloalkyl ring and 1-6 carbon atoms in the alkyl
moieties; phenyl; aralkyl, aralkenyl and aralkynyl
wherein the aryl moiety is phenyl and the aliphatic
portion has 1-6 carbon atoms; heteroaryl, heteroaralkyl,
heterocyclyl and heterocyclylalkyl wherein the hetero
a~om or atoms in the above-named heterocyclic moieties
are selected from the group consisting of 1-~ oxygen,
nitrogen or sulfur atoms and the alkyl moieties
-
-
. ~
.
.
~l2~
9(a)
associated with said heterocyclic moieties have 1-6
carbon atoms; wherein the above-named R5 radicals are
optionally substituted by 1-3 substitu~nts independently
selected from:
~2843~
-- 10 --
Cl-C6 21kyl o~'ionzlly substi~u'ed by 2~no,
fluoro, chloro, ca-boxyl, ~ydro~ or c~ aoyl;
fluoro, chloro or brom~;
_ 0~.3
~ C2 ~3
- ocoR3
.
-oSo2R3;
--ox~
NR R ; .
R3 C o~R4
-~:R C02R ; . ..
-NR3 CoNR3 R
~NR3502R4;
-sR3 i
~ ,.
-S-R';
O O
9 :
-S-R
-S03:R3 ~
-C02~ ;
-CoNR3R4;
-CN; or
phenyl optionzlly sl~'os~ituted ~y 1-3 'luoro,
chloro, b:romo, Cl-C6 21kyl, -oR3 ~ 3:R4,
-S03R, -C02"~ or -CoNR3R4, wherein R.3,. R 2~
R in such ~ subs ~ituen~s ~re 2S de ~ined æ~ove;
.
~ , . ' .
,
3~L
or RS may represent a di~alent phenylene or Cl-C4 alkylene group
joined to the
J
ring so as to form 2 bridged polycyclic group; A is cyclopentylene,
or cyclohexylene or C2-C6 alkylene optionally substituted ~y one
or more Cl-C4 alkyl gxoups; ~ is hydrogen,, an anionic charge o~
a.con~entional readily removable carboxyl protecting group, provid-
ing that when R2 is hydrogen or a protecting group, there is also
present 2 counter ion; and
, N J
represents a substituted or unsubstituted mono-, bi- or polycyclic
non-aromatic heterocyclic radical containing at least one nitrogen
in the ring and attached to A through a ring nitrogen, thereby
formlng' a quaternary ammonium group; and pharmaceutically accept-
able salts thereof, by the process shown in the following reaction
scheme:
R8 ~ ` ~
dipXenyl chlorophosphate
0~ --COOR2
~ III
.
- 12 -
r R ~ ~ P(C6H5)2~
HS - A - OH
N COOR
A=alkylene or
IV
C5-C6 cycloalkylene
R
Rl ~ S -A - OH
methanesulfonyl chloride
N ____I__ COOR
R H
Rl ~ S-A-OSO2CH3 I~
~ N 2'
O COOR
VI
~ ~ S-A-I
O . COOR Ag X~
`
R8 HR ~
Rl 5-A ~ N J optional
N ~ COOR2' de-blocking
I'B
. .
-
. , .
~ 13 -
R8 H R5
Rl ~ S-A-~ N
COOR2
To elaborate on the prior art scheme, starting
material III is reacted in an inert organic solvent with
diphenyl chlorophosphate in the presence of a base to give
intermediate IV. Intermediate IV is then reacted with a
mercaptan reagent of the formula HS-A-OH in an inert organic
solvent and in the presence of a base to give intermediate V.
Intermediate V is then acylated with methanesulfonyl chloride
in an inert organic solvent and in the presence of base to
give intermediate ~I which is reacted with a source of iodide
ions in an inert organic solvent to give intermediate II.
Intermediate II is reacted with the desired a~ine in an inert
organiC solvent and in the presence o silver ion to produce
the quaternized product I'A or I'B which may then be de-
blocked to give the corresponding de-blocked carbapenem of
Formula IA or IB.
,
The abo~e-described process has several disadvantages.
Thus, for example, the process involves several steps which
advantageously could be reduced in number. The overall reaction
yield is also quite low and the quaternizatio~ step is performed
on the entire carbapenem compound. It would be desirable to have
a new process for producing compounds of Formula IA or IB which
(1) involves fewer reaction steps, (2) gives higher yields,(3~ allows
the quaternized ~mine to be formed first and then attached to the
carbapenem nucleus at a later stage in the synthesis and (4) can be
used to more easily form quaternary amine products with a wide ~~
variety of amines, i.e. amines with steric hindrance and those with
low PKb values. ~ ~
~ , .
.' '
32~L
- 14 -
The present inven~ion provides a novel process
for preparation of carbapenem deri~-atlves of the formula
Rl~S_A_R14
O COOR
wherein R is hydrogen and Rl is selected from the group con~
sisting o~ hydrogen; substituted and unsubstituted: alkyl,
alkenyl and alkynyl, having from 1-10 carbon atoms; cycloalkyl
and cycloalkylalkyl, having 3-6 carbon atoms in the cycloalkyl
ring and 1-6 carbon atoms in the alkyl moieties; phenyl;.aralkyl,
aralkenyl and aralkynyl wherein the aryl moiety is phenyl and
the aliphatic portion has 1-6 carbon atoms; heteroaryl, hetero-
aralkyl, heterocyclyl and heterocyclylalkyl wherein the hetero
atom or atoms in the above-named heterocyclic moieties are
selected from the group consisting of 1-4 oxygen, nitrogen or
sul~ur atoms and the alkyl moieties associated with said hetero-
cyclic moieties have 1-6 carbon atomsi wherein the substituent:
or substituents relative to the above-named radicals are selected
from the group consisting o~:
:
:
: : :
.. .
_ .
r ' '
' ' ' . ' '
- : ' . , :, ', .
. " ' ~ , ' ' . ' ' I "
,
', ' ., ' ' , '
~!L2~
-- 15 --
Cl-C6 21~;yl c~p.io~21~y s~sti ,u~e~ by
æ-nino, halo, hydroxy or car;)oxyl
3 ~ 4
R3 ~4
-NR R
~R3 R4
3 4
-S02~R ~
: ~I 3 4
-~nC~2 R
3 ll 4
Z
=~
ll 3
- OCR
~, ~
~ ~ 5~
o
I g
-SR
- ~ R
: : O ,, ,
:
3 ~ ~
: ~ :
- 16 -
-oSo3R3
-oso2-~3
-NR3-So2R4
-NR3~=NR4
R3
-NR3Co2R4
-N02
wherein, relative to the above-name~ substituents, the
groups R3 and R4 are independently selected from
hydrogen; alkyl, alkenyl and alkynyl, having from 1-10
carbon atoms; cycloalkyl, cycloalkyalkyl and
alkylcycloalkyl, having 3-6 carbon atoms in the
cycloalkyl ring and 1-6 carbon atoms in the alkyl
moieties; phenyl; aralkyl, aralkenyl and aralkynyl
wherein the aryl moiety is phenyl and the aliphatic
portion has 1-6 carbon atoms; and heteroaryl,
heteroaralkyl, heterocyclyl and heterocyclylalkyl wherein
the hetero atom or atoms in the above-named heterocyclic
moieties are selected from the group consisting of 1-4
oxygen, nitrogen or sulfur atoms and the alkyl moieties
associated with said heterocyclic moieties have 1-6
carbon atoms, or R3 and R4 taken together with the
nitrogen to which at least one is attached may form a 5-
or 6-membered nitrogen-containing heterocyclic ring; R9
is as defined for R3 except that it may not be hydrogen;
or wherein Rl and R8 taken together represent C2-C10
alkylidene or C2-C10 alkylidene substituted.by hydroxy; A
is cyclopentyl.ene, cyclohexylene or C2-C6:alkylene~
optionally substituted by one or more C1-C4~alkyl groups;
R2 is hydrogen, an anionic charge or a conventional
readily removable carboxyl protecting group, providing
that when R2 is hydrogen or a protecting group, there is
also present a counter
'
~1 ,
i
. - . . ,:
.~ ....
.
. . :
- 17 -
anion; and Rl4 is a quaternized nitrogen-con-aining aromatic
or non-aromatic heterocycle attached to A through a ring
nitrogen, thereby forming a quaternary ammonium group, or a
pharmaceutically acceptable salt thereof, which process comprises
reacting an intermediate of the formula
R8 H
Rl ~--.
o N ~~ ~cooR2'
IV
wherein Rl and R8 are as defined above, R2 is a conventional,
readily removable carboxyl protecting group and L is a conventional
leaving group)with a thiol compound of the formula
~S-A-Rl~2 X~3
VII
wherein A and Rl4 are as defined above and X~ is a counter anion
in an inert solvent and in ~he presence of base~ to produce a
carbapenem product of the formula
8 H
R~ S_A_R14
~ N ~ 2' xa
o COOR
wherein Rl, R8, R2 , A, Rl4 and X3 are as defined above and, if
desired, removing the carboxyl protecting group R to give the
corresponding de-blocked compound of formula I, or a pharma-
ceutically acceptable salt thereof. --
~2~
~ 18 -
Also provided by the present invention are
intermediates of Formula VII and processes for preparing
such intermediates.
The carbapenem compounds of Formula I are
potent antibacterial agents or intermediates userul in the
preparation of such agents.
The compounds of general Formula I above contain
the carbapenem nucleus
6 ~ 2
~ N 1 3
0 4
and may thus be named as l-carba-2-penem-3-carboxylic acid
derivatives. Alternatively, the compounds may be considered
to have the basic structure 4
` O ~ ~ 2
and named as 7-oxo-l-azabicyclot3.2.0)hept-2-ene-2-carboxylic
acid derivatives. While the present invention includes compounds
wherein the relative stereochemistry of the 5,6-protons is cls
as well as transr the preferred compounds have the 5R,6S (trans)
stereochemistry as in the case of thienamycin.
The compounds of Formula I may be unsubstituted in
the 6-position or substituted by~substituent groups previously
disclosed for other carbapenem derivatives. More specifically,
.
.
R8 may be hydrogen and R1 may be hydrogen or a non-hydrogen
substituent disclosed, for example, in European Pa~ent Application
38,869 (see definition of R6). Alternatively, R8 and Rl taken
together may be C2-C10 alkylidene or C2-C10 alkylidene substituted,
for example, by hydroxy.
To elaborate on.the definitions for Rl and R8:
(a) The aliphatic "al~yl", "alkenyl" and "alkynyl" groups
may be straight or branched chain having 1-10 carbon atoms;
preferred are 1-6, mos~ preferably 1-4, carbon groups;
when part of another substituent, e.g. as in cycloalkyl-
alkyl, or heteroaralkyl or aralkenyl, the alkyl, alkenyl
and alkynyl group preferably contains 1-6, most preferably
1-4, carbon atoms.
.,
(b) "heteroaryl" includes mono-, bi- and.polycyclic aromatic
heterocyclic groups containing 1-4 O, N or S atoms; pre-
ferred are 5- or 6- membered heterocyclic rings such as
thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl,
isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, tetrazolyl,
oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl,
pyrrolyl, pyrazolyl, etc.
(c) "heterocyclyl" includes mono-, bi- and polycyclic
saturated or unsaturated non-aromatic heterocyclic groups
con~aining 1-4 O, N or S atoms; preferred are 5- or 6-
membered heterocyclic rings such as morpholinyl, piperazinyl,
piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl,
imidazolidinyl, pyrrolinyl, pyrrolidinyl, etc.
(d) "halo" includes chloro, bromo, fluoro and iodo and i5
preferably chloro or bromo.
~4.~2~
- 20 -
The term "conventional readily removable carboxyl
protecting group" refers to a known ester group which has been
employed to block a carboxyl group dùring the chemical reaction
steps described below and which can be removed, if desired, by
methods which do not result in any appreciable destruction of
the remaining portion of the molecule, e.g. by chemical or
enzymatic hydrolysis, treatment with chemical reducing agents
under mild conditions, irradiation with ultraYiolet light or
catalytic hydrogenation. Examples of such ester protecting
groups include benzhydryl, p-nitrobenzyl, 2-naphthylmethyl,
allyl, benzyl, trichloroethyl, silyl such as trimethylsilyl,
phenacyl, p-methoxybenzyl, acetonyl~ o-nitrobenzyl, 4-pyridyl-
methyl and Cl-C6 alkyl such as methyl, ethyl or t-butyl.
Included within such protecting groups are those which are
hydrolyzed under physiological conditions such as pivaloyloxy-
methyl, acetoxymethyl, phthalidyl, indanyl'and methoxymethyl.
Particularly advantageous carboxyl protecting groups are p-
nitrobenzyl which may be readily remoYed by catalytic hydro-
genolysis and allyl which can be removed by Pd(P~3)4-catalyzed
reaction.
The pharmaceutically acceptable salts referred to
above include the nontoxic acid addition salts, e.g. salts with
mineral acids such as hydrochloric, hydrobromic, hydroiodic,
phosphoric, sulfuric, etc. and salts with organic acids such as
maleic, acetic, citric, succinic, benzoic, tartaric, fumaric,'
mandelic, ascorbic, lactic, gluconic and malic. Compounds of
Formula I in the form of acid addition salts may be written as
R8 H
R~ ~ A R14
N COOR2
R - H or protecting group
- 21 -
where X9 represents the acid anion. The counter anion X~
may be selected so as to provide pharmaceutically acceptable
salts for therapeutic administration but, in the case of
intermediate compounds of Formula I, X3 may also be a toxic
anion. In such a case the ion can be subsequently removed or
substituted by a pharmaceutically acceptable anion to form
active end product for therapeutic use. When acidic or basic
groups are present in the Rl group or on the quaternized Rl4
radical, the present invention may also include suitable
base or acid salts of these functional groups, e.g. acid
addition salts in the case of a basic group and metal salts
(e.g. sodium, potassium, calcium and aluminum), the ammonium
salt and salts with nontoxic amines (e.g. trialkylamines, procaine,
dibenzylamine, l-ephenamine, N-benzyl-~-phenethylamine, N,N'-
dibenzylethylenediamine, etc.) in the case of an acidic group.
Compounds of Formula I wherein R2 is hydrogen, an
anionic charge or a physiologically hydrolyzable ester group
together with pharmaceutically acceptable salts thereof are
useful as antibacterial agents. The remaining compounds of
Formula I are valuable intermediates which can be conver~ed
into ~he above-mentioned biologically active compounds.
A preferred embodiment of the present invention
comprises compounds of Formula I wherein R8 is hydrogen and
is hydrogen, CH3CH2-
` CH CH OH OH
~ 1
~ CH-, j C- or CH3CH-
CH3 CH3
Among this subclass, the preferred compounds are those in
which Rl is ---
O~
CH3CH-, most preferably compounds having the
': '
,
.
- 22 -
absolute conriguration 5R, 6S, 8R.
Another preerred embodiment comprises compounds of
Formula I in which R~ and R8 taXen together form an alkylidene
radical of the formula
C- -
. C~3
The alkylene or cycloalkylene radical A in the compounds
of Formula I may be
cyclopentylene \ 7
~ ~2C CH2 .
cyclohexylene ( ~ ' ) f
or C2-C6 alXylene optionally substituted by one or more Cl-C4
alkyl substituents. Preferred A substituents are cyclopentylene,
cyclohexylene or alkylene of the formula
~,10 R12
--C~ ~
~ 13
i hich R10 Rll R12 and R13 are each independently hydrogen or
Cl C4 alkyl. A preferred embodiment comprises those compounds of
Formula I in which-substituent A is -CH2CH2-, -ICHCH2-,
CH3
or -CH~CH-
In the case of certain compounds of Formula I having acycloalkylene or branched alkylene A substituent, one or more --
additional assymmetric carbon atoms may be created which resul~
in formation of diaster~éolsomers. The present invention includes
mixtures of such diastereoisomers as well as the individual
ourified diastereoisomers.
,.~, ~f ' .
- 23 -
The quaternized ~14 substituent may be an optionally
substituted mono-, bi- or polycyclic aromatic or non-aromatic
heterocyclic radical containing at least one nitrogen in the
ring and attached to A through a ring nitrogen, thereby forming
a quaternary ammonium group.
One preferred class of R14 substituents may be
represented by the general formula
. ~I J
which is meant to define a substituted or unsubstituted mono-,
bi- or polycyclic heteroaryl radical containing at least one
nitrogen in the ring and attached to a carbon atom of substituent
A through a ring nitrogen, thereby forming a quaternary ammonium
group. The heteroaryl radical may be optionally substituted by
such substituents as Cl-C4 alkyl, Cl-C4 alkyl substituted by
hydroxy, amino, carboxy or halo, C3-C6 cycloalkyl, Cl-C4 alkoxy,
Cl-C4 alkylthio, amino, Cl-C4 alkylamino, di(Cl-C4 alkyl)amino,
halo, Cl-C4 alkanoylamino, Cl-C4 alkanoyloxy, carboxy,
R
-C-OCl-C4 alkyl,
hydroxy, amidino, guanidino, trifluoromethyl, phenyl, phenyl
substituted by one, two or three amino, halo, hyd~roxyl, trifluoro-
methyl, Cl-C4 alkyl or Cl-C4 alkoxy groups, heteroaryl and hetero-
aralkyl in which the hetero a~om or atoms in the above-named
heterocyclic moieties are selected from the group consisting of
1-4 O, N or S atoms and the alkyl moiety associated with hetexo-
aralkyl has I-6 carbon atoms.
The heteroaryl radical attached to substituent A is
preferably a 5- or 6- membered aromatic heterocyclic radical
containing a quaternized nitrogen atom (which is directly bonded
to a carbon atom o' the alkylene or cycloalkylene radical) and,
optionally, one or more additional hetero atoms selected .rom O,
N or S. While, in general, any heteroaryl radical bonded to A
.
,
via a quaternized nitrogen atom is found to produce biologically
ac~ive carbapenem derivatives, a preferred embodiment comprises
compounds in which
- N J
/
represents a radical selected from the group consisting of
R7
(a) \ ~ ~ R6
R
wherein R5, R6 and R7 are independently selected from hydrogen;
Cl-C4 alkyl; Cl-C4 alkyl substituted by hydroxy, amino, carboxy
or halo; C3-C6 cycloalkyl; Cl-C4 alkoxy; Cl-C4 alkylthio; amino;
Cl-C4 alkylamino; di(Cl-C4 alkyl)amino; halo; Cl-C4 alkanoylamino;
Cl-C4 alkanoyloxy; carboxy;
, -C-OCl-C4 alkyl; hydroxy, amidino;
guanidino; trifluoromethyl; phenyl; phenyl substituted by one,
two or three amino, halo, hydroxyl, trifluoromethyl, Cl-C4 alkyl
or Cl-C4 alkoxy groups; and heteroaryl and heteroaralkyl in which
the hetero atom or atoms in the above-named heterocyclic moieties
are selected from the group consisting or 1-4 oxygen, nitrogen or
sulfur atoms and the alkyl moiety associated with said hetero-
aralkyl moiety has 1-6 carbon atoms; or wherein two of R5, R6 or
R taken together may be a fused saturated carbocyclic ring, a
fused aromatic carbocyclic:ring, a fused saturated heterocyclic
ring or a fused heteroaromatic ring,
~b) ~ ~ or ~3
, ,
optionally su~stituted on a carbon atom by one or more sub-
stituents independently selected from Cl-C4 alkyl; Cl-C4 alkyl
substituted by hydroxy, amino, carboxy or halogen; C3-C6 cyclo-
alXyl; Cl-C4 alkoxy; Cl-C4 alkylthio; amino; Cl-C4 alkylamino;
`di(Cl-C4 alkyl)amino; halo; Cl-C4 alkanoylamino; Cl-C4 alkanoyloxy;
carboxy; -C-OCl-C4 alkyl; hydroxy; amidinoi guanidino; trifluoro-
methyl; phenyl; phenyl substituted by one, two or three amino,
halo, hydroxyl, trifluoromethyl, Cl-C4 alkyl or Cl-C4 alkoxy
groups; and heteroaryl or heteroaralkyl in which the heteroatom
or atoms in the above-named heterocyclic moieties are selected
from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms
and the alkyl moiety associated with said heteroaralkyl moiety
has 1-6 carbon atoms, or optionally substituted so as to form a
fused carbocyclic or heterocyclic ring;
(C) ~ ~N N~
N ~ N
~ 3 ~ 3 ~ 3
optionally substituted on a carbon atom by one or more substituents
independently selected from C1-C4 alkyl; Cl-C4 alkyl substituted
by hydroxy, amino, carboxy or halogen; C3-C6 cycloalkyl; C1-C~.
alkoxy; Cl-C4 alkylthio; amino; Cl-C4 alkylamino; di(Cl-C~ alkyl)-
amino; halo; Cl-C4 alkanoylamino, Cl-C4 alkanoyloxy; carboxy;
q _.
-C-OCl-C4 alkyl; hydroxy; amidino; guanidino; trifluoromethyl
phenyl; phenyl substitated by one, two or three amino, halo,
.
.
'
- 26 -
hydroxyl, trifluoromethyl, Cl-C4 alkyl or Cl-C4 alkoxy groups;
and heteroaryl or heteroaralkyl ln whlch the hetero atom or
atoms in the above-named heterocyclic moieties are selected
from the group consisting of 1-4 oxygen, nitrogen or sulfur
atoms and the alkyl molety assoclated with said heteroaralkyl
moiety has 1-6 carbon atoms, or optionally substituted so as
to form a fused carbocyclic or heterocyclic ring;
(d) ~ ~ ~ ~ N
~ N ~ N N ~
~N 3 ~ ~N N N ~N
optionally substituted on a carbon atom by one or more substituents
j independently selected from Cl-C4 alXyl; Cl-C4 alXyl substituted
; by hydroxy, amino, carboxy or halogen; C3-C6 cycloalkyl; Cl-C~
alkoxy; Cl-C4 alkylthio; amino; Cl-C4 alkylamino; di(Cl-C4 alXyl)-
amino; halo; Cl-C4 alkanoylamino; Cl-C4 alkanoyloxy; carboxy;
-C-OCl-C4 alkyl; hydroxy; amidino; guanidino; trifluoromethyl;
l~ phenyl; phenyl substituted by one, two or three amino, halo,
I hydroxyl, trifluoromethyl, Cl-C4 alkyl or Cl-C4 alkoxy groups;
and heteroaryl or heteroaralkyl in which the hetero atom or atoms
in the above-named heterocyclic moieties are selected from the
group consisting of 1-4 ox~gen, nitrogen or sulfur atoms and the
alkyl moiety associated with said heteroaralkyl moiety has 1-6
carbon atoms, or optionally substituted so as to form a fused --
carbocyclic or heterocyclic ring,
3~
~ or ~ ~
wherein X is 0, S or NR in which R is Cl-C4 alkyl or phenyl,
said radical being optionally substituted on a carbon atom by
one or more substituents independently selected from Cl-C4 alkyl;
Cl-C4 alkyl substituted by hydroxy, amino, carboxy or halogen;
6 1 C4 alkoxy; Cl-C4 alkylthio; amino; C C
alkylamino; ~i(Cl-C4 alkyl)amino; halo; Cl-C4 alkanoylamino;
-Cl-C4 alkanoyloxy; carboxy;
-C-OCl-C4 alkyl; hydroxy; amidino; guanidino; trifluoro~ethyl;
phenyl; phenyl substituted by one, two or three amino, halo,
hydroxyl, trifluoromethyl, Cl-C4 alkyl or Cl-C4 aIkoxy grou2s;
and he~eroaryl or heteroaralkyl in which the hetero atom or atoms
in the abo~e-named heterocyclic moieties are selected from the
group consisting of 1-4 oxygen, nitrogen or sulfur atoms and
the alkyl moiety associated with said heteroaralkyl moiety has
1-6 carbon atoms, or optionally substituted so as to form a
~used carbocyclic or heterocyclic ring;
N- , \X ~
- or ~ -X >
_ .
:
.
- ~ . ::, ... . .
- 28 -
wherein X is 0, S or NR ln which R is Cl-C4 alkyl or phenyl,
said radical being optionally substituted on a carbon atom by
one or more substituents independently selected from Cl-C4
alkyl; Cl-C4 alkyl substituted by hydroxy, amino, carboxy or
halogen; C3-C6 cycloalkyl; Cl-C4 alkoxy; Cl-C4 alXylthio; amino;
Cl-C4 alkylamino; di(Cl-C4 alkyl)amlno; halo; Cl-C4 alkanoylamino;
Cl-C4 alkanoyloxy; carboxy;
o
-C-OCl-C4 alkyl; hydroxy; amidino;
guanidino; trifluoromethyl; phenyl; phenyl substituted by one,
two or three amino, halo, hydroxyl, trifluoromethyl, Cl-C4 alkyl
or Cl-C4 alkoxy groups; and heteroaryl or heteroaralkyl in which
the hetero atom or atoms in the abo~e-named heterocyclic moieties
are selected from ~he group consisting of 1-4 oxygen, nitrogen
or sulfur atoms and the alkyl moiety associated with said hetero-
aralXyl moiety has 1-6 carbon atoms; and
N _ N\ j - ~
N ~ N-R N ~ N-R
N - N ~ N\-R
- ~ -R N ~ N-
~~N ~-R or ~R
~8~
29 -
wherein R is Cl-C4 alkyl or phenyl, said radical being optionally
substituted on ~he carbon atom by a substituent selected from Cl-C~
alXyl; Cl-C4 alkyl substituted by hydroxy; amino, carboxy or halogen
C3-C6 cycloalkyl; Cl-C4 alkoxy; Cl-C4 alkylthio; amino; Cl-C4
alkylamino; di(Cl-C4 alkyl)amino; C1-C4 alkanoylaminoi carboxy;
11 .
-C-OCl-C4 alkylj hydroxy; amidino; guanidino; trifluoromethyl;
phenyl, phenyl substituted by one, two or three amino, halo,
hydroxyl, trifluoromethyl, C1-C4 alkyl or C1-C4 alkoxy groups;
and heteroaryl or heteroaralkyl in which the hetero atom or
.atoms in the above-named heterocyclic moieties are selected
from the group consisting of 1-4 oxygen, nitrogen or sulfur
atoms and the alkyl moiety associated with said heteroaralXyl
moiety has 1-6 carbon atoms.
Within the above subclass, the preferred compounds
are those in which substituent A is -CH2CH2-, -~HCH2 , ~ or
1 8 CH3 ~
-CH~CH- and wherein either (a) R- and R taken together represent
CH3 HOC
C 3
or (b) R is hydrogen and R represents hydrogen, CH3CH2-,
C ~ ~ I OH
", CH- , ," C- or CH3CH-
CH3 C 3
Particularly preferred are the compounds wherein R8 is hydrogen
and Rl is
OH
CH3CH-, preferably compounds having the absolute -.
configuration SR, 65, 8R.
:;
,
- ~ '
- 30 -
A particularly preferred embodiment of the present
invention comprises preparation of compounds wherein
- ~ )
represents a radical of the formula
I ~ R7
R ~
in which R5, R6 and R7 are independently selected from
the group consisting bf hydrogen, C1-C4 alkyl, C1-C4
alkoxy, Cl-C4 alkyl substituted by a hydroxy group, Cl-C4
alkylthio, amino, carboxy and carbamoyl. Within this
subclass, the preferred compounds are those wherein
substitu~nt A is
-CH CH -, -CHCH2- ' ~ or CH2CH
and wherein either (a) R1 and R8 taken together represent
HOC ~
CH C=
or (b) R8 is hydrogen and Rl represents hydrogen,
CH3CH2-
CH CH OH OH
~ ~1. .......... I
CH- , C- o~ CH3CH
` CH3 3
Particularly preferred are the compounds wherein R8 is
.
~ .
,~
~, .
'. ' ' ' : . : '' . '
'.. :' . ' .
' ' ' ' ' ~'; '' '
.
- 30(a) -
hydrogen and R1 is
~H
CH3CH-, preferably compounds having the absolute
configuration 5R, 6S, 8R.
:
.
-" ~2~32~
~.
- 31 -
A most preerred embodi~ent of the present invention
comprises preparation or compounds wherein
represents a radical of the formula
--~R7
in which R5, R6 and R7 are independently selected from the
, group consisting of hydrogen, Cl-C4 alkyl, Cl-C4 alkoxy, Cl-C4
alkyl substituted by a hydroxy group, C1-C4 alkylthio and amino.
Within this subclass, the preferred compounds are those wherein
substituent A is -CH2CH2-, -ICHCH2 ~ ~ or -CH2CH-
and wherein either (a) Rl and R8 taken together represent
HOC~
~ C=CH
or (b) R8 is hydrogen and Rl represents hydrogen, CH3CH~-,
CH~ CH ~ OH OH
CX- , C- or CH3CH~
CH3 CH3
Particularly preferred are the compounds wherein R8 is hydrogen
and Rl is
~H
CH3CH-, preferably compounds having the absolute
configuration SR, 6S, 8R.
Another preferred,embodiment of the present inventlon
comprises preparation of compounds wherein
~ ~0 ' ' '
,
.
.
,
' ~ ' -
.
~ 8 ~
represents a radical of the formula
-N - \ SCH3
~ -CH3
Within this subclass, the preferred compounds are those wherein
substi~uent A is -CHCH2-, -CH2CH2- , ~ or -CH2CH-
3 ~ CH3and wherein either (a) Rl and ~8 taXen together represent
HOC~
C=
C~'
or (b) R8 is hydrogen and Rl represents hydrogen, CH3CH2-,
CX~ CH ~ OH OH
CH- C- or CH3CH-
CH3 CH3
Particularly preferred are the compounds wherein R8 is
hydrogen and Rl is
OH
CH3CH-, preferably compounds having the
absolute con~iguration 5R, 6S, 8R.
Another preferred em~odiment of the present invention
comprises preparation of compounds wherein
~ ~ .
- N
represents a pyridinium radical. Within this subclass, the
j preferred compounds are those wherein substituent A is -ICHCH2-,
CH3
' -CH2CH2- ~ ~ or -C~2C~- and wherein either (a) --
¦1 Rl and R8 taken together represent
.
.. . .
. .
. - . .
:. .. .
., . , , .
. - . . .
~8~3~
- 33 -
HOC\2
~C=
C 3
or (b) R8 is hydrogen and Rl represents hydrogen, ~H3CH2-,
CH~ CH ~ fH OH
CH- , C- or CH3CH-
CH3 CH3
~articularly preferred are the compounds wherein R8 is hydrogen
and R is
~H
CH3CH-, preferably compounds ha~ing the absolute
configuration 5R, 6S, 8R.
A most preferred embodiment of the present invention
comprises the preparation of compounds of the formula
OH H
(R) ~ ~ ~ 5 - A-N
9 COOR
~ .
wherein
- A -N
represents
... ~
- 34 -
OE~ 3 ~
H2CH2 N~3 ( 2? -CH CH ~N~3
t 3 ) - CH 2 C~l 2 eN~ ( 4 ) 2 . 2
H
S) -Ch' CH ~N~ 3 ~ ~) SC~3
(7) -C1~2C~2 ~3C,a2oH ( 8 ) ~3
(9 ) C~2C~2 ~ I (iO ) CH2C~
( 11 ) - C~ CR2 e~N~3 , 3
. 3. ~ (12 ) -CR2C~2 N~
R or S di~stexeoisom~s ~N~3
N~ 2 ~ . . . ..... , .. _ ,
( 13) ~ -CH2CH2~N~ o_ (14) 1~3J
R,R or S ,S
~J dizstereoisorners
at two a~ylTmetric
carbons of the
cyclohexy l grou~
,
.
~L~8~
and R2 is hydrogen, an anionic charge ox a conventional readily
removable carboxyl protecting group, providlng that when R2 is
hydrogen or a protecting group, there is also present a counter
anion, and pharmaceutically acceptable acid addition salts thereof
Another preferred class of quaternized R14 substituents
may be represented by the general formula
--N/~)
in which R~6 is selected rom the group consisting of substitu.ed
and unsubstituted: alkyl, alkenyl and alkynyl., having from 1-10
carbon atoms; cycloalkyl and cycloalkylalkyl, having 3-6 ca~bon
atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl
moieties; phenyl; aralkyl, aralXenyl and aralkynyl wherein the
aryl moiety is phenyl and the aliphatic portion has 1-6 carbon
atoms; heteroaryl, he~eroaralXyl, heterocyclyl and heterocyclyl-
alkyl wherein the hetero atom or atoms in the above-named hetero-
cyclic moieties are selected from the group consisting of 1-4
oxygen, nitrogen or sulfur atoms and the alkyl moieties associated
with said heterocyclic moieties have 1-6 carbon atoms; wherein
the above-named R15 radicals are optionally substituted by 1-3
substituents independently selected from:
~, ~ ' ' .
: - :' . '
-- 36 --
Cl-C6 alkyl optionall~ subs~ilu~ed by ~ o,
^ uoro, chloro, c:a~boxyl, hydroxy or cæba~oyl,
luoro, chloro or bromo ;
_ o~.3
-oco2~3
o23
.' ,, . C~S2R ; , '
--ox~
-NR :R ; .
_~. 3
' ' -NR3CoNR3 ~ ; '
-NR3 S 02R
-S~.3;
O
S-R9;
,0 ~0
-S-R
` ~ -503R3;
.
--C2 R3 . ; , '.
3 4
- CON2 3~ ;
-CN; or
phenyl option?lly substituted by 1-3 ~'luoro,
chlo_o, bromo, Cl-C6 al~cyl, -oR3, -N~3R4~, ;
-~03R3 , ~ 2i?.3 or -coNx3~4 , wherei2l ~3, R4 .an
R9 in such- ~16 sub5-t:i tuents are 2S de~ined 2~ove, ::.
:
; .
: , . .
, ~ .. . ,
. ' .
2~
- 37 -
or Rl6 may represent a divalent phenylene or C1-C4
alkylene group joined to the
\~
-N
ring so as to form a bridged polycyclic group and
\~
. - N J
represents a substituted or unsubstituted mono-, bi-or
polycyclic non-aromatic (which may be fused to another
aromatic o~ non-aromatic ring) heterocyclic radical
containing at least one nitrogen in the ring and attached
to A through a ring nitrogen, thereby forming a
quaternary ammonium group. The heterocyclic radical may
be saturated or unsaturated (with 1-2 double bonds) and
may contain up to two additional hetero atoms in addition
to the quaternary nitrogen, such additional hetero atoms
being selected from O, S(O)m/ N, NR15 or NR17R18 wherein
m is O, 1 or 2, R15 is hydrogen, optionally substituted
C1-C6 alkyl or optionally substituted phenyl and R17 and
R18 are each independently optionally substituted C1-C6
alkyl or optionally substituted phenyl.
In a preferred embodiment
\~ ~
. . ,
represents a non-aromatic 4-7 membered, preferably 5- or
6- membered, N-containing heterocyclic ring containing 0-
2 double bonds and 0-2 additional hetero-ato~s selected
from O, S(O~m/ N, NR15 or NR17R18 wherein m is 0, 1 or 2,
Rl5 is hydrogen, Cl-C6 alkyl optionally substituted by 1-
.. ~, . .... ~ ...
_ 37(a) ~
2 substituents independently selected from -oR3, -NR3R4,
-Co2R3, oxo, phenyl, fluoro, chloro, bromo, -S03R3 and
-CoNR3R4 or phenyl optionally substituted by 1-3
substituents independently selected from C1-C6 alkyl,
-oR3, -NR3R4, fluoro, chloro, bromo, -$03R3, -C02R2 and
-CoNR3R4 and Rl7 and R18 are each independently C1-C6
alkyl optionally substituted by 1-2 substituents
independently selected from -oR3, -NR3R4, -C02R3, oxo,
phenyl, fluoro, chloro, bromo, -S03R3 and -CoNR3R4 or
phenyl
. . .
-
- 38 -
o~tionally substituted by 1-3 substituents independ~ntly selec~d
from Cl-C6 al~yl, -oR3~ -NR3R4, fluoro, chloro, bromo, SO3R ,
-CO R~ and -coNR3R4~ wherein ~3 and R4 in such heterocyclic N~15 or
NRl~R18 gro-~ps are as defined above in connection with the R16
substituent. In such preferred embodiment the
\1~
N
ring may be optionally substituted by 1-3 substituents independently
selected from
~a) Cl-C6 alXyl optionally substituted by 1-2 substituents
inde~endently selected from fluoro, chloro, bromo, -oR3,
OCOR , -oCoNR3~4, oxo, -N~3R4 NR3CoR4 3 3 4
-NR3So2R4~ -SR , -S03R , -C02R and -CON~
(b) C2-C6 alkenyl optionally substituted by 1-2 substituents
independently selected from fluoro, chloro, bromo, -oR3
-OCOR , -oCONR3R4, oxo, -NR3R4, -NR3CoR4, -NR3CoN~3R4
-NR3So2R4, -SR3 r -So3R3, -Co2R3 and -CoNR3R ;
tc) C2-C6 alXynyl optionally substituted by 1-2 substituents
independently selected f~om fluoro, chloro, bromo, -OR,
CoNR3~4 o~o -N~3R4r -NR3CoR , -NR CONR
-NR So2R4, -SR3, -So3R3, -C02R and -CONR R ;
(d) C3-C6 cycloalXyl optionally substituted by 1-2 substituents
independently selected from fluoro, chloro, bromo, -oR3,
-oCoR3, -oCoN~3R4, oxo, -NR3R4, -NR3CoR , -N~ CON~ R ,
-NR3So2R4, -SR3, -S03R3, -C02R and -CONR R ~
(e) cycloalkylalkyl ha~ing 3-6 carbon a,oms in the cycloalXyl
¦ ring and 1-6 carbon atoms in the alkyl moiety~ optionally
~ substituted by 1-2 substituents independently selected from
! fluoro, chloror bromo ~ -oR3 / -ocoR3 ~ -OCONR R , oxo, -N~3R4,
-NR3CoR4, NR3CoNR R4, -NR3S02R , -SR , -SO3R , -CO2~ and
~C5NR R;
!
.
- 39 -
(r~ hete-oaryl wherein ~e hetero.atom or a~oms are selected
from the group consisting of l-4 oxygen, n-t-ogen or
sulfur atoms, optionally substituted by 1-2 substituents
independently selected from fluoro, chloro, bromo, -oR3,
-ocoR3~ -oCoNR3R4, oxo, -NR3R4, -N~3CoR4; -NR3coNR3R4
-N~3so2R , -SR3, -S03R3, -C02R3 and -coN~3~ ; pre'erred
heteroaryl radicals are 5- or 6-~embered aromztic hetera-
cyclic xings;
(g) heteroaralkyl wherein the hetero atom or atoms are selected
from the sroup cons1sting of 1-4 oxygen, nitrogen or sulrur
atoms and the alkyl moiety has 1-6 carbon ato~s, optionally
substituted by 1-2 substituents independently selec,ed f om
fluoro, chloro, bromo, -oR3r -OCOR, -OCONR R, oxo,
-NR3R4, -NR3coR4r -N~3CoNR3R4, -N~3So2~4, -S~3, -S03R3,
-C02R and -CoNR3R4; preferred heteroarai~yl are those
in which t~e heteroa~yl _adical is a 5- or 6-membe~ed
aromatic heterocyclic ring and the alkyl moiety has
1-2 carbon atoms;
(h) heterocyclyl wherein the hetero atom or atoms are selected
from t~e group consisting of 1-4 oxygen, nitrogen or
sulfur atoms, optionally substituted by 1-2 substituents
independently selected from 'luoro, chloro, bromo, -oR3,
-OCOR , -oCoNR3R4, oxo, -NR3R4, -NR3Co~4, -NR3CoNR3R4
-NR S02R , -SR , -S03R., -C02R and -CONR R ; preferred
heterocyclyl are S- or 6-membered saturated or u~saturated
rings; - -
(i) heterocyclylalky.l wherein the hetero atom or atoms areselected from the group consisting o 1-4 oxygen, nitrogen
or sulfur atoms and the al~yl moiety has 1-6 carbon atoms,
optionally su~stituted by 1-2 substituents.independently
selected from ~luoro, chloro, bromo, -oR3, -oCoR3, -oCoNR3R4,
ox~, -NR3R4, -NR3CoR4, -NR3Co~R3R4, -N~3So2R4, -sR3, -S~3R3,
-Co2R3 and -CoNR3R; prererred heterocyclylalkyl are those
in which the heterocyclyl moiety is a S- or 6-mem~ered
s aturated or unsaturated ring;
.
- 40 -
~j) fluoro, chloro or bromo;
(k) -oR3;
~ oCO~R3 ;
(m) -ocoR3;
(n) -oCoNR3R
( o) -oso2R3
(P) oxo;
(q) -NR3R
(r) R3CoN~4_;
(s) NR3Co2R4 ;
.(t) -NR3CoNR3R4 ; ~ - - -.... . _
(u) -N~3502R4 ;
(v) -SR3 ;
(w) o
-S-R9 ;
(~) O O
~ 9 ,
(~) -503R3 ;
( Z) -Co2R3
(aa) -CONR R
(bb) -CN; or
(cc) phenyl optionally substituted by 1-3 fluoro, chloro, bromo,
.~ Cl-C6 alkyl, -OR , -NR R , -503R , C02R or CON~ R .
The R3, ~4 and R9 substituents mentioned a~o~e are as defined i~
connection wi',h substituent Rl.
The -N
. , ~
~8~3~
- 41 -
ring as defined above is a non-aromatic hetexocycle group.
This ring, however, may be fused to another ring which may be
a saturated or unsaturated carbocyclic ring, preferably a C4-C7
carbocyclic ring, a phenyl ring, a 4-7 membered heterocyclic
~saturated or unsaturated) ring containing 1-3 hetero atoms
selected from 0, N, S(O)~, NR15 or NX17R18 or a 5-6 membered
heteroaromatic ring containing 1-3 hetero atoms selected 0, S(O) ,
N~ NR15 or NR17R18 in which m, R 5, Rl and R are as defined
above.
The R16 substituent of the non-aromatic R14 radical
may be either (a) an optionally substituted Cl-C6 alXyl, C2-C10
alkenyl, C2-C10 alkynyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl~-
Cl-C6 alkyl, phenyl, phenyl-Cl-C6 alkyl, phenyl-C2-C6 alkenyl,
phenyl-C2-C6 alkynyl, heteroaryl, heteroaralkyl in which the
alkyl moiety has 1-6 carbon atoms, heterocyclyl or hetero-
cyclylalkyl in which the alkyl moiety has 1-6 carbon atoms or (b)
a divalent phenylene or Cl-C4 alkylene group joined to the
:
~2~
- 42 -
ring so as to form a bridged rlng polycyclic sroup, e.g. a
quinuclidine grou?. The heteroaryl (or he~eroaryl portion o'
heteroaralkyl) substituent may be a mono-, bi- or polycyclic
aroma,ic heterocyclic ~roup containing 1-4 O, N or S atoms;
preLerred are 5- or 6-membered heterocyclic rinss such as
~hienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl,
~hia~olyl, imidazolyl, isoxazolyl, tetrazolyl, oxazolyl, pyridyl,
pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl znd pyrazolyl. The
heterocyclyl ~or heterocyclyl portion of heterocyclylalkyl) sub-
stituen. may be a mono-, bi- or polycyclic saturated or unsatu-
ra~ed non-aromatic heterocyclic group containing 1-4 O, N or S
atoms; preferred are 5- or 6-membered heterocyclic rinss such 2S
mor~holinyl, piperazinyl, piperidyl, pyrazolinyi, pyrazolidinyl,
imidazolinyl, imidazolidinyl, pyrrolinyl and pyrrolidinyl.
In the case where ~he R16 su~stituent is ~ alkyl,
alkenyl, 21Xynyl, cycloalkyl, cycloaIXylalXyl; phenyl,_ phe~.yl-
alXyl, phenylalkenyl, phenylalkynyl, hete_oaryl, hetero~ralXyl,
he,erocyclyl or heterocyclylalkyl group, such s_oups may be
optionally subs~ituted by 1-3 substituents independently
selected rrom:
(a) Cl-C6 alXyl optionally substituted by, preferably 1-3,
amino, 'l~oro, chloro, c2rboxy1, hydroxy or c~rb2moyl
srou~s;
~b~ Lluoro, chloro or bromo;
( c) -oR3
(d~ -oC02R3 ;
:
~e) -oCoR3 ;
- ocoN~3 ~4 ;
( s) -oso2R3
(h) -oxo ;
(i) -NR3R4 ;
( j ~ R3CoNR4_ ;
(X~ -NR3C02R ;,
( 1~ -NR3CoNR3R4 ; - -
- 43 -
(m) -~R3so2R4;
(n) -SR3;
(o) -SOR9;
(p) -S02R9;
( q) -S03R3;
(r) -C02R3;
( s ) -CoNR3R4;
(t) -CN; or
(u) phenyl optionally substituted by 1-3 substituents
independently selected from fluoro, chloro, bromo, Cl-C6
alkyl, -oR3~ -NR3R4, -S03R3, -Co2R3 or -CoNR3R4~ wherein
relative to the above-named R16 substituents, the groups
R3 and R4 are independently selected from hydrogen;
alkyl, alkenyl and alkynyl, having 1-10 carbon atoms;
cycloalkyl, cycloalkylalkyl and alkylcycloalkyl, having
3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon
atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl
and aralkynyl wherein the aryl moiety is phenyl and the
aliphatic portion has 1-6 carbon atoms; and heteroaryl,
heteroaralkyl, heterocyclyl and heterocyclylalkyl wherein
the heteroaryl and heterocyclyl group or portion of a
group is as defined above for R16 and the alkyl moieties
associated with said heterocyclic moieties have 1-6
carbon atoms; or R3 and R4 taken together with the
nitrogen to which at least one is attached may form a 5-
or 6-membered nitrogen-containing heterocyclic (as
defined above for R5) ring; and R9 is as defined above
for R3 except that it may not be hydrogen. A most
preferred R16 substituent is C1-C6 alkyl, especially
methyl.
In the case where R16 is a divalent phenylene or C1-
C6 alkylene group, ~uch group is bonded to another atom
of the
\1~
.;.
"
.
3~
_ 43(a) -
ring so as to form a bridged polycyclic ring, e.g. a
quaternized quinuclidine ring of the formula
.. . ..
!i
- 44 -
A particularly preferred embodiment of the present
invention comprises preparation o~ compounds of Formula I wherein
R16
-\N
reDresents
\ ~ y , _ ~ CH
\ ~,r~ \ ~ ~
y y Y
\ /f \ ~ ~N/f \N r
-~N ~ _CH3 , -~N ~ ~ ' ~ ~ ~ CH3
CH~ ~ ~ C\ ~
wherein Y is hydrogen, Cl-C6 alkyl, hydroxy, -SCl-C6 alkyl,
carboxyl, carbamoyl, chloro, bromo, iodo,~ fluoro or phenyl.
Within this subclass, the pre~erred compounds are those
_.
.
~ fp ~
- 45 -
wherein A is - ~ CH2- , ~ \CH
-~CH2)n- in which n is 2, 3 or 4, more preferably those
in which A is -CH2CH2-, -CH2CH2CH2-, -IHcH2
CH3
or c~2l~
CH3
and most preferably those in which A is -CH2CH2-, and
wherein either
(a) R1 and R8 taken together represent
~C~2
C
~ 3
or (b) ~8 is hydrogen and Rl represents hydrogen, CH3CH2- --
O~E
C3 ~C C~3C3
3 C~3
Particularly preferred are the compounds wherein R8 is
hydrogen and Rl is
~H
CH3~H-
especially compounds having the absolute configuration
5R, 6S, 8R.
A still more preferred embodiment ~f the present
invention comprises preparation of compounds of Formula I
wherein ~6
I -N~
represents /
.. ..... ....... _ .; .. . ....
'
,
- 46 -
CH3 ~ S-~0 , ~ ~ N-CH3 , ~ N
CH ~ ~ ~
Within ~his preferred subclass, the preferred compounds are those
wherein A is -ICHCH2- , ~ , 21 or
CH3 ~ CH3
-(CH2)n- in which A is 2, 3 or 4, more preferably those in which
A is -CH2CH2-, -CH2CH2CH2-, -C~CH2- , ~ or -CH2CH- , and
CH3 ~ C 3
most preferably those in which A is -CH2C~2-, and where~n eithe~
(a) Rl and R8 taken together represent ..
~OC~
C~
C~3
or (b) R8 is hydroge~ and Rl repres~n.s hydrogen, C~3C~2-,
C~3~ ~ C~3 qH
~ C~ or C~3C~
CR3 . C~3
Pa~~icularly preferred a-e the com?ounds whe~ei~ ~8 is
hydrogen and ~1 is
C~3C~- , especially compounds
having '~he absolute configuration 5R, 6S, 8R.
A still more preferred embodLment o~ the present inYen~;on
comprises preparation of compound.s of Formula I wherein
. ~ ~6 ~ _.
-N
\~ ~ .
- 47 -
rep_esen.s
'~J ' . .
in which Y is hydrogen, Cl-C6 aIkyl, hyd,oxy, -S-Cl-C6 alkyl,
carboxyl, carbamoyl, chloro, bromo, iodo, fluoro or phenyl.
Uithin this preferred subclas~, the p~eferred compounds ære
those wherein ~ is -(C~2)~ in which n is 2, 3 or 4, ~ost
pre-erably those in which A is -C~2~..2- and wherein ei,her
(a) Rl æna R8 taXen togethe~ represent
.
~C~2
~ 3
or ~b) R8 is hydrogen 2nd Rl represents hydrogen, C~3C~2-,
c~3 C~
C~_ , ~C- or C~3C~-
C~3 C~3
Particula~ly preferred are the compounds wherein o8 is hyd~oge~
and ~1 is
' CH3C~ ,
i especially compounds having the 2b501ute configuhation 5~ 65
00
_ .
.
.~ :
,:
. .
~ *
- 48 -
A most preferred embodiment of ~.he present invention
comprises preparation of compounds of Formula I wherein
Rl 6~ ,.
-N
represen,s
.. Wi~hin ~his pre erred subcl~ss, the preferred com~ounds ar2
~,hose wherein A i5 ~(C~2)n i~ which n is 2, 3 or 4, most
preferably those in which A is -C$2C~2- and whe ein either
(à) Rl ~nd R8 t~ken together represent
~OCE~ z
~C
. C~3
or (~) R8 is hydrogen ænd Rl re~resents hydrogen, C~3C~2-,
3~ C~
C~- ~ C- o~ ~3C~-
C~3 C~3
~ ~ Particularly preferred are the compounds whexein R8 is hyd ogen
¦ and Rl is 0
C~3C~ ~
especially compounds having the absolute con,igura.ion 5R, 6S,
. ., .` . , . ~ ;, . ,
?~i~L
- 49 ~
A most preferred embodiment of the present invention
comprises preparation of the compounds of the formula
'- ~ ---S-C~.2CH2
O oOR2
wberein ~ ~ represents
`C~ C~ ~'
(both ~- and B~
di2stereoisomers)
-~ N-C~3 or ~ N~/C~3
and R2 is hydrogen, an anionic charge or a conventional
readily removable carboxyl protecting group, providing
that when R2 is hydrogen or a protecting group, there is
also present a counter ion, and pharmaceutically
acceptable acid addition salts thereof.
It will be appreciated that certain products within
the scope of formula I may be formed as optical isomers
as well as epimeric mixtures thereof. It is intended
that the present invention include within its scope all
such optical isomers and epimeric mixtures. For example,
when the 6-substituent is hydroxyethyl, such substituent
may be in either the R or S configuration and the
resulting isomers as well as epimeric mixtures thereof
are encompassed by the present invention.
. ~, ~ . . ,
-,, :..... . ~ . ~ - . . . , , "
- 50 -
The process of the present invention utilizes the
intermediate of the formula
1 ~ b ~
O COOR
IV
which has been disclosed, for example, in European Patent
Application 38,~69 and which may be prepared by the general
methods described therein. L represents a conventional leaving
group (defined as "X" in European Patent Application 38,869)
such as chloro, bromo, iodo, benzenesulfonyloxy, p-toluene-
sulfonyloxy, p-nitrobenzenesulfonyloxy, methanesulfonyloxy,
trifluoromethanesulronyloxy, diphenoxyphosphinyloxy or di(tri-
chloroethoxy)phosphinyloxy. The preferred leaving group is
diphenoxyphosphinyloxy Intermediates of Formula IV are
generally ~ormed ln situ by reacting an intermediate of the
formula
R8 H
R~ o
O ~ N COOR
III
wherein Rl, R8 and R2 are as defined above with a suitable
acylating agent R -L. The preferred intermediate IV where L
is diphenoxyphosphinyloxy may be prepared by reacting keto ester
~III in an inert organic $olvent such as methylene chloride,
acetonitrile or dimethylformamide wlth about an equimolar amount
of diphenyl chlorophosphate in the presence of a base such as
diisopropylethylamine, triethylamine, 4-dimekhylaminopyridine
or the like at a temperature of from about ~20C to +40C, most
preferably at about 0C. ;Intermediate IV may be isolated, if
desired, but is conveniently used as the starting material for
the process of the present invention without isolation or
purification. : -.
, ~
. - :
In the prPsent process, carhapenem intermediate IV
is reacted with a quaternary amine thiol compound of the
formula
HS -A -R14
xe
VII
wherein A is cyclopentylene, cyclohexylene or C2-C6
alkylene optionally substituted by one or more C1-C4
alkyl groups, most preferably cyclopentylene,
cyclohexylene or :-
R10 R12
--C
` ~~ lll l13
in whieh R10, R11, R12 and R13 are each independently
hydrogen or C1-C4 alkyl, X~ is a counter anion associated
with a strong acid such as Cl-, Br~, CH3S03-, CF3S03- or
C~3 ~ 3 . ~nd
R14 is a quaternized nitrogen-containing aromatic or non-
aromatic heterocycle as defined above. The reaction is
earried out in an inert solvent sueh as acetonitrile,
acetonitrile-dimethylformamide, tetrahydrofuran,
tetrahydorfuran-H20, acetonitrile-H20 or acetone in the
presence of base. The nature of the base is not
critical. Best results, however, have been obtained when
a non-nucleophilic tertiary amine base sueh as
diisopropylethylamine, 1,8-diazabieyelo-[5.4.0]undec-7-
ene, l,5-diazabicyclo[4.3.0]non-5-ene or tri~Cl-
C4)alkylamine sueh as triethylamine, tributylamine or
X~
~8~
tripropylamine is employed. Reaction of intermediate IV
with thiol VII may be carried out over a wide temperature
range, e.g. -15C up to room temperature, but is
preferably done at a temperature in the range of from
about -15C to +15C, most preEerably at around 0C.
The carbapenem product produced by reaction of the
quaternary amine thiol VII with intermediate IV will have
a counter anion associated with it [e.g.(C6H5O)2PO2~, Cl
or the anion associated with the quaternary thiol] which
may at this stage be substituted by a different counter
anion, e.g. one which is more pharmaceutically
acceptable, by conventional procedures. Alternatively,
the counter anion may be removed during the subsequent
de-blocking step. Where the quaternized carbapenem
compound and counter anion form an insoluble product, the
product may crystallize out as it is formed and be
collected pure by filtration.
Following formation of the desired carbapenem
product, the carboxyl protecting group R2 of Compound
I may be optionally removed by conventional procedure
such as solvolysis, chemical reduction or hydrogenation.
Where a protecting group such as p-nitrobenzyl, benzyl,
benzhydryl or 2-naphthylmethyl is used which can be
removed by catalytic hydrogenation, intermediate I in a
suitable solvent such as dioxane-water-ethanol,
tetrahydrofuran-diethylether-buffer, tetrahydrofuran-
aqueous dipotassium hydrogen phosphate-isopropanol or the
like may be treated under a hydrogen pressure of from 1
to 4 atmospheres in the presence of a hydrogenation
catalyst such as palladium on charcoal, palladium
hydroxide, platinum oxide or the like at a temperature of
from 0 to 50C for from about 0.24 to 4 hours. When R2
is a group such as o-nitrobenzyl, photolysis may also be
used for deblocking. Protecting groups such as 2,2,2-
trichloroethyl may be removed by mild zinc reduction.
~r
- 52(a) -
The allyl protecting group may be removed by using a
catalyst comprising a mixture of a palladium compound and
triphenyl phosphine in a suitable aprotic solvent such as
tetrahydrofuran, methylene chloride or diethyl ether.
Similarly, other conventional carboxyl protecting groups
may be removed by methods known to those skilled in the
art. Finally, as mentioned above, compounds of formula
I' where R2 is a physiologically hydrolyzable ester such
as acetoxymethyl, phthalidyl, indanyl, pivaloyloxymethyl,
methoxymethyl, etc. may be administered directly to the
host without de-blocking since such esters are hydrolyzed
n vivo under physiological conditions.
It will be understood that where the R1 andtor R8
substituent or the quaternized nucleophile R14 attached
to substituent ~ contain a functional group which might
interfere with the intended course of reaction, such
group may be protected by a conventional blocking yroup
and then subsequently de-blocked to regenerate the
desired functional group. Suitable blocking groups and
procedures for introducing and removing such groups are
well know to those
; ~ .,
i
- . . - ~ ,
'. `: '. ~ : ~ . ,
~ ~ ,
- : . ~ . :
` _ 53 _
sXilled in the art.
As in the case of o~her ~-lactam antibiotics, com?o7m ~s
of general Formula I may be converted by known proceduIes to
pharmaceutically acceptable salts which, for purposes of the
present invention, are substantially equivalent ~o the non-salted
compounds. Thus, for example, one may dissolve a compound o.
Formula I wherein R2 is an anionic charge in a suitable inert
solvent and then add an equivalent of a pharmaceutically accep,aDle
acid. The desired acid addition salt may be recovered by
conventional procedures, e.g. solvent precipitation, lyophiliza-
tion, etc. Where other basic or acidic functional sroups are
present in the compound of ~ormula I, ~harmaceutically acceptable
base addition salts and acid addition salts may be similarly
oreoared by known methods.
A compound of Formula I where R2 is ~ydrogen or an
anionic charge, or a pharmaceu~ically acceptable salt thereof may
also be converted by conventional procedures to a corresponding
compound where R2 is a physiologically hydrolyzable ester grouo, or
a compound of Formula I wherein R2 is a conven~ional carboxyl
pro~ecting group may be converted to the corresponding compound
where R2 is hydrogen, an anionic charge or a physiologically
hydrolyzable ester grou~, or a pharmaceutically acceptable salt
thereof.
Certain of the thiol intermediates of ~orm~la VII may be
prepared, for example, by reacting a sulfide of the formula
; ~ or
VIIIa VIIIb
Ri 7~ C--R 12
Rll R13 _.
VIIIc
wherein R10, Rll, R12 and R13 are each lndependently hydrogen or
Cl-C4 alkyl with a heteroaromatic amine (as defined above) of the
formula
~ . ~ ' ~' ''' '
, ~ .
~ 54 --
or a non-aroma~ic heterocyclic amine (as defi~ed above) of
the formula
\ f ~
N J
and a strong acid. The reaction may be carried out in ~he
presence or absence of an inert organic solvent which is prerer-
ably a non-polar organic solvent such as methylene chloride,
ben2ene, xylene, toluene or the liXe. Where tne am~ne a~d
sulfide reagen~s are liquids or where a solid amine is soluble
in a liquid sulfide reagent, it is prefe_red to carry out ~he
reaction without use of an additional solvent.
The particular strong acid used in the reaction is
not critical and may be, for example, such strong inorganic or
organic acids as hydrochloric, hydrobromic, methanesulfonic,
p-toluenesulfonic, tri~luoromethanesulfonic, etc.
Formation of the quaternary amine thiol intermediate
VII may be carried out at a temperature in ~he range of from
about -20DC to about lOODC. Preferred te~peratures are ge~erally
in the range of about 50-70DC.
The su~fide reagent, aromatic amine and acid are
preferably employed so that the sulfide and acid are used ~n
approxLmately equimolar amounts wi~th the amine being use~ ln
excess, e.g. two to three moles of amine per mole of sulfide or
acid.
The quaternary ~mine thiol intermediate will have a
counter anion assoaiated with it which wiIl be determined by the
particular acid employed. It is, of course,~ possible to --
substitute at this point a different counter anion by conventi~nal
procedures for use in the subsequent reaction with carbapenem
intermediate IV.
'
The carbapenem derivatives of general Formula I
where R2 is hydrogen, an anionic charge or a
physiologically hydrolyzable carboxyl protecting group,
or the pharmaceutically acceptable salts thereof, are
potent antibiotics active against various gram-positive
and gram-negative bacteria and they may be used, for
example, as animal feed additives for promotion of
growth, as preservatives in food, as bacteriocides in
industrial applications, for example in waterbased paint
and in the white water of paper mills to inhibit the
growth of harmful bacteria and as disinfectants for
destroying or inhibiting the growth of harmful bacteria
on medical and dental equipment. They are especially
useful, however, in the treatment of infectious disease
in humans and other animals caused by gram-positive or
gram-negative bacteria~
The pharmaceutically active compounds provided by
the novel process of this invention may be used alone or
formulated as pharmaceutical compositions comprising, in
addition to the active carbapenem ingredient, a
pharmaceutically acceptable carrier or diluent. The
compounds may be administered by a variety of means;
those of principal interest include: orally, topic or
parenterally (intravenous or intramuscular injection).
The pharmaceutical compositions may be in solid form such
as capsules, tablets, powders, etc., or in liquid form
such as solutions, suspensions or emulsions.
Compositions for injection, the preferred route of
delivery, may be prepared in unit dose form in ampules or
in multidose containers and may contain formulatory
agents such as suspending, stabilizing and dispersing
agents. The compositions may be in ready to use form or
in powder form for reconstitution at the time of delivery
with a suitable vehicle such as sterile water.
~1 '
.
.
The dosage to be administered depends to a large ex,ent
on the particular compound being used, the pax,icular composition
rormulated, the route of ad~nistration, the nature and condition
of the host and the particular situs and organism beins t-eated.
Selection of the particular pre~erred dosage and route of
application, then, i5 left to the discretion of the therapist.
In general, however, the compounds may be administered parenterally
ox orally to mamm21ian hosts in an amo~nt of from about 5 to 200
mg/kg/day. Ad~inistration is generally carried out in divided
doses, e.g. three to four times 2 day.
The following examples illustrate but do not limit ~he
scope OL =he present inventicn.
~ ~ :
:
:
.
.- .
: - ~ ~` ' ` '- ' ' `
:~
- 57 -
Exam~le 1
Preparation of 3-(2-~1-pyridinium~ethYlthio~-
6 ~ ( R ) -hydroxyethyl)-7-oxo-1-
azabicyclo(3.2.0)-hept-2-ene-2-carboxylate
A. 1-(2-mercaptoethyl)pyridinium methanesulfonate
t ~ ~. 55~C, 1- ~
To a suspension of pyridinium methanesulfonate in
pyridine prepared by the dropwise addition of
methanesulfonic acid (1.95 mL, 0.03 mol) to pyridine (8.0
mL, 0.099 mol) with cooling, was added ethylene sulfide
(1.96 mL, 0.033 mol). The resulting mixture was stirred
at 55C for 16 h and concentrated under reduced pressure
to a thick syrup which was mixed with few mL of water.
The solution was poured on top of a column (40 x 16 cm)
of ~-"Bondapak"* C-18 which was eluted with water.
Lyophilization of the appropriate fractions gave a
colorless syrup 6.5 g (91%), ir (film.)~max: 2300-2600
(br, SH), 1635 (pyridinium), 1490, 1200 (sulfonate),
1068, 1060, 1045, 791, 780 cm 1, lHmr (DMSO-d6) ~:2.32
(3H, s, CH3SO3-); 2.61, 2.70, 2.73, 2.82 (lH, B part of
A2B system, SH), 3.07 (2H, m [with D2O, 3008 (2H, t,
J=6.5 Hz~], CH2S), 4.76 (2H, t, J=6.5 Hz, CH2N+), 8.19
(2H, m, Hm of pyridinium), 8.6 (lH, m, Ho of pyridinium),
9.08 (2H, dd, J=6.8 Hz, J=1.4 Hz Ho of pyridinium), uv
(H2O) ~maX:206 (65230), 258 (~3760) m~.
* Trademark
,~1
.'' ..
.' . ' '
~4;~
- 58 -
METHOD A
B. 1-(2-mercaptoethyl)~yridinium chloride
e~uti "S-l Cl
An aqueous solution of crude l-(Z-
mercaptoethyl)pyridinium methanesulfo~ate (9.4 g, 0.04
mol) was ~oured on top of column (2.5 x 41 cm) of
"Permutit S l"* Cl-. The column was eluted with water at
a rate of 0.5 mL per min and the appropriate fractions
were combined and lyophilized giving a yellowish syrup
7.0 g (100%) which was used as it was for the next step,
Hmr (D2O) ~:3.22 (2H, m, CH2S), ~.88 (m, CH2N+), 8.18
(2H, m; Hm of pyridinium), 8.7 (lH, m, Hp of pyridinium),
9.0 ppm (2H, m. Ho of pyridinium).
METHOD B
~ ~ ~S ~> Cl . .
To a precooled (ice bath) pyridine (5.6 mL, 70 mmol)
was added pyridine hydrochloride (4.05 g, 35 mmol) and
ethylene sulfide (2.1 mL, 35 mmol). The mixture was
heated at 65C and stirred for 75 min to give a two
phases system. The lighter phase was removed. The
remaining oil was washed with ether (5 x 10 mL) and
pumped under high vacuum to give the title compound (90-
100%) which was used as such for the next step
*Trademark
- 59 -
C. Paranitrobenzvl 3-~2-(1-pyridinium)ethYlthiol-6~- r 1-
(~)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0~hept-2-
ene-2-carboxylate chloride
1- ~E~ r)2
~"~C 2~ tOP~)
3-- XS~ N~
C00?~3 4- ~ (i~)2
0
~ S~
` ~O?N3 Cl
A solution of p-nitrobenzyl 6~-[1-(R)-hydroxyethyl]-
3,7-dioxo-1-azabicyclo (3.2.0) heptane-2-carboxylate
(6.09 g, 17.5 mmol) in acetonitrile (20 mL) was cooled to
+5OC under a nitrogen atmosphere and treated successively
with diisopropylethylamine (3.65 mL, 21.0 mmol) and
diphenyl chlorophosphate (4.34 mL, 21.0 mmol). The
resulting mixture was stirred for 30 min at 5C, cooled
to -5C and treated successively with a solution of
crude 1-(2-mercaptoethyl)pyridinium chloride (4.3 g, 24
mmol) in N,N-dimethylformamide (1.0 mL)~and dropwise with
diisopropylethylamine (3.65 mL, 21.0 mmol). The reaction
mixture was stirred at 0C for 1 h, cooled t~ -30C and
stirred for 15 min. more. ~he solid was filtered off and
washed with cold (-30C ) acetonitrile 5.77 g (65%), ir
("Nujol")* vmax:3300 (OH), 1775 (c=o of ~-lactam~ 90
(c=o of PNB ester), 1630 (pyridinium), 1605 (phenyl of
PNB ester), 1515 (N02), 1335 cm~l (N02), 1Hmr (DMSO-d6)
:
.
'
'. '
,
~8~i~
- 59(a) -
~:1.17 (3H, d, J=6.1 Hz, CH3 CHOH), 3.2-3.75 (5H, H-4, H-
6, CH2S), 3.75-4.5 (2H, H-5, CH3CHOH), 4.92 (2H, brt,
J=6.5 Hz, CH2N+), 5.18 (lH, d, J=4.9 Hz, OH), 5.37
(center of ABq, Ja b=l4.2 Hz, CH2 of PNB), 7.69 (2H, d,
J=8.7 Hz, Ho of PNB), 8.24 (d, Jk=8.7 Hz, Hm of PNB),
8.0-8.4 (4H, Hm of PNB, Hm
* Trademark
:
:
:
~ ~ :
:
~ ~ :
:~ :
.
:
..
:, ~ i,'' ~ ' ', "
' ' `, ~; , ' '
..
- 60 -
of pyridinium), 8.66 (lH, m, Hp of pyridinium), 9.17 ~2H,
brd, J=5.5 Hz, Ho of pyridinium). The filtrate and
washing were combined and diluted with ether (150 mL).
The supernatant was decanted and the gum was dissolved in
water (40 mL) containing enough acetonitrile to have a
solution which was poured on top of a column (3 x 10 cm)
of ~-"Bondapak"* C-lg. The column was eluted with 10%
acetonitrile - 90% water (150 Ml) and 50% acetonitrile -
50% water (100 mL) mixtures. The appropriate fractions
were combined and lyophilized after the acetonitrile has
been removed under vacuum giving a yellowish powder. An
NMR of it showed the presence of the title compound mixed
with some p-nitro-benzyl 3-[2-(1-pyridinium)ethylthio]-
6~-[1-(_)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-
ene-2-carboxylate diphenylphosphate (2:1). The powder
was dissolved in water (minimum amount) and passed
through a column (1.5 x 21 cm) of "Permutit"* S-lC1- with
water. Lyophilization of the appropriate fractions gave
1.8 g (20%) of the title compound.
* Trademark
'
'
.
'~ ~' : ' ','' ' ' ' '
- - : ' , ' ' :
. ' ' '
3~
- 61 -
D. Paranitrobenzyl 3-~2-(1-pyridinium)ethylthiol-6~-rl-
)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2 O)hept-2-
ene-2-carhoxylate diphenYlphos~hate
1~ ~E~ )2.
J ~ 2) C~O~2
O ~ N ~ 3) ~S
0~
J~s ~ -
~,~ ~
~hO)2?o
A solution of p-nitrobenzyl 6~[1-(R)-hydroxyethyl]-
3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-carboxylate (0.174
g, 0.50 mmol) in acetonitrile (2 mL) was cooled to 0C
under a nitrogen atmosphere and treated successively with
diisopropylethylamine (0.105 mL, 0.60 mmol) and diphenyl
chlorophosphate (0.124 mL, 0.60 mmol). The resulting
solution was stirred for 30 min at 0C and treated
successively with a solution of 1-(2-
mercaptoe~hyl)pyridinium methanesulfonate (0.170 g, 0.72
mmol) in acetonitrile (0.6 mL) and diisopropylethylamine
(0.105 mL, 0.60 mmol). The reaction mixture was stirred
at 0C for 15 min, diluted wlth cold (0C) water (7 mL)
and poured on top of a column (1.5 x 6.4 cm) of ~-
"Bondapak*" C-18. The column was eluted with a mixture
of acetonitrile (25%-50%) in water (75%-50~). The
appropriate fractions were combined and lyophilize.d after
the acetonitrile has been removed under vacuum giving a
yellowish powder 0.33 g (92%), ir (KBr) ~max: 3600-3000
.;, .
: . `
.
~;
- 61a -
(OH), 1765 (C=O of B-lactam), 1690 (C=O of PNB ester),
1625 (pyridinium), 1585 (phenyl), 1510 (NO2), 1330 (NO2),
885 cm~1 (NO2),
* Trademark
' ~ '
: ,
;
;
: '~ , ' , .
.: ' ' ' . ' ' ' ' '
- '' ' , : '
', . .
~8~
- 62 -
Hmr (DMSO-d6) : 1.16 (3H, d, J=6.2 Hz CH3CHOH), 4.87
(2H, brt, J=6.6 Hz, CH2S), 5.37 (center of Abq, Ja b=14 3
Hz, CH2 of PNB), 6.7-7.5 (phenyl), 7.68 (d, J~8.8 Hz, Ho
of PNB), 8.23 (d, J=8.8 Hz, Hm of PNB), 8.0-~.3 (m, Hm of
pyridinium), ~.4-8.8 (lH, Hp of pyridinium), 9.09 (2H,
dd, J=6.7 Hz, J=1.3 Hz, Ho of pyridinium).
E. 3-~2-(1-pyridinium~ethylthio]-6~-~1-(R)-
hydroxyethyl]-7-oxo-1-azabicyclot3.2.0)hept-2-ene-2-
` carboxylate
M
~ s N~
O N ~ ~-:~, E~-
C00~3 t?~O~2~o
O~;S ~ ..,
To a solution of p-nitrobenzyl 3-[2-(1-
pyridinium)ethylthio]-6~-[1-(R)-hydroxyethyl]-7-oxo-1-
azabicyclo (3.2.0) hept-2-3n3-2-carboxylate
diphenylphosphate (0.16 g, 0.22 mmol) in wet
tetrahydrofuran (10 mL) was added ether (10 mL),
potassium phosphate monobasic-sodium hydroxide buffer pH
7.4 (16 mL, 0.05 M) and 10% palladium on charcoal (0.16
g). The resulting mixture was hydrogenated under 40 psi
for 1 h at 25C. The two phases were separated and the
organic phase was extracted with water (2 x 3 mL). The
aqueous solutions were combined, washed with ether (2 x
10 mL) and poured on top of a column (1.5 x 6.2 cm) of ~-
. : ' . .,
,
~8~3;;~
- 62a -
"Bondapak"* C-18 after the traces of organic solvents
have been removed
* Trademark
.
.
:
;,
'
~ : -
- ` ' , ' , .
'.
:,
~2~
- 63 -
under vacuum. Elution of the column with water gave
after lyophilization of the appropriate fractions a
yellowish powder 0.062 g (84%), ir (KBr) vmaX : 3700-
3000 (OH), 1755 (C=O of ~-lactam), 1630 (pyridinium),
1590 cm~l (carboxylate), lHmr (D2O): 1.22 (3H, d, J=6.4
Hz, C_3CHOH), 2.92 (d, J=9.1 Hæ, H-4), 2.97 (d, J=9.1 Hz,
H-4), 3.20 (dd, J=2.5 Hz, J=6.1 Hz, H-6), 3.44 (t, J=6.0
Hz, CH2S), 3.93 (dd, J=9.1 Hz, J=2.5 Hz, H-5), 4.82 (t,
J=6.0 Hz, CH2N+), 8.04 (m, Hm of pyridinium), 8.5 (m, Hp
of pyridinium), 8.82 (dd, J=3.2 Hz, J=1.1 Hz, Ho of
pyridinium), uv (H2O) max : 259 (~5800), 296 (~7030) m~
T~=13.5 h (measured at a concentration of 10 4 M in
phosphate buffer pH 7.4 at 36.8C).
.
`:
'
- 64 -
Method B
0~
~S~N~ ~
o ~u~_er 7 2
~OO~
'J"~ ~ N~
COO ' .
-To a solution of-p-nitrobenzyl 3-[2-(1-
pyridinium)ethylthio]-6~[1-(R)-hydroxyethyl]-7-oxo-1-
azabicyclo (3.2.0) hept-2-ene-2-carboxylate chloride
(5.77 g, 11.4 mmol) in potassium phosphate monobasic-
~odium hydroxide buffer (170 mL, 0.2 M, pH 7.22) was
added tetrahydrofuran (30 mL), ether (30 mL) and 10~
palladium on charcoal (5.7 g). The resulting mixture was
hydrogenated at 22C under 40 psi for 1 h and filtered on
a "Celite"* pad. The pad was washed with water t2 x 15
mL~. The filtrate and washings were combined and diluted
with ether (100 mL). The aqueous phase was separated,
washed with ether (3 x 100 mL) and poured on top of a
column (4.5 x 20 cm) of ~-"Bondapak"* C-18 after the
organic solvents have been removed under vacuum. Elution
of the column with water followed by a mixture of 1%
acetonitrile in water gave after lyophilization of the
appropriate fractions 2.48 g (65%) of the title compound
as a yellowish powder. The analytical data were
identical to those reported for the compound prepared in
the method A.
* Trademark
X
~;.
- 65 -
Example 2
Preparation of 3-~2-(1-(3.5-dimethy~YridiniumL
ethylthiol]-6~ [1- (R) -hydroxyethyl]-7-ox~
azabicyclo(3.2.0!hept-2-ene-2-carboxylate
~-- C~3
~ S ~ ~
COO C~3
A. 1-(2-merca~toethyl)-3 5-dimethylpyridinium
~ethanesulfonate
1 N~ I MsO~ S ~ ~
To a suspension of 3,5-lutidinium methanesulfonate
in 3,6-lutidine prepared by the addition of
methanesulfonic acid (0.65 mL, 0.010 mol) to cold 3,5-
lutidine (2.51 mL, 0.022 mol) was added ethylene sulfide
(0.655 mL, 0~011 mol). The resulting mixture was stirred
under a nitrogen atmosphere at 55C for 24 h, cooled to
23C and diluted with water (5 mL) and ether t5 mL). The
organic layer was separated and the aqueous solution was
washed with ether (6 x 4.mL). The traces of ether were
removed under vacuum and the solution was applied on top
of a column (2.5 x 6.0 cm) of ~-"Bondapak"* C-18. The
column was eluted with water and lyophilization of the
* Tradem~rk
~7
,
,
- 66 -
appropriate fractions gave a colourless syrup 2.4 g
(91~; ir (film) VmaX:2520 (SH), 1628 (pyridinium),
1600, 1495, 1325, 1305, 1283, 1200 (sulfonate), 1040,
938, 765, 680 cm~1, lHmr (DMSO d6) ~:2.31 (3H, s,
CH3SO3-), 2.47 (6H, s, CH3 on pyridinium), 2.57, 2.66,
2.69, 2.78 (lH, B part of A2B system, SH), 3.06 (2H, m
[with D2O added (2H, t, J-6.5 Hz)~, CH2S), 4.65 (2H, t,
J=6.5 Hz, CH2N+), 8.34 (lH, s, Hp of pyridinium), 8.79
~2H, s, Ho of pyridinium); uv (H2O) ~max 271 (~4
m~. Anal. calcd. for CloH17NO3S2 0.5H2O: C 44
6.66, N 5.14, S 23.54; found: C 44.26, H 6.49, N 5.17, S
24.18.
B. Paranitrobenzyl 3- r 2~ (3 5-
dimethylpyridinium))ethylthio]-6~k-~1-(~)-
hvdroxyethyll-7-oxo-1-azabicyclo(3.2.0)he~t-2-ene-2-
carboxylate diphenylphosphate
0~ t(~?r)2 'o~y ~_~
~.~ 2J Cl~OPb)2 ~~S ~ ~>
O f ~_~ o~ N~
CO~?N3 3) ~ ~ N ~ M~0 \CCo~ R
4~ ~E~ r)2 (?~0)2~o
To a cold (0C; solution of p-nitrobenzyl 6~-(1-(R)-
hydroxyethyl)-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-
carboxylate (0.523 g, 1.50 mmol) in acetonitrile (6.0 mL)
kept under a nitrogen atmosphere was added
diisopropylethylamine (0.314 mL, 1.8 mmol) followed by
diphenyl chlorophosphate (0.373 mL, 1.8 mmol). The
reaction mixture was stirred for 30 min and treated with
~2~3~
- 66a -
a solution of 1-(2-mercaptoethyl)-3,5-dimethylpyridinium
methanesulfonate (0.493 g, 1.87 mmol) in acetonitrile
~'
,
- ' : ' . ~ - ;
~. . .
: . . ,' ,.
(l.9 mL) followed by diisopropylethylamine (0.314 mL, 1.8
mmol). The reaction mixture was stirred at 0c for lh
diluted with cold (0c) water (26 mL) and poured on top
of a column (7.0 x 3.5 cm) of ~-I'Bondapak'' C-18. Elution
of the column with 25-50% acetonitrile - 75-50% water
mixture gave after lyophilization of the appropriake
fractions 1.01 g (90%) of the title compound as yellowish
powder, ir (KBr) ~max : 3700-3100 (OH), 1778 (C=O of ~-
lactam), 1700 (C=O of PNB ester), 1635 (pyridinium), 1595
(phenyl), 1521 (NO2), 1335 (NO2), 895 cm~l (NO2), 1Hmr
(DMSO d6) ~:1.16 (3H, d, J=6.1 Hz, CH3CHOH), 2.43 (s, CH3
on pyridinium), 4.75 (2H, m, CH2N+), 5.38 (center of ABq,
Ja b=14-3 Hz, CH2 of PNB), 6.6-7.5 (lOH, m, phenyl), 7.70
(2H, d, J=8.7 Hz, Ho of PNB), 8.0-8.5 (3H, m, Hp of
pyridinium, Hm of PNB), 8.82 (2H, s, Ho of pyridninium),
uv (H2O) ~max : 270 (~11570), 306 (~7343) m~. anal.
calcd. for C37H38N310SP-H2O C 58
4.18; found: C 57.98, H 5.05, N 5.22, S 4.34
;_
.
.
,
3'~
- 68 -
C. 3-[2-(1-(3,5-dimethylpyridinium))ethvlthiol-6~- r 1-
(~)-hvdroxyethyll-7-oxo-1-azabicvclo(3.~.0) he~t-2-
ene-2-carboxylate
O;~
10~ ~d/C, ~2
S ~ ~ ~ ethe-, bu~-e-
C~O NB (~h~) ~o~
To a solution of p-nitrobenzyl 3-[2-(1-(3,5-
dimethylpyridinium))ethylthio]-~-[1-(R)-hydroxyethyl]-7-
oxo-l-azabicyclo (3.2.0) hept-2-ene-2-carboxylate
diphenylphosphate (0.600 g, 0.80 mmol) in wet
tetrahydrofuran (36mL) was added ether (36 mL), potassium
phosphate monobasicsodium hydroxide buffer (0.05M, pH
7.4, 44mL) and 10% palladium on charcoal (0.60 g). The
resulting mixture was hydrogenated under 40 psi at 23C
for 1.25 h. The organic layer was separated and
extracted with buffer (2 x 5 mL). Water layers were
combined, filtered through a "Celite" pad, washed with
ether (40 mL, pumpe~ to eliminate traces of organic
solvents and poured on top of a column (2.5 x 10.0 cm) of
~-"Bondapak" C-18. Elution of the column with water and
lyophilization of the appropriate fractions gave the
title compound 0.186 g (64%) as a yellowish powder, ir
(XBr).~max:3700-3100 (OH), 1760 (C=O of B-lactam), 1595
cm~l (carboxylate), 1Hmr (D2O) ~ 1.21 (3H, d, J=6.3 Hz,
.
,
- 69 -
CH3CHOH), 2.45 (6H, s, CH3 on pyridinium), 2.81 (d, J-9 2
Hz, H-4), 2.96 (d, J-9.2 Hz, H-4), 3.22 (dd, J-2.6 Hz,
J=6.2 Hz, H-6), 3.40 (t, J-6.2 Hz, CH2S), 3.84 (dd, J-9.2
Hz, J-2.6 Hz, H-5), 4.15 (m, CH3CHOH), 4.71(t, J=6.2 Hz,
CH2N+), 8.21 (lH, s, Hp of pyridinium), 8.46 (2H, s, Ho
of pyridinium), uv (H2O) ~max 279 ( 8345),
m, [~]~ + 40.7 (c 0.53, H2O), T~ ~ 16.9 h (measured at a
concentration of 10-4 M in phosphate buffer pH 7.4 at
36.8C).
Example 3
Preparation of (5~,6~)-3-[[2-(3-hydroxymethylpyrid-
inio)ethyl]thiol-6-[~ )-hydroxyethyl]-7-oxo-1-
azabicyclo[3.2.01hept-2-ene-2-carboxylate
0~ .
J ~ ~ ~2~
- 70 -
A. 3-HYdroxymethyl-1-(2-merca~toethvl)pyridinium
trifluoromethanesulfonate
0X
+ OE3So
~2~
XS~32CH2N ~>
~ ~3S03~
Trifluromethanesulfonic acid (1.327 mL, 0.015 mol)
was added dropwise to 3-pyridinemethanol (2.gl mL, 0.030
mol), followed by ethylene sulfide (0.89 mL, 0.015 mol).
The resulting homogeneous mixture was heated (oil bath)
at 50-70C under N2 for 20 h. The reaction mixture was
then taken up in H2O (15 mL) and extracted with CH2Cl2
(5 x 5 mL). The aqueous phase was concentrated in vacuo
and then applied to C18 reverse-phase column. Elution
with H2O followed by evaporation of the rele~ant
fractions gave a pale yellow oil. This material was
rechromatographed to give a nearly colourless oil. After
drying in vacuo (P2O5) this afforded the product (4~50 g,
94%) as a viscous oil. ir (film) ~m~x:3450 (s, OH), 2560
(w, SH) cm~l; lHmr (d6-acetone) ~:9.10-8.05 (m, 4H,
aromatic), 5.01 (t, J=5.5 Hz, 2H, N-CH2) 4.93 (s, 2H, -
CH2OH), 4.43 (br S, lH, -OH), 3.43-3.18 (m, 2H, S-CH2),
2.34-2.l0 (m, lH, SH).
:: :
`,, ' ' ' '
': . ` : ` ' .
: . . . ~ , .
- , ~ . . . . . . .
.
.
'~ ' ': ' ; ` . ',": ,, ',
- 71 -
B. p-Nitrobenzyl (5B 6~)-3-~2-(3-hYdroxymeth~
pyridinio~ethyl thiol-6-[1-(B)-hydroxyethyll~7-oxo-
azabicyclo[3.2.0]hept-2-ene-2-carpoxylate
diphenylphosphate
d~ ~-`<
CO ~3
~2
'",cr~s~N~
,_ p
To a solution of p-nitrobenzyl (5R, 6S)-6-[1-(R)-
hydroxyethyl]-3,7-dioxo-1-azabicyclo[3.2.0]heptane-2-
carboxylate (0.174 g, 0.50 mmol) in 2 mL of dry
acetonitrile was added diisopropylethalimine (0.096 mL,
0.55 mmol) at 0C under N2. Diphenyl chlorophosphate
(0.114 mL, 0.55 mmol was then added dropwise and the
reaction mixture was stirred at 0C for 30 min. A
solution of 3-hydroxymethyl-1-(2-mercaptoethyl)pyridinium
trifluoromethanesulfonate (0.223 g, 0.70 mmol) in 0.50 mL
of acetonitrile was then added, followed by
diisopropylethylamine (0.122 mL, 0.70 mmol). After being
kept at 0C for 30 min the reaction mixture was
concentrated in vacuo and the residual yellow gum was
taken up in H2O (enough acetonitrile was added to aid in
dissolving the gum). This solution was applied to a C18
reverse-phase column which was eluted with 15%
acetonitrile-H2O. Lyophilization of the relevant
fractions afforded the product (0~305 g, 81%) as a beige-
coloured solid. ir (KRr) ~max:3420 (br, OH), 1775 (B-
lactam CO), 1695 (-CO2PNB) cm~1; 1Hmr (d6-acetone)
- 71(a) -
~:9.44-7.72 ~m, 8H, aromatic), 7.22-6.91 (m, lOH,
diphenylphosphate), 5~53, 5.27 (ABq, J=14Hz, 2H,
benzylic), 5.04 (t, J=7.4 Hz, 2H, N-CH2), 4.75 (s, 2H,
CH20H), 4.5-3.1 (m, 8H), 1.21 (d, J=6.3 Hz, 3H, CHMe).
~ -
'
C. (5B~6~)-3-~2-(3-hydrox~methylpYridinio)ethYll
thiol-6-[1-(~-hydroxyethyll-7-oxo-1-azabicvclo-
~3.2.0]hept-2-ene-2-carboxylate
C~
o~ 4~ 2
~ ~ ~ 2~ ~
0~ C32O~
;
To a solution of p-nitrobenzyl (5R,6S)-3-[2-(3-
hydroxymethylpyridinio)ethyl thio]-6-[1-tR)-
hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]-hept-2-ene-2-
carboxylate diphenylphosphate (0.145 g, 0.194 mmol) in 10
mL of THF containing 5 drops of H2O, was added 6.0 mL of
phosphate buffer (0.05 M, pH 7.4), 0.145 g of I0%
palladium-on-charcoal and 10 mL of ether. The mixture
was hydrogenated (Parr) at 40 psi of 1 h and then
filtered through a pad of "Celite". The filter cake was
washed with a little H2O and ether and the aqueous phase
was separated and extracted with ether (3x). The aqueous
solution was then cooled at 0C and the pH was adjusted
to 7.0 with pH 7.4 buffer. After removing residual
volatiles in vac~o the aqueous solution was applied to a
C18 reverse-phase column which was eluted with H2O.
Lyophilization:of the relevant fractions gave the product
(36 mg, 51%) as a light yellow solid. Further
purification by reverse-phase hplc gave the pure product
(31 mg, 41~) as a solid- ir (KBr)~maX:330o (br~ OH)~
1755 (B-lactam CO)j 1590 (-CO2-) cm~1; lHnmr (D2O)
:. :
'
~8~3~
- 72(a) -
~:8.78-7.94 (m, 4H, aromatic), 4.83 ~t, J-6.0 Hz, 2H, N-
CH2), 4.83 (s, 2H, CH2OH),
;
:~ .
: ~ ~
: ` ~' , '' '
.' ' - ;. ' ~
32~
- 73 -
4.16 (d of q, J=J'=6.2 Hz, lH, H-l'), 3.9B (d, of t, J=9.1 Hz,
J'=2.6 Xz, lH, X-5), 3.75-3.20 (m, 3X), 3.20-2.65 (m, 2X), 1.22
(d, ~=6.4 Hz, 3X, CHMe); ~v (H20) ~ : 294 (~7614), 266 (~6936)
n~; t~ (pH 7.4, 36.8C) 14.0 h.
Exzm~le 4
Preparation of (52,6S)-3-[2-(4-hydroxymethylpyridinio)ethyl-
thio]-6-[1-(R)-hydroxye~hyl]-7-oxo-1-azabicyclo[3.2.0]hept-
.
2-ene-2-carboxylate
.
0~ . . ' ', .
'1~S~2C~ , ,
2
3~
- 74 -
A. 4-Hydroxymethyl-1-(2-mercaptoethvl)pYridinium
trifluoromethanesulfonate
/ ~ C3 0~ + -C~3SO3~
32Q ~ 3203
C~ , .
To a solution of 4-pyridinemethanol (1.635 g, 0.015
mol) in 10 mL of CH2Cl2, at 0C under N2, was added
dropwise trifluoromethanesulfonic acid (1.327 mL, 0.015
mol~. A yellow-brown oil rapidly separated out. An
additional equivalent of 4-pyridinemethanol (1.635 g,
0.015 mol) was added to this mixture and the solvent was
removed under reduced pressure to give an oil. To this
oil was added ethylene sulfide (0.891 mL, 0.015 mol) and
the resulting homogeneous mixture was heated (oil bath)
at about 60C for 3 h. The reaction mixture was then
taken up in 15 mL of H2O and the aqueous solution was
washed with CH2Cl2~(5 x 5 mL3. After removing residual
organic solvent in vacuo the aqueous solution was applied
to a Cl8 reverse-phase column. Elution with H2O and
subsequent evaporation of the relevant fractions afforded
an oil which was further dried in vacuo over P2O5 to give
the product (4.64 g, 97%) as a colourless oil. ir (film)
max:3455 (s, OH), 2565, (w, SH) cm~1; 1Hnmr~(d6~acetone)
~:9.07, 8.18 (ABq, 3=6.8 Hz, 4H, aromatic), 5.03~(s, 2H,
CH2OH), 4.96 (t, J=6.5 Hz, 2H, N-CH2), 4.09 (br s, lH, -
OH), 3.5-3.1 (m, 2H, S-CH2~, 2.25 (brs, lH, -SH).
~ .
~ ' '
.
,~
. . .
37~
- 75 ~
B. p-Nitrobenzyl (5~!6~)-3-r2-(4-hYdroxymethYlPyrid-
inio)ethyl thiol-6- r~ -hvdroxyethyll-7 oxo-1-aza-
bicyclo r 3.2.0lhept-2-ene-2-carboxylate
diphenylE?hosphate
0:~
0~ . -~
CO2 ~3
20=
~ 3 (~~2 ~
To a solution of p-nitrobenzyl (5R,6S)-6-[1-(R)-
hydroxvethyl~-3,7-dioxo-1-azabicyclo[3.2.0]heptane-2-
carboxylate (0.34~ g, 1.0 mmol) in 5 mL of dry
acetonitrile, at 0C under N2, was added dropwise
diisopropylethylamine ~0.191 mL, 1.1 mmol) followed by
diphenyl chlorophosphate (0.228 mL, 1.1 mmol). The
resulting golden-yellow solution was stirred at 0C for
40 min. To this solution was added a solution of 4-
hydroxymethyl-l-(2-mercaptoethyl)pyridinium trifluoro-
methanesulfonate (0.447 g, 1.4 mmol) in 1 mL of
acetonitrile, followed by diisopropylethalamine (0.191
mL, 1.1 mmol). A reddish-black gum separated from the
reaction mixture. After 20 min at 0C the reaction
mixture was filtered and concentrated in vacuo. The
residue was taken up in a minimum volume of acetonitrile-
H2O (1:1) and applied to a C18 reverse-phase column.
Elution with 25% acetonitrile-H2O and subsequent
lyophilization of the relevant fractions gave the product
X~ . :
`
.
z~
- 75(a) -
~0.353 g, 47%) as a cream-coloured solid. ir (XBr)
maX:3240 (br, OH~, 1775 (B-lactam CO), 1695 (-CO2PNB)cm
1; lHnmr (d6-acetone) ~: 9.24-7.84 (m, 8H, aromatic),
7.4-6.9 (m, 10H, diphenylphosphate), 5.52, 5.24 (ABq, J-
14 Hz, 2H, benzylic), 5.15 4.80 (m, 4H), 4.45-3.05 (m,
7H), 1.35 (d, J=6.6 Hz, 3H, CHMe).
.
'
. ,~: '
,
...
.: , : .. ,
. . .
.
C. (5~,6~)-3-[2-(4-hydroxymethvl~vridinio)ethvl thiol-
6-[1-(B)-hydroxyethyll-7-oxo-1-
azabicvclo[3.2.olhept-2-ene-2-carboxylate
0
J'~ ~ ~ 2~
C02~3 (~)2
0'~
~ r~ ~ ~ C
A mixture of p-nitrobenzyl (5R,6S)-3-[2-(4hydroxy-
methylpyridinio)ethyl thio]-6-[1-(R)-hydroxyethyl]-7-oxo-
1-azabicyclo-[3.2.0]hept-2-ene-2-carboxylate
diphenylphosphate (0.348 g, 0.465 mmol) and 10%
palladium-on-charcoal (0.35 g) in 11 mL of phosphate
buffer (0.05 M p~ 7.~), 5 mL of THF and 10 mL of ether
was hydrogenated at 4~ psi for 1.25 h. The mixture was
then filtered through a "Celite" pad and the aqueous
phase was washed with ether (3 x). The pH of the aqueous
solution was then adjusted to 7.0 using additional pH
7.4 buffer. After removing residual volatiles in vacuo
the aqueous solution was applied to a C18 reverse-phase
column. Elution with 2% acetonitrile-H2O and subsequent
lyophilization gave a yellow-brown solid. This material
was rechromatographed (Cl~ reverse-phase/H2O) to give the
desired product (0.060 g, 36%) as a light yellow solid.
ir (KBr) ~max:3400 (br, OH), 1755 (B-lactam CO), 1590 (-
CO2-) cm~1; 1Hnmr (D2O) ~: 8.73, 7.96 (ABq, J=6.8 Hz, 4H,
aromatic), 4.93 (s, 2H, CH2OH), 4.77 (t, J=6.0 Hz, 2H, N-
- 76 (a) -
CH2), 4.15 (d of q, J=JI=6.3 Hz, lH, H-l'), 3.96 (d of
t, J=9.2 Hz, Jl=2.6 Hz, lH, h-5), 3.65-3.20 (m, 3H),
3.13-2.62 (m, 2H), 1.21 (d, J-6.3 Hz , 3H, CHMe); uv (H20)
~maX:295 (~6880), 256 (~5595), 224 (~8111) nm; ~ (pH
7.4, 36.8C) 14.5 h
; :
." :
- ` '
`
~2~3~3;~
Exam~le 5
Preparation of 3-[2-(1-(2-methylPYridinium))
ethylthio]-6~-[1-(RL=~ydroxyethyll- -oxo-1-
azabicyclo(3.2.0)hept-2-ene-carboxy]ate
A. 1-(2-mercaptoethyl)-2-methylpyridinium
methanesulfonate
N~ 55~C, 21 h N~ .~30
To a suspension of 2-methylpyridinium
methanesulfonate in 2-methylpyridine prepared by the
addition of methanesulfonic acid (0.65 mL, 0.010 mol) to
cold 2-methylpyridine (2.17 mL, 0.022 mol) was added
ethylene sulfide (0.655 mL, 0.011 mol). The reaction
mixture was stirred under a nitrogen atmosphere at 55C
for 21 h, cooled to 23C and diluted with water (5 mL).
The aqueous solution was washed with ether (6 x 4 mL)
pumped to remove traces of organic solvents and poured on
top of a column (2.5 x 10.0 cm)
-
... . ~ . ... .. -- . -, . ~ --
- ~ ,
~, ` .
- 7~ -
of ~-"Bondapak~" C-18. The column was eluted with water
and lyophilization of the appropriate fractions gave 2.13
g (85%) of the title compound, ir (film max:2520 (SH)~
1623 (pyridinium) 1574, 1512, 1485, 1412, 1195
(sulfonates), 1038 cm~1, 1Hmr (DMSO-d6 ~ D2O) ~:2.37 (3H,
s, CH3SO3-), 2.83 (3H, s, CH3 on pyridinium), 3.09 (2H,
J=6.9 Hz, CH2S), 4.71 (2H, t, J-6.9 Hz, CH2N+), 7.93 (2H,
m, Hm of pyridinium), 8.44 (lH, m, Hp of pyridinium),
8 89 (lH, m, Ho of pyridinium), uv (H2O) ~max 266
(~3550) m~.
B. Paranitrobenævl 3-[2-(1-(2-methylpyridinium))-
ethylthio)-6~-[1-(R)-hydroxyethyl]-7-oxo-1-
azabicyclo(3 2.0)hept-2-ene-2-carboxylate
di~henylphosphate
~, ~ 2~ Cl~(O~'O 2
C00~3 3) ~ ~5
4) ~Et(i~r)2
i~
C00~
(PhOt2Po
To a cold (0C) solution of p-nitrobenzyl 6~-[1-
(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-
carboxylate (0.523 g, 1.50 mmol) in acetonitrile (6 mL)
kept under a nitrogen atmosphere was added
diisopropylethylamine t0.314 mL, 1.80 mmol) followed by
diphenyl chlorophosphate (0.373 mL, 1.80 mmol). The
... .. _. . . . .. ~
- 78(a) -
reaction mixture was stirred for 30 min at 0C and
treated with solution of 1-(2-m~rcaptoethyl~-2-
methylpyridinium methanesulfonate (0.530 g, 2.16 mmol) in
acetonitrile (18 mL) followed by diisopropylethylamine
~0.314 mL,
* Trademark
. .
1.8 mmol). The reaction mixture was stirred at 0C for 1
h diluted with cold (0C) water (26 mL) and poured on top
of a column (3.5 x 7.0 cm) of ~-"Bondapak"* C-18.
Elution of thee column with 25% acetonitrile ~ 75% water
and with 50% acetonitrile - 50% water gave after
lyophilization of the appropriate fractions 1.06 g, (96%)
of the title compound as a yellowish powder, ir (KBr)
~max 3650-3100 (OH), 1770 (c=o of ~-lactam), 1695 and
1690 (C=O of PNB ester), 1630 (pyridinium), 1595
(phenyl), 1518 (NO2), 1335 (NO2), 890 cm~l (NO2), lHmr
(DMSO, d6) ~: 1.15 (3H, d, J=6.1 Hz, CH3CHOH), 2.87 (s,
CH3 on pyridinium), 3.6-4.4 (2H, m, H-5, CH3C_OH), 4.75
(2H, m, CH2N+), 5.37 (center of ABq, J=14 Hz, CH2 of
PNB), 6.5-7.4 (10H~ m, phenyl), 7.70 (2H, d, J=8.8 Hz, Ho
of PNB), 8.0 (2H, m, Hm of pyridinium), 8.24 (2H, d,
J=8.8 Hz, Hm of PNB), 8.50 (lH, m, Hp of pyridinium),
8.95 (lH, brd, J=6.1 Hz, Ho of pyridinium), uv (H2O)
:265 (E11990), 314 (~8020)m~
C. 3-~2-(1-(2-methylpyridinium)lethylthiol-6~ (B~-
hvdroxyethyl~-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-
carboxylate
OR
~,~ ~N~ --- ~ shér buSrer
~Coo?NB (P~O)~O
*Trademark
,
3~
- 80 -
To a solution of p-nitrobenzyl 3-[2-(1-(2-
methylpyridinium)) ethylthio]-6~-[1-(R)-hydroxyethyl]-7-
oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate
diphenylphosphate (0.66 g, 0.90 mmol) in wet
tetrahydrofuran (34 mL) was added ether (34 mL),
potassium phosphate monobasic-sodium hydroxide buffer
(0.15 M, 16.5 mL, pH 7.22) and 10% palladium on charcoal
(0.66 g). The resulting mixture was hydrogenated under
40 psi at 23C for 1.25 h. The organic layer was
separated and extracted with buffer (2 x 6 mL). Water
layers were combined, filtered through a "Celite" pad,
washed with ether (40 mL), pumped to eliminate traces of
organic solvents and poured on top of a column (2.5 x 10
cm) of ~-"Bondapak"* C-18. Elution of the column with
water and lyophilization of the appropriate fractions
gave the title compound 0.098 g (31%) as a yellowish
powder, ir (KBr) vmax: 3650-3100 (OH), 1755 (C=O of ~-
lactam), 1630 (pyridinium), 1595 cm~1 (carboxylate), lHmr
(D2O) S: 1.20 (3H, d, J=6.3 Hz, CH3CHOH), 2.83 (s, CH3 on
pyridinium), 2.7-3.1 (5H, H-4, CH3 on pyridinium), 3.1-
3.7 (3H, m, CH2S, H-6), 3.90 (dd, J=9.1 Hz, J-2.6 Hz, H-
5), 3.1 (m, CH3CHOH), 4.78 (t, J=6.2 Hz, CH2N+), 7.8
(2H, m, Hm of pyridinium), 8.3 (lH, m, Hp of pyridinium),
8.65 (lH, m, Ho of pyridinium), uv (H2O) ~ max:268
(~9350), 296 (~8840) m~, [~]D23 +41 (C 0.5, H2O), ~ =
15.0 h (measured at a concentration of 10-4 M in
phosphate buffer pH 7.4 at 36.8C)~
* Trademark
.
- : ' ,. ... .
, . .
'
": ' ' '
- 81 -
Example 6
Preparation of ;~iL -(1-(4-methYlPYridinium~)-
ethylthio]-6~-~1(~)-hydroxYethyl1-7-oxo-1-
azabicy~lo(3.2.0)hept-2-ene-2-carboxYlate
o~ ~ S~ N~3
-
A. 1-t2-mercaptoethyl)-4-methylpyridinium
methanesulfonate
.sG~ ~ 55C, 24 h
~N~ M5 -
To a suspension of 4-picolinium methanesulfonate in
4-picoline prepared by the addition of methanesulfonic
acid (0.65 mL, 0.010 mol) to 4-picoline (2.14 mL, 0.022
mol) in cooling was added ethylene sulfide (0.655 mL,
0.011 mol). The reaction mixture was stirred under a
nitrogen atmosphere at 55C for 24 h, cooled to 23C and
diluted with water (5 mL) and ether (10 mL) . The organic
layer was separated and the aqueous layer was washed with
ether (5 x 5 mL) and applied on top of a column (2.5 x 10
cm) of ~-"Bondapak"* C-18 after traces of ether have been
removed under reduced pressure. Elution of the column
with 15~ acetonitrile 85% water mixture gave after
lyophilization of the appropriate fractions a colorless
* Trademark
' ,
,, , ' : :
.
"' : '
- 82 -
syrup 2.66 g (100%), ir (film) vmax:2500 (SH~, 1540
(pyridinium), 1572, 1520, 1478, 1200 (sulfonate), 1040,
833 and 768 cm~l, 1Hmr (DMSO-d6) ~: 2.31 (3H, s,
CH3S03-), 2.62 (s, CH3 on pyridinium), 2.2-2.9 (4H, SH,
CH3 on pyridinium), 3.04 (2H, m, CH2S), 4.68 (2H, t,
J=6.4 Hz, CH2N+), 8.01 (2H, d, J=6.6 Hz, Hm of
pyridinium), 8.89 (2H, d, J=6.6 Hz, Ho of pyridinium), uv
(H2O) ~max: 256 (~4100), 221 (~7544) m~.
B. 1-(2-mercaptoethyl)-4-methyl~yridinium p-
toluenesulfonate
N~ i 1~s~l ~_ 5' ,~N~ ~55
To a suspension of p-toluenesulfonic acid (1.72 g,
0.01 mol) in benzene (6.5 mL) was added 4-picoline (1.17
mL, 0.012 mol). The resulting mixture was stirred under
a nitrogen atmosphere at 23C for 30 min, treated with
ethylenesulfide (0.65 mL, 0.011 mol) and stirred at 75
for 24 h. More ethylenesulfide (0.65 mL, 0.011 mol) was
added and the stirring was continued at 75~C for 24 h
more. The reaction mixture was cooled to 23C and
diluted with water (5 mL) and ether (8 mL). The aqueous
layer was separated and washed with ether (3 x 8 mL).
The traces of organic solvents were removed under vacuum
and the compound was chromatographed on ~-"Bondapak"* C-
18 with water as eluting solvent to give 2.94 g (90%) of
the title compound as a colorless syrup; ir (film) vmaX:
2510 (SH) 1640 (pyridinium), 1595, 1582, 1475, 1200
(sulfonate), 1031, 1010, 818 cm~1, lHmr
* Trademark
'
.
- 83 -
(DMSO, d6) ~: 2.29 (3H, s, CH3 on pyridinium), 2.61 15,
CH3 Ph), 2.4-2.8 (4H, SH, CH3Ph), 3.03 (2H, m[addition of
D2O gave a t, J=6.4 Hz, at 3.04], CH2S), 4.68 (2H, t,
J=6.4 Hz, CH2N~), 7.11, 7.49 (4H, 2d, J=7.9 Hz, Phenyl),
8.00 (2H, d, J=6.5 Hz, Hm of pyridinium), 8.89 (2H, d,
J=6.5 Hz, Ho of pyridinium), uv (H2O) ~max:256 (~4315),
222 (~17045) m~.
c. Paranitrobenzyl 3-[2-(1-4-methylpyridinium)~-
ethylthiol-6~-~1-(~)-hydroxyethyl1-7 oxo-l-
azabicyclo(3.2.0)hept-2-ene-2-carboxylate
diphenylphosphate
~t (~3 2
2~ c1~(0~)2
I ~0 _
O ~XL~3 3)
0~
J"~ 2~
C~0~ ),
Top a cold (0C) solution of p-nitrobenzyl 6~-[1-
(R)-hydroxyethyl]-3 ! 7-dioxo-1-azabicyclo(3.2.0)heptane-2-
carboxylate (0.522g, 1.5 mmol) in acetonitrile (6 mL)
kept under a nitrogen atmosphere was added
diisopropylethlamine (0.314 mL, 1.8 mmol) followed by
diphenyl chlorophosphate (0.373 mL, 1.9 mmol). The
reaction mixture was stirred for 45 min and treated
dropwise with a solution of 1-(2-mercaptoethyl)-4-
methylpyridinium methanesulfonate (0.539 g, 2.16 mmol) in
~_ ,
,
-
- 83(a~ -
acetonitrile (1.8 mL) followed by diisopropylethylamine
(0.314 mI" 1.8 mmol). The reaction mixture was stirred
at 0C for 1 h, diluted with cold (0C) water (24 mL) and
poured on top of a column
.
,
- 84 -
(2.5 x 8.5 cm) of ~-"Bondapak"*C-18. Elution of the
column first with 25~ acetonitrile -75% water mixture
(100 mL) then with 50% acetonitrile 50% water mixture
(100 mL) afforded after lyophilization of the appropriate
fractions 0.91 g (83%) of ~he title compound as a
yellowish powder, ir (KBr~ vmaX:3700-2800 (OH), 1770 (c=o
of ~-lactam), 1700 (c=o of PNB ester), 1640 (pyridinium),
1595 (phenyl), 1520 (NO2), 1340 (NO2), 890 cm~1 (NO2),
Hmr (DMSO, d6) ~:1.16 (3H, d, J=6.2 Hz, CH3 CHOH), 2.61
(s, CH3 on pyridinium), 3.1-3.7 (3H, m, H-6, CH2S), 3.7-
4.4 (2H, m, H-5, CH3CHOH), 4.79 (2H, brt, J=6.3 Hz,
CH2N+), 5.17 (d, J=4.9 Hz, OH), 5.37 tcenter of ABq,
J=14.1 Hz, CH2 of PNB), 6.7-7.4 (lOH, m, phenyl), 7.69
(2H, d, J=8.8 Hz, Ho of PNB), 8.00 (2H, d, J=6.5 Hz, Hm
of pyridinium), 8.23 (2H, d, J=8.8 Hz, Hm of PNB), 8.92
(2H d J=6.5 Hz, Ho of pyridinium), uv (H2O ~maX:262
(~10835), 311 (~9670) m~. Anal. calcd. for
-36H36-3QloSP 1 5 H2O: C 56-84, H 5.17, N 5. 52. S 4.21:
found: C 56.89 H 5.13, N 5.19. S 4.41.
D. 3- r 2-(l-(4-methylpyridinium))ethylthio]-6~- r 1- (R)-
hydroxyethyl1-7-oxo-1-azabicvclo(3.2.0)hept-2-ene-2-
carboxylate
JD~ S~_~_~N ~ ~ 2 _~
~hO) ~O
CD5~3 2
0~
,J"~ ~
-
C~
*Trademark
;_
3~
- 85 -
To a solution of p-nitrobenzyl 3-[2-(1-(4-
methylpyridinium))-ethylthio~-6~-[1-(R)-hydroxyethyl]-7-
oxo-l-azabicyclo(3.2.0)hept-2-ene-2-carboxylate
diphenylphosphate (0.587 g, 0.80 mmol) in wet
tetrahydrofuran (30 mL) was added ether (30 mL) potassium
phosphate mono basic-sodium hydroxide buffer (0O15 M,
14.7 mL, pH 7.22) and 10% palladium on charcoal (0.59 g).
The resulting mixture was hydrogenated under 40 psi at
23~C for 1.25 h. The organic layer was separated and
extracted with the buffer (2 x 6 mL). The aqueous
extracts were combined, filtered through a "Celite"* pad,
washed with ether (3 x 20 mL), pumped to remove traces of
organic solvents and poured on top of a column (2.5 x 10
cm) of ~-"Bondapak*" C-18. Elution of the column with
water and lyophilization of the appropriate fractions
gave 0.136 g (49%) of the title compound as a yellowish
powd~r, ir (KBr) vmaX:37oo-3ooo (OH), 1770 (c=o of 8-
lactam), 1642 (pyridinium) 1592 cm~1 (carboxylate), 1Hmr
(D2O) ~:1.19 (3H, t, J=6.3 Hz, C_3CHOH), 2.59 (3H, s, CH3
on pyridinium), 2.84 (d, J=9.1 Hz,H-4~, 2,90 (d, J- 9.1
Hz, H-4), 3.0-3.6 (3H, m, CH2S, H-6), 3.86 (dd, J-9.1 Hz,
J=2.6 Hz, H-5), 4.12 (m, CH3CHOH), 4.5-4.9 (CH2N+ masked
by HOD), 7.80 (2H, d, J=6.6 Hz, Hm of pyridinium), 8.58
(2H, d, J=6.6 Hz, Ho of piperidinium), uv (H2O) ~maX:256
(~5510), 262 (~5360), 296 (~7050) m, [~]D23 +20.8 (C
0.48, H2O), ~=12.8 h (measured at a concentration of 10-
4 M in a phosphate buffer pH 7.4 at 36.8C).
* Trademark
~ .
.
. .
- 86 -
Exam~le 7
Preparation of (5R) 3-[2~(4-methyl-thioPYridinio)-
ethylthio~-(6~ ! - [ ( 1g~ oxyethyl~-7-oxo-1-
azabicyclo[3.2.0]hept-2-ene-2-carboxYlate
0~ ' '
J ~ 5 ~ ~ C~3
~2
A. 4-Methylthiopyridine*
N ~ H - ~ N ~ Me
4-Mercaptopyridine (5.55 g, 50.0 mmol; Aldrich*) was
dissolved in boiling abs. EtOH(50 mL). The insoluble
material was removed by filtration over "Celite"**. The
filtrate was heated to xe-dissolve, and when it cooled to
ca. 50C, methyl iodide (3.17 mL, 51.0 mmol; Aldrich) was
added at once. The mixture was cooled to crystallize.
Filtration of the solid gave 6.77 g (26.7 mmol, y. 53.5%)
of the title compound as the hydroiodide: 1Hmr (D2O)
~: 2.70 (3H, s, -SCH3) and 7.65 -7.77-8.35-8.48 ppm (4H,
A2B2 type, aromatic Hs); ir ("Nujol")** ~max l6l5, 1585
(aromatic) and 780 cm~1; uv (H2O)
* Trademark
** Trademark
~ ~ ,
. . :, ' . . ,~ ' :
; ' '
': ' : ' ' . ' ' ~ ~ '
~8~
- 87 -
~max 227 (~2.02 x 104) and 298 nm 1~1.64 x 104).
The hydroiodide (6.33 g, 25.0 mmol) was dissolved in
H2O (40 mL) and the insoluble material was removed and
washed with H2O (10 mL). To the filtrate was added at 0-
5 NaOH pellet (5 g) and extracted with ~t2O (3 x 25 mL),
saturating the aqueous layer with NaCl. The combined
organic extracts were washed with brine (x 2), dried
(MgSO4) and evaporated, yielding 2.92 g (23.4 mmol,
overall yield 50%) of the title compound as an oil: lHmr
(CDC13) ~: 2.4~ (3H, s, -SCH3) and 7.03-7.13-8.38-8.48
ppm (4H, A2B2type, aromatic-Hs); ir (film) ~max 1580 and
800 cm~l.
*Preparation of this compound was reported by Xing and
Ware, J. Chem. Soc., 873(1939). The procedure, described
in this ref~rence was followed.
B. 4-Methylthio-N-(2-mercaptoethyl)pyridinium
methànesulfonate
+ N ~ 50-60
yS ~ ~ C335 ~
4-Methylthiopyridine (2.75 g, 22.0 mmol) was added
slowly to methanesulfonic acid* (0.65 mL, 10.5 mmol) by
cooling in an ice-bath. To this solid was added ethylene
sulfide* (0.66 mL! 11.0 mmol, Aldrich) and the mixture
was heated at 50-60C for 21 h. As reaction proceeds the
solid went to solution. After cooling,
~;~
: ' '', ' '
'- . .
.
', , ' 1 ' ~ '
'
- 88 -
the reaction mixture was dissolved in H2O (5 mL) and
washed with Et2O (5 x 4 mL). The cloudy aqueous layer
was filtered over "Celite~' and the ~iltrate was purified
by reverse phase silica gel column chromatography (C18
"Microbondapak"1 10 g) eluting with H2O. Each fraction
of 10 mL was collected. Fractions 2 and 3 were combined
and repurified by the reverse phase column. Fraction 2
gave 1.1258 g (4.48 mmol, y. 42.6%) of the title compound
as a viscous oil: 1Hmr (DMSO-d6, CFT-20) ~: 2.32 (3H, s,
MESO3~), 2.72 (3H, s, -SMe), 2.68 (lH, m, SH), 2.9-3.2
(3H, m, -CH2S-), 4.59 (2H, t, J=6.4 Hz, -CH2 ~, 7.97 (2H,
"d", J=7.2 Hz, aromatic-Hs) and 8.72 ppm (2H, "d", J=7.2
Hz, aromatic-Hs); ir (neat) vmax:1630, 1200 (br, -SO3~),
7.85 and 770 cm~l.
* These reagents were distilled prior to use.
C. (5R) p-Nitrobenzyl 3-[2-(4-Methyl-
thiopyridino~ethylthio]-(5S)-[(lB)-hydroxyethyl~-7
oxo-l-azabicyclo~3.2.0lhept-2-ene-2-carboxylate
chloride OH - -
J'~'~o ~
~O2P~3 Ms
J ~ ~ ~,~_~ ~ ~.'e
C~2~
To a solution of (SR3 p-nitrobenzyl 3,7-dioxo(6S)-
[(lB)-hdyroxyethyl~ azabicyclo[3.2.0]heptane-(2B)-
carboxylate (475 mg, 1.36 mmol) and diisopropylethlamine
(0.24 mL, 1.4 mmol) in CH3CN (5 mL) was added at 0-5C
under a nitrogen atmosphere diphenyl chloro-
1 Trademark
,
. .
.
- 89 -
phosphate (0.29 mL, 1.41 mmol). The mixture was stirred
at 0-5, for 30 min. To this mixture was added an oily
suspension of 4-methylthio-N-(2-mercaptoethyl)pyridiniUm
methanesulfonate (678 mg, 1.45 mmol; 60% pure) in CH3CN
(1.5 mL) followed by diisopropylethylamine (0.24 mL, 1.4
mmol). The mixture was stirred at 0-5C for 1 h.
Immediately after addition of the base, yellowish
precipitate formed. The precipitate was filtered and
washed with cold CH3CN (3 mL), yielding 413 mg of
yellowish solid. This was triturated from 10% MeOH in
H2O (5 mL) to obtain 341 mg (0.618 mmol, y. 45.4%) of the
title compound as white crystals: mp 118-120C; lHmr
(DMSO-d6, CFT-20) ~: 1.16 (3H, d, J=601 Hz, 1'-CH3), 2.72
(3H, s, -SCH3), 3.1-3.7 (5H, m), 3.7-4.3 (2H, m), 4-71
(2H, t, J=6.3 Hz, -CH2 ~), 5.15 (lH, d, J=4.9 Hz, OH),
5.20-5.35-5.40-5.55 (2H, Abq, CO2CH2-Ar), 7.70 (2H, "d"
J=8.8 Hz, nitrophenyl-Hs), 7.97 (2H, "d", J=7.0 Hz,
pyridinio-Hs), 8.25 (2H, "d", J=8.8 Hz, nitrophenyl-Hs),
and 8.76 ppm (2H, "d", J=7.1 Hz, pyridinio-Hs); ir
("Nujol") vmaX: 3250 (OH), 1775 (~-lactam), 1700 (ester)
and 1625 cm~1 (pyridinio); uv (abs, EtOH) ~maX:308 nm
(~4.47 x 104); [~]n3 ~ 24.8 (c 0.5, MeOH); Anal. calcd.
for C24H26N3o6s2cl-H2o: C 50-56, H 4.95, N 7.37; found
C 50.63, H 4.72, N 6.89.
D. (5R) 3-[2-(4-Methylthiopyridinio)ethylthio~-(6~)-
[(l~)-hydroxyethyl)~7-oxo-l~azabicyclo[3.2.0~hept-2-
ene-2-carboxylate
'
.
.
.
- 89 (a)
0:~
J~ 2jPd C,
:. 7. 4 buf'e~
Co2~1;3
OB
F~S ~5~
C02~'
:
:
: : : ' ~ : :
X~
- :
,
. ' ', ~ ' ' , ' ,,
~: : , `
- 9o -
(5R) p Nitrobenzyl 3-[2-(4-
methylthiopyridinio)ethylthio~-(6S)-[(1_)-hydroxyethyl]-
7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate
chlorlde (380 mg, 0.68~ mmol) was dissolved in THF (31.5
mL) and pH 7.40 phosphate buffer (31.5 mL; 0.05M Fisher)
and diluted with Et2O (31.5 mL). rrhis solution was mixed
with 10% Pd-C (380 mg, Engelhard) and hydrogenated at 35
psi on the "Parr"* shaker at room temperature for 1 h.
The aqueous layer was filtered over "Celite"* to remove
the catalyst and the "Celite"* pad was washed with H2O (2
x 5 mL). The filtrate and washing were combined and
washed with Et2O (2 x 30 mL). The aqueous layer was
pumped off to remove any organic solven~s and purified by
reverse phase column chromatography (C18 'IMicrobondapak'l*
13 g, Waters Associates) eluting with H2O. Fractions
having a uv absorption at 307 nm were collected (ca. 1 L)
and lyophilized to obtain 127 mg (0.334) mmol, y. 48.5~)
of the title compound as a yello~ish powder; 1Hmr (D2O,
CFT-20) ~: 1.20 (3H, d, J=6.4 Hz, 1'-Ch3), 2.64 (3H, 5, -
SCH3), 2.81 (2H, m, -SCH2-), 3.19 (lH, dd, J6-l' =6.1 Hz,
J6_5=2.6 Hz, 6-H), 3.32 (2H, dd, J=11 Hz, J=5.5 Hz, 4-
Hs), 3.92 (lH, dt, J=9.2 Hz, J5_6-2.6 Hz, 5-H), 4-1 (lH,
m, 1'-H), 4.61 (2H, t, J=5.~ Hz, -CH2 ~), 7.70 (2H, "d",
J=7.1 Hz, aromatic-Hs), and 8.40 ppm (2H, "d", J=7.1 Hz,
aromatic-Hs); ir (KBr, disc) Vmax: 3400 (OH~ 1750 (~-
lactam), 1630 (pyrindinium) and 1590 cm~1 (carboxylate);
uv (H2O) ~max 231 (~9800) and 307 nm (~25000); [~]n3
3.14 (c-0.5, H2O)-
* Trademark
. " .
,. . ~ .
.
. .
, ' ~
.
,
~2~2~
-- 91 --
Example 8Pr~paration of 3-~-(3-methoxv-l-Pyridinium2
ethylthiol-6~-[.l'-(R~-hydroxyethyll-7-oxo-1-
azabicyclo)3.2.0)-hept-2-ene-2-carboxylate
0~
A. 1-(2-mercaptoethyl)-3-methoxypyridinium
methanesulfonate
~eO~> ~_
~O~
To precooled (5C) 3-methoxypyridine (698 mg, 6.4
mmol) was added dropwise methanesulfonic acid (0.216 mL,
3.05 mmol) and ethylene sulfide (0.19 mL, 3.2 mmol). The
mixture was then heated at 60C for 18 h, cooled to 20C,
diluted with water (10 mL) and washed with ether (3 x 10
mL). The aqueous phase was pumped under high vacuum for
15 min and poured on a C18 reverse phase column. The
title compound was eluted with water. The appropriate
fractions were combined and evaporated under high vacuum
to give the desired thiol (61.6 mg, yield 76.3%); ir
(CH2C12) VmaX:2550 (w, SH) and I620, 1600, 1585 cm~1 (m,
,
.
- 91(a) -
aromatic; lHmr (DMSO d6) ~: 8.90-7.90 (4H, m, aromatic C-
H), 4.72 (2H, t, J-6.6 Hz, CH2N+), 4.01 (3H, s, OCH3),
3.5-3.0 (m, hidden CH2S), 2.66 (lH, dd, J=9.5 Hz, J=7.5
Hz, SH) and 2.31 ppm (3H, s, CH3SO3-).
X'
:
.
''
~g~
- 92 -
B. ~aL~-Nitrobenzyl 3~2-(3-methox~-1-pvridinium
chloride?ethylthiol-6~ hydroxyethy~l-7-oxo
l-azabicyclo(3.2.0)-hept-2-ene-2-carbox~late
O~i ,1 ) ~ ~N ~--<) 2
J" ~ Z) ClPO(O~)2
O N ~ 3) ~ ~ ~` ~ ~sO
CO P~3 , -
2 4~ N~2
~ C~
0~
2~x3
A cold (0C) solution of p-nitrobenzyl 6a-[1'-(R)-
hydroxyethyl]-3,7-dioxo-1-azabicyclo-(3.2.0)-heptane-2-
carboxylate (1.04 g, 3 mmol), in acetonitrile (12 mL) was
treated dropwise with diisopropylethylamine (0.63 mL, 3.6
mmol) and diphenylchlorophosphate (0.75 mL, 3.6 mmol)
and stirred at 0C for 30 min. The resulting enol
phosphate was treated with 1-(2-mercaptoethyl)-3-
methoxypyridinium methanesulfonate (1.14 g, 4.30 mmol) in
CH3CN(7mL), diisopropylethylamine (0.63 mL, 4.30 mmol),
stirred for 30 min. and cooled at -10C for 30 min. The
solid that precipitated out of the mixture was filtered,
washed with cold acetonitrile (2 mL) and dried to give
the title compound (1.32 g, yield 82%); ir (''Nujol'')Vmax
3320 (m, OH), 1780, 1765 (s, ~-lactam c=o), 1700, 1695
(m, ester c=o) and 1520 cm 1 (s, NO2); 1Hmr (DMSO d6) ~:
9.01 (lH, bs, H-3 aromatic), 8.75 (lH, bd, J-5.4 Hz, H-6
aromatic), 8.35-7.95 (4H, m, H-aromatic), 7.70 (2H, d,
J-7.7 Hz, H-aromatic), 5.37 (2H, center of Abq, J=13 Hz,
CH2PNB), 5.17 (lH, d, -4.9 Hz, OH), 4.87 (2H, t, J=6.3
Hz, CH2-N~), 4.35-3.75 (2H, m, H-5 and H-1'), 4.00 (3H,
s, OCH3) 3.56 (part of a t, J=6.3 Hz, CH2S), 3.5-3.20
(3H, m, H-6, H-3) and 1.16 ppm (3H, d, J-6.1 Hz, C_3CHO).
~1 ~
.
- 93 -
C. 3-[2-(3-methoxy-1-pyridinium)ethylthiol-6~-[1'-(~)-
hydroxyethyl~-7-oxo-1-azabicyclor3.2.0)-hept-2-ene-
2-carboxylate
0~ Q~ ,0.~e
~2-N3
O~ Q~
. j~S
A solution of Para-nitrobenzyl 3[2-(3-methoxy-1-
pyridinium chloride)ethylthio]-6~-[1'-(R)-hydroxyethyl]-
7-oxo-1-azabicyclo(3.2.0)-hept-2-ene-2-carboxylate (600
mg, 1.12 mmol) in THF (25 mL), ether (25 mL) and pH 7.4
phosphate buffer (0.lM, 25 mL) was hydrogenated in a
"Parr"* shaker over 10% PD/C (1.1 g) for 1 h at 40 psi.
The mixture was diluted with ether and the aqueous phase
was filtered through a #52 hardened filter paper. The
aqueous layer was washed with ether (2 x 20 mL), pumped
under vacuum and poured on a silica gel reverse phase
column. The title compound was eluted with water
containing 2 and 5% acetonitrile. The appropriate
fractions were combined and lyophilized to give a yellow
solid that was repurified by hplc to give the penem
carboxylate (150 mg, 38%); ir "(Nujol"*) vmaX 1750 (s, ~-
lactam C=O) and 1580 cm~1 (s, carboxylate); 1Hmr (D2O) ~:
8.55-8.30 (2H, m, H-2, H-6 aromatic), 8.17-7.75 (2H, m,
H-3~ H-4 aromatic), 4.77 (2H, t, J=5.9 Hz, CH2 ~), 4.10
(lH, part of 5 lines, J=6.3 Hz, H-l'), 3.97 (3H, s,
OCH3), 3.85, 3.82 (2 lines, part of dt, J=2.6 Hz, part of
;_
.
_ 93(a) -
H-5), 3.42 (2H, t, J=5.9 Hz, CH2-S), 3.25 (lH, dd, J=6.1
Hz, J=2.6 Hz, H-6), 2.g9-2.60 (2H, 6 lines, part of H-4)
and 1.20 ppm (3H, d, J=6.4 Hz, CH3); uv (H20, c 0.05)
~ma~: 290 (~10517), 223 (~6643); ~% (0.1 M pH 7.4
phosphate buffer, 37C) 20 h.
* Trademark
. ~ . . _ . ...
,
- 94 -
Example 9
Preparation of (5 ,6~)-3-~2-(3-methylthiopyridinio~ethyl
thiol-6- r 1_ (~) -hydroxyethyl]-7-oxo-1-
azabicyclo[3 2.0]hept-2-ene-2-carboxylate
0~ S~
A. 3-Methylthio-1-(2-mercaptoethyl)pyridinium chloride
5~
N~ A
~2C,~
To a solution of 3-methylthiopyridine 1 (2.00 g,
0.016 mol) in 10 mL of ether was added 15 mL of 1 N HCl
and the mixture was well shaken. The aqueous phase was
separated, washed with 10 mL of ether and then
evaporated. The residual hydrochloride was then dried in
vacuo (P2O5) to give a white solid. To this solid
hydrochloride was added 3-methylthiopyridine (1.88 g,
0.015 mol) and ethylene sulfide (0.89 mL, 0.015 mol) and
the rustling mixture was heated (oil bath) at 55-65C
under N2 for 15 h. This gave a slightly turbid oil which
was taken up in 125 mL of H2O and washed with CH2Cl2.
The aqueous solution was concentrated to about 25 mL and
then a few drops of acetonitrile were added to make the
mixture homogeneous. The resulting aqueous solution was
applied to a C18 reverse phase column. Elution with H2O
and subsequent evaporation
,: ' , ,
~8~
- 35 -
of the relevant fractions afforded the product (2.66 g
80%) as a pale yellow, viscous oil. ir (film) ~max
2410 (br, ~SH) cm~1; lHnmr (d6-DMSO+D2O) ~: 8.88-7.88
(m, 4H, aromatic), 4.70 (t, J=6.5 Hz, 2H, N-CH2), 3.08
(skewed t, J-6.5 Hz, 2H, S-CH2), 2.64 (s, 3H, S-Me).
1Prepared by the method of J.A. Zoltewiez and C. Nisi,
J. Org. Chem. 34, 765 (1969).
B. p-Nitrobenzyl (5B,6~)-3- r 2-(3-
methylthiopyridinio)ethyl thiol-6- r 1- (~) -
hvdroxyethyl~-7-oxo-l-azabicyclo[3.2.o~hepto-2-ene
2-carboxylate chloride
O N
CO222
o~ ~S4Ccl~
A solution of p-nitrobenzyl t5R,6S)-6-[1-(R)-
hydroxyethyl]-3,7-dioxo-1-azabicyclo[3.2.0]heptane-2-
carboxylate (0.522 g, 1.50 mmol) in 7 mL of dry
acetonitrile was cooled at 0C and then
diisopropylethylamine (0.287 mL, 1.65 mmol) was added
dropwise. To the resulting yellow~brown solution was
added dropwise diphenyl chlorophosphate (0.342 mL, 1.65
mmol) and the reaction mixture was kept at 0C for 30
min. Diisopropylethylamine (0.313 mL, 1.80 mmol) was
then added, followed by a solution of 3-methylthio-1-(2-
mercaptoethyl)pyridinium chloride (0.398 g, 1.80 mmol) in
0.70 mL of dry DMF. About a minute after the addition
- 95(a) -
was completP a precipitate separated from the reaction
mixture and further cooling at -10C for 10 min gave a
solid orange-coloured mass. This solid was subsequently
triturated with acetonitrile and the residue was
collected by filtration. The residue
' ' '
.
~, . .
'
'. . :'
d~L,~
- 96 -
was washed with acetonitrile, then acetone and finally
dried in vacuo to give the product (0.455 g, 55%) as a
cream-coloured solid. The combined filtrate was
evaporated to give a yellow oil which was taken up in a
minimum volume of acetonitrile and cooled at 0C for 30
min. Filtration of this mixture afforded an additional
0.139 g of the product as a light yellow solid. The
combined yield was 0.594 g (72%). ir (KBr) ~max 3345
(br, -OH), 1770 (B-lactam CO), 1680 (-CO2PNB) cm~l; 1Hnmr
(d6-DMSO) ~: 8.98-7.96 (m, 4H, pyridinium aromatic),
8.20-7.65 (ABq, J=7.0 Hz, 4H, PNB aromatic), 5.53-4.80
(m, 4H), 4.3-3.7 (m, 2H), 3.6-3.25 (M, 6H), 2.66 (s, 3H,
S-Me), 1.16 (d, J-6.0 Hz, 3H, CHMe).
C. (5B.6S~-3-[2-(3-methylthiopyridiniolethyl thio~-6-
r 1- (R) -hydroxyethyll-7-oxo-1-azabicYclo r 3.2.0]hept-2-
ene-2-carboxylate
~ SMe
. ~ ~
~ ~ \~Cl~ '
22~
OH ~s
~'~S
C~;2
To a mixture of p-nitrobenzyl (5R,6S)-3-[2-(3-
methylthiopyridinio)ethyl thio]-6-[1-(B)-hydroxyethyl]-7-
oxo-l-azabicyclo[3.2.0]-hept-2-ene-2-carboxylate chloride
(0.551 g, 1.0 mmol) and 10% palladium-on-charcoal (0.55
g) in 25 mL of phosphate buffer ~0.05 M, pH 7.4) was
~ L 'p~
- 9~(a) -
added 5 mL of THF and 25 mL of ether. This mixture was
hydrogenated (Parr) at 40 psi for 1 h, The reaction
mixture was then filtered through "Celite" and the filter
cake was washed with H20 and ether. The aqueous phase
was separated and washed with additional ether (3 x).
.
'; . , : ,
.
- 97 -
After removing residual organic solvents in vacuo the
aqueous solution was cooled at 0C and the pH was
adjusted to 7.0 with saturated aqueous NaHCO3. This
solution was immediately applied to a C18 reverse-phase
column. Elution with H2O and subsequent lyophilization
of the relevant fractions afforded 0.25 g of a bright
yellow solid. This material was repurified by reverse-
phase hplc to give the product (0.210 g, 55%) as a light
yellow solid. ir (KBr) vmaX:3400 (br, -OH), 1755 (~-
lactam CO), 1590 (-CO2-) cm~1; lHnmr (D2O) ~:8.60-7.76
(m, 4H, aromatic), 4.76 (t, J=5.8 Hz, 2H, N-CH2), 4.13 (d
of q, J=J'=6.3 Hz, lH, H-l'), 3.95 (d of t, J=9.0 Hz,
J'-2.8 Hz, lH, H-5), 3.45-2.75 (M, 5H), 2.59 (s, 3H, S-
Me), 1.20 (d, J=6.4 Hz, 3H, CHMe); uv (H2O) ~maX:296
(~8509), 273(~13005), 231(~11576) nm; ~ (ph 7.4, 36.8C)
20 h.
Example 10
Preparation of 3-[2-(1-(2,6-dimethylpyridinim)-
ethylthiol-6~ -(~)hydroxyethyll-7-oxo-1-
azabicyclo(3.2.0)hept-2-ene-2-carboxylate
~ C~3
J,,c;~
COo C~3
._
.
- 98 -
A. 1-(2-mercaptoethyl-2!6-dimethylpyridinium
methanesulfonate
N~ ~ ~eO3 100~ ~ 42 ~ ~ y,5o
A mixture of 2,6-diemthylpyridine tl9.2 mL, 0.165
mol) and methanesulfonic acid (3.27 mL, 0.050 mol) was
stirred for 15 min, treated with ethylene sulfide (4.17
mL, 0.070 mol) and stirred at 100C for 42 h under a
nitrogen atmosphere. After cooling to 25C, the
reaction mixture was diluted with ether (45 mL) and water
(30 mL). The two layers were separated and the organic
layer was extracted with water (2 x 5 mL). The aqueous
layers were combined, filtered through a "Celite" pad,
washed with ether (2 x 15 mL), pumped to remove the
traces of organic solvents and poured on top of a column
(3.0 x 12 cm) of ~-"Bondapak"C-18. Elution with 3%
acetonitrile 97% water mixture gave after lyophilization
of the appropriate fractions 2.5 g of the impure title
compound as a syrup. It was repurified by hplc (~-
"Bondapak" C-18) to give 0.90 g t7%) for the title
compound. ir (film) VmaX:2520 (SH), 1640 and 1625
(pyridinium), 1585, 1490, 1200 cm~1 (sulfonate), lHmr
(DMSO-d6 + D2O) ~: 2.36 (3H, s, CH3SO3-), 4.62 (2~, m,
CH2N+), 7~74 (2H, m, Hm of pyridinium), 8.24 (lH, m, Hp
of pyridinium), uv (H2O) ~maX:272 (4080)m~
r
- 99 -
B . Paranitrobenzyl 3-[2-rl-(2 6-
~ thvlro~ yn~ethylthiol-6~-~l-(~-
hydroxyethyl]-7-oxo-1-azabicyclot3.2.0)he~t-2-ene-
carboxYlate diphenylphosphate
~ 1) N:~, (i~._)
J~= 2) cl~(a~?h).
3) ~(~
~sO
4) ~ ~i~)2
CC0~3 ~hO)2~o
To a cold (0C) solution o~ p-nitrobenzyl 6~-[1-(~-
hdyroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-
carboxylate (0.658 g, 1.89 mmol) in acetoni~rile (6 mL)
kept under a nitrogen atmosphere was added
diisopropylethylamine (0.394 mL, 2.26 mmol) and diphenyl
chlorophosphate (0.~68 mL, 2.26 mmol). The reaction
mixture was stirred 30 min and treated with a solution of
1-(2-mercaptoethyl)-2,6-dimethylpyridinium
methanesulfonate (0.720 g, 2.73 mmol) in acetonitrile (3
mL) followed by diisopropylethylamine (0.394 mL, 2.26
mmol). The reaction mixture was stirred at 0C for 2 h,
diluted with cold (0C) water (27 mL) and poured on top
of a column (2.5 x 9.0 cm) of ~-"Bondapak" C-18. Elution
with acetonitrile-water mixtures and lyophilization of
the appropriate fractions gave 0.92 g (65~) of the title
compound, ir (KBr) ~maX:3700-3000 (OH), 1765 (C=O of B-
lactam), 1690 (c=o of PNB ester), 1620 (pyridinium), 1590
(phenyl), 1517 (NO2), 1330 ~NO2), 880 cm~l (NO2~, lHmr
(DMSO, d6) ~: 1.15 (3H, d, J=6.2 Hz, CH3CHOH), 2.7-3.7
~r
~L~89
- 99 (a
(11 H, CH2S, 2-CH3 on pyridinium, H-4, H-6)j 3.7-4.4 (2H,
CH3CHOH, H-5), 4.7 (2H, m, CH2N~), 5.14 (lH, d, J=4.5 Hz,
OH), 5.37 (center of ABq, J=13.2 Hz, CH2 of PNB), 6.7-
7.5 (lOH, m, phenyl), 7.5-8.7 (7H, pyridinium, H's of
PNB), uv (H2O) ~maX 274 (~14150), 319 (~9445) m~
: : : :
,
- ~ :
: .
8~
-- 100 --
C. 3- r 2-(1-(2 6-dimethylpyridinium))ethylthio~-6~-rl-
~ hYdroxyethyl]-7-oxo-1-azabicyclo(3.2.0!he~t-2-
ene-2-carboxylate
r~F, e~ er !
COO?~3 ~b~)2~O
OX
o5C~S~~
To a solution of p-nitrobenzyl 3-[2-(1-(2,6-
dimethylpyridinium))-ethylthio]-6~-[1-(R)-hydroxyethyl]-
7-oxo-1-azabicyclo(3.2.0)hept 2 ene-2 carboxylate
diphenylphosphate (0.80 g, 1.07 mmol) in wet
tetrahydrofuran (42 mL) was added ether (42 mL),
potassium phosphate monobasic-sodium hydroxide buffer
(0.15M, pH 7.22, 21 ML) and 10% palladium on charcoal
(0.80 g). The result.ing mixture was hydrogenated for 1 h
under 40 psi at 23C and filtered through a "Celite" pad.
The two layers were separated and the organic layer was
extracted with the buffer (3 x 8 mL). The aqueous phase
were combined, washed with ether (50 mL), pumped to
remove traces of organic solvent and poured on top of a
column (3.0 x 10.2 cm) of ~-"Bondapak" C-18. Elution of
the column with 5% acetonitrile - 95% water mixture and
lyophilization of the appropriate fractions gave the
title compound 0.246 g (63%) as a yellowish powder, ir
(KBr) ~maX:3700-2800 (OH), 1750 (c=o of the ~-lactam),
1620 (pyridinium), 1585 cm 1 (carboxylate~, lHmr (D2O)
~:1.23 (3H, d, J-6.4 Hz, CH3CHOH), 2.5-3.5 (llH, H 4, H-
6, CH2S, 2CH3 on pyridinium), 3.8-4.4 (2H, CH3CHOH, H-5),
~i
,
~.~84
- loo (a)
4.5-4.9 (CH2N+, HOD), 7.64 and 7.74 (2H, A part of A2B
system, Hm of pyridinium), 8.07, 8.16, 8.18 and 8.27 (lH,
B part of A2B system, Hp of pyridinium), uv (H2O) ~max
277 (~9733), 300 (E8271) m~, [a] 23 +50.7'(C 0.48, H2O),
Anal. calcd.for C18H22N~o4s-l-5 H2O: C 55-51
7.19; found: C 55.14, H 6.23, N 6.46
. ` . "'
.
'
- 101 -
Exa~n?le 11
Pre~ar2tio~ of (;~, 6~)-3-[2-(2-~ io-3-
me.hylimidazolio)ethyl-thio~-6~ )-hydroxy-
ethyl] -7-oxo-1-azabi~clo [ 3 . 2 . O ~ he~t-2-ene-
2- caxboxy 12 .e
0~ ~Me
~ S ~ ~ N-Me
o C023
. 2-Methyl~hio-3-methyl-1-(2-merc2ptoe'hyl)imud2zO1i~
tri'luoromethanesulfonate ' SMe
~3S3~ 5C~2c~2 ~ ~ ~-Me
Me C~3S03 .
Trifluoromethanesul'onic acid (1.38 mL, 0.015 mol)
was added dropwise to 2-methylthio-i-me.hylimidazolel (4.0 g~,
0.03 mol) at 0C under,N2. Ethylene su}.ide (0.9 mL, 0.015
mol) wzs then added and the mix~ure was heated at 5;C under
N2 ror 24 h. The,reaction mixture was tri.urated with e~he~
~3x) and the residue was taXen up in aceto~e, filtered and
evaporated. This gave the product (4.2 g, 8~2~) as a semi-
c_Ysta~ e solid which was used.as such without rurther
purification. ir( ilm),vmax:. 2550 (w, sh) c~ ; 'Hnmr
(d6-acetone) ~: 7.97 (s, 2H)~ 4.66'(t, J=7 Hz , 2~, methylene),
~.17 ~s, 3~, N-Me), 3.20 (d of t, J=7 Hz, Jt~9 Hz, 2H,
meehylene), 2.72 (s, 3H, S-Me), 2.20 (t,' J=9 Bz, l~ SH).
l. ?repared as ,Per A. Wohl and W. Mzrc~wzld, Chem. Ber. 2 ,
1353 ~188
.-
~\ '\
~ ~ :
3~
-102-
B. p-Nitrobenzyl (5~ 6 )-3_L_-(2-methylthio-3-methYl-
imidazolio)ethyl thiol-6-[1-(~-hydroxyethyl~-?-oxo-1-
azabicyclo~3.2.0lhept-2-ene-2-carboxyla~e
diphenylphosphate
0~ 0~ ~e
~0 , r~ S~ h _
b ~ ~ ~ W
C02~N~ C02?~3 (~)2~~
To a solution of p-nitrobenzyl (5R,-6S)-6-[1-
(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo[3.2.0]heptane-2-
carboxylate (1.40 g, 4.0 mmol) in 50 mL of dry
acetonitrile, at 0C under N2, was added dropwise
diisopropylethylamine (0.76 mL, 4.4 mmol) followed by
diphenyl chlorophosphate (0.91 mL, 4.1 mmol). After
stirring the reaction mixture at room temperature for 1
h, diisopropylethylamine (0.76 mL, 4.4 mmol) was added
and then a solution of 2-methylthio-3-methyl-1-(2-
mercaptoethyl)imidazolium trifluoromethanesulfonate (2.0
g, 5.9 mmol) in 5 mL of acetonitrile was added dropwise.
The reaction mixture was kept at room temperature for 1.5
h and was then concentrated ln vacuo to give a gum.
This gum was taken up in H2O and applied to a C18
reverse-phase column. Eution with H2O, then 20%
acetonitrile-H2O finally 30% acetonitrile-H2O, followed
by lyophilization of the appropriate fractions afforded
the product (0.90 g, 30%) as a light yellow solid. ir
(KBr) VmaX:3380 (br, OH), 1770 (B-lactam CO) cm~1; ~Hnmr
(d6-acetone) ~:8.35 (br,s, lH), 8.24, 7.78 (AB q, J=8.8
Hz, 4H, aromatic), 7.89 (br s, lH), 7.25-6.91 (m, 10H,
diphenylphosphatej, 5.50, 5.25 (AB q, J=12 Hz, 2H,
benzylic), 4.75-4.27 (m,3H), 4.03 (s, 3H, N-Me), 4.15-
2.75 (m, 8H), 2.59 (s, 2H, S-MMe), 1.22 (d, J=6.2 Hz, 3H,
~CHMe).
r
3r~c
-103-
C. (5R, 6S)-3-~2-(~-Methvlthio-3-methYlimidazolio)ethylthiol-6-
Cl-iB)-hvdroxveth~ 7-oxo-l-azabicy~lor3.2.olhept-2-ene-2
carboxylate
0~ e
-- - C 2 ~ 0 ) 2 ? 0
0~
Y SM.
r ~ -5 ~ ~ e
co2~
To a solution of p-nitrobenzyl (5R, 6S)-3-[2-(2-methylthio-3-
methylimidazolio)ethyl thio[-6-[1-(B)-hydroxyethyl]-7-oxo-1-
azabicyclo[3.2.0]hept-2-ene-2-carboxylate diphenylphosphate (1.20
g, 1.56 mmol) in a mixture of 70 mL of THF, 70 mL of ether and 31
mL of phosphate buffer (0.05M, pH 7.4) was added 1.2 g of 10%
palladium-on-charcoal. This mixture was hydrogenated (Parr) at 35
psi for 55 min. The reaction mixture was then filtered through
"Celite" and the filter cake was washed with H2O and ether. The
aqueous phase was separated, cooled at 0C and the pH was adjusted
to 7.0 with saturated aqueous NaHCO3. After removing residual
organic solvents in vacuo the aqueous solution was applied to a
C18 reverse-phase column. Elution with H2O and then 8%
acetonitrile-H2O and subsequent lyophilization of the relevant
fractions gave 0.25 g of a solid. This material was repurified by
reverse-phase hplc to give the product (0.114 g, 19%) as an off-
white solid. ir(KBr),~max:3420 (OH), 1750 (B-lactam CO), 1590 (-
CO2O) cm~l; 1Hnmr (D2O) ~: 7.58 (s, 2H), 4.52 (t, J=6 Hz, 2H),
4.28-3.82 (m, 2H), 3.90 (s, 3H, N-Me), 3.40-2.87 (m, 5H), 2.40 (s,
3H, S-Me), 1.20 (d, J=6.4 Hz, 3H, -CHMe); uv (H2O) ~maX:297 (E
7572), 262 (E6259), 222 (E 7955) nm.
.~
. ~
- 104 -
~xam~le 12
Pre~aration of (5~, 6S)-3-~ 2-(3-amino~r~dinio)e.hyl -
thio]-6-[l~(R)-hydroxyethvl]-7-oxo-1-azabicyclo[3 2 0]-
hept- 2-ene-2-c2rboxylate
= 023 ~ 2
A 3-~mino-1-(2-merca?toethyl)pyridin~um chloride
N2N~ 1~2N~Q 5 1~5C32C~I~C~ 2
3-Aminopyridine (1 50 g, 0 016 mol) W25 taXe~ U?
in 15 mL of 1 N meth~nolic HCl and the resulting solution
w2s evapor2ted to give the hydrochloride 2s an oil To ~his
oil was added 3-~mlnoDyridine (1.32 g, 0 015 mol) and
ethylene sul'ide (0 89 ml, 0 01; mol) and the resulting
mix!ure was heated (oil bzth) 2t 60-65C under N2 ~or 2 h
Ano her equivalent of ethylene sul'ide (0 89 ml, O OlS mol)
W25 added and heating was contlnued 2t 55-65C or 65 h.
The reaction mixture W2S washed wi'.h CH2C12 and hen tzken
` up in ~2 (25 ml). The zqueous solution was applied to a
C18 reverse-phase column which was eluted with ~2
Evaporation of the relevant fractions gave the produc~ (1 26 g,
~4%) as a colorless, viscous oil ir(~ilm) ~max 3180 ~NP2)
cm l; ~ mr (d6-D~5O) o: 8 19-7 59 (m, 4H, aromatic), 4 59
(t, J-6 2 Hz, 2H, N-C~ ); 3 5 (br s, 2H, -NH~), 3 20-2 7?
(m, 3~) _
j
.~ :
-105-
B. p-Nitrobenzyl (5~ 65~-3-(2-(3-aminopyridinio)ethyl-thio~-6-
( 1- ( R ) -hydroxyethyl)-7-oxo-1-azabicyclo(3.2 0)-hept-2-ene-2-
carboxylate diphenylphosphate
~ ~2~3 (~ )2
To a solution of p-nitrobenzyl (5B, 6S)-6-(1-(B)-
hydroxyethyl)-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-carboxylate
(0.696 g, 2.0 mmol) in 10 mL of dry acetonitrile, at 0C under N2,
was added dropwise diisopropylethylamine (0.382 mL, 2.2 mmol)
followed by diphenyl chlorophosphate (0.457 m~, 2.2 mmol). After
stirring at ooc for 30 min. a solution of 3-amino-1-(2-
mercaptoethyl)pyridinium chloride (0.475 g, 2.5 mmol) in 1 ml of
dry DMF was added, followed by additional diisopropylethylamine
(0.435 mL, 2.5 mmol). The reaction mixture was kept at 0C for 1.5
h and was then concentrated in vacuo. The resulting gum was taken
up in acetonitrile-H2O (1:1) and applied to a C18 reverse-phase
column. Elution with H2O , followed by 20% acetonitrile-H2O and
subsequent lyophilization of the relevant fractions afforded the
product (0.730 g, 50%) as a beige-colored solid. ir(KBr) ~max:
3330 (br, OH), 3180 (br, NH2), 1770 (B-lactam CO), 1690 (-
CO2PNB)cm~l; lHnmr (d6-DMSO) ~: 8.29-7.63 (m, 8 aromatic), 7.2-
6.7 (m, 10H, diphenylphosphate), 5.47, 5.18 (AB q, J=14 H2, 2H,
benzylic), 4.73-4.45 (m, 3H) 4.2-3.8 (m, lH), 3.6-2.6 (m, 8H),
1.15 (d, J-6.2 Hz, 3H, CHMe).
--10~--
C. (5B. 6~)-3-(2-(3-Aminopyridinio)ethyl thio)-6~
hydroxyethyl)-7-oxo-l-azabicyclo(3.2.0)hept-2-ene-2-carboxylate
~ O
~ ~ 11 ~
C02 ( ~ 2PO
N
CO
To a mixture of p-nitrobenzyl (5R, 6S)-3(2-(3-
aminopyridinio)ethyl thio)-6-(1-(R)-hydroxyethyl)-7-oxo-1-
azabicyclo(3.2.0)hept-2-ene-2-carboxylate diphenylphosphate (0.730
g, l.0 mmol) and 10~ palladium-on-charcoal (0.7 g) in 25 mL of a
phosphate buffer (0.05 M, pH 7.4) was added 8 mL of THF and 20 mL
of ether. This mixture was then hydrogenated (Parr) at 40 psi for
l h. The resulting mixture was filtered through a pad of
"Celite" and the filter cake was washed with H2O and ether. The
aqueous phase was separated, washed with ether (2 x) and then
residual volatiles were removed ln vacuo. The aqueous solution
was immediately applied to a C18 reverse-phase column which was
eluted with H2O. Lyophilization of the relevant fractions
afforded 0.45 g of an off-white solid. This material was
repurified by reverse-phase hplc to give the desired product
(0.123 ~, 35%) as an ivory-colored solid.
ir(KBr)~max:3340 (br), 1750 (br, B-lactam CO), 1S80 (br, -CO20)
cm l; 1Hnmr (D2O~ ~-8.0?-7.59 (m, 4H, aromatic), 4.61 (t, J= 5~8
Hz, 2H, N-CH2), 4.14 (d of q, J=J'=6.3 Hz, lH, H 1'), 3.97 (d of
+, J-9.2 Hz, J'=2.6 Hz, lH, H-5), 3.38 (t, J=5.8 Hz, 2H, S-CH2),
3.24 (d of d, J=6.0 Hz, J'=2.6 Hz, lH, H-6), 3.17-2.57(m, 2H, H-
4) 1.21 (d, J=6.3 Hz, 3H, CHMe); UV(H2) ~max:299 ( 7949),
(8822) nm; ~1/2 (pH 7.4, 36.8C) 18.5 h.
. ~
- 107 -
Ex2m~le 13
PR~P~RATION O~
N
.
~5~,6S)3~ S)-methyl-2-(1-~y_idiniu~)e'hylthioj-6~ (R~-
hy~roxyethyl~-7-oxo-1-zzabicyclo(3.2.0?hept-2- ene- 2-c2rboxylate
a~
'"r;~
COO~ ' ' '
.. .
.
5~,6S)3~ tR)-methyl 2-(1-pyridinium)ethylthio]-6-[1~
hydroxyethyl~-7~ox=~ zablcyc~o~3.2.0~h-ot 2-ene-2-~- Osxyl~e
, ' .`' " ~ ` .
''
' ,
-108-
_ -t2-mercapto-2-methylethYl)pyridinium methanesulfonate d -1-
(2-mercapto-1-methylethyl)pyridinium methanesulfonate
~ + ~ sO~ ~
~ N ~ MsO
~S~r ~ ~sO
Methanesulfonic acid (1.95 mL, 0.030 mol) was added slowly to
cold pyridine (7.83, 0.097 mol) and the resulting mixturP was
stirred at 40C for 15 min, treated with d -propylenesulfide (2.59
mL, 0.033 mol) and stirred at 60~C under a nitrogen atmosphere for
90 h. Pyridine was removed under vacuum; the residue was mixed
with water and purified by chromatography (hplc, Prep. Bondapak C~
18). The appropriate fractions were combined and lyophilized
giving d -1-(2-mercapto-2-methylethyl)pyridinium methanesulfonate
1.14 g (15~) as a colorless syrup, ir (film) Vmax:2520 (SH)~ 1640
(pyridinium), 1180 (s, CH3SO3-), 1040 (CH3SO3~)cm~l, 1Hmr (DMSO
d6) ~:1.35 (d, J=6.8 Hz, 3H, CH3CHS), 2.30 (s, 3H, CH3SO3-), 2.90
(d, J=8.5 Hz, lH, SH) 3.2-3.7 (m, CHSH), 4.52 (dd, Jgem=12~9 Hz,
J=8.4 Hz, CHCH2N+), 4.87 (dd, Jgem=12~9 Hz, J=6.0 Hz, CHCH2N+),
8.0-8.4 (m, 2H, Hm of pyridinium), 8.5-8.8 (m, lH, Hp of
pyridinium), 9.04 (dd, J=1.4 ~z, J=6.7 Hz, 2H, Ho of pyridinium)
uv (H2O) ~max: 208 (~5267), 259 (3338), Anal. calcd. for
CgH15NO3S2.2H2O; C 37.88, H 6.71, N 4.91, S 22.47; found: C 37.49,
H 6.85, N 4.86, S 22.09 and d -1-(2-mercapto-1-
' . , , ~,
.: , . .
,
-109--
methylethyl)pyridinium methanesulfonate 0.82 y (11%) as a
colourless syrup; ir (film) ~max:2500 (S~), 1628 (pyridinium),
1180 (sulfonate), 1035 (sulfonate) cm~1, 1Hmr (DMSO d6) ~ :1.69
(d, J=6.8 Hz, 3H, CH3CHN+), 2.31 (s, 3H, CH3SO3 ), 3.0-3.3 (m, 2H,
CH2S), 4.2-5.2 (m, lH, CHN+), 8.0-8.4 (m, 2H, Hm of pyridinium),
8.5-8.8 (m, lH, Hp of pyridinium), 9.0-9.2 (m, 2H, Ho of
pyridinium)~ uv (H2O) ~max 209 ( 4987), 258 (E3838) . Anal.
calcd- for C9H15N3S2-1-5H2O: C 39.11, H 6.56, N 5.07; found C
39.13, H 5.92, N 5.20.
B. (5R!6S) paranitrobenzyl 3-[1-(R,S)methyl-2-(1-
pyridinium)ethylthio~6-[1-(R)-hydroxyethyl]-7-oxo-1-
azabicyclo(3.2.0)hept-2-ene-2-carboxylate diphenylphosphate
1) NE~ r)2
O _ 2 ~ _~
CCOPN~3 _ 3) RS~>
4) N2~(iPr)2
C)R
~S ~ ~
(?hO) 2gO
To a cold (0C) solution of (5R,6S)paranitrobenzyl 6-[1-(R)-
hydroxyethyl]-3,7~dioxo-1-azabicyclo(3.2.0)heptane-2-carboxylate
(0.523 g, 1.5 mmol) in acetonitrile (6 mL) kept under a nitrogen
atmosphere was added diisopropylethylamine (0.314 mL, 1.8 mmol)
followed by diphenyl chlorophosphate (0.373 mL, 1.8 mmol). The
reaction mixture was stirred for 30 min and treated with a
solution of d -1~(2-mercapto-2-methylethyl)pyridinium
methanesulfonate (0.539 g, 2.16 mmol) in acetonitrile (2 mL) and
diisopropylethylamine (0.314 mL, 1.8 mmol). The reaction mixture
was stirred at 0C for 1 h, diluted with cold (0C)
~r
'~ ~' " ' .
--110--
water (24 mL) and chromatographed over a "PrepBondapak" C-18 column
(2.5 x 8.5 cm) with 25-50~ acetonitrile in water as eluting
solvents to give 1.07 g (97~) of the title compound as a yellowish
powder after lyophilization; ir (KBr) v max 3700-3100 (OH), 1770
(C=O of ~-lactam), 1695 (C=O of PNB ester), 1630 (pyridinium), 1590
(phenyl), 1518 (NO2), 1348 (NO2), 885 (NO2) cm~1, lHmr (DMSO d6) ~
1.14 (d, J=6.1 Hz, 3H, CH3CHO), 1.33 (d, J=6.3 Hz, 3H, CH3CHS), 4.6-
5.0 (m, CH2N+), 5.14 ~d, J=5.2 Hz, lH, OH), 5.37 (center of ABq,
J=12.4 Hz, 2H, CH2 of PNB), 6.6-7.5 (m, 10H, phenyl of phosphate),
7.69 (d, J=8.7 Hz, 2H, Ho of PNB), 8.0-8.4 (m, 4H, Hm of PNB, Hm or
pyridinium), 8.4-8.8 (m, lH, ~p of pyridinium) 9.08 (d, J=5.6 Hz,
2H, Ho of pyridinium), uv (H2O) ~max 263 (~13325), 308 (E8915).
Anal calcd. for C36H36N3olosp-H2o C57-52~ H 5
4.27; found: C 57.76, H 4.96, N 5.36, S 4.35.
C. (5B, 6S)3-[1-(B and S)-methYl-2-(1-pyridiniumlethylthio)-6-[1-
(~)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene~2-
carboxylate
0~3
I ~5~ r,-~, et~ er
CCOp~3 ~hO)2~O
,N~
-
To a solution of (5B,6S)paranitrobenzyl 3-[1-(R,S)methyl-2-(1-
pyridinium)ethylthio]-6-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo-
(3.2.0)hept-2-ene-2-carboxylate diphenylphosphate (0.60 g, 0.82
mmol) in wet tetrahydrofuran (33 mL) was added ether (33 mL),
potassium phosphate mono basic-sodium hydroxide buffer (17 mL,
0.15N, pH 7.22) and 10% palladium on charcoal (0.60 g). The
resulting mixture was hydrogenated for 1 h under ~0 psi at 23C.
The two layers were separated and the organic layer was extracted
with water (3x7 mL).
`;~
': . '
. ' ' , ' ,
.
3~
The aqueous layers were combined, filtered through a "Celite" pad,
washed with ether (3 x 20 mL) and chromatographed on a
"PrepBondapak"* C-18 column (2.5 x 9.5 cm) with water as eluting
solvent to give 0.18 g (63%) of mixture of diastereoisomers. The
two diastereoisomers were separated by hplc ("PrepBondapak"* C-18)
with water as eluting solvent: isomer with lower retention time,
0.068 g (23%) compound "B",
ir (XBr) vmax: 1770 (C=O of ~-lactam), 1633 (pyridinium), 1593
(carboxylate)cm~l, lHmr(D2O) ~:1.20 (d, J=6.3 Hz, 3H, CH3C~0), 1.42
(d, J=6.9 Hz, 3H, CHCHS), 2.3-3.2 (m, 3H, H-4, H-6), 3.5-3.9 (m, lH,
SCH), 3.9-4.2 (m, 2H, H-5, CH3CH0), 4.3-5.1 (m, CH2N~), 7.8-8.2 (m,
2H, Hm of pyridinium), 8.4-8.7 (m, lH, Hp of pyridinium), 8.7-9.0
(m, 2H, Ho of pyridinium uv (H2O) ~max 260
( 6727), 300 ( 8245), [~]D23-39.3O (c, H2O), -~=12.6 h (measured
at a concentration of 10-4M in phosphate ~uffer pH 7.4 at 36.8C);
isomer with higher retention time, 0.081 g (28%), compound "A",
ir(KBr) max 1755 (C=O of ~ lactam), 1630 (pyridinium), 1590
~carboxylate) cm~1, 1Hmr (D20) :1.18 (d, J=6.3 Hz, 3H, C_3CH0),
1.40 (d, J=7.0 Hz, 3H, CH3CHS), 2.84 (d, J=9.3 Hz, 2H, H-4), 3.26
(dd, J=2.7 Hz, J=5.9 Hz, lH, H-6), 3.4-4.2 (m, 3H, SCH, CH3CHO, H-
5), 4.2-5.1 (m, CH2N+), 7.7-8.1 (m, 2H, Hm of pyridinium), 8.3-
8.65 tm, lH, Hp of pyridinium~, 8.65-8.9 (m, 2H, Ho of pyridinium),
uv (H20) maX:259 (~5694), 296 (~6936), [~]D23 ~96.9 (_ 0.56, HaO)~
~=15.6 h (measured at a concentration of 10-4 M in phosphate buffer
pH 7.4 at 36.8C).
*Trademark
~43~a~
-112-
Example 14
PREPARATION OF
OH
S~
COO~ [~
(5-~6s)3-r2-[ts)-(l-pyridinium)]-l-(s) cyclohexylthio]-6-rl-(R)-
hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate
and
o}~
5""'
~ ~ '
(5_,6S)3-[2-[(R)-(l-pyridinium)]-1-(R)-cyclohexylthio]-6-[1-(R)-
hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate.
:'
, .
,-
.: . : .
.
: , , ,
-113-
A. d -1-~2-mercapto-1-cyclohexyl)pyridinium methanesulfonate
C~ ~ N~> f ~5 03
SO-
Methanesulfonic acid (0.65 mL, 0.01 mol) was added dropwise to
pyridine (2.42 mL, 0.03 mol) with cooling. The mixture was stirred
under a nitrogen atmosphere for 10 min, treated with d -
cyclohexene-sulfide [1.377 g (85% pure), 0.0102 mol] and stirred at
72C for 25 h. The excess of pyridine was removed under vacuum and
the traces were codistilled with water. The residue was mixed with
water and chromatographed through a "PrepBondapakl'*C-18 column (5 x
13 cm) with 0~2% acetonitrile in water as~eluting solvent giving
after lyophilization a colourless syrup 1.57 g (53~), ir (film)
vmax: 2500 (SH), 1625 (pyridinium), 1190 (SO3 ), 1Hmr (DMSO d6)
~:1.2-2.5 (m, 8H, cyclohexyl H), 2.32 (s, 3H, CH3S03-), 2.82 (d,
J=9.8 Hz, SH), 3.0-3.5 (m, lH, CHSH), 4.2~4.9 (m, lH, CHN+), 8.0-8.3
(m, 2H, Hm of pyridinlum), 8.4-8.8 (m, lH,~Hp of pyridinium), 8.9-
9.3 (m, 2H, Ho of pyridinium), uv (H20) ~max 214 (E5365)~ 258 (E
3500). Anal. calcd. for C12HlgNO3S2OH2O: C 46.88, H 6.88, N 4.56;
found: C 46.61, H~6.46, N 4.65.
*Trademark
' ,
.
''
v~
-114-
B. (5B 6~)~aranitrobenzyl 3-[2-L~ or S)-(1-pyridinium)l_l_(R or
S)-cyclohexvlthiol-6-(1-(~)-hydroxyethyll-7-oxo-1-
azabicyclo(3.2.0)hept-2-ene-2-carboxylate diphenvlPhosphate
1) NEt~i~r)
.. ~ 2) Cl~(OPh)2
,~c o ~
O~ N 3) ~ M~O
COOPNB N
4) N~:t(iPr)2 ~H
J~C~
COO~ hO)2~O
To a cold (0C) solution of (5R,6S)paranitrobenzyl 6-[1-tR)-
hydroxyethyl]-3~7-dioxo-l azabicyclo- (3.2.0)heptane-2-carboxylate
(1.37 g, 3.93 mmol) in acetonitrile (15 mL~ kept under a nitrogen
atmosphere was added diisopropylethylamine (0.822 mL, 4.7 mmol) and
diphenyl chlorophosphate (0.979 mL, 4.7 mmol). The resulting
solution was stirred for 30 min and treated with a solution of d -1-
(2-mercapto l cyclohexyl)- pyridinium methanesulfonate (1.64 g, 5.66
mmol) in acetonitrile (4.7 mL) followed by diisopropylethylamine
(0.822 mL, 4.7 mmol). The reaction mixture was stirred at 0C for 1
h, diluted with cold (0C) water (75 mL) and chromatographed on
"PrepBondapak"*C-18 with 25-50% acetonitrile in water as eluting
solvent giving after lyophilization of the appropriate fractions 1.9
g (53%) of the title compound, ir (KBr) vmax: 3700-3000 (OH), 1770
(C=O of B-lactam), 1700 (C=O of PNB ester), 1628 (pyridinium), 1590
(phenyl), 1515 (NO2), 1345 (NO2) 880 (NO2) cm~1, 1Hmr (D2O) ~:1.13
(d, J=6.1 Hz, 3H, CH3CHO), 1.2-2.5 (m, 8H, cyclohexyl H), 2.7-3.5
(m, 4H, H-4, H-6, CHS), 3.5-4.4 (m, 2H, CH3CHO, H-5), 4.4-5.0 (m,
lH, CHN+), 5.30 (center of ABq, J=12.8 Hz, CH2 of PNB), 6.7-7.4 (m,
10H, phenyl), 7.65 (d, J=8.6 Hz, 2H,
*Trademark
. .
' , ' '.
.
-115-
Ho of PNB), 7.9-8.4 (m, 4H, Hm of PNB, Hm of pyridinium), 8.4-8.8
(m, lH, Hp of pyridinium), 9.0-9.4 (m, 2H, Ho of pyridinium), uv
(H20) ~max: 263 (~9038), 309 (~6394). Anal. calcd for
C39H40N3010SP.H2o: C 59.16, H 5.35, N 5.31; found: C 58.95, H 5.15,
N 5.57.
C; (5R. 6S) 3-~2-~(R or S)-(1-pyridinium)l-1-(R or S)-
cyclohexylthio~-6-[1-(~)-hYdroxyethyl~-7-oxo-1-
azabicyclo(3.2.0)hept-2-ene-2-carboxylate
0
J` r l lO~ 2d~C,
.~-, e ~e~, bUCr~
Oo~Ng ~ N~ O
~ (PhO)2~0
~ -s.~
To a solution of (5R, 6S) paranitrobenzyl 3-[2-[(B or S)-(1-
pyridinium)]-1-(R or S~-cyclohexylthio]-6-[1-(R)-hydroxyethyl]-7-
oxo-l-azabicyclo(3.2.0)hept-2-ene-2-carboxylate diphenylphosphate
(1.85 g, 2.34 mmol) in wet tetrahydrofuran (96 mL) was added ether
(96 mL), potasslum phosphate monobasic-sodium hydroxide buffer
(0.15M, pH 7.22, 50 mL) and 10% palladium on charcoal (1~9 g). The
resulting mixture was hydrogenated at 23 under 40 psi for 1.25 h.
The organic layer was separated and washed with water (3 x 20 mL).
The aqueous solutions were filtered through a "Celite" pad, washed
with ether (2 x 60 mL), pumped to remove the traces of organic
solvents and chromatographed on a "PrepBondapak" C-18 column ~4.5 x
9 cm) with 0-5% acetonitrile in water as eluting solvent giving
after lyophilization 0.705 g (76%) of a mixture of diastereoisomers.
The separation of the diastereoisomers was done by hplc
("PrepBondapak" C-18) with 4% acetonitrile in water as eluting
.;.
-116-
solvent; diastereoisomer with lower retention time, compound "A",
(0-29 g, 31~), ir (KBr) vmaX: 1750 (C=O of ~-lactam), 1620 (sh,
pyridinium), 1685 (carboxylate) cm~l, lHmr (D2O) ~:1.21 (d, J=6.3
Hz, 3H, CH3CHO), 1.4-2.5 (m, 8H, cyclohexyl H), 2.5-3.05 (m, 2H, H-
4), 3.05-3.25 (m, lH, H-6), 3.3-3.7 (m, lH, CHS), 3.9-4.3 (m, 2H, H-
5, CH3CHO), 4.3-4.8 (m, CHN+), 7.8-8.2 (m, 2H, Hm of pyridinium),
8.3-8.7 (m, lH, Hp of pyridinium), 8.8-9.1 (m, 2H, Ho of pyridinium)
uv (H2O) ~max 260 (~7123), 300 (~8685), [~]D23 ~6.2 (c 0.63, H2O),
~=16.6 h (measured at a concentration of 10-4M in phosphate buffer
pH 7.4 at 36.8C); Anal. calcd- for C20H24N2O4S 2H2o C 56-59
H 6.65, N 6.60, S 7.55; found: C 56.83, H 6.47, N 6.59,
S 7.43; diastereoisomer with higher retention time, compound "B",
(0.35 g, 38%) ir (KBr) vmax: 1750 (C-O of ~-lactam), 1622 (sh,
pyridinium), 1588 (carboxylate) cm~l, lHmr (D2O) ~:1.19 (d, J=6.4
Hz, 3H, CH3CHO), 1.3-2.5 (m, 8H, cyclohexyl H), 2.5-3.1 (m, 2H, H-
4), 3.1-3.3 (m, lH, H-6), 3.3-3.8 (m, 2H, H-5, CHS), 4.1 (center of
m, lH, CH3CHO), 4.25-4.7 (m, lH, CHN+), 7.8-8.1 (m, 2H, Hm of
pyridinium), 8.3-8.7 (m, 1~, Hp of pyridinium), 8.75-9.0 (m, 2H, Ho
of pyridinium), uv (H2O) ~max 259 (~5992), 296 (~7646), [~]D23
+65.3 (c 0.43, H2O) T~=20.2 h (measured at a concentration of 10 4
M in phosphate buffer pH 7.4 at 36.8C).
,
-117-
Exam~le 15
A. (5E~ Allyl 3-[(2-pyridinioethyl~thiol-(6~-r(1~)-
hydroxyethyl~-7-oxo-1-azabicyclo[3 2.0]hept-2-ene-2-carboxylate
diphen~lphosphate
O~H
O ~ 2) ~ ~ ~ Cl~
OH
o'l~S ~-
C2 ~'~G~ ~30~0(0~)2
To a solution of (sR) allyi, 3,7-dioxo-~6S)-[(1g)-hydroxyethyl]-1-
azabicyclo[3~2~o]heptane-(2~)-carboxylate (473 mg, 1.87 mmol) in
CH3CN (6 mL) was added at ca. -10C under a nitrogen atmosphere
diisopropylethylamine (0.42 mL, 2.4 mmol) followed by diphenyl
chlorophosphate (0.50 mL, 2.4 mmol). The mixture was stirred at 0C
for 30 min. and then cooled to -15C. To this was added an oily
suspension of N-(2-mercaptoethyl)-pyridinium chloride (527 mg, 3.00
mmol) in CH3CN (1 mL) containing 5 drops of DMF, followed by
diisopropylethylamine (0.42 mL, 2.4 mmol). The mixture was stirred
at -15 for 30 min and then diluted with H2O (20 mL). This mixture
was directly purified on a reverse phase silica gel (C18 PrePAK*,
12. g, Waters Associates~ eluting with H2O (200 mL), 10% CH3CN/H2O
(100 mL), 20~ CH3CN/H2O (100 mL), 30% CH3CN/H2O (100 mL) and then
40% CH3CN/H2O (100 mL). Appropriate fractions were collected, the
organic solvent removed by a vacuum pump and lyophilized to obtain
786 mg (1.26 mmol. y. 67.3%) of the title compound as brownish
powder: 1Hmr (DMSO-d6. CFT-20) ~ (3H, d J=6 Hz, l'-CH3), 2.6-
3.7 (m), 3.75-4.3 (2H, m, 5-H and l'-H), 4.65 (2H, m, -CO2CH2-),
4.87 (2H, t, J=6 Hz, -CH2-N+),
*Trademark
'
., , ~ .~ .
, ' , ~ ' '
'. ,'
- 118 -
5-6.2 (3H, m, olefinic protons), 6.6-7.4 (m, aromatic protons), 8.1;
(2H, "c`', J~7 ~z, aromatic protons meta tq the nitrogen), 8.63 (lr~, "t",
J~7 Hz, aromatic proton para to the nitrogen) and 9.07 ppm (2:H, ~d",
J~7 Hz, aromatic protons ortho to the nitrogen); ir (film) v: 3~00 ~Orl),
1770 (B-lactam), 1690 (ester), 1625 (pyridinio).
B. (5R) 3-[(~-~yridinioethyl)thio]-(65)-1(lR)-hydroxyethyl]-7-oxo-1-
azabicyclo~3.2.0~hept-2-ene-2-car~oxylate
0~ ' ' 0
o ~C3 S ~ Pd ( P~ 3 ) 4 _ ~ J~L~_ ~ ~
` C02 ~ (~)OPO(0~)2 ~ C02~C co G
To a solution of (5~) allyl 3-[(2-pyridinioethyl)thio~ - (6S) -
[(lR)-hyd~oxyethylj-7-oxo-1-a~abicyclo~3.2.0]hept-2-ene-2-carboxylate
diphenylphosphate (156 mg, 0.25 mmol) in CH3CN (2 ~L) W25 successively
~dde~ ~t ca. 22C a solution of potassiu~ 2-ethylhexanoate in EtOAc
(0.5 M, 0.6 m~; 0.3 mmol), triphenylphosphine (15 mg, 0 057 mmol)
and tetrakistriphenylphosphine palladiu~ (15 mg, 0.013 ~mol). The
mixture was stirred at-ca. 22C under a nitrogen atmosphere for 2 h.
A~ter addition of anhy2rous Et20 (7 mL), the precipitate was filtered,
washed with anhydrous ~t20 (7 mL) and dried in vac~o to yield 101 mg
of brownish solid.~ This was purified by reverse phase column c~roma-
i tography (cla PrepPAK, 12 g, ~iaters Assoc{ates) eluting ~ith H~O.
Appropriate fractions (fr. 7-12, each 20 mL) were collected and lyo-
philized to ~btain 53 mg (0.16 mmol, y. 6~) of the title compound as;
yellowish powder. This material was eontaminated with potassium diphe-
ny phosphaee and potassium 2-cthylhs~noate: l8mr (D20, CFT-20~ ~: 0.80
" "~' ' .
.
3~4L
- 119 -
(t, J-6.4 Hz, Me fro~ ethylhexanoate), 1.21 ~3H, d, J=6.3 Hz, l'-Me),
2-93 (211, dd, Jl 5-9 ~z, ~ =4 Hz, l-Hs), 3.28 ~lH, dd, ~6 1,=6-2 Hz,
J6 5=2.5 Hz, 6-H), 3.42 (2H, t, J=6 Hz, ~CH2S), 3.98 (lH, td, J5 1=
9 Hz, J5 6=2.5 Hz, S-H), 4.15 (lH, q, J=6.2 Hz, l'-H), 4.80 (2H, t,
J=6.0 Hz, -CH2N ), 7-7.5 (m, phenyl protons from diphenyl phosphate),;
8.03 (2H, m, Hm of pyridinium), 8.56 (lH, m, Hp of pyridinium) and
8.81 ppm (2H, "d", ~-6.5 Hz, ~o of pyridinium) .
` : :
-~
- ~ : . .
. . ...
-. . . . .
32a~
- 120 -
Example 16
Preparation or 3-[2-(N-MethYl-thiomorpholinium)ethylthio]-
6~-tl'-~R)-hydroxyethyl]-7-oxo-l--azabicyclo[3.2~o]-hept
2-ene-2-carboxylate
OH o e Me
A. N-methy~-N-(2-merca~toethyl)thiomorpholinium methanesulfonate
~ \ 1) MsOH HS Me
Me ~ 2)
~ To precooled (ice bath) N-methylthiomorpholine*
(5.00 g, 42.7 mmol) was added methanesulfonic acid (1.47 mL,
20.5 mmol) and ethylene sulfide ~1.30 mL, 21.4 mmol). The
mixture was heated at 65C for 24 h and diluted with water
(25 mL). The aqueous solution was washed with diethyl ether
(3 x 25 mL), pumped under~vacuum and ~oured over a silica gel
reverse phase column; the title compound being eluted with
water. The appropriate fractions were combined and eYaporated
to afford the thiol as an oil (4.~80 g, yield 86%); ir (film)
~max 2550 cm 1 (w, SH);~lHmr (DMSO d6)~: 3.25-2.95 (6~, m,
CH~N~), 3j32 (3H, s, CH3N~)~; 3.20-2.65 (7H, m, CH2S, SH) and
2.32 ppm (3H, s~, CH3SO3).
*J.M. Lehn and J. Wagner. Tetrahedron, 26, 4227 (1970)
:
.
,
- ' ' , ' , .
~. : .
- 121 -
B. para-~itrobenzyl 3-[2-(N-methyl-thiomorpholiniUm diphenyl
phosphate) ethylthio]-6~-[l'-tR)-hydroxyethyl]~7-o
azabicyclo[3.2.0]-hept- 2-ene-2-carbo~ylate
N ~) 2
J ~ 2 ) Cl~O ( ¢') 2 ~ 5 ~ \S
od~ N ~/ 3 ? ~5~ o N ~ ,~3 /
CO ~3 ~(e . C2P~ )2
2 4 ) E'~N (~ ) 2
A cold (ice bath) solution of para-nitrobenzyl
6~-[1'-(R)-hydroxyet~yi]-3,7-dioxo-1 azabicyclo[3.2.0]heptane-
2-car~oxylate (557 mg, 1.60 mmol) in CH3CN (8 mL) was treated
dropwise with diisopropylethyl amine (0.336 mL, 1.92 mmol) &nd
diphenylchlorophosphate (0.400 mL, 1.92 mmol) and stirred for
30 min. The reaction mixture was treated with N-methyl-N-t2-
mercaptoethyl)thiomorpholini~m methanesulfonate (~3 mg, 2,29mmol) in CH3CN (4 mL) and diisopropylethyl amine (0.336 mL,
1.92 mmoi) and stirred for 30 min. The solution W25 diluted
with water (20 mL) a~d poured over a silica gel reversed phase
column. The desired compound was eluted with a 50% acetonitrile-
water mixture. The appropriate fractions were combined, pumped
under ~acuum for 2 h. ~nd lyophilized to afford the title
compound (1.01 g, yield 85%): ir t"Nujol~)vmax: 1760 (s, ~-lactam
C=O) and 1510 cm 1 ~5, NO2); lHmr (DMSO-d6) ~: 8.25 (2H, d,
i J=8.8 Hz, H-aromatic), 7.70 (2R, d, J=8.8 Hz, H-aromatic ), 7.33-
6.84 (10 H, m, H-aromatic), 5.37 (2H, center of ABq, J=14.2 Hz,
I CH2), 5.14 (lH, d, J=4.5 Hz, OH1, 4.35-3.80 (2H, m, H-l' and
i H-5), 3.75-3.45 (6H, m, CH2N ), 3.31 (3H, s, CH3N ), 3.45-2.75
(9H, m, CH25, H-6 and H-4) and 1.15 ppo ~3H, d, J-6.2 Hz, CH3~.
`` ..~
~ :
.
.'
3~
- 122 -
C. 3-[2-(N-methyl-thiomor2holinium)ethylthio]-6~-~1'-(R)-
hydroxvethyl]-7-oxo-1-azabicyclo[3.2.0]-he~t-2-ene-2-
carboxylate
. . .
. ~ , . 0~'
U7 ~ Ue
~)2
, C)' .
A solution of ara-nitrobenzyl 3-[2-(N-methyl-
thiomorpholinium diphenylphosphate)ethylthio]-6~-[1'-(R)-
hydroxyethyl]-7-oxo-1-azabicyclo[3.2.03hept-2-ene-2-carboxylate
(1.31 g, 1.76 mmol) ln O.lM pH 7.4 phosphate buffer (48.8 mL),
tetrahydrofuran (20 mL) and diethyl ether ~20 ml) wzs hydrogenated
over 10~ pd/C (1.5 g) in a"P ~ " ~ r for 1 h a~ 40 psi. The
reaction mixture was diluted with diethyl ether ~40 mL) and the
phases were separated~ The organic phase was washed with water
~2 x 5 mL). The aqueous phases were combined, riltered ~hrough
a ~52 hardened filter paper, washed with diethyl ether (2 x 20 mL)
and pumped under vacuum. The a~ueous solution was poured on a
silica gel reverse phase column and the desired carbapenem was
eluted with 5% acetonitrile-water. The appropriate fractions
were combined, a~d lyophilized to gi~e title compound as an
amorphous solid ~205 mg, 31%); ir ("~ujol'l)~max: 1750 (s, B-lactam
C=O) and 1590 cm (s, C=O); lHmr (D2O) ô:4.25-3.95 (2H, m, H-l',
H-5), 3.70-3.40 (6H, m, CH2N ), 3.35 (lH, dd, J-6.1 Hz, J=2.6 Hz,
H-6)`, 3.08 (3H, s, CH3N ), 3.25-2.75 (8H, CH2S, H-4), znd 1.24 ppm
(3H, d, J=6.4 Hz, C~3); uv (H2O, c 0.062)~max: 299 (~10,962) Tl/2
17.7 h (O.lM pH 7 phosphate buffer, 37C).
*Trademark
,~,
- 123 -
Example 17
Preparation of (5R,6S)-3-[2-(1-me.hylmorpholino)ethylthio]-
6-[(R)-l-hydroxyethyl]-7-oxo-1-azabicyclo~3.2.0]hept-2-ene-
2-carboxylate
JY Me ~
N
C02~
A. l-Methyl 1-(2-mercaptoethyl)morpholinium tri'].uoromethane-
sulfonate
C~3S03H ~ ~ HSC~2C~2N ~ O
Me C~3S039
To N-methylmorpholine (3.29 mL, 0.030 mol) was added
dropwise trifluoromethanesulfonic acid (1.327 mL, 0.015 mol) at
10C, followed by ethylene sulfide (0.8g mL, 0.015 mol). The
resulting yellow-brown solution was heated (oil bath) at 50-60C
under N2 for 18 h. Volatile material was then removed in vacuo
and the~ residual oil was taXen up in 10 mL of H20. The aqueous
solution was washed with diethyl ether ~3 x 5 mL) and then
residual organic solvent was removed ln vacuo. The resulting
aqueous solution was applied to a C18 reverse-phase column which
was eluted with H2O, then 5~ acetonitrile-H2O and finally 10~
acetonitrile-H2O. Evaporation of the relevant fractions afforded
i a white solid which was dried ln vacuo (P2O5) to give the produc~
(1.92 g, 41~). ir (KBr)~max: 25~0 ~SH) cm l; lHnmr (d6-acet~ne)
~: 4.25-3.6 (m, 8H), 3.49 (s, 3H, N-Me), 3.35-2.7 (m, 5H).
- 124 -
B. p-Nitrobenzyl (5R,6S)-3-[2-(1-methylmorpholino~ethylthio]-
6-[(R)-l-hydroxyethyl]-7~oxo-1-azablcycloL3.2.0]hept-2-ene-
2-carboxvlate diphenvl~hosphate
OH OH M~
" ~ ~ " ~ N
L ~co ,> I />~ ~~ /
~ CO2PNB CO2PNB1l 3
(00) 2PO
To a solution of p-nitrobenzyl (5R,6S)-6-[(R)-l-
hydroxyethyl]-3,7-dioxo-1-azabicyclo[3.2.0]heptane-2-carboxylate
(0.348 g, 1.0 mmol) in 25 mL of dry acetonitrile was added drop-
wise diisopropylethylamine t0.191 mL, l.l.mmol) and then diphenyl
chlorophosphate (0.228 mL, 1.1 mmol) at 0C under ~2. A'ter
stirring at 0C for 1 h diisopropylethylamine (0.226 mL, 1.3 mmol)
was added to the resulting enol phosphate, followed by 1-me',hyl-
1-(2-mercaptoethyl)morpholinium trifluoromethanesulfonate (O.373 g,
1.2 mmol). The reaction mixtuxe was stirred at room temperature
for 1.5 h and then concentrated ln vacuo. The residual material
was taken up in H2O and applied to a C18 reverse-phase column.
Elution with H2O, then 20~ acetonitrile-H2O and ~inally 30%
acetonitrile-H2O followed by lyophilization of the relevant
fractions gave the product (0.360 g, 40%) as an amorphous solid.
ir (Lilm) 3300 (-OH), 1770 (~-lactam CO), 1700 (-CO2PNB) cm l;
Hnmr (d6-acetone) ~: 8.25, 7.80 (ABq, J=8.6 Hz, 4H, aromatic),
7.4-6.8 (m, 10H, diphenylphosphate), 5.56, 5.27 ~ABq, J=14.2 Hz,
2H, benzylic), 4.42 (d of t, J=9.2 Hz, J'=2.7 Hz, lH, H-5), 4.1-
2.7 ~m, 17H), 3.4G (s, 3H, N-Me), 1.22 (d, J=6.2 Hz, 3H, -CHMe~.
''
.
- 125 -
C. (5R,6S)-3-~2-(1-methylmorpholino)ethylthio]-6-[(R)-1-
hydroxvethyl~-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-
carboxylate
~3
2 ~) 2$~ C~2 -
To a solution of p-nitrobenzyl (5R,6S)-3-[2~
methylmorpholino)ethylthio]-6-[(R)-l-hydroxye'~hyl]-7-oxo-1-
azabicyclo[3.2.0]hept-2-ene-2-carboxylate diphenylphosphate
~0.360 g, 0.49 mmol) in 13 mL of phosphate buîfer (0.05 M pH
7.4) was added 0.36 g of 10% palladium-on-charcoal, 20 mL of
tetrahydrofuran and 20 mL of diethyl ether. This mixture was
hydrogenated (Parr) at 32 psi for 1 h. The mixture was
filtered through Celite and the filter pad was washed with H20
and diethyl ether. The aqueous phase W2S separated and the pH
was adjusted to 7.0 with additional pH 7.4 phosphate buf~er.
A~ter removing residual organic solvents in vacuo the aqueous
solution was applied to a C18 reverse-phase column. Elution
with H2O and lyophilization of the relevant f-actions afforded
0.130 g of an amorphous solid. This ma~erial was repurified
by reverse-phase hplc to give the pure product (0.058 g, 34%)
as an amorphous solid. ir (KBr)vm~x: 3420 (br, OH), 1750
(~-lactam CO), 1590 (-CO2 ) cm 1; Hnmr (D2O) ~: 4.35-2.77
(m, 17~, 3.18 (s, 3H, N-Me), 1.23 (d, J=6.3 Hz, 3H, CHMe);
uv (H~O)~maX: 300 (~6344) nm; tl/2 (pH 7.4, 36.8C) 18.5 h.
.
- ~26~
ExamDle 18
Preparation of (5R,6S)3-[2-(1,4-dimethyl-1-piperazinium)-
ethylthio]-6-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo-
[3.2.0]hept-2-ene-2-carboxylate
OH
~ C~ ~
A. 1-(2-acetylthioethyl)-1,4-dimethylDi~erazinium bromide
C~3CS r + _ ~ ~ _ Acetone C~ CS ~ N N-,8r
A solution of 2-bromoethyl thiolacetate *(2.20 g,
0.012 mol) and 1,4-dimethylpiperazine (1.95 mL, 0.014 mol) in
acetone (4 mL) was stirred at 50C for 65 h. After cooling to
25C, the liquid phase was decanted from the gum which was
triturated twice in diethyl ether; a hygroscopic yellowish
powder, 3.2 g (90%) was obtained; ir (Nujol) vmax: 1685 (C=O of
thioester) cm l; lHmr (D2O) ~:
2.37, 2.39 (2s, 6H, CH3CO, ~ N-CH3), 3.18 (s, 3H, N ~ 3 ).
*B. Hansen, Acta Chem. Sca~d. 11, 537-40 (1957)
3~:~
- 127 -
B. 1 4-dimethyl-l-t2-mercaptoethyl)piperazinium bromide hydro-
chloride
CH3CS~'~_~'N ~ N - , Br ~ ~S ~ ~ - , Br , HCl
A solution of 1-(2-acetylthioethyl)-1,4-dimethyl-
piperazinium bromide (1.1 g, 3.7 mmol) in 6N hydrochloric acid
(4 mL) was heated at 80C under a nitrogen atmosphere for 1 h.
The solution was concentrated under reduced pressure to give
a white powder, 0.41 g (38~), lHmr (DMSO, d6) ~: 2.90 (s,
~H '~ CH3
~ N~CH )~ ~ ~
Anal. calcd. for C8H20N2SBrCl~H20: C 31.03, H 7.16, N 9.05,
S 10.35; found: C 31.62, H 7.46, N 9.19, S 10.19.
''
` ~
' ~. ' ' '
-128-
C. (5~,6O Paranitrobenzyl 3- r 2-(1 4-dimethvl-1-pi~erazinium)-
ethylthio~-6-~l-(~)-hydrox~ethyl~-7-oxo-l-azabicyclo r 3.2.01-hept-
2-ene-2-carboxylate di~henYlphosPhate
O~ 1) NEt(iP~) 2
2) C~tOPh)2
' COOPNB 3) ~S~~--, ~r , ~Cl
4) NEtli~r) 2
hO) 2
CQOPN23
To a cold (0~C) solution of ~5R, 6S) paranitrobenzyl 6-~1-(R)-
hydroxyethyl]-3,7-dioxo-l-azabicyclo[3.2.0]hePtane-2- (B) -
carboxylate (0.465 g, 1.33 mmol) in acetonitrile (2 mL) kept under
a nitrogen atmosphere was added diisopropylethylamine (0.278 mL,
1.59 mmol) and diphenyl chlorophosphate (0.33 mL, 1.59 mmol). The
reaction mixture was stirred for 30 min and treated with a
suspension of 1,4-dimethyl-1-(2-mercaptoethyl)-piperazinium bromide
hydrochloride (0.40 g, 1.37 mmol) in acetonitrile (3 mL)-water (1
mL) mixture and diisopropylethylamine (0~278 mL, 1.59 mmol). After
stirring for 18 h at 5C, cold water (15 mL) was added to the
mixture. The resulting solution was chromatographed over a
"PrepPak"-500/C-18 (Waters Associates) column (2.5 x 7.5 cm) with
25-35% acetonitrile in water as eluting solvents to give a yellowish
powder 0.50 g (50%) after lyophilization; ir (KBr) ~max 1765 (C=O
of B-lactam), 1690 (C=O of PNB ester), 1585 (phenyl), 1512 (NO2),
875 (NO2)cm~l, lHmr (DMSO, d6) ~:1.16, 1.18 ~2d~, J=6.1 Hz, 3H,
C_3CHOH), 2.44 (s, ~ N~-CH3), 3.14 (s, ~ ~ 3 ), 5.31 (d, J=6 Hz,
OH), 5.39 (center of ABq, J=13 Hz, CH2 of PNB), 6.6-7.4 (m, 10H,
phenyl of phosphate), 7.71 (d, J=8.8 Hz, 2H, Ho of PNB), 8.26 (d,
J=8.8 Hz, Hm of PNB).
~ 2~2~
- 129 -
D. ~5R,6S)3-[2-(1,4-dimethyl-1-piperazinium) ethylthio] -6-
[l-(R)-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0~hept-2-
ene-2-carboxylate
~\N5~ PhO~ 2~ THI~ De~
COO
To a solution of (SR,6S)paranitrobenzyl 3-[2-~1,4-
dimethyl-l-piperazinium)ethyl'hio]-6-[1-(R)-hydroxyethyl]-7-
oxo-l-azabicyclo[3.2.0]hept-2-ene-2-carboxylate diphenylphosphate
~0.47 g, 0.623 mmol) in wet tetrahydrofuran (25 mL) was added
diethyl ether (25 mL), potassium phosphate monobasic-sodi~m
hydroxide buffer (13 mL, pH 7.22~ and 10% palladium on chzrcoal
(0.47 g). The resulting mixture W2S hydrogenated at 23C under
40 psi for 1 h. The two layers were separated and the orsanic
layer was extracted with water (2 x 7 mL). The a~ueous layers
were combined, filtered through a Celite pad, washed with diethYl-
ether (2 x 15 mL) and chromatographed on~PrepPak-500/C18 ~Waters
Associates) column (2.5 x 9~5~cm~ ~lt fwater as eluting solvent
to give, 0.097 g (43~) after ~y~ LLz~tiDn; ir (~Br) vmax:
3000-3700 (OH), 1750 (C-O of B-lactam), 1585 (carboxylate)cm 1,
Hmr (D2O) ~: 1.24 (d, J=6.4 ~z, 3H, CH3CHOH), 2.33 (s, 3H,
~J N--CH3), 3.15 (s, ~ ~ ), 4.0-4.5 (m, H-5, CH3CHOH), uv
(H2O) ~max 296 (~9476), ~a]D 61.1 (C 0.26, H2O), tl/2=12.4 h
(measured at a concentration of 10 4 M in phosphate buffer pH 7.4
at 36.8C).
3%~
- 130 -
Example 19
Preparation o~ (SR,6S)-3-[2-(~-methyl-thiomorpholinium-
oxide)ethylthio]-6-[1-(~)-hydroxyethyl]-7-oxo-l-azabicy
clo(3.2.0)-hept-2-ene-2-carboxylate
OH
~ ~ " /N ~ ~ -O
OH OH
\ mC~BA ~~\ (~
$S co
~ .
To~a cold (-10~C) solution of (SR,6S)-3-
[2-(N-methyl-thiomorpholinium)ethylthio]-6-[1-(R)-hydroxyethyl]-
7-oxo-1-azabicyclo(3.2.0)-hept-2-ene~2-carboxylate (608 mg,
1.65 mmol) in a 1:1 mixture of acetonitrile-water (9 mL) w2s
added m-chloroper~enzoic acïd (334.8 mg, 1.65 mmol) in small
portion over a l hour period. The mixture was then diluted ~~
.
,
~21~
- 131 -
with water (15 mL) and washed with diethyl ether (3 x 15 mL).
The aqueous phase W2S pumped under vacuum and passed through
a reversed phase silica gel column (H2O) ~o give a solid which
consisted of a mixture of compcunds. This mixture was separated
by reversed phase HPLC and afforded fraction A 52.4 mg (yield 12%)
and fraction B 23.6 mg (yield 6%) as diastereomers of the title
com~ound; Fraction A: ir (nujol) vmax: 1750 (s, ~-lact~m C=O)
and 1580 cm (s, C=O); Hmr (D2O) ~: 4.26-2.91 (20 H, m, H-4,
H-5~ ~-6~ H-l', CH2S, CH2S-O, CH3-N and CH2N ) and 1.24 ppm
(3H, d, J=6.4 Hz, CH3); uv (~2~ c 0-06) ~max 302 (~10425)i
T 1/2`12 h ~0.065 M, pH 7.4 phosphate bufCer~ 37C). Fractio~ B:
ir (nujol) vmax: 1750 (s, 3-lactam C=O) and 1585 cm 1 (5, C=O);
Hmr (D2O) ~: 3.36-2.90 (17 H, m, H-4, H-5, H-6, H-l',.CH2S,
CH2S-O, CH2N ), 3.25 (3H, s, CH3N ) and 1.24 ppm (3H., d, J=6.4 Hz,
CH3); uv (H2O, c 0 05) ~max 2-99 (~6517); T 1/2:10.75 h tO.06; M,
pH 7.4 buffer solution, 37C).
- 132 -
Exam~le 20
-
Preparation of (5R,6S)-3-[2-(1,4,4-Trimethyl-1-piperazinium)-
ethylthio~-6-[lR-hydroxyethyl]-7-oxo-l-azaDicyclo(3.2.o)hept
2-ene-2-carboxvlate chloride
U~ \ ~ , C 1
o coo3
A~ 1-(2-acetylthioe hyl)-1,4,4-trimethYl~i~erazlniu~ bromideiodide
2'.eI ~ ~ ~ + N ', B~, I
/~ N + ~, Br ~ t CH3 S
CH3~S ~ \J ,~--OH, 55-60C
A suspension of 1-(2-acetylthioethyl)-~,4-dimethyl-
piperazinium bromide (1.48 g, 5.0 mmol) in isopropyl alcohol (10 mL)
was treated with methyliodide (0.373 mL, 6.0 mmol) and heated at
5S-60C for 30 h. The solvents were evaporated under reduced pressure,
the residue was triturated in hexane and the solid was filtered,
1.85 g. The solid was dissolved in hot water (8 mL~ and the solution
was dilu~ed with acetone until turbidity (70-80 mL). Two successive
crystallizations gave 1.5 g, mp 220-5C dec., 68~ of the title
com~ound; ir (XBr)~max: 1692 cm (C=O); Hmr (D2O) ~: 2.40 (s, 3H,
CH3CO0), 3.37 (s, N-CH3), 3.39 (s, N-CH3), 3.99 (s); uv (H2O)~maX
226 (~13144). Anal. calcd for CllH24N20SBrI: C 30.08, H 5.51,
N 6.38; round: C 30.48, H 5.53, N 6.86.
,
2~
- 133 -
B. l-t2-merca~toethyl)-1,4,4-trimethvlDiperaziniu~ b~schloride
\ ~ / .Hcl 6N Per~utit ~ ~ /+
C~3CS ~ /N +, gr , I 5-1 Cl ~.S N~ , 2C:L
A mixture of 1-(2-acethylthioe~hyl)-1,4,4-trimethyl-
piperazinium bromideiodide ~1.84~g, 4.19 mmol) and 6N hydrochloric
acid ~15 mL) was heated at 57C under a nitrogen a~osphere for
2.5 h. The solution was concentrated under reduced pressure to
dryness. The solid was"s~spended in water (10 mL) and the well-
stirred suspension was treated with ~Permutit~ S-l Cl u~til a
solution was obtained. The solution was pour.ed on a column (1.2 x
60 cm) ~ "Penmutit" S-l Cl . The column was eluted with water
(1.5 mL/min). The appropriate fractions were combined and lyo?hilized
to give a white powder, 0.93 g, mp l90-lglC, 85%; lr(''Nujol'')vmax:
2460 (SH); lHmr (D2C) ~: 3.4 (s, N-CH3), 3.45 (s, N-CH3), 4.07 (s).
Anal. calcd for CgH22N~SC12 0~75 H20: C 39.34, H 8.62, N 10.20,
S 11.67; found: C 39.48, 3 8.39, N 10.55, S 11.15.
* trade mark.
. .
_ .
'
~9 :
.
':
.
, ~ .
-134-
C. (5~ 6~) paranitrobenzyl 3-[2-(1 4,4-trimethyl-1-
piperazinium)-ethylthiol-6-(1~-hydroxyethyl1-7-oxo-1-
azabicyclo(3.2.0)hept-2-ene-2-carboxylate bischloride
0~ Et(i?-)2
~, ~ 2) Cl~(O?h)2 ~er~utit S-l Cl
I ~--
o6 ~ / , 3) N~ r?2~
.00?1~3 4~ XS~;~ N ~, 2Cl
0
~" ~ ~ N~
0 N ~ 2Ci
To a cold (5~C) solution of (5R,6S) paranitrobenzyl 6-[lR-
hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2R-carboxylate
(0.94 g, 2.7 mmol) in acetonitrile (3mL) kept under a nitrogen
atmosphere was added diisopropylethylamine (0.557 mL, 3.2 mmol) and
diphenyl chlorophosphate (0.653 mL, 3.2 mmol). The reaction mixture
was stirred at 5C for 30 min and treated with
diisopropylethylamine (0.599 mL, 3.44 mmol) and an aqueous solution
(4 mL) of 1-(2-mercaptoethyl)-1,4,4 trimethylpiperazinium
bischloride (0.90 g, 3.44 mmol). After 1.25 h, diisopropylethylamine
(0.1 mL, 0.57 mmol) was added and the stirring was continued for 2h.
A part of the acetonitrile was eliminated under reduced pressure
and the resulting red mixture was chromatographed on a
"PrepPak"*500/C-18 (Water Associates) column with 25-75%
acetonitrile in water as eluting solvent to give a yellowish powder
(1.4 g) after lyophilization. The powder was solubilized in water
and the solution was passed on a column (1.2 x 58 cm) of "Permutit"*
S-1 Cl- using water as an eluting solvent. Lyophilization of the
appropriate fractions gave 1.17 g of powder that was repurified on a
column of "PrepPak"*-500/C-18. Lyophilization of the appropriate
fractions gave a yellowish powder, 0.80 g (53%); ir (KBr) vmax: 3400
~8~
-134a-
(br, OH), 1770 (C=O of the ~-lactam), 1690 (C=O of PNB ester), 1605
(aromatic), 1515 (NO2, 1345 (NO2) cm~1; lHmr (D2O) ~:1.26 (d, J=6.3
Hz, 3H, CH3CHOH), 3.39 (s, NCH3), 4.00 (s), 5.37 (br, s, CH2 of
PNB), 7.60 (d, J=8.6 Hz, 2H, Ho of PNB), 8.20 (d, J=8.7 Hz, 2H, Hm
of PNB); uv (H2O) ~max 276 (~12094), 306 (E10752). Anal. calcd.
for C25H36N4O6SCl2-3H2O: C 46.51, H 6.56, N 8.68, S 4.97, Cl 10.98;
found: C 46.31, H 6.18, N 8.57, S 5.36, Cl 11.37.
*Trademark
. . . . . .
3~
- 135 -
D. (SR,6S)-3-[2-(1,4,4-trime~lyl-1-piperazinium)ethylthio~-6-
~lR-hydroxyethyl]-7-oxo-l-az2bicyclo(3.2.0)hept-2-ene-2-
carboxvlate chloride
, 2Cl ~ ---+ ~ Cl
CCOP~3 . COO
. ~
A mixture of (5~,6S) paranitrobenzyl 3-[2-(1,4,9-
trimethyl-l-piperazini~)ethyl~hlo]-6-[lR-hydroxyethyl]-7-oxo-1-
azabicyclo(3.2.0)hept-2-ene-2-carboxylate bischloride (0.40 g,
0.68 mmol), phosphate bu fer (30 mL, 0.0;M, pH 7.0), tetrahy~rofu~an
(10 mL), ether (30 mL) and 10% palladium on charcoal (0.40 g) W25
hydrogenated at 23C under 35 psi for 1 h. The two phases were
separated. The organic phase was extracted with wa.er (10 ~).
Th~ aqueous phases were filtered on a~Celite~ pad, washed wi~ e.he-
(10 mL), concentrated under ~acuum to 10 mL and chromatogra?hed on a
"PrePak" -SOO/C18 column (2.2 x 11 cm) with water 25 eluting solvent
to give 70 mg (25~) after lyophilization n; ir (KBr)v~a~: 3400 (br, O~),
17;5 (C=O of the B-lactam), 1585 (carboxylate) cm 1; Hmr (D2O) ô:
1.24 ~3H, d, J=6.3 H2, CH3CHOH), 3.36 (s, NCH3), 3.98 ~s); uv (H20)
~max 296 (~7987); [~]D 35 9 (c, 0.30, H2O), Tl/2= 9.8 h (measured
at a concentration of 10 M in phosphate buffer pH 7.4 at 36.8C).
~2~ ~ rk
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