U.S. patent application number 13/558989 was filed with the patent office on 2012-11-15 for monocarbams.
This patent application is currently assigned to Pfizer Inc.. Invention is credited to Steven Joseph Brickner, Mark Edward Flanagan, Manjinder Singh Lall.
Application Number | 20120289455 13/558989 |
Document ID | / |
Family ID | 41727494 |
Filed Date | 2012-11-15 |
United States Patent
Application |
20120289455 |
Kind Code |
A1 |
Brickner; Steven Joseph ; et
al. |
November 15, 2012 |
Monocarbams
Abstract
The invention relates to compounds of formula (I): ##STR00001##
wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, and R.sup.6 as
defined herein. The invention also relates to pharmaceutical
compositions and methods of treating bacterial infections using
compounds of formula (I).
Inventors: |
Brickner; Steven Joseph;
(Ledyard, CT) ; Flanagan; Mark Edward; (Gales
Ferry, CT) ; Lall; Manjinder Singh; (East Lyme,
CT) |
Assignee: |
Pfizer Inc.
|
Family ID: |
41727494 |
Appl. No.: |
13/558989 |
Filed: |
July 26, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12641343 |
Dec 18, 2009 |
8252782 |
|
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13558989 |
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61139159 |
Dec 19, 2008 |
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Current U.S.
Class: |
514/2.4 ;
514/21.91; 540/363 |
Current CPC
Class: |
C07D 417/14 20130101;
A61P 31/04 20180101 |
Class at
Publication: |
514/2.4 ;
540/363; 514/21.91 |
International
Class: |
A61K 38/05 20060101
A61K038/05; A61P 31/04 20060101 A61P031/04; C07K 5/078 20060101
C07K005/078 |
Claims
1. -20. (canceled)
21. A compound of formula (I), ##STR00085## or pharmaceutically
acceptable salt thereof, wherein R.sup.1 is (C.sub.1-C.sub.6)alkyl
substituted with an NR.sup.7R.sup.8; R.sup.2 is hydrogen; R.sup.3
is hydrogen; R.sup.4 is methyl; R.sup.5 is methyl; R.sup.6 is
--C(.dbd.O)OH; R.sup.7 and R.sup.8 are independently
(C.sub.1-C.sub.6)alkyl; and X is C(H).
22. The A compound of formula (I), ##STR00086## or pharmaceutically
acceptable salt thereof, wherein R.sup.1 is (C.sub.1-C.sub.6)alkyl
substituted with --C(.dbd.O)NH.sub.2; R.sup.2 is hydrogen; R.sup.3
is hydrogen; R.sup.4 is methyl; R.sup.5 is methyl; R.sup.6 is
--C(.dbd.O)OH; and X is C(H).
23.-29. (canceled)
30. A compound of formula (IB): ##STR00087## wherein R.sup.1 is
selected from the group consisting of ##STR00088## and
pharmaceutically acceptable salts thereof.
31. A compound of formula (IB): ##STR00089## wherein R.sup.1 is
selected from the group consisting of ##STR00090## and
pharmaceutically acceptable salts thereof.
32. A compound of formula (IB): ##STR00091## wherein R.sup.1 is
selected from the group consisting of ##STR00092## and
pharmaceutically acceptable salts thereof.
33. The compound of claim 29 A compound of formula (IB):
##STR00093## wherein R.sup.1 is selected from the group consisting
of ##STR00094## and pharmaceutically acceptable salts thereof.
34. A pharmaceutically acceptable salt of any one of claims 30 to
33, wherein said salt is a potassium or sodium salt.
35. The pharmaceutically acceptable salt of any one of claims 30 to
33 wherein said salt is a bis-potassium or bis-sodium salt.
36. A pharmaceutical composition comprising the compound of any one
of claims 30 to 33, or pharmaceutically acceptable salt thereof,
and a pharmaceutically acceptable carrier.
37. -46. (canceled)
47. A composition comprising a compound of formula (I),
##STR00095## or pharmaceutically acceptable salt thereof; wherein
R.sup.1 is (C.sub.1-C.sub.6)alkyl substituted with 1 to 3
substituents selected from the group consisting of halo, hydroxy,
(C.sub.1-C.sub.6)alkoxy, --NR.sup.7R.sup.8,
--C(.dbd.O)NR.sup.7R.sup.8, and a 3 to 7 membered heterocycle,
wherein R.sup.7 and R.sup.8 are independently hydrogen or
(C.sub.1-C.sub.6)alkyl, wherein said heterocycle contains 1 to 3
heteroatoms independently selected from O, N, or S; R.sup.2 is
hydrogen or methyl; R.sup.3 is hydrogen or methyl; R.sup.4 is
hydrogen, deuterium, or methyl optionally substituted withl to 3
substituents independently selected from F or Cl; R.sup.5 is
hydrogen, deuterium or methyl optionally substituted with 1 to 3
substituents independently selected from F or Cl; R.sup.6 is H or
--C(.dbd.O)OH; and X is C(H), C(F), C(Cl), or N, and an additional
antibacterial agent selected from the group consisting of
beta-lactams, aminoglycosides, polymyxins, penicillins, and
lincosamides.
48. The composition of claim 47, or pharmaceutically acceptable
salt thereof, wherein said additional antibacterial agent is a
beta-lactam selected from the group consisting of cephalosporins,
carbapenems, and beta-lactamase inhibitors or
beta-lactam/beta-lactamase inhibitor combinations.
49. A composition comprising a compound of formula (I) ##STR00096##
or pharmaceutically acceptable salt thereof; wherein R.sup.1 is
(C.sub.1-C.sub.6)alkyl substituted with 1 to 3 substituents
selected from the group consisting of halo, hydroxy,
(C.sub.1-C.sub.6)alkoxy, --NR.sup.7R.sup.8,
--C(.dbd.O)NR.sup.7R.sup.8, and a 3 to 7 membered heterocycle,
wherein R.sup.7 and R.sup.8 are independently hydrogen or
(C.sub.1-C.sub.6)alkyl, wherein said heterocycle contains 1 to 3
heteroatoms independently selected from 0, N, or S; R.sup.2 is
hydrogen or methyl; R.sup.3 is hydrogen or methyl; R.sup.4 is
hydrogen, deuterium, or methyl optionally substituted withl to 3
substituents independently selected from F or Cl; R.sup.5 is
hydrogen, deuterium or methyl optionally substituted with 1 to 3
substituents independently selected from F or Cl; R.sup.6 is H or
--C(.dbd.O)OH; and X is C(H), C(F), C(Cl), or N and an additional
antibacterial agent selected from the group consisting of
clindamycin, metronidazole, imipenem, meropenem, doripenem,
ertapenem, cefotetan, cefepime, and cefpirome, or a third
generation cephalosporin.
50. The composition of claim 49 wherein the additional
antibacterial agent is cefepime.
51. The composition of claim 49 wherein the additional
antibacterial agent is meropenem.
52. The composition of claim 47 wherein said compound is
##STR00097## or a pharmaceutically acceptable salt thereof.
53. The composition of claim 47 wherein said compound is
##STR00098## or pharmaceutically acceptable salt thereof.
54. The composition of claim 47 wherein said compound is
##STR00099## or pharmaceutically acceptable salt thereof.
55. The composition of claim 47 wherein said compound is
##STR00100## or pharmaceutically acceptable salt thereof.
56. The composition of claim 47 wherein said compound is
##STR00101## or pharmaceutically acceptable salt thereof.
57. The composition of claims 52 to 56 wherein said additional
anti-bacterial agent is cefepime.
58. The composition of claims 52 to 56 wherein said additional
anti-bacterial agent is meropenem.
59.-60. (canceled)
Description
FIELD OF THE INVENTION
[0001] The present invention relates to Monocarbam compounds and
their use as antibacterial agents in animals, including humans. The
invention also relates to methods of preparing compounds,
intermediates useful in preparing compounds, and pharmaceutical
compositions containing compounds. The present invention further
includes methods of treating disease, e.g., bacterial infections by
administering compounds or compositions to subjects in need of such
treatment.
BACKGROUND OF THE INVENTION
[0002] Monocarbams are a class of synthetic monocyclic beta-lactam
antibacterial agents which have as their salient feature, a
substituted sulfonylaminocarbonyl activating group at the N-1
position. The early studies in this area were conducted by workers
at the Squibb Institute for Medical Research, Cimarusti, C. M.
& R. B. Sykes: Monocyclic .beta.-lactam antibiotics. Med. Res.
Rev. 4: 17-20, 1984. Monocarbams have also been previously
discussed in EP 0281289, published Sep. 7, 1988. These and all
documents cited herein are fully incorporated in their entirety by
reference herein.
[0003] Although not limiting to the present invention, it is
believed that monocarbams of the present invention exploit the iron
uptake mechanism in bacteria through the use of
siderphore-monobactam and siderphore-monocarbam conjugates.
Barbachyn, M. R., Tuominen, T. C.: Synthesis And Structure-Activity
Relationships of Monocarbams Leading to U-78608. Journal of
Antibiotics Vol. XLIII No. 9: 1199-1203, 1990. Thus, at least in
general terms, the activity and mechanism of action of monocarbams
are generally known, although the present invention is not bound or
limited by any theory.
[0004] There is a continuing need for new antibiotics, such as
monocarbams, in response to the increasing emergence of resistant
organisms and to improve safety, among other reasons.
SUMMARY OF THE INVENTION
[0005] The present invention relates to certain compounds of
formula (I), their preparation and useful intermediates,
pharmaceutical compositions thereof, and methods of treating and
preventing bacterial infections therewith. In many embodiments, the
compounds are active and effective against organisms that are
resistant to other antibiotics.
[0006] In particular, the present invention relates to a compound
of formula (I):
##STR00002##
or pharmaceutically acceptable salt thereof; wherein
[0007] R.sup.1 is (C.sub.1-C.sub.6)alkyl substituted with 1 to 3
substituents selected from the group consisting of halo, hydroxy,
(C.sub.1-C.sub.6)alkoxy, --NR.sup.7R.sup.8,
--C(.dbd.O)NR.sup.7R.sup.8, and a 3 to 7 membered heterocycle,
wherein R.sup.7 and R.sup.8 are independently hydrogen or
(C.sub.1-C.sub.6)alkyl, wherein said heterocycle contains 1 to 3
heteroatoms independently selected from O, N, or S;
[0008] R.sup.2 is hydrogen or methyl;
[0009] R.sup.3 is hydrogen or methyl;
[0010] R.sup.4 is hydrogen, deuterium, or methyl optionally
substituted with 1 to 3 substituents independently selected from F
or Cl;
[0011] R.sup.5 is hydrogen, deuterium or methyl optionally
substituted with 1 to 3 substituents independently selected from F
or Cl;
[0012] R.sup.6 is H or --C(.dbd.O)OH; and
[0013] X is C(H), C(F), C(CI), or N.
In one embodiment, the compound of formula (I) has the formula
(IA):
##STR00003##
or pharmaceutically acceptable salt thereof. In one embodiment
R.sup.4 is methyl optionally substituted with 1 to 3 substituents
selected from F or Cl. In another embodiment R.sup.4 is hydrogen.
In another embodiment R.sup.5 is methyl optionally substituted with
1 to 3 substituents selected from F or Cl. In another embodiment
R.sup.5 is hydrogen. In another embodiment R.sup.6 is
--C(.dbd.O)OH. In another embodiment R.sup.6 is hydrogen. In
another embodiment X is C(F). In another embodiment X is C(H).
[0014] In another embodiment X is C(CI). In another embodiment X is
N. In another embodiment R.sup.2 is hydrogen. In another embodiment
R.sup.2 is methyl. In another embodiment R.sup.3 is hydrogen. In
another embodiment R.sup.3 is methyl.
[0015] In yet another embodiment, R.sup.2 is hydrogen; R.sup.3 is
hydrogen; R.sup.4 is methyl; R.sup.5 is methyl; R.sup.6 is
--C(.dbd.O)OH; and X is C(H). In another embodiment, additionally
R.sup.1 is (C.sub.1-C.sub.6)alkyl substituted with 1 to 3
substituents selected from the group consisting of halo, hydroxy,
--NH.sub.2, --C(.dbd.O)NH.sub.2, and a 3 to 7 membered heterocycle,
wherein said heterocycle contains 1 to 3 heteroatoms independently
selected from O, N, or S. Alternatively, in another embodiment,
R.sup.1 is (C.sub.1-C.sub.6)alkyl substituted with 1 to 3 halo.
Alternatively, in another embodiment, R.sup.1 is
(C.sub.1-C.sub.6)alkyl substituted with 1 to 3 hydroxy.
Alternatively, in another embodiment, R.sup.1 is
(C.sub.1-C.sub.6)alkyl substituted with 1 to 3 NH.sub.2.
[0016] Alternatively, in another embodiment, R.sup.1 is
(C.sub.1-C.sub.6)alkyl substituted with --C(.dbd.O)NH.sub.2.
Alternatively, in another embodiment, R.sup.1 is a 3-7 membered
heterocycle, wherein said heterocycle contains 1 to 3 heteroatoms
independently selected from O, N, or S.
[0017] In one particular embodiment, the invention is:
##STR00004##
or a pharmaceutically acceptable salt thereof.
[0018] In another particular embodiment, the invention is:
##STR00005##
or a pharmaceutically acceptable salt thereof.
[0019] In another particular embodiment, the invention is:
##STR00006##
or a pharmaceutically acceptable salt thereof.
[0020] In another particular embodiment, the invention is:
##STR00007##
or a pharmaceutically acceptable salt thereof.
[0021] In another particular embodiment, the invention is:
##STR00008##
or a pharmaceutically acceptable salt thereof.
[0022] In another embodiment, the compound of formula I has the
formula (IB):
##STR00009##
wherein R.sup.1 is selected from the group consisting of:
##STR00010## ##STR00011## ##STR00012##
and pharmaceutically acceptable salts thereof.
[0023] In one embodiment the invention is a compound of formula
(IB) wherein R.sup.1 is selected from the group consisting of:
##STR00013##
and pharmaceutically acceptable salts thereof.
[0024] In another embodiment the invention is a compound of formula
(IB) wherein R.sup.1 is selected from the group consisting of:
##STR00014##
and pharmaceutically acceptable salts thereof.
[0025] In another embodiment the invention is a compound of formula
(IB) wherein R.sup.1 is selected from the group consisting of:
##STR00015##
and pharmaceutically acceptable salts thereof.
[0026] In another embodiment the invention is a compound of formula
(IB) wherein R.sup.1 is selected from the group consisting of:
##STR00016##
and pharmaceutically acceptable salts thereof.
[0027] In one embodiment, the pharmaceutically acceptable salt of
compounds of the invention is a potassium or sodium salt.
[0028] In one embodiment, the pharmaceutically acceptable salt of
compounds of the invention is a bis-potassium or bis-sodium
salt.
[0029] In another embodiment, the invention is a pharmaceutical
composition comprising the compounds described above, or
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0030] In another embodiment, the invention is a method for the
treatment of a bacterial infection in a mammal comprising
administering to said mammal an amount of a compound of formula (I)
or pharmaceutically acceptable salt thereof that is effective in
treating a bacterial infection. In one particular embodiment the
bacterial infection is resistant or susceptible. In another
particular embodiment, the bacterial infection is MDR (multi-drug
resistant). In one embodiment, the bacterial infection is selected
from the group consisting of respiratory tract infections, lung
infection in cystic fibrosis patients, complicated urinary tract
infections, burn infections, wound infections, blood infections,
complicated skin and soft tissue infections, nail infections, ear
infections, infections caused from medical devices, infections
caused from a catheter, noscomial pneumonia, ventilator-associated
pneumonia (VAP), community-acquired pneumonia (CAP), bacteremia,
hot-tub rash (dermatitis), and post-operative infection in radial
keratotomy surgery in humans. In another embodiment, the bacterial
infection is selected from the group consisting of Nosocomial
pneumonia, ventilator-associated pneumonia (VAP), complicated UTI
(urinary tract infection), complicated skin and skin structure, and
bacteremia. In another embodiment, the bacterial infection is a
burn infection. In another embodiment, the bacterial infection is a
lung infection in cystic fibrosis patients.
[0031] The present invention also relates to a method of treating
infection caused by Pseudomonas aeruginosa, Escherichia coli, a
Klebsiella species, or an Acinetobacter species, comprising
administering a therapeutically effective amount of the compound of
formula (I) or pharmaceutically acceptable salt thereof to a
mammalian subject in need thereof. In one particular embodiment,
the infection is caused by Pseudomonas aeruginosa.
[0032] The present invention also relates to a method of treating
infection by Pseudomonas aeruginosa that is resistant to doripenem,
meropenem or piperacillin comprising administering a
therapeutically effective amount of the compound of formula (I) or
pharmaceutically acceptable salt thereof to a mammalian subject in
need thereof.
[0033] The present invention also relates to a composition
comprising a compound of formula (I), or pharmaceutically
acceptable salt thereof, and an additional antibacterial agent
selected from the group consisting of beta-lactams,
aminoglycosides, polymyxins, penicillins, and lincosamides. In one
embodiment, the additional antibacterial agent is a beta-lactam
selected from the group consisting of cephalosporins, carbapenems,
and beta-lactamase inhibitors or beta-lactam/beta-lactamase
inhibitor combinations. In another embodiment, the additional
antibacterial agent is selected from the group consisting of
clindamycin, metronidazole, imipenem, meropenem, doripenem,
ertapenem, cefotetan, cefepime, and cefpirome, or a third
generation cephalosporin. In one particular embodiment, the
additional antibacterial agent is cefepime. In another embodiment,
the addition antibacterial agent is meropenem. In another
embodiment of the composition, the compound is:
##STR00017##
or a pharmaceutically acceptable salt thereof. In another
embodiment of the composition, the compound is:
##STR00018##
or pharmaceutically acceptable salt thereof. In another embodiment
of the composition, the compound is:
##STR00019##
or pharmaceutically acceptable salt thereof. In another embodiment
of the composition, the compound is:
##STR00020##
or pharmaceutically acceptable salt thereof. In another embodiment
of the composition, the compound is:
##STR00021##
or pharmaceutically acceptable salt thereof.
[0034] In another embodiment, the composition is one of the
specific compounds shown above and the additional anti-bacterial
agent is cefepime.
[0035] In another embodiment, the composition is one of the
specific compounds shown above and the additional anti-bacterial
agent is meropenem.
[0036] The present invention also relates to a complex or chelate
comprising a compound of formula (I) or a pharmaceutically
acceptable salt thereof, as a ligand and an iron (+3) cation,
wherein the ratio of ligand to iron cation is from about 1:1 to
about 3:1, respectively. In one embodiment, the ratio is about
3:1.
[0037] The present invention includes methods of treatment of the
human or non-human animal body, e.g., to combat or treat (including
prevention) bacterial infections, comprising administering to
subjects a useful or effective amount of a compound of the
invention, including a physiologically acceptable salt or solvate
thereof, and including compositions.
[0038] The compounds of the invention can also be combined with
other active ingredients as desired to attain combination therapy
for more than one condition or biological target. For example, the
compounds of the invention can be combined with other
anti-infectives, or agents that increase the efficacy or other
properties of the anti-infective, e.g., efflux inhibitors.
[0039] The compounds of formula (I) are useful for treating a
patient suffering from a disorder such as, e.g., a bacterial
infection.
[0040] Bacterial infections amenable to treatment by compounds of
formula (I), pharmaceutical compositions, and methods of the
present invention include those caused by Acinetobacter baumannii,
Acinetobacter spp., Bacteroides fragilis, Citrobacter diversus,
Citrobacter freundii, Enterobacter aerogenes, Enterobacter cloacae,
Escherichia coli, Haemophilus influenzae .beta.-lactamase negative,
Haemophilus influenzae .beta.-lactamase positive, Klebsiella
oxytoca, Klebsiella pneumoniae (including those encoding
extended-spectrum .beta.-lactamases (hereinafter "ESBLs"),
Legionella pneumophila, Moraxella catarrhalis
.beta.-lactamase-negative, Moraxella catarrhalis .beta.-positive,
Morganella morganii, Neisseria meningitidis, Prevotella spp. (and
members of the Enterobacteriaceae that express ESBLs and AmpC-type
beta-lactamases that confer resistance to currently available
cephalosporins, cephamycins and beta-lactam/beta-lactamase
inhibitor combinations), Proteus mirabilis, Pseudomonas aeruginosa,
Salmonella/Shigella, and Serratia marcescens.
[0041] The compounds of formula (I) may, in one embodiment, be used
to treat a variety of hospital and community acquired infections
such as respiratory tract infections (including lung infection in
cystic fibrosis patients), complicated urinary tract infections,
burn infections, wound infections, blood infections, complicated
skin and soft tissue infections, nail and ear infections,
infections caused from medical devices (e.g., catheter, etc.),
noscomial pneumonia (including ventilator-associated pneumonia
(VAP)), community-acquired pneumonia (CAP), bacteremia, "hot-tub
rash" (dermatitis), and post-operative infection in radial
keratotomy surgery in humans (hereinafter "the infections").
[0042] In one embodiment, the infection is selected from the group
consisting of noscomial pneumonia, ventilator-associated pneumonia,
complicated urinary tract infections, complicated skin & skin
structure infections, and bacteremia.
[0043] In one embodiment, the composition of the invention
comprises a therapeutically effective amount of a compound of
formula (I) of the invention.
[0044] The invention also relates to compositions of the invention
which comprise any combination of one or more compounds of formula
(I) and at least one additional ingredient (hereinafter "the
compositions of the invention").
[0045] Non-limiting examples of the at least one additional
ingredient include impurities (e.g., intermediates present in the
unrefined compounds of formula (I)), active or pharmaceutical
agents as discussed below (e.g., another antibacterial agent),
pharmaceutically acceptable excipients, or one or more solvents
(e.g., a pharmaceutically acceptable carrier as discussed
herein).
[0046] Compositions of the invention that are suitable for
administration to a patient in need thereof (e.g., a human) are
also referred to herein as "pharmaceutical compositions of the
invention."
[0047] Administration of the compounds of the present invention
(hereinafter the "active compound(s)") can be effected by any
method that enables delivery of the compounds to the site of
action. These methods include oral routes, intraduodenal routes,
parenteral injection (including intravenous, subcutaneous,
intramuscular, intravascular or infusion), topical, and rectal
administration. In one particular embodiment, the method of
administration is intravenous.
[0048] The pharmaceutical composition may, for example, be in a
form suitable for oral administration as a tablet, capsule, pill,
powder, sustained release formulation, solution, suspension, for
parenteral injection as a sterile solution, suspension or emulsion,
for topical administration as an ointment or cream or for rectal
administration as a suppository. The pharmaceutical composition may
be in unit dosage forms suitable for single administration of
precise dosages. The pharmaceutical composition will include a
conventional pharmaceutical carrier or excipient and a compound
according to the invention as an active ingredient. In addition, it
may include other medicinal or pharmaceutical agents, carriers,
adjuvants, etc.
[0049] Exemplary parenteral administration forms include solutions
or suspensions of active compounds in sterile aqueous solutions,
for example, aqueous propylene glycol or dextrose solutions. Such
dosage forms can be suitably buffered, if desired.
[0050] Suitable pharmaceutical carriers include inert diluents or
fillers, water and various organic solvents. The pharmaceutical
compositions may, if desired, contain additional ingredients such
as flavorings, binders, excipients and the like. Thus for oral
administration, tablets containing various excipients, such as
citric acid may be employed together with various disintegrants
such as starch, alginic acid and certain complex silicates and with
binding agents such as sucrose, gelatin and acacia. Additionally,
lubricating agents such as magnesium stearate, sodium lauryl
sulfate and talc are often useful for tableting purposes. Solid
compositions of a similar type may also be employed in soft and
hard filled gelatin capsules. Preferred materials, therefore,
include lactose or milk sugar and high molecular weight
polyethylene glycols. When aqueous suspensions or elixirs are
desired for oral administration the active compound therein may be
combined with various sweetening or flavoring agents, coloring
matters or dyes and, if desired, emulsifying agents or suspending
agents, together with diluents such as water, ethanol, propylene
glycol, glycerin, or combinations thereof.
[0051] Methods of preparing various pharmaceutical compositions
with a specific amount of active compound are known, or will be
apparent, to those skilled in this art. For examples, see
Remington's Pharmaceutical Sciences, Mack Publishing Company,
Easter, Pa., 15th Edition (1975).
[0052] The minimum amount of the compounds of formula (I) to be
administered is a therapeutically effective amount. The term
"therapeutically effective amount" means the amount of compound
which prevents the onset of, alleviates the symptoms of, stops the
progression of, and/or eliminates a bacterial infection in a
mammal, e.g., a human.
[0053] Typically, an effective dosing schedule of the compounds of
formula (I) of the invention for adults is about 50 mg to about
3000 mg of a compound of formula (I) in a single dose; in another
embodiment, an effective single dose is about 100 mg to about 2000
mg. In another embodiment, an effective single dose is about 800 mg
to about 1000 mg. Typically the dosages are given 1 to 4 times per
day. In one embodiment, the dosages are given 3 times per day. In
some cases, it may be necessary to use dosages outside these
limits.
[0054] The compounds of formula (I) of the invention may be
administered in combination with one or more additional medicinal
or pharmaceutical agents ("the additional active agent"). Such use
of the compounds of formula (I) in combination with an additional
active agent may be for simultaneous, separate or sequential
use.
[0055] In one embodiment, the additional active agent is an
antibacterial agent.
[0056] In one embodiment the antibacterial agent is a
.beta.-lactam. Non-limiting examples of .beta.-lactams include
cephalosporins (e.g., cefepime, ceftazidime, cefpirome, cefditoren
pivoxil (Spectracef.RTM.), cefoperazone, ceftazidime, cefdinir,
cefotaxime, cefpodoxime, cephalothin, cefaclor or cefixime),
cephamycins (e.g., cefotetan), carbapenems (e.g., imipenem,
meropenem, ertapenem, doripenem), beta-lactamase inhibitors and
beta-lactam/beta-lactamase inhibitor combinations such as
sulbactam, clavulanic acid, tazobactam and piperacillin in
combination with tazobactam (Zosyn.RTM.), and sulopenum.
[0057] In another embodiment the antibacterial agent is may be
selected from aminoglycosides (e.g., amikacin, gentamicin,
kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin,
streptomycin, tobramycin, apramycin, etc.), polymyxins (e.g.,
polymyxin B, colistin), fluoroquinolones (norfloxacin,
ciprofloxacin, levofloxacin (Levaquin.RTM.), moxifloxacin
(Avelox.RTM.), or enoxacin), penicillins (e.g., amoxicillin,
ampicillin, etc.), and lincosamides (e.g., clindamycin, lincomycin,
etc.).
[0058] In another embodiment the additional anti-bacterial agent is
selected from metronidazole, glycopeptides (e.g., vancomycin,
dalbavancin, telavancin, oritivancin), oxazolidinones (e.g.,
linezolid), lipeopetides (e.g., daptomycin), and tetracyclines
including gylcylcyclines (e.g., tigecycline).
[0059] Other non-limiting examples of additional antibacterial
agents can be found in Walsh and Wright, Chemical Reviews
105(2):391-394 (2005); and Bush et al., Current Opinion in
Microbiology 7:466-476 (2004).
[0060] In one embodiment, the additional antibacterial agent is
used in combination with compounds or pharmaceutically acceptable
salts of the invention to lower the frequency of resistance.
Examples include cefepime, cefpirome, imipenem, meropenem,
ertapenem, doripenem, sulopenem, ceftazidime,
piperacillin/tazobactam, ciprofloxacin, levofloxacin, moxifloxacin,
polymyxin B, and tigecycline.
[0061] In another embodiment, the additional antibacterial agent
may be a standard anti-anaerobe drug used in combination with
compounds or pharmaceutically acceptable salts of the invention to
treat intra-abdominal infections. Examples include clindamycin,
metronidazole, imipenem, meropenem, doripenem, ertapenem,
cefotetan, cefepime, cefpirome, and third generation
cephalosporins.
[0062] In another embodiment, the additional antibacterial agent
may be an acceptable anti-Gram positive agent used in combination
with compounds or pharmaceutically acceptable salts of the
invention for empiric therapy to treat P. aeruginosa and all
Enterobateriaceae. Examples include vancomycin, linezolid,
daptomycin, dalbavancin, telavancin, and oritivancin.
[0063] In one embodiment, the one or more additional active agents,
when used, are administered prior to administration of a compound
of formula (I). In another embodiment, the one or more additional
active agents, when used, are administered after administration of
a compound of formula (I). In another embodiment, the one or more
additional active agents, when used, are administered at about the
same time as administration of a compound of formula (I).
[0064] The additional active agent may be administered by any route
useful to administer said additional active agent.
[0065] In one embodiment, the one or more additional active agents
are present in the pharmaceutical composition of the invention.
Accordingly, in another embodiment, the invention relates to a
method of treating a patient with a pharmaceutical composition of
the invention further comprising one or more additional active
agents.
[0066] It is to be understood that any section headings and
subheadings herein are for the convenience of the reader and are
non-limiting. For example, the subject matter in the Summary of the
Invention has no special status solely as a result of its placement
in that section.
[0067] Unless otherwise indicated, the language and terms used in
this document are to be given their broadest reasonable
interpretation as understood by the relevant skilled artisan. In
addition, in descriptions and claims in which the subject matter
(e.g., substitution at a given molecular position) is recited as
being selected from a group of possibilities, the recitation is
specifically intended to include any subset of the recited group.
In the case of multiple variable positions or substituents, any
combination of group or variable subsets is also contemplated.
[0068] Unless otherwise stated, the following abbreviations have
the following meaning: "L" means "liter", "mL" means "milliliter",
"mol" means "moles", "mmol" means "millimoles", "Ac" means
"acetyl", "Ph" means "phenyl", "Bz" means "benzoyl", "DCM" or
"CH.sub.2Cl.sub.2" means "dichloromethane", "DMSO" means
"dimethylsulfoxide", "MIC" means "minimum inhibitory
concentration", "MS" means "Mass Spectrometry" (all samples herein
were analyzed either by LCMS-electrospray (gradient elution using
acetonitrile, water, formic acid mixtures) or probe APCI methods),
"LCMS" means "liquid chromatography mass spectrometry", "NMR" means
"nuclear magnetic resonance spectroscopy" (All samples herein were
run at 400 MHz on Varian instruments, unless otherwise indicated),
"THF" means "tetrahydrofuran", "spp." means "species" and "cfu"
means "colony-forming unit".
[0069] As used herein, the term "(C.sub.1-C.sub.6)alkyl" refers to
linear or branched hydrocarbons (e.g., methyl, ethyl, n-propyl,
isopropyl) of 1 to 6 carbon atoms in length.
[0070] Unless otherwise indicated, the term "heterocycloalkyl", as
used herein, refer to non-aromatic cyclic groups containing one or
more heteroatoms, preferably from one to four heteroatoms, each
preferably selected from oxygen, sulfur and nitrogen. The
heterocycloalkyl groups of this invention can also include ring
systems substituted with one or more oxo moieties. Examples of
non-aromatic heterocycloalkyl groups are aziridinyl, azetidinyl,
pyrrolidinyl, piperidinyl, azepinyl, piperazinyl,
1,2,3,6-tetrahydropyridinyl, oxiranyl, oxetanyl, tetrahydrofuranyl,
tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl,
morpholino, thiomorpholino, thioxanyl, pyrrolinyl, indolinyl,
2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl,
dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl,
imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl,
3-azabicyclo[4.1.0]heptanyl, quinolizinyl, quinuclidinyl,
1,4-dioxaspiro[4.5]decyl, 1,4-dioxaspiro[4.4]nonyl,
1,4-dioxaspiro[4.3]octyl, and 1,4-dioxaspiro[4.2]heptyl.
[0071] Unless otherwise indicated, the term "heteroaryl", as used
herein, refers to an aromatic ring containing one or more
heteroatoms (preferably oxygen, sulfur and nitrogen), preferably
from one to four heteroatoms. Examples of 5 to 6 membered
heteroaryls are pyridinyl, pyridazinyl, imidazolyl, pyrimidinyl,
pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl,
isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, triazinyl,
purinyl, oxadiazolyl, thiadiazolyl, furazanyl.
[0072] The term "heterocycle" includes heteroaryl and
heterocycloalkyl rings as well as non-aromatic heterocyclic rings
containing zero or more double bonds.
[0073] Unless otherwise apparent or indicated, the compounds of the
invention and term "compound" in the claims embraces any
pharmaceutically acceptable salts or solvates, and any amorphous or
crystal forms, or tautomers, whether or not specifically recited in
context. Similarly, a recitation is open to any material or
composition containing the recited compound (e.g., a composition
containing a salt of a racemic mixture of compounds, tautomers,
epimers, stereoisomers, impure mixtures, etc.).
[0074] The compounds of formula (I) may exist in unsolvated and
solvated forms. Thus, it will be understood that the compounds of
the invention also include hydrate and solvate forms as discussed
below.
[0075] The term "solvent" as it relates to the compositions of the
invention includes organic solvents (e.g., methanol, ethanol,
isopropanol, ethyl acetate, methylene chloride, and
tetrahydrofuran) and water. The one or more solvents may be present
in a non-stoichiometric amount, e.g., as a trace impurity, or in
sufficient excess to dissolve the compound of the invention.
Alternatively, the one or more solvents may be present in a
stoichiometric amount, e.g., 0.5:1, 1:1, or 2:1 molar ratio, based
on the amount of compound of the invention.
[0076] The term "solvate" is used herein to describe a noncovalent
or easily reversible combination between solvent and solute, or
dispersion means and disperse phase. It will be understood that the
solvate can be in the form of a solid, slurry (e.g., a suspension
or dispersion), or solution. Non-limiting examples of solvents
include ethanol, methanol, propanol, acetonitrile, dimethyl ether,
diethyl ether, tetrahydrofuran, methylene chloride, and water. The
term `hydrate` is employed when said solvent is water.
[0077] A currently accepted classification system for organic
hydrates is one that defines isolated site, or channel
hydrates--see Polymorphism in Pharmaceutical Solids by K. R. Morris
(Ed. H. G. Brittain, Marcel Dekker, 1995). Isolated site hydrates
are ones in which the water molecules are isolated from direct
contact with each other by intervening organic molecules. In
channel hydrates, the water molecules lie in lattice channels where
they are next to other water molecules.
[0078] When the solvent or water is tightly bound, the complex will
have a well-defined stoichiometry independent of humidity. When,
however, the solvent or water is weakly bound, as in channel
solvates and hygroscopic compounds, the water/solvent content will
be dependent on humidity and drying conditions. In such cases,
non-stoichiometry will be the norm.
[0079] Unless otherwise indicated, the term "pharmaceutically
acceptable salt(s)", as used herein, unless otherwise indicated,
includes salts of acidic or basic groups which can be present in
the compounds. Compounds that are basic in nature are capable of
forming a wide variety of salts with various inorganic and organic
acids. The acids that can be used to prepare pharmaceutically
acceptable acid addition salts of such basic compounds are those
that form non-toxic acid addition salts. The compounds can form,
e.g., sulfates, phosphates, citrates, acetates, tosylates,
succinates, besylates, mesylates, lactates, and hydrochlorides.
Basic salts can be mono or dibasic. In one preferred embodiment,
the salt is a fumarate.
[0080] Unless otherwise indicated, the terms "treat," "treatment,"
and "treating", as used herein in the context of using the
compounds of the present invention, unless otherwise indicated,
means reversing, alleviating, inhibiting the progress of one or
more symptoms of such disorder or condition.
[0081] As used herein the term "patient" refers to a mammal such
as, e.g., a human, dog, cat, horse, pig, cow, and the like. In one
embodiment, the patient is a human.
[0082] Unless otherwise indicated, the term "pharmaceutical
composition" refers to an active compound in any form suitable for
effective administration to a subject, e.g., a mixture of the
compound and at least one pharmaceutically acceptable carrier.
[0083] Unless otherwise indicated, the term "pharmaceutically
acceptable carrier" refers to a material that can be administered
to a subject together with a compound in a pharmaceutical
composition. The carrier should not destroy the pharmacological
activity of the compound and should be non-toxic when administered
in doses sufficient to deliver a therapeutic amount of the
compound.
[0084] The term "excipient" means an inert material that is
combined with the compounds of formula (I) to produce a
pharmaceutical composition or oral drug dosage form. The term
"pharmaceutically acceptable excipient" means that the excipient
must be compatible with other ingredients of the composition, and
not deleterious to the recipient thereof. The pharmaceutically
acceptable excipients are chosen on the basis of the intended
dosage form.
[0085] Compounds of the present invention have asymmetric centers
and therefore can exist in different enantiomeric and
diastereomeric forms. The invention includes all optical isomers
and stereoisomers, and mixtures thereof in all ratios, and to all
pharmaceutical compositions and methods of treatment that can
employ or contain them. Although specific compounds exemplified in
this application can be depicted in a particular stereochemical
configuration, compounds having either the opposite stereochemistry
at any chiral centers or mixtures thereof are also envisioned. The
foregoing can be present as mixtures or enriched in any component
to any degree. Where stereochemistry at a position is not
specified, such is intended to encompass either configuration or a
mixture of any ratio.
[0086] Compounds of this invention include pharmaceutically
acceptable derivatives or prodrugs thereof. A "pharmaceutically
acceptable derivative or prodrug" means any pharmaceutically
acceptable salt, ester, salt of an ester or other derivative of a
compound that, upon administration to a recipient, is capable of
providing (directly or indirectly) a compound of this invention or
a metabolite or residue thereof. Particularly favored derivatives
and prodrugs of the invention are those that increase the
bioavailability of the compounds when such compounds are
administered to a patient (e.g., by allowing an orally administered
compound to be more effectively absorbed into the blood), enhance
delivery of the parent compound to a given biological compartment,
increase solubility to allow administration by injection, alter
metabolism or alter rate of excretion.
[0087] The compounds of formula (I) may exhibit polymorphism.
Polymorphic compounds of formula (I) may be prepared by
crystallization of a compound of the present invention under
various conditions. For example, there may be employed various
solvents (including water) or different solvent mixtures for
recrystallization; crystallization at different temperatures;
various modes of cooling ranging from very fast to very slow
cooling during crystallization. Polymorphs may also be obtained by
heating or melting a compound of the present invention followed by
gradual or fast cooling. The presence of polymorphs may be
determined by solid probe NMR spectroscopy, IR spectroscopy,
differential scanning calorimetry, powder X-ray diffraction or
other such techniques.
[0088] The present invention includes compounds wherein one or more
hydrogen, carbon or other atoms are replaced by different isotopes
thereof. Such compounds can be useful as research and diagnostic
tools in metabolism pharmacokinetic studies and in binding assays.
These isotopically-labeled compounds are identical to those
compounds of formula (I), but for the fact that one or more atoms
are replaced by an atom having an atomic mass or mass number
different from the atomic mass or mass number predominantly found
in nature. Examples of isotopes that can be incorporated into
compounds of the invention include isotopes of hydrogen, carbon,
nitrogen, oxygen, and sulfur, such as, but not limited to, .sup.2H,
.sup.3H, .sup.13C, .sup.14C, .sup.15N, .sup.18O, .sup.17O, and
.sup.35S, respectively. The compounds of formula (I) of the
invention containing the aforementioned isotopes and/or other
isotopes of these atoms are within the scope of this invention.
Certain isotopically-labeled compounds of formula (I), for example
those into which radioactive isotopes such as .sup.3H and .sup.14C
are incorporated, are useful in drug and/or substrate tissue
distribution assays. Tritiated, i.e., .sup.3H, and carbon-14, i.e.,
.sup.14C, isotopes are particularly preferred for their ease of
preparation and detectability. Further, substitution with isotopes
such as deuterium, i.e., .sup.2H, can afford certain therapeutic
advantages resulting from greater metabolic stability, for example
increased in vivo half-life or reduced dosage requirements and,
hence, may be preferred in some circumstances. Isotopically-labeled
compounds of the invention can generally be prepared by carrying
out the procedures disclosed in the Schemes and/or in the Examples
and described below, by substituting a readily available
isotopically-labeled reagent for a non-isotopically-labeled
reagent.
[0089] The term "protecting group" refers to a suitable chemical
group that can be attached to a functional group and removed at a
later stage to reveal the intact functional group. Examples of
suitable protecting groups for various functional groups are
described in T. W. Greene and P. G. M. Wuts, Protective Groups in
Organic Synthesis, 2nd Ed., John Wiley and Sons (1991 and later
editions); L. Fieser and M. Fieser, Fieser and Fieser's Reagents
for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette,
ed. Encyclopedia of Reagents for Organic Synthesis, John Wiley and
Sons (1995). The term "hydroxy protecting group", as used herein,
unless otherwise indicated, includes Ac, Bz, and various hydroxy
protecting groups familiar to those skilled in the art, including
the groups referred to in Greene.
[0090] The examples and preparations provided below further
illustrate and exemplify the compounds of the present invention and
methods of preparing such compounds. It is to be understood that
the scope of the present invention is not limited in any way by the
scope of the following examples and preparations.
[0091] All patents, patent applications, publications, test
methods, literature, and other materials cited above and below
herein are hereby incorporated herein by reference in their
entireties.
DETAILED DESCRIPTION OF THE INVENTION
[0092] As noted above, in one embodiment, the present invention
relates to compounds of formula (I) and pharmaceutically acceptable
salts thereof, as described above. The compounds of formula (I) are
depicted structurally in the Summary of the Invention an elsewhere
herein for the convenience of the reader.
General Preparation Methods
[0093] The compounds of the present invention may be prepared
according to the descriptions, schemes, and examples herein, which
are non-limiting, in combination with the knowledge of the skilled
artisan.
##STR00022##
[0094] The compounds of the present invention can be prepared as
outlined in schemes A through C. Compounds of the general formula I
(Scheme A), prepared as described in Yamawaki, K., et al.,
Bioorganic & Medicinal Chem., (2007), 15, 6716 and Yamamoto,
H., et al., Bioorganic and Medicinal Chem., (2002), 10, 1535, can
be reacted with hydroxylamines of the general formula II (prepared
as described in WO 2007/065288, published Jun. 14, 2007) in a
solvent such as methanol at ambient temperature for approximately 2
hours to form carboxylic acids of the formula III. Activated esters
of the formula IV can be prepared by reaction of compounds of the
formula III with N-hydroxysuccinamide in the presence of coupling
reagent such as dicyclohexylcarbodiimide or diisopropylcarbodiimide
in a solvent such as dichloromethane at ambient temperature. A
compound of formula V, prepared as described by Waulte, S. R. et
al. J. Org. Chem (1986), 51, 3133; Paloma, C., et al., J. Org.
Chem. (1997), 62, 2070; Lall, M. S., et al., J. Org. Chem. (2002),
67, 1536 and Chhabra, S. R., et al., J. Org. Chem. (2002), 67,
4017, can be generated by deprotection of the corresponding
N-benzyloxycarbonyl (Cbz) protected compound by hydrogenolysis at
ambient temperature in the presence of palladium on carbon, under
approximately two to four atmospheres of hydrogen gas, in a solvent
such as methanol, ethanol, tetrahydrofuran, toluene or acetic acid
sometimes requiring a binary combination thereof. If the
hydrogenation is done in the presence of acetic acid, the
intermediate aminoazetidinone can be isolated as the acetate salt
and subsequently reacted with compounds of the formula IV in
solvents such as methanol, ethanol or acetonitrile in the presence
of a base such as triethylamine to form amides of the general
formula VI. When acetic acid is not used in the hydrogenation, the
aminoazetidinones produced, once the catalyst is removed by
filtration, can be reacted in situ with compounds of the formula IV
to generate amides of the formula VI.
##STR00023##
[0095] Triazolones of the formula XI can be prepared as outlined in
Scheme B. Starting from commercially available Kojic acid (CAS
number: 501-30-4), compound VII can be prepared in five steps as
described in EP 0281289, published February 19, 1988. Reaction of
VII with phosgene or a phosgene equivalent such as
carbonyldiimidazole in a solvent such as dichloromethane or
tetrahydrofuran at ambient temperature produces compound VIII.
Reaction of compound VIII with a primary amine of formula IX in a
solvent such as tetrahydrofuran at elevated temperature such as
between 40.degree. C. and 60.degree. C. will produce compounds of
the general formula X. Compounds of the formula X can be cyclized
to form compounds of the formula XI by reaction in water at reflux
in the presence of a base such as sodium hydroxide or potassium
hydroxide. Alternatively, in some cases, a similar cyclization
reaction can be accomplished by reaction of compounds of the
formula X in N-trimethylsilyl-N-methyltrifluoroacetamide (MSTFA) at
approximately 150.degree. C. resulting from microwave
irradiation.
##STR00024##
[0096] The coupling of compounds VI and XI and the final
construction of the compounds of the present invention can be
accomplished as outlined in Scheme C. The coupling of VI and XI to
generate compounds of the general formula XII can be accomplished
by first reacting compounds of the formula XI with excess MSTFA in
tetrahydrofuran at approximately 40.degree. C. for one to two hours
followed by removal of the tetrahydrofuran, excess MSTFA and
byproducts under vacuum. Separately, compounds of the formula VI
can be reacted with chlorosulfonylisocyanate (CSI) in
dichloromethane at 0.degree. C. for approximately 45 minutes. The
adduct of the compound of formula XI can then be re-dissolved in
tetrahydrofuran and to this mixture added the adduct from reaction
of the compound of formula VI with CSI. Stirring of these two
components at 0.degree. C. for approximately two hours followed by
stirring at ambient temperature for up to 18 hours produces the
compounds of the general formula XII. Alternatively, this coupling
reaction can be accomplished as follows: the CSI adduct prepared as
described above is mixed with the compound of formula XI, which can
be silylated by reaction with excess hexamethyldisilizide (HMDS) in
the presence of a catalytic amount of trimethylsilylchliride
(TMS-CI) at approximately 140.degree. C. After cooling to ambient
temperature, this material can be dissolved in a solvent such as
dichloromethane and mixed the CSI adduct generating the compound of
formula XII. Removal of the benzyl protecting groups from compounds
of the formula XII can be accomplished by reaction with between two
and four atmospheres of hydrogen gas in a binary solvent system
consisting of tetrahydrofuran and acetic acid at ambient
temperature and in the presence of a palladium catalyst such as
palladium black. Following removal of the catalyst and solvent,
treatment of the crude material with an acid such as
trifluoroacetic acid in a solvent such as dichloromethane at
ambient temperature removes the tertiary-butyloxycarbonyl
protecting group and the tertiary-butylester if contained within
R.sup.6. The crude material of the present invention (XIII) can
then be purified by reverse-phase chromatography using a C18 resin
with a gradient mobile phase consisting of acetonitrile and water,
buffered with formic acid. The sodium salts with the general
formula XIV can then be generated from the compounds of formula
XIII by treatment with sodium bicarbonate in water followed by
lyophilization. If a second acidic site is present within R.sup.6
such as a carboxylic acid, the bis-sodium salts of the formula XIV
can be produced following the same procedure, but adding a second
equivalent of sodium bicarbonate prior to lyophilization.
[0097] Without further elaboration, it is believed that one skilled
in the art can, using the preceding description, practice the
present invention to its fullest extent. The following detailed
examples describe how to prepare the various compounds and/or
perform the various processes of the invention and are to be
construed as merely illustrative, and not limitations of the
preceding disclosure in any way whatsoever. Those skilled in the
art will promptly recognize appropriate variations from the
procedures both as to reactants and as to reaction conditions and
techniques.
EXAMPLES
[0098] The Examples below were generally carried out under inert
atmosphere (nitrogen or argon), particularly in cases where oxygen
or moisture-sensitive reagents or intermediates were employed.
Commercial solvents and reagents were generally used without
further purification. All products were dried before
characterization or use in subsequent chemistry. Chemical shifts
for nuclear magnetic resonance (NMR) data are expressed in parts
per million (ppm, .delta.) referenced to residual peaks from the
deuterated solvents employed.
Example 1
Preparation of
2-({[(1E)-1-(2-amino-1,3-thiazol-4-yl)-2-{[(3S)-1-({[4-ethyl-3-(5-hydroxy-
-4-oxo-1,4-dihydropyridin-2-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl]su-
lfonyl}carbamoyl)-2-oxoazetidin-3-yl]amino}-2-oxoethylidene]amino}oxy)-2-m-
ethylpropanoic acid (1).
##STR00025##
[0100] Compound 1 was prepared by the procedures depicted in
schemes 1 to 6 and outlined in detail below.
##STR00026##
Step 1. Preparation of benzyl
4,5-bis(benzyloxy)pyridine-2-carboxylate (C4)
[0101] A. Preparation of
5-(benzyloxy)-2-(hydroxymethyl)-4H-pyran-4-one (C1).
5-Hydroxy-2-(hydroxymethyl)-4H-pyran-4-one (300 g, 2.11 mol) was
dissolved in methanol (9 L) and treated with potassium carbonate
(439 g, 3.18 mol), followed by slow addition of benzyl chloride
(433 g, 3.42 mol). The reaction mixture was stirred at 65.degree.
C. for 8 hours. After cooling to room temperature, it was stirred
for an additional 16 hours, then concentrated in vacuo to a thick
paste. This residue was cooled to 10.degree. C. and diluted with
ice water, resulting in a precipitate that was gathered by
filtration to provide C1 as a solid. Yield: 325 g, 1.40 mol, 66%.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.29 (s, 2H), 4.94 (s,
2H), 6.32 (s, 1H), 7.33-7.42 (m, 5H), 8.17 (s, 1H).
[0102] B. Preparation of 5-(benzyloxy)-4-oxo-4H-pyran-2-carboxylic
acid (C2). A solution of chromium(VI) oxide (64.6 g, 0.646 mol) in
water (90 mL) was cooled to -5.degree. C. and treated drop-wise
with concentrated sulfuric acid (56 mL). This was diluted with
additional water (40 mL), and then added drop-wise to a cold
(-5.degree. C.) solution of C1 (100 g, 0.43 mol) in acetone (4.5
L). The reaction mixture was stirred at 20.degree. C. for 3 hours,
then filtered through a pad of Celite. Concentration of the
filtrate provided a residue, which was washed with hexane to
provide C2. Yield: 80 g, 0.325 mol, 76%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 4.97 (s, 2H), 6.93 (s, 1H), 7.34-7.42 (m,
5H), 8.37 (s, 1H).
[0103] C. Preparation of
5-(benzyloxy)-4-oxo-1,4-dihydropyridine-2-carboxylic acid (C3). A
mixture of C2 (100 g, 0.406 mol) and aqueous ammonium hydroxide
solution (25%, 1 L) was stirred in an autoclave for 1 hour, and
then heated at 83.degree. C. for 7 hours at atmospheric pressure.
After cooling slowly over about 18 hours, the reaction mixture was
acidified to pH 3 with concentrated hydrochloric acid. The
resulting precipitate was collected by filtration, washed with
water, and dissolved in saturated aqueous sodium bicarbonate
solution. The solution was washed with dichloromethane, then
acidified with concentrated hydrochloride acid. The resulting solid
was collected by filtration, washed with water and dried at
50.degree. C. to provide C3. Yield: 85 g, 0.347 mol, 85%. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 5.17 (s, 2H), 7.17 (br s, 1H),
7.33-7.49 (m, 7H).
[0104] D. Preparation of C4. Benzyl chloride (105.6 mL, 0.918 mol)
was added to a solution of C3 (90 g, 0.367 mol) in
dimethylformamide (1.25 L). Potassium carbonate (124.8 g, 0.903
mol) was added, and the mixture was stirred at 80.degree. C. for 16
hours. After cooling to room temperature, the reaction was treated
with ice water, and the resulting solid was collected by filtration
and purified by silica gel chromatography to afford C4. Yield: 50
g, 0.118 mol, 32%. MS m/z 426 (M+1). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 5.32 (s, 6H), 7.33-7.46 (m, 15H), 7.76 (s,
1H), 8.37 (s, 1H).
##STR00027##
Step 2. Preparation of 5-(4,5-bis(benzyloxy)pyridin-2-yl)-1
,3,4-oxadiazol-2(3H)-one (C6).
[0105] A. Preparation of
4,5-bis(benzyloxy)pyridine-2-carbohydrazide (C5). Hydrazine
monohydrate (47.5 mL, 978 mmol) was added drop-wise over 10 minutes
to a suspension of C4 (20 g, 47.0 mmol) in methanol (100 mL). The
resulting mixture was heated to 65.degree. C. for 2 hours, then
cooled to room temperature and filtered under vacuum. The collected
solids were washed with methanol to provide C5 as a white solid.
Yield: 15.4 g, 44.1 mmol, 94%. LCMS m/z 350.1 (M+1). .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 4.47 (d, J=4.6 Hz, 2H), 5.30 (s,
2H), 5.32 (s, 2H), 7.31-7.48 (m, 10H), 7.67 (s, 1H), 8.23 (s, 1H),
9.65 (t, J=4.5 Hz, 1H).
[0106] B. Preparation of
5-[4,5-bis(benzyloxy)pyridin-2-yl]-1,3,4-oxadiazol-2(3H)-one (C6).
Carbonyl diimidazole (97%, 2.87 g, 17.2 mmol) was added to a
suspension of C5 (5.00 g, 14.3 mmol) in tetrahydrofuran (75 mL).
The reaction mixture was stirred at room temperature for 3 hours,
during which time the white suspension became a homogeneous
solution, and then a white suspension. The solid was collected by
filtration and washed with tetrahydrofuran (3.times.5 mL) to
provide C6 as a white solid. Yield: 4.92 g, 13.1 mmol, 92%. LCMS
m/z 376.1 (M+1). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 5.31
(s, 2H), 5.33 (s, 2H), 7.32-7.48 (m, 10H), 7.56 (s, 1H), 8.38 (s,
1H), 12.64 (br s, 1H).
##STR00028##
Step 3. Preparation of
5-(4,5-bis(benzyloxy)pyridin-2-yl)-4-ethyl-2H-1,2,4-triazol-3(4H)-one
(C8)
[0107] A. Preparation of
1-(3,4-bis(benzyloxy)picolinoyl)-4-ethylsemicarbazide (C7). To a
solution of C5 (0.75 g/0.215 mol) in 5 mL N,N-dimethylformamide at
5.degree. C. was added slowly a solution of 229 mg (3.22 mmol) of
ethylisocyanate in 5 mL of tetrahydrofuran and the resulting
mixture stirred at room temperature for 2 hours, at which point,
the reaction mixture was transferred to a solution containing 0.161
g (0.0032 mol) hydrazine monohydrate in 10 mL of tetrahydrofuran at
a rate such to maintain a temperature less than 15.degree. C. Once
added, the resulting mixture stirred at room temperature for 1
hour, at which point the mixture was poured into 50 mL of ice-water
forming a precipitate, which was collected by filtration and dried
in vacuo affording C7 as a white solid. LCMS m/z 421 (M+1). .sup.1H
NMR (400 MHz, CDCL3-d.sub.6) .delta. 1.25 (t, J=6.2 Hz, 3H), 4.25
(br. d, 2H), 5.24 (br. s, 4H), 7.31-7.46 (m, 10H), 7.55 (br. s,
1H), 8.18 (s, 1H).
[0108] B. Preparation of
5-(4,5-bis(benzyloxy)pyridin-2-yl)-4-ethyl-2H-1,2,4-triazol-3(4H)-one
(C8) (Cyclization Method 1). To a stirred solution of 0.42 g (0.999
mmol) of C7 was added 8 equivalents (0.448 g/7.99 mmol) of
potassium hydroxide dissolved in 2 mL of water and the resulting
mixture heated to reflux for 24 hours at which point the reaction
mixture was concentrated to dryness in vacuo. The crude product was
then purified by column chromatography (silica-gel, 3 to 5%
methanol in ethylacetate producing 0.683 g (46%) of compound C8.
LCMS m/z 403.3 (M+1). .sup.1H NMR (400 MHz, CDCL3-d.sub.6) .delta.
1.26 (t, J=7.3 Hz, 3H), 4.26 (q, J=6.6 Hz, 2H), 5.24 (s, 4H),
7.29-7.47 (m, 10H), 7.57 (s, 1H), 8.16 (s, 1H), 9.86 (br. s,
1H).
##STR00029##
Step 4. Preparation of (3S)-3-aminoazetidin-2-one (C10)
[0109] Benzyl [(3S)-2-oxoazetidin-3-yl]carbamate (C9, 13.37 g, 60.7
mmol) was mixed with degassed ethanol (500 mL) and toluene (125
mL). For synthesis of C9, see M. J. Miller et al., Tetrahedron,
1983, 39, 2571-2575, and M. S. Lall et al., Journal of Organic
Chemistry 2002, 67, 1536-1547. The reaction mixture was sonicated
until all the solids dissolved, then purged with nitrogen.
Palladium on carbon (10%, 4.45 g) was added, and the reaction
mixture was hydrogenated on a Parr shaker for 1 hour at 15 psi. The
palladium was removed by filtration through Celite under nitrogen,
and rinsed with degassed ethanol. The filtrate, containing C10, was
carried directly into the coupling reaction with C12, Step 4B.
Yield: assumed quantitative. Material from a similar experiment was
concentrated to dryness to obtain NMR data: .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 2.12 (br s, 2H), 2.78 (dd, J=5.1, 2.3 Hz,
1H), 3.31 (dd, J=5.3, 5.3 Hz, 1H), 3.97 (m, 1H), 7.69 (br s,
1H).
##STR00030##
Step 5. Preparation of tert-butyl
2-({[(1Z)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-4-yl}-2-oxo-2-{[(-
3S)-2-oxoazetidin-3-yl]amino}ethylidene]amino}oxy)-2-methylpropanoate
(C13)
[0110] A. Preparation of tert-butyl
24({(1Z)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-4-yl}-2-[(2,5-diox-
opyrrolidin-1-yl)oxy]-2-oxoethylidene}amino)oxy]-2-methylpropanoate
(C12). 1-Hydroxypyrrolidine-2,5-dione (N-hydroxysuccinimide, 8.84
g, 76.8 mmol) was added to a suspension of
(2Z)-{2-[(tert-butoxycarbonyl)ami
no]-1,3-thiazol-4-yl}[(2-tert-butoxy-1,1-dimethyl-2-oxoethoxy)imino]aceti-
c acid (C11, 30 g, 70 mmol) in dichloromethane (400 mL). For
synthesis of C11, see K. Yamawaki et al., Bioorganic and Medicinal
Chemistry 2007, 15, 6716-6732. The mixture was cooled to 0.degree.
C., N,N'-dicyclohexylcarbodiimide (97%, 15.6 g, 73.3 mmol) was
added, and the reaction was stirred at 0.degree. C. for 30 minutes
and then at room temperature for 3 hours. The mixture was filtered
through Celite and concentrated in vacuo to afford C12 as a white
solid. Yield: 36.17 g, 68.7 mmol, 98%. LCMS m/z 527.2 (M+1).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 1.43 (s, 9H), 1.54 (s,
9H), 1.61 (s, 6H), 2.91 (br s, 4H), 7.50 (s, 1H), 8.31 (br s,
1H).
[0111] B. Preparation of C13. A solution of C10 (5.23 g, 60.7 mmol)
in ethanol/toluene (900 mL, solution obtained in Step 4) was
treated with compound C12 (26.6 g, 50.6 mmol), and the reaction
mixture was slowly concentrated under reduced pressure, over the
course of an hour, to one-third of its original volume. The
resulting suspension was stirred at 35.degree. C. under nitrogen
for about 18 hours. Removal of solvent in vacuo afforded a crude
product, which was dried under vacuum for 30 minutes. The resulting
solids were partitioned between 1:1 ethyl acetate/tetrahydrofuran
(1 L) and aqueous sodium bicarbonate solution (500 mL). Additional
water was required to dissolve solids observed during the
separation. The aqueous layer was extracted with 1:1 ethyl
acetate/tetrahydrofuran (2.times.300 mL), and the combined organic
layers were filtered and concentrated in vacuo. The crude solid was
triturated with 3:2 ethyl acetate/heptane (60 mL) for 30 minutes,
and the solids were collected by filtration, rinsing with heptane,
to provide C13 as a white solid. Yield: 22.08 g, 44.4 mmol, 88%.
LCMS m/z 498.6 (M+1). .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
1.47 (s, 9H), 1.52 (s, 6H), 1.54 (s, 9H), 3.39 (dd, J=5.7, 2.5 Hz,
1H), 3.65 (dd, J=5.5, 5.5 Hz, 1H), 5.10 (dd, J=5.3, 2.5 Hz, 1H),
7.34 (s, 1H).
##STR00031##
Step 6. Preparation of Preparation of
2-({[(1E)-1-(2-amino-1,3-thiazol-4-yl)-2-{[(3S)-1-({[4-ethyl-3-(5-hydroxy-
-4-oxo-1,4-dihydropyridin-2-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl]su-
lfonyl}carbamoyl)-2-oxoazetidi
n-3-yl]amino}-2-oxoethylidene]amino}oxy)-2-methylpropanoic acid
(1).
[0112] A. Preparation of C14 (Coupling Method 1). To a stirred
solution of C8 (180 mg/0.45 mmol) in 3 mL of tetrahydrofuran was
added 6.0 equivalents of
N-trimethylsilyl-N-methyltrifluoroacetamide (MSTFA, Aldrich
ampoule) forming a pale yellow solution, which stirred at room
temperature for 45 minutes. The mixture was then concentrated under
reduced pressure and heated under vacuum (high vac) at 45.degree.
C. for 1 hour. Separately, to 0.223 g (0.45 mmol) of C13 dissolved
in 4 mL dichloromethane and cooled to 0.degree. C. was added 0.082
g (0.58 mmol/0.051 mL) of chlorosulfonylisocyanate and the
resulting solution stirred at 0.degree. C. for 30 minutes. At this
point, tetrahydrofuran (3 mL) was added to the adduct of C8 and the
resulting solution introduced to the CSI reaction via cannula. The
resulting solution stirred at 0.degree. C. for 1 hour then brought
to room temperature and concentrated under reduced pressure. The
crude material was cleaned up by column chromatography (silica-gel,
45 to 60% ethylacetated in heptane) affording 332 mg of C14 in a
mixture that was carried forward without additional purification.
LCMS m/z 1003.2 (M-1).
[0113] B. Preparation of compound 1. In a Parr bottle was placed
0.190 g (0.19 mmol) of C14 dissolved in 20 mL of methanol and the
solution degassed with nitrogen gas. Palladium-black (0.063 g) was
then added and mixture agitated under an atmosphere of 13 psi
hydrogen at room temperature for 40 min (reaction complete by
LCMS). The reaction mixture was then filtered through Celite and
concentrated to dryness in vacuo. The material was then carried on
crude by dissolving in 10 mL of dichloromethane. To this solution
was then added 10 mL of trifluoroacetic acid and the resulting
mixture stirred at room temperature for 2 hours, at which time the
reaction mixture was concentrated in vacuo. The crude product (1)
was then purified by preparative HPLC (Symmetry C8, 3 to 23%
acetonitrile in water with 0.1% formic acid modifier).
Approximately 5 mg of 1 were collected following concentration to
dryness in vacuo. LCMS m/z 669.4 (M+1). .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 1.0-1.2 (m, 6H), 1.3-1.5 (br. s, 5H), 3.97 (d,
J=6.8 Hz, 2H), 4.88 (br. s, 1H), 6.81 (s, 1H), 7.32 (s, 1H), 7.99
(s, 1H), 9.05 (d, J=4.2 Hz, 1H).
Example 2
Preparation of
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-[({4-[2-(dimethylamino)-
ethyl]-3-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)-5-oxo-4,5-dihydro-1H-1,-
2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethyli-
dene]amino}oxy)-2-methylpropanoic acid (2)
##STR00032##
[0115] Compound 2 was prepared by the procedures depicted in scheme
7 and outlined in detail below.
##STR00033##
Step 1. Preparation of
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-[({4-[2-(dimethylamino)-
ethyl]-3-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)-5-oxo-4,5-dihydro-1H-1,-
2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethyli-
dene]amino}oxy)-2-methylpropanoic acid (2) and diasteriomeric diol
mixture (Example 10)
[0116] A. Preparation of C16. Compound C15 was prepared in an
analogous manner to C14 in example 1 using coupling method 1. LCMS
m/z 991.8 (M+1). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
1.33-1.39 (m, 9H), 1.41-1.45 (br. s, 6H), 3.32-3.38 (m, 1H), 6.67
(t, J=8.2 Hz, 1H), 4.67 (m, 1H), 4.85-4.92 (m, 1H), 4.98 (d, J=5.8
Hz, 1H), 5.26 (d, J=9.3 Hz, 2H), 5.73-5.86 (m, 1H), 7.27-7.49 (m,
10H), 7.58 (s, 1H), 8.31 (s, 1H), 8.99 (d, J=9.34 Hz, 1H). To a
stirred solution of C15 (0.30 g/0.3 mmol) in 5 mL of 9:1
acetone/water was added 0.138 g (1.18 mmol)
N-methylmorpholine-N-oxide (NMO) followed by 0.746 g (0.09 mmol) of
osmium tetroxide and the resulting mixture stirredl6 hours at room
temperature. Another addition of NMO and osmium tetroxide were then
added and mixture stirred an additional 24 hours, at which time the
reaction was complete by LCMS. The reaction was filtered through
Celite, concentrated in vacuo then purified by column
chromatography (silica; 30 to 100% ethylacetate in heptane then
switched to 3 to 7% methanol in dichloromethane collecting 0.302 g
of C16. LCMS m/z 1051.3 (M+1). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 1.33-1.38 (m, 9H), 1.42 (s, 6H), 3.20-3.26 (m, 2H),
3.31-3.37 (m, 1H), 3.58-3.69 (m, 2H), 3.94-4.11 (m, 2H), 4.49-4.58
(m, 1H), 4.89 (br. s, 1H), 4.94 (dd, J=5.4 Hz, J=10.1 Hz, 1H),
5.24-5.29 (m, 4H), 7.27-7.48 (m, 10H), 7.52 (s, 1H), 8.31 (s, 1H),
8.98 (d, J=8.7 Hz, 1H). This material was then deprotected in an
analogous manner to that described for C14 for preparing compound 1
in example 1 in order to prepare example 10.
[0117] B. Preparation of C17. In a flame dry flask was placed C15
(0.486 g, 0.48 mmol) dissolved in 8 mL of 3:1 dioxane/water and to
this mixture was then added sodium periodate (0.311 g, 1.43 mmol)
and osmium tetroxide (0.025 g, 0.003 mmol) and the resulting
mixture stirred for 16 hours at room temperature (reaction complete
by LCMS). The reaction mixture was then partitioned between
saturated sodium bicarbonate and ethylacetate. The organic layer
was washed with saturated brine, dried over sodium sulfate,
filtered and concentrated to dryness in vacuo. The crude product
was then purified by column chromatography (silica-gel, 30 to 100%
ethylacetate in heptane then switched to 3 to 7% methanol in
dichloromethane collecting 0.230 g of C17, which appears by .sup.1H
NMR to exist as a hydrate. LCMS m/z 1019.9 (M+1). .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 1.38-1.56 (m, 24H), 3.72 (t, J=4.7 Hz,
1H), 3.84 (t, J=4.1 Hz, 1H), 3.89-3.56 (m, 1H), 4.16-4.22 (m, 2H),
4.45 (t, J=5.3 Hz, 1H), 4.72-4.78 (m, 1H), 5.00-5.09 (m, 1H),
5.19-5.29 (m, 4H), 7.19-7.49 (m, 10H), 7.77 (s, 1H), 8.11 (s, 1H),
8.22 (d, J=9.4 Hz, 1H).
[0118] C. Preparation of C18 and compound 2. To a stirred solution
of C17 (1.22 g, 1.20 mmol) in 10 mL tetrahydrofuran was added
dimethylamine (0.098 g, 1.2 mmol) and 3 equivalents of glacial
acetic acid (0.215 mL, 3.59 mmol) and the resulting solution
stirred at room temperature for 2 hours. Sodium
triacetoxyborohydride (0.532 g, 2.51 mmol) was then added and the
resulting mixture stirred at room temperature for 18 hours. The
reaction mixture was partitioned between saturated sodium
bicarbonate and ethylacetate, the organic layer washed with
saturated brine, dried over sodium sulfate, filtered and
concentrated to dryness in vacuo. The crude product was purified by
column chromatography (silica-gel, 30 to 100% ethylacetate in
heptane then switched to 5 to 10% methanol in dichloromethane)
collecting 0.340 g of C18. LCMS m/z 1048.9 (M+1). .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 1,38-1.54 (m, 24H), 2.44 (s, 1H), 2.85 (s,
6H), 3.40 (br. s, 1H), 3.71-3.77 (m, 1H), 3.90 (t, J=6.0 Hz, 1H),
4.46 (br. s, 2H), 5.03 (s, 2H), 5.18-5.26 (m, 4H), 7.20-7.35 (m,
9H), 7.40 (d, J=7.0 Hz, 2H), 7.65 (s, 1H), 8.21 (s, 1H). Compound 2
was then prepared from C18 by deprotection and HPLC purification in
an analogous manner to that described for compound 1 of Example 1.
LCMS m/z 712.5 (M+1). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
1.39 (s, 6H), 2.50 (s, 6H), 3.30-3.45 (m, assumed 3H, obscured by
water peak), 3.68 (m, 1H), 4.37 (m, 2H), 6.70 (s, 1H), 7.31 (br s,
2H), 7.42 (s, 1H), 8.03 (s, 1H), 8.98 (d, J=7.0 Hz, 1H), 9.99 (br
s, 1H), 10.88 (br s, 1H).
Example 3
Preparation of
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-[({4-[(2R)-2,3-dihydrox-
ypropyl]-3-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)-5-oxo-4,5-d
ihydro-1H-1,2
,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethyli-
denelamino}oxy)-2-methylpropanoic acid, disodium salt (3)
##STR00034##
[0120] Compound 3 was prepared by the procedures depicted in
Schemes 8 to 10 and described in detail below.
##STR00035##
Step 1. Preparation of
544,5-bis(benzyloxy)pyridin-2-yl]-4-[(2R)-2,3-dihydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one (C20)
[0121] A. Preparation of
2-{[4,5-bis(benzyloxy)pyridin-2-yl]carbonyl}-N-[(2R)-2,3-dihydroxypropyl]-
hydrazinecarboxamide (C19). (2R)-3-Aminopropane-1,2-diol (0.291 g,
3.19 mmol) was added to a suspension of C6 (1.0 g, 2.66 mmol) in
tetrahydrofuran (50 mL), and the mixture was heated to 60.degree.
C. for 20 hours. After cooling to room temperature, the suspension
was filtered, and the solid was washed with tetrahydrofuran
(3.times.5 mL) to afford C19 as a white solid. Yield: 1.07 g, 2.29
mmol, 86%. LCMS m/z 467.2 (M+1). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 2.93 (m, 1H), 3.19 (m, 1H), 3.27 (m, 2H),
3.44 (m, 1H), 4.53 (t, J=5.8 Hz, 1H), 4.77 (d, J=4.8 Hz, 1H), 5.33
(s, 4H), 6.31 (t, J=5.5 Hz, 1H), 7.31-7.48 (m, 10H), 7.69 (s, 1H),
8.01 (br s, 1H), 8.28 (s, 1H), 10.04 (br s, 1H).
[0122] B. Preparation of C20. A solution of C19 (3.00 g, 6.43 mmol)
in aqueous potassium hydroxide (1.6 M, 40.2 mL, 64.3 mmol) was
heated at 100.degree. C. for 13 hours, after which it was cooled to
0.degree. C., diluted with water (100 mL) and acidified to pH 7
with concentrated hydrochloric acid. The resulting solid was
filtered and washed with water (3.times.10 mL) to afford C20,
contaminated with about 30% of the hydrolysis product
4,5-bis(benzyloxy)pyridine-2-carboxylic acid. Yield: 2.66 g,
<5.93 mmol, <92%. LCMS m/z 449.2 (M+1) and 336.1 (M+1 for the
hydrolysis product). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
3.28 (m, 2H), 3.70 (m, 1H), 4.05 (dd, half of ABX pattern, J=13.7,
5.0 Hz, 1H), 4.12 (dd, half of ABX pattern, J=13.7, 8.0 Hz, 1H),
4.61 (v br s, 1H), 5.01 (br s, 1H), 5.28 (s, 2H), 5.31 (s, 2H),
7.32-7.48 (m, 10H), 7.58 (s, 1H), 8.32 (s, 1H), 12.03 (br s, 1H).
Selected peaks for hydrolysis product: 5.29 (s), 7.70 (s), 8.28
(s).
##STR00036##
Step 2. Preparation of tert-butyl 2-({[(1
Z)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-4-yl}-2-({(3S)-1-[({4-[(-
2R)-2,3-di hydroxypropyl]-3-(5-hydroxy-4-oxo-1 ,4-di
hydropyridin-2-yl)-5-oxo-4,5-di
hydro-1H-1,2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-
-2-oxoethylidene]amino}oxy)-2-methylpropanoate (C22)
[0123] A. Preparation of tert-butyl
2-({[(1Z)-2-({(3S)-1-[({3-[4,5-bis(benzyloxy)pyridin-2-yl]-4-[(2R)-2,3-di-
hydroxypropyl]-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl}sulfonyl)carbamoyl]-
-2-oxoazetidin-3-yl}amino)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-4-
-yl}-2-oxoethylidene]amino}oxy)-2-methylpropanoate (C21). A mixture
of C20 (4.00 g, 8.92 mmol) in tetrahydrofuran (35 mL) was treated
with 2,2,2-trifluoro-N-methyl-N-(trimethylsilyl)acetamide (MSTFA,
98%, 10.2 mL, 53.7 mmol). After 45 minutes of stirring, the light
yellow milky mixture was concentrated in vacuo at 60.degree. C. for
1 hour, then dried under vacuum at 60.degree. C. for 1.5 hours. In
a separate flask, a suspension of C13 (4.88 g, 9.81 mmol) in
dichloromethane (32 mL) was cooled to 0.degree. C., treated
drop-wise with isocyanatosulfuryl chloride (chlorosulfonyl
isocyanate, 95%, 0.929 mL, 10.7 mmol) and allowed to stir for 30
minutes under ice-cooling. The material derived from C20 was
dissolved in tetrahydrofuran (8 mL), cooled to 0.degree. C. The
ice-cooled C13-containing reaction mixture was then transferred
into this solution via cannula. After stirring at 0.degree. C. for
1 hour, then at room temperature for 1.5 hours, the reaction
mixture was quenched with methanol (5 mL), stirred for 10 minutes
and concentrated in vacuo. The residue was purified by silica gel
chromatography (Gradient: 40-100% ethyl acetate in heptane, then
0-12% methanol in ethyl acetate) to afford C21 as a solid. Yield:
3.85 g, 3.66 mmol, 41%. LCMS m/z 1051.4 (M+1). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 1.38 (s, 9H), 1.39 (s, 6H), 1.46 (s,
9H), 3.3 (obscured by HOD signal), 3.66 (m, 1H), 3.70 (dd, J=6.3,
6.3 Hz, 1H), 4.00-4.13 (m, 2H), 4.56 (m, 1H), 4.93 (m, 2H), 5.29
(s, 2H), 5.30 (s, 2H), 7.25 (s, 1H), 7.31-7.50 (m, 10H), 7.57 (s,
1H), 8.35 (s, 1H), 9.02 (d, J=8.5 Hz, 1H), 11.84 (br s, 1H).
[0124] B. Preparation of C22. A solution of C21 (0.460 g, 0.438
mmol) in tetrahydrofuran (10 mL) and acetic acid (0.1 mL) was
degassed and flushed with nitrogen (3.times.) and treated with Pd
black (134 mg). The mixture was hydrogenated using a Parr shaker
under 36 psi hydrogen at room temperature for 4 hours (reaction
complete by LCMS). The sample was filtered through acid washed
cellulose powder and washed with THF to give a pale red filtrate,
which was concentrated to dryness in vacuo affording 0.382 g (100%)
as a red solid. LCMS m/z 871.8 (M+1). .sup.1H NMR (500 MHz,
DMSO-d.sub.6) .delta. 1.39 (s, 9H), 1.40 (s, 6H), 1.46 (s, 9H),
3.29 (m, 2H), 3.39 (dd, J=6.3, 3.3 Hz, 1H), 3.65 (HOD lump obscures
signal), 3.71 (m, 1H, estimated), 3.94 (m, 2H, estimated), 4.92 (m,
1H), 7.26 (s, 1H), 7.39 (s, 1H), 8.02 (s, 1H), 9.01 (d, J=8.0 Hz,
1H), 11.82 (br s, 1H).
##STR00037##
Step 3. Preparation of
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-[({4-[(2R)-2,3-dihydrox-
ypropyl]-3-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)-5-oxo-4,5-dihydro-1H--
1,2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethy-
lidene]amino}oxy)-2-methylpropanoic acid, disodium salt (3)
[0125] A. Preparation of compound 3. Trifluoroacetic acid (13 mL)
was added to a cooled (0.degree. C.) solution of C22 (2.54 g, 2.91
mmol) in 13 mL of dichloromethane. The reaction mixture was stirred
at room temperature for 2 hours and then transferred slowly via a
teflon cannula to another round bottom flask containing 186 mL of a
2:1 mixture of heptane/methyl-t-butyl ether (MTBE) resulting in a
fine precipitate. The solids were collected by filtration, washed
with heptane/MTBE (2:1) and dried in vacuo affording 1.82 g (88%)
of the trifluoroacetic acid salt of 3 as a rose colored solid. A
portion of this material (2.42 g) was then purified by reverse
phase chromatography using an Isco Rf Chromatography system
employing a RediSep Rf C18 column (130 g), loading the crude
trifluoroacetic acid salt as a solution in dimethylsulfoxide (1.5
mL) in two batches. The gradient was 5% to 30% water (0.1% Formic
acid)/acetonitrile (0.1% Formic acid). The product came off the
column at 15-18% acetonitrile. The fractions were pooled and the
solvent was removed under reduced pressure affording 0.847 g (35%)
of material as a white solid. The solid was sonicated in methanol
(4 times) and solvent was removed (done to remove formic acid). The
.sup.1H NMR confirms the free-form product with a minimal amount of
formic acid. LCMS m/z 715.0 (M+1). .sup.1H NMR (500 MHz,
DMSO-d.sub.6) .delta. 1.42 (s, 3H), 1.43 (s, 3H), 3.28 (m, 2H),
3.38 (dd, J=6.3, 3.4 Hz, 1H), 3.65 (m, 1H), 3.70 (m, 1H), 3.95 (br
d, J=6.5 Hz, 2H), 4.91 (m, 1H), 6.79 (s, 1H), 7.36 (s, 1H), 8.01
(s, 1H), 9.03 (d, J=8.3 Hz, 1H). To a slurry of 1.20 g (1.65 mmol)
of the free-form acid in 30 mL of deionized water at 0.degree. C.
was slowly added 0.277 g (3.30 mmol) of sodium bicarbonate
dissolved in 6 mL of deionized water (solids completely dissolve
upon addition of the sodium bicarbonate solution). The resulting
solution was then frozen and lyophilized affording 1.12 g of the
disodium salt as a light pink lyophile. LCMS m/z 715.6 (M+1).
.sup.1H NMR (500 MHz, D.sub.2O) .delta. 1.31 (s, 3H), 1.32 (s, 3H),
3.44 (dd, 1/2 ABX, J=12.1 Hz, 4.8 Hz, 1H), 3.48 (dd, 1/2 ABX,
J=11.8 Hz, 4.0 Hz, 1H), 3.65 (dd, J=7.3 Hz, 3.3 Hz, 1H), 3.73-3.92
(m, 3H), 4.90 (dd, J=3.2 Hz, 3.2 Hz, 1H), 6.79 (s, 1H), 6.97 (s,
1H), 7.72 (s, 1H).
Example 4
Preparation of
2-({[(1Z)-2-{[(3S)-1-({[4-(2-amino-2-oxoethyl)-3-(5-hydroxy-4-oxo-1,4-dih-
ydropyridin-2-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl]sulfonyl}carbamo-
yl)-2-oxoazetidin-3-yl]amino}-1-(2-amino-1,3-thiazol-4-yl)-2-oxoethylidene-
]amino}oxy)-2-methylpropanoic acid, disodium salt (4)
##STR00038##
[0127] Compound 4 was prepared by the procedures depicted in
Schemes 11 to 13 and described in detail below.
##STR00039##
[0128] Step 1. Preparation of
{3-[4,5-bis(benzyloxy)pyridin-2-yl]-5-oxo-1,5-dihydro-4H-1,2,4-triazol-4--
yl}acetonitrile (C24)
[0129] A. Preparation of
2-{[4,5-bis(benzyloxy)pyridin-2-yl]carbonyl}-N-(cyanomethyl)hydrazinecarb-
oxamide (C23). Aminoacetonitrile (0.11 g, 1.92 mmol) and
triethylamine (0.162 g, 1.60 mmol) were added drop-wise over one
minute to a suspension of C6 (0.60 g, 1.6 mmol) in tetrahydrofuran
(5 mL), and the mixture was heated to 55.degree. C. for 20 hours.
Additional aminoacetonitrile (0.108 g, 1.92 mmol) and triethylamine
(0.162 g, 1.60 mmol) were added and heating was continued at
55.degree. C. for 20 hours. After being cooled to 0.degree. C. with
an ice-bath, the suspension was filtered, and the solid was washed
with tetrahydrofuran, and dried under vacuum to afford C23 as a
solid. Yield: 0.550 g, 1.27 mmol, 79%. LCMS m/z 430.3 (M-1).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.00 (d, J=5.5 Hz, 2H),
5.33 (s, 2H), 5.34 (s, 2H), 7.30-7.48 (m, 10H), 7.70 (s, 1H), 8.27
(s, 1H), 8.38 (br s, 1H), 10.18 (br s, 1H).
[0130] B. Preparation of C24 (Cyclization Method 2).
2,2,2-Trifluoro-N-methyl-N-(trimethylsilyl)acetamide (MSTFA, 98%, 8
mL, 37 mmol) and C23 (0.310 g, 0.728 mmol), were combined in a
microwave tube and heated to 150.degree. C. for 15 minutes. This
process was repeated six times for a combined total of 2.20 g of
C23 employed. The reactions were combined and concentrated in
vacuo, and the residue was purified by silica gel chromatography
(Gradient: 30-50% ethyl acetate in heptane) to afford C24 as a
solid. Yield: 1.1 g, 2.66 mmol, 52%. LCMS m/z 414.2 (M+1). .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 5.17 (s, 2H), 5.32 (s, 2H),
5.34 (s, 2H), 7.29-7.50 (m, 10H), 7.64 (s, 1H), 8.36 (s, 1H), 12.34
(s, 1H).
##STR00040##
Step 2. Preparation of tert-butyl
2-({[(1Z)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-4-yl}-2-{[(3S)-1--
({[4-(cyanomethyl)-3-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)-5-oxo-4,5-d-
ihydro-1H-1,2,4-triazol-1-yl]sulfonyl}carbamoyl)-2-oxoazetidin-3-yl]amino}-
-2-oxoethylidene]amino}oxy)-2-methylpropanoate (C26)
[0131] A. Preparation of tert-butyl
2-({[(1Z)-2-({(3S)-1-[({3-[4,5-bis(benzyloxy)pyridin-2-yl]-4-(cyanomethyl-
)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl}sulfonyl)carbarnoyl]-2-oxoazetid-
in-3-yl}amino)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-4-yl}-2-oxoet-
hylidene]amino}oxy)-2-methylpropanoate (C25). Compound C25 was
prepared according to the general procedure for the synthesis of
C21 in Example 3, except that C24 was used in place of C20. The
crude material was purified by silica gel chromatography (Gradient:
35-75% ethyl acetate in heptane) to afford C25.
[0132] Yield: 1.64 g, 1.61 mmol, 21%. This material was used in the
next step without further purification. LCMS m/z 1016.5 (M+1).
[0133] B. Compound C26. Compound C26 was prepared according to the
general procedure for the synthesis of C22 in Example 3, except
that C25 was used in place of C21, and the reaction was
hydrogenated at 25 psi for 1.5 hours to afford C19 as a brown
solid. Yield: 0.635 g, 0.759 mmol, 98%. LCMS m/z 836.3 (M+1).
##STR00041##
Step 3. Preparation of 4
[0134] A. Preparation of
2-({[(1Z)-2-{[(3S)-1-({[4-(2-amino-2-oxoethyl)-3-(5-hydroxy-4-oxo-1,4-dih-
ydropyridin-2-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl]sulfonyl}carbamo-
yl)-2-oxoazetidin-3-yl]amino}-1-(2-amino-1,3-thiazol-4-yl)-2-oxoethylidene-
]amino}oxy)-2-methylpropanoic acid (C27). Compound C27 was prepared
according to the general procedure for the synthesis of 3 in
Example 3, except that C26 was used in place of C22. Also, the
crude trifluoroacetic acid salt of compound 4 was generated by
evaporation of the trifluoroacetic acid/dichloromethane solution as
opposed to using the precipitation technique employed in example 3.
The crude product was dissolved in dimethyl sulfoxide to a
concentration of 100 mg/mL, filtered, and purified by preparative
HPLC (column: Waters Symmetry C8, 5 .mu.m, 30.times.50 mm; Solvent
A: 0.1% aqueous formic acid; Solvent B: 0.1% formic acid in
acetonitrile. Gradient: 3% to 22% B). The fractions that pertained
to the desired product were combined, cooled to -78.degree. C. and
lyophilized to provide C27 as a pink solid. Yield: 0.078 g, 0.11
mmol, 12%. LCMS m/z 698.9 (M+1). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 1.38 (br s, 6H), 3.33 (m, 1H), 3.65 (m, 1H),
4.61 (s, 2H), 4.88 (m, 1H), 6.74 (br s, 1H), 7.03 (br s, 1H), 7.30
(s, 1H), 7.89 (s, 1H), 8.99 (d, J=7.42 Hz, 1H).
[0135] B. Preparation of 4. A solution of C27 (78 mg, 0.11 mmol) in
a mixture of acetonitrile (5 mL) and water (45 mL) was cooled to
0.degree. C. and sodium bicarbonate (18.8 mg, 0.224 mmol) was
added. The mixture was vigorously stirred for ten minutes at
0.degree. C. The suspension was then cooled to -78.degree. C.
(using a dry ice/acetone bath) and lyophilized to afford 4 as a
pink solid. Yield: 0.079 g, 0.106 mmol, 95%. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 1.42 (s, 3H), 1.50 (s, 3H), 3-3.5 ppm
obscured by water peak, 3.78 (m, 1H), 4.57 (d, J=16.4 Hz, 1H), 4.72
(d, J=16.4 Hz, 1H), 5.15 (m, 1H), 6.78 (s, 1H), 6.99 (br. s, 1H),
7.18 (br s, 3H), 7.38 (br s, 1H), 7.41 (s, 1H), 7.81 (s, 1H).
Example 5
Preparation of 2-({[(1
Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-1-[({4-[(2S)-2,3-dihydroxvprop-
yl]-3-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)-5-oxo-4,5-dihydro-1H-1,2,4-
-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethyliden-
e]amino}oxy)-2-methylpropanoic acid, disodium salt (5)
##STR00042##
[0137] Compound 5 was prepared by the procedures depicted in
Schemes 14 to 16 and described in detail below.
##STR00043##
Step 1. Preparation of
5-[4,5-bis(benzyloxy)pyridin-2-yl]-4-[(2S)-2,3-dihydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one (C29)
[0138] A. Preparation of
2-{[4,5-bis(benzyloxy)pyridin-2-yl]carbonyl}-N-[(2S)-2,3-dihydroxypropyl]-
hydrazinecarboxamide (C28). Compound C28 was prepared according to
the general procedure for the synthesis of C19 in Example 3, except
that (2S)-3-aminopropane-1,2-diol was used in place of
(2R)-3-aminopropane-1,2-diol. Compound C28 was obtained as a yellow
solid. Additional product was obtained by removing the solvent from
the filtrate in vacuo to afford a yellow solid (8.58 g), which was
slurried in tetrahydrofuran (50 mL), heated to reflux and then
filtered to afford a second crop of C28. The combined yield for C28
was 16.73 g, 35.88 mmol, 90%. LCMS m/z 467.2 (M+1). .sup.1H NMR
(500 MHz, DMSO-d.sub.6) .delta. 2.91 (m, 1H), 3.20 (m, 1H), 3.28
(m, 2H), 3.44 (m, 1H), 5.31 (s, 2H), 5.32 (s, 2H), 6.49 (m, 1H),
7.31-7.48 (m, 10H), 7.69 (s, 1H), 8.25 (s, 1H),
[0139] B. Preparation of C29. Compound C29 was prepared according
to the general procedure for the synthesis of C20 in Example 3,
except that C28 was used in place of C19. The crude product was
heated with methanol (100 mL), the hot mixture was filtered, and
the filtrate concentrated to 20 mL. The resulting solid was
collected by filtration to afford C29. Yield: 150 mg, 0.334 mmol,
22%. LCMS m/z 449.2 (M+1). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 3.28 (m, 2H), 3.70 (m, 1H), 4.09 (t, J=5.8 Hz, 1H), 5.01
(d, J=5.4 Hz, 1H), 5.27 (s, 2H), 5.31 (s, 2H), 7.32-7.49 (m, 10H),
7.58 (s, 1H), 8.32 (s, 1H), 12.03 (br s, 1H).
##STR00044##
Step 2. Preparation of tert-butyl 2-({[(1
Z)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-4-yl}-2-({(3S)-1-[({4-[(-
2S)-2,3-dihydroxypropyl]-3-(5-hydroxy-4-oxo-1
,4-dihydropyridin-2-yl)-5-oxo-4,5-dihydro-1H-1
,2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethy-
lidene]amino}oxy)-2-methylpropanoate (C31)
[0140] A. Preparation of tert-butyl
2-({[(1Z)-2-({(3S)-1-[({3-[4,5-bis(benzyloxy)pyridin-2-yl]-4-[(2S)-2,3-di-
hydroxypropyl]-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl}sulfonyl)carbarnoyl-
]-2-oxoazetidin-3-yl}arnino)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-
-4-yl}-2-oxoethylidene]amino}oxy)-2-methylpropanoate (C30).
Compound C30 was prepared according to the general procedure for
the synthesis of C21 in Example 3, except that C29 was used in
place of C20. After the reaction was quenched with methanol and
concentrated in vacuo, the residue was purified by silica gel
chromatography (Gradient: 25-100% ethyl acetate in heptane, then
0-7% methanol in ethyl acetate) to afford C30 as a solid. Yield:
5.41 g, 5.14 mmol, 53%. LCMS m/z 1051.7 (M+1). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 1.38 (s, 9H), 1.40 (s, 3H), 1.40 (s,
3H), 1.46 (s, 9H), 3.28 (m, 2H), 3.39 (dd, J=6.1, 3.2 Hz, 1H), 3.68
(m, 2H), 4.03 (m, 1H), 4.11 (m, 1H), 4.92 (m, 1H), 5.28 (s, 2H),
5.30 (s, 2H), 7.25 (s, 1H), 7.31-7.50 (m, 10H), 7.58 (s, 1H), 8.35
(s, 1H), 9.00 (d, J=8.3 Hz, 1H).
[0141] B. Preparation of C31. Compound C31 was prepared according
to the general procedure for the synthesis of C22 in Example 3,
except that C30 was used in place of C21, and the reaction was
hydrogenated at 25 psi for 1.5 hour to afford C31 as a red solid.
Yield: 3.49 g, 4.00 mmol, 95%. LCMS m/z 871.6 (M+1).
##STR00045##
Step 3. Preparation of 2-({[(1Z)-1-(2-amino-1
,3-thiazol-4-yl)-2-({(3S)-1-[({4-[(2S)-2,3-dihydroxypropyl]-3-(5-hydroxy--
4-oxo-1,4-dihydropyridin-2-yl)-5-oxo-4,5-dihydro-1 H-1
,2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethy-
lidene]amino}oxy)-2-methylpropanoic acid, disodium salt (5)
[0142] A. Preparation of
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-[({4-[(2S)-2,3-dihydrox-
ypropyl]-3-(5-hydroxy-4-oxo-1
,4-dihydropyridin-2-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl}sulfonyl)-
carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethylidene]amino}oxy)-2-methylpr-
opanoic acid C32. Compound C32 was prepared according to the
general procedure for the synthesis of 3 in Example 3, except that
C31 was used in place of C22. The crude product was dissolved in
dimethyl sulfoxide to a concentration of 100 mg/mL, filtered, and
purified by preparative HPLC (column: Waters Symmetry C8, 5 .mu.m,
30.times.50 mm; Solvent A: 0.1% aqueous formic acid; Solvent B:
0.1% formic acid in acetonitrile. Gradient: 3% to 23% B). The
fractions that pertained to the desired product were concentrated
in vacuo, keeping the water bath <30.degree. C., to provide a
solid. This solid was dissolved in a mixture of acetonitrile (10
mL) and water (100 mL), cooled to -78.degree. C. and lyophilized to
provide C32 as a pink solid. Yield: 0.155 g, 0.217 mmol, 9%. LCMS
m/z 715.2 (M+1). .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 1.42
(s, 3H), 1.43 (s, 3H), 3.27 (m, 2H), 3.37 (dd, J=6.1, 3.1 Hz, 1H),
3.65 (m, 1H), 3.70 (m, 1H), 3.95 (m, 2H), 4.91 (m, 1H), 6.81 (s,
1H), 7.37 (s, 1H), 8.01 (s, 1H), 9.04 (d, J=8.3 Hz, 1H). HPLC
analysis: Hewlett Packard 1100; Column: Waters Symmetry C8, 5
.mu.M, 4.6.times.50 mm; Flow rate 1.2 mL/min; Solvent A: 0.1%
aqueous formic acid; Solvent B: 0.1% formic acid in acetonitrile;
Gradient: 5% to 100% B over 6 minutes; Injection volume: 15 uL;
Detection: 254 nm; Retention time: 3.46 min.
[0143] B. Preparation of compound 5. Compound 5 was prepared
according to the general procedure for the synthesis of 4 in
Example 4, except that C32 was used in place of C27, to afford 5 as
a pink solid. Yield: 0.155 g, 0.204 mmol, 97%. LCMS m/z 715.2
(M+1). .sup.1H NMR (500 MHz, D.sub.2O) .delta. 1.40 (s, 3H), 1.42
(s, 3H), 3.49 (dd, half of an ABX pattern, J=12.2, 4.9 Hz, 1H),
3.57 (dd, half of an ABX pattern J=12.2, 3.7 Hz, 1H), 3.74 (m, 1H),
3.88 (m, 1H), 3.98 (m, 3H), 5.03 (m, 1H), 6.90 (s, 1H), 7.02 (s,
1H), 7.80 (s, 1H). HPLC analysis: Hewlett Packard 1100; Column:
Waters Symmetry C8, 5 .mu.M, 4.6.times.50 mm; Flow rate 1.2 mL/min;
Solvent A: 0.1% aqueous formic acid; Solvent B: 0.1% formic acid in
acetonitrile; Gradient: 5% to 100% B over 6 minutes; Injection
volume: 15 uL; Detection: 254 nm; Retention time: 3.44 min.
Example 6
Preparation of
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-[({3-(5-hydroxy-4-oxo-1-
,4-dihydropyridin-2-yl)-4-[(2R)-2-hydroxypropyl]-5-oxo-4,5-dihydro-1H-1,2,-
4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethylide-
nelamino}oxy)-2-methylpropanoic acid, disodium salt. (6)
##STR00046##
[0145] Compound 6 was prepared by the procedures depicted in
Schemes 17 to 19 and described in detail below.
##STR00047##
Step 1. Preparation of
5-[4,5-bis(benzyloxy)pyridin-2-yl]-4-[(2R)-2-hydroxypropyl]-2,4-dihydro-3-
H-1,2,4-triazol-3-one (C34)
[0146] A. Preparation of
2-{[4,5-bis(benzyloxy)pyridin-2-yl]carbonyl}-N-[(2R)-2-hydroxypropyl]hydr-
azinecarboxamide (C33). Compound C33 was prepared according to the
general procedure for the synthesis of C19 in Example 3, except
that (2R)-1-aminopropan-2-ol was used in place of
(2R)-3-aminopropane-1,2-diol, and the reaction was heated for 60
hours, to provide C33 as a white solid. Yield: 4.54 g, 10.1 mmol,
84%. LCMS m/z 451.2 (M+1). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 1.00 (d, J=6.2 Hz, 3H), 2.91 (m, 1H), 3.01 (m, 1H), 3.61
(m, 1H), 4.64 (d, J=4.7 Hz, 1H), 5.33 (br s, 4H), 6.28 (dd, J=5.8
Hz, 1H), 7.31-7.49 (m, 10H), 7.69 (s, 1H), 7.93 (br s, 1H), 8.28
(s, 1H), 9.97 (br s, 1H).
[0147] B. Preparation of C34. Compound C34 was prepared according
to the general procedure for the synthesis of C20 in Example 3,
except that C33 was used in place of C19. After the solid was
filtered, it was recrystallized from methanol (250 mL) to obtain
two combined crops of C34. Yield: 36.5 g, 84.4 mmol, 74%. LCMS m/z
433.6 (M+1). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 0.98 (d,
J=6.3 Hz, 3H), 3.81 (m, 1H), 3.96 (dd, half of an ABX pattern,
J=13.3, 5.3 Hz, 1H), 4.05 (dd, half of an ABX pattern, J=13.3, 7.4
Hz, 1H), 4.84 (d, J=5.1 Hz, 1H), 5.28 (s, 2H), 5.31 (s, 2H),
7.32-7.49 (m, 10H), 7.59 (s, 1H), 8.32 (s, 1H), 11.96 (br s,
1H).
##STR00048##
Step 2. Preparation of tert-butyl
2-({[(1Z)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-th
iazol-4-yl}-2-({(3S)-1-[({3-(5-hyd
roxy-4-oxo-1,4-dihydropyridin-2-yl)-4-[(2R)-2-hydroxypropyl]-5-oxo-4,5-di-
hydro-1H-1,2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)--
2-oxoethylidene]amino}oxy)-2-methylpropanoate (C36)
[0148] A. Preparation of tert-butyl
2-({[(1Z)-2-({(3S)-1-[({3-[4,5-bis(benzyloxy)pyridin-2-yl]-4-[(2R)-2-hydr-
oxypropyl]-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-o-
xoazetidin-3-yl}amino)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-4-yl}-
-2-oxoethylidene]amino}oxy)-2-methylpropanoate (C35). A suspension
of C34 (1 g, 2.31 mmol) in 1,1,1,3,3,3,-hexamethyldisilazane (2.54
mL, 11.6 mmol) was treated with trimethylsilyl chloride (0.002 mL,
0.012 mmol), and the mixture was heated at 140.degree. C. for 2
hours. The yellow solution was then cooled to room temperature and
concentrated in vacuo to afford a yellow gum. In a separate flask,
a suspension of C13 (1.15 g, 2.31 mmol) in dichloromethane (2 mL)
under nitrogen at 0.degree. C. was treated with carbonylsulfamoyl
chloride (0.211 mL, 2.31 mmol) and stirred for 1.5 hours at
0.degree. C. The mixture became a homogenous solution. The material
derived from C34 was treated with dichloromethane (2 mL), and the
resulting yellow solution was cooled to -40.degree. C. and stirred
under nitrogen. The ice-cooled C13-containing reaction mixture was
transferred into this solution via syringe. The mixture was stirred
at -40.degree. C. for 30 minutes, warmed to room temperature over 1
hour and stirred for 2 hour at room temperature. The mixture was
quenched by the addition of methanol (5 mL), the solvent was
removed in vacuo, and the crude material was purified by silica gel
chromatography (Gradient: 0-3% methanol in ethyl acetate) to afford
C35 as a solid. Yield: 1.42 g, 1.37 mmol, 59%. LCMS m/z 1035.7
(M+1) .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 0.95 (d, J=5.8
Hz, 3H), 1.33-1.43 (m, 15H), 1.46 (s, 9H), 3.40 (m,1H), 3.71 (m,
1H), 3.77 (m, 1H), 3.95 (m, 1H), 4.06 (m, 1H), 4.84 (d, J=5.1 Hz,
1H), 5.29 (s, 2H), 5.31 (s, 2H), 7.25 (s, 1H), 7.31-7.51 (m, 10H),
7.60 (s, 1H), 8.36 (s, 1H), 9.02 (d, J=8.3 Hz, 1H), 11.85 (br s,
1H).
[0149] B. Preparation of C36. Compound C36 was prepared according
to the general procedure for the synthesis of C22 in Example 3,
except that C35 was used in place of C21, and the reaction was
hydrogenated at 25 psi for 1.5 hour, to afford C36 as a red solid.
Yield: 3.84 g, 4.49 mmol, 88%. LCMS m/z 853.0 (M-1).
##STR00049##
Step 3. Preparation of
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-[({3-(5-hydroxy-4-oxo-1-
,4-di
hydropyridin-2-yl)-4-[(2R)-2-hydroxypropyl]-5-oxo-4,5-dihydro-1H-1,2-
,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethylid-
ene]amino}oxy)-2-methylpropanoic acid, disodium salt (6)
[0150] A. Preparation of
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-[({3-(5-hydroxy-4-oxo-1
,4-dihydropyridin-2-yl)-4-[(2R)-2-hydroxypropyl]-5-oxo-4,5-dihydro-1H-1
,2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethy-
lidene]amino}oxy)-2-methylpropanoic acid C37. Compound C37 was
prepared according to the general procedure for the synthesis of 3
in Example 3, except that C36 was used in place of C22. The crude
product was dissolved in dimethyl sulfoxide to a concentration of
100 mg/mL, filtered, and purified by preparative HPLC (column:
Waters Symmetry C8, 5 .mu.m, 30.times.50 mm; Solvent A: 0.1%
aqueous formic acid; Solvent B: 0.1% formic acid in acetonitrile;
Gradient: 6% to 26% B). The fractions that pertained to the desired
product were concentrated in vacuo to provide a solid, which was
dissolved in a mixture of acetonitrile (10 mL) and water (100 mL),
cooled to -78.degree. C. and lyophilized to provide C37 as a pink
solid. Yield: 0.130 g, 0.186 mmol, 15%. LCMS m/z 699.0 (M+1).
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 0.97 (d, J=6.1 Hz, 3H),
1.43 (s, 3H), 1.43 (s, 3H), 3.38 (dd, J=6.3, 3.2 Hz, 1H), 3.69 (dd,
J=6.1, 6.1 Hz, 1H), 3.78 (m, 1H), 3.86 (m, 1H), 4.91 (m, 1H), 6.83
(s, 1H), 7.39 (s, 1H), 8.02 (s, 1H), 9.08 (d, J=8.3 Hz, 1H).
[0151] B. Preparation of 6. Compound 6 was prepared according to
the general procedure for the synthesis of 4 in Example 4, except
that C37 was used in place of C27, and that the starting material
C37 was dissolved in methanol (20 mL), sonicated for five minutes,
and concentrated in vacuo. This process was repeated three times
before the reaction was run. Compound 6 was obtained as a pink
solid. Yield: 0.150 g, 0.202 mmol, 96%. LCMS m/z 699.8 (M+1).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 0.95 (d, J=5.3 Hz, 3H),
1.41 (s, 3H), 1.49 (s, 3H), 3.30-3.40 (m, 1 H, assumed; obscured by
water peak) 3.82 (m, 1H), 3.97 (m, 3H), 5.11 (m, 1H), 6.78 (s, 1H),
7.19 (br s, 1H), 7.36 (s, 1H), 7.88 (s, 1H).
Example 7
Preparation of
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-[({3-(5-hydroxy-4-oxo-1-
,4-di
hydropyridin-2-yl)-4-[(2S)-2-hydroxvpropyl]-5-oxo-4,5-dihydro-1H-1,2-
,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethylid-
ene]amino}oxy)-2-methylpropanoic acid, disodium salt (7)
##STR00050##
[0153] Compound 7 was prepared by the procedures depicted in
Schemes 20 to 22 and described in detail below.
##STR00051##
Step 1. Preparation of
5-[4,5-bis(benzyloxy)pyridin-2-yl]-4-[(2S)-2-hydroxypropyl]-2,4-dihydro-3-
H-1,2,4-triazol-3-one (C39)
[0154] A. Preparation of
2-{[4,5-bis(benzyloxy)pyridin-2-yl]carbonyl}-N-[(2S)-2-hydroxypropyl]hydr-
azinecarboxamide (C38). Compound C38 was prepared according to the
general procedure for the synthesis of C19 in Example 3, except
that (2S)-1-aminopropan-2-ol was used in place of
(2R)-3-aminopropane-1,2-diol, and the reaction was heated for 12
hours, to provide C38 as a white solid. Yield: 12.56 g, 27.88 mmol,
87%. LCMS m/z 451.6 (M+1). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 1.00 (d, J=6.2 Hz, 3H), 2.91 (m, 1H), 3.01 (m, 1H), 3.61
(m, 1H), 4.65 (d, J=4.7 Hz, 1H), 5.33 (s, 4H), 6.28 (dd, J=5.8 Hz,
1H), 7.31-7.49 (m, 10H), 7.69 (s, 1H), 7.93 (br s, 1H), 8.28 (s,
1H), 9.99 (br s, 1H).
[0155] B. Preparation of C39. Compound C39 was prepared according
to the general procedure for the synthesis of C20 in Example 3,
except that C38 was used in place of C19 and the reaction was
heated for 18 hours to afford C39 as a red solid. Yield: 4.25 g,
9.82 mmol, 95%. LCMS m/z 433.3 (M+1). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 0.98 (d, J=6.3 Hz, 3H), 3.85 (m, 1H), 3.98
(m, 1H), 4.07 (m, 1H), 5.28 (s, 2H), 5.31(s, 2H), 7.31-7.49 (m,
11H), 7.60 (s, 1H), 8.32 (s, 1H).
##STR00052##
Step 2. Preparation of tert-butyl
2-({[(1Z)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-4-yl}-2-({(3S)-1--
[({3-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)-4-[(2S)-2-hydroxypropyl]-5--
oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3--
yl}amino)-2-oxoethylidene]amino}oxy)-2-methylpropanoate (C41)
[0156] A. Preparation of tert-butyl
2-({[(1Z)-2-({(3S)-1-[({3-[4,5-bis(benzyloxy)pyridin-2-yl]-4-[(2S)-2-hydr-
oxypropyl]-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl}sulfonyl)carbarnoyl]-2--
oxoazetidin-3-yl}amino)-1-{2-[(tert-butoxycarbonyl)amino]-1,3-thiazol-4-yl-
}-2-oxoethylidene]amino}oxy)-2-methylpropanoate (C40). Compound C40
was prepared according to the general procedure for the synthesis
of C35 in Example 6, except that C39 was used in place of C34.
After the mixture was quenched by the addition of methanol (3 mL),
the solvent was removed in vacuo and the crude material was
purified by silica gel chromatography (Gradient: 0-3% methanol in
ethyl acetate) to afford C40 as a yellow solid. Yield: 0.71 g,
0.685 mmol, 44%. LCMS m/z 1035.6 (M+1). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 0.96 (d, J=6.2 Hz, 3H), 1.38-1.47 (m, assumed
24H), 3.40 (dd, J=6.2, 3.1 Hz, 1H), 3.71 (m, 1H), 3.78 (m, 1H),
3.95 (m, 1H), 4.07 (m, 1H), 4.83 (d, J=5.5 Hz, 1H), 4.91 (m, 1H),
5.29 (s, 2H), 5.31 (s, 2H), 7.25 (s, 1H), 7.31-7.51 (m, 10H), 7.60
(s, 1H), 8.36 (s, 1H), 9.01 (d, J=8.6 Hz, 1H), 11.82 (br s,
1H).
[0157] B. Preparation of C41. Compound C41 was prepared according
to the general procedure for the synthesis of C22 in Example 3,
except that C40 was used in place of C21, and the reaction was
hydrogenated at 30 psi for 1 hour. Additionally, in this case
filtration was carried out through a 1 cm bed of iron-free Celite
(Celite was pre-washed with 1N aqueous hydrochloric acid, then with
deionized water, then with acetone, and then dried). Compound C41
was obtained as a red solid. Yield: 0.630 g, 0.7 mmol, 100%. LCMS
m/z 855.1 (M-1).
##STR00053##
[0158] Step 3. Preparation
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-[({3-(5-hydroxy-4-oxo-1-
,4-dihydropyridin-2-yl)-4-[(2S)-2-hydroxypropyl]-5-oxo-4,5-dihydro-1H-1,2,-
4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino)-2-oxoethylide-
ne]amino}oxy)-2-methylpropanoic acid, disodium salt (7). A solution
of C41 (0.630 g, 0.76 mmol) in dichloromethane (0.5 mL) was cooled
to 0.degree. C. and treated with trifluoroacetic acid (3.4 mL). The
mixture was warmed to room temperature and stirred for 18 hours;
the reaction mixture was then slowly added drop-wise to a stirring
solution of methyl-tert-butyl ether (10 mL) and heptane (20 mL).
The resulting solid was filtered, dried in vacuo, dissolved into
dimethyl sulfoxide (1 mL) and purified via reverse phase
chromatography (RediSep RF C.sub.18 Column, 65 g; Solvent A: 0.1%
aqueous formic acid; Solvent B: 0.1% formic acid in acetonitrile;
Gradient: 5% to 25%B). The fractions that pertained to the desired
product were concentrated in vacuo to provide a solid. The solid
was sonicated in methanol and the solvent was removed (this was
carried out 4 times) to give free form material as a white solid.
Yield: 0.103 g, 0.147 mmol, 19%. LCMS m/z 699.0 (M+1). .sup.1H NMR
(500 MHz, DMSO-d.sub.6) .delta. 0.95 (d, J=6.1 Hz, 3H), 1.43 (s,
3H), 1.44 (s, 3H), 3.38 (dd, J=6.5, 3.3 Hz, 1H), 3.70 (dd, J=6.1,
6.1 Hz, 1H), 3.77 (m, 1H), 3.86 (m, 2H), 4.91 (m, 1H), 6.82 (s,
1H), 7.38 (s, 1H), 8.02 (s, 1H), 9.06 (d, J=8.8 Hz, 1H). Combined
batches of free form (0.676 g, 0.92 mmol) were placed round bottom
flask with 10 mL of deionized water. The suspension was cooled to
0.degree. C. in an ice bath and to this mixture added (dropwise) a
solution of 0.154 g of sodium bicarbonate in 1.0 mL of water. The
suspension was stirred until all the solids were dissolved. The
solution was then frozen and lyophilized affording 0.680 g of
compound 7 as a light pink solid. LCMS m/z 699.6 (M+1). .sup.1H NMR
(500 MHz, D.sub.2O-d.sub.6) .delta. 1.01(d, J=8.5 Hz, 3H), 1.32 (d,
J=6.0 Hz, 6H), 3.61-3.70 (m, 2.5H), 3.77 (dd, 1/2 ABX, J=18.5 Hz,
4.0 Hz, 0.5H), 3.88 (t, J=8.0 Hz, 1H), 4.90 (dd, J=7.5 Hz, 4.5 Hz,
1H), 6.80 (s, 1H), 6.93 (s, 1H), 7.71 (s, 1H).
Example 8
Preparation of
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-{(3S)-1-[({4-[(1,5-dimethyl-1H-p-
yrazol-3-yl)methyl]-3-(5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)-5-oxo-4,5--
dihydro-1H-1,2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-oxoazetidin-3-yl}amino-
)-2-oxoethylidenelamino}oxy)-2-methylpropanoic acid (8)
##STR00054##
[0160] Compound 8 was prepared by the procedures depicted in scheme
23 and outlined in detail below.
##STR00055##
[0161] A. Preparation of C43 (Coupling Method 2). Compound C42 was
prepared in an analogous manner to that described for the
preparation of C8 in Example 1 affording 0.67 g (0.72 mmol) of
triazalone C42 as a white solid. LCMS m/z 483.4 (M+1). A suspension
of C42 (0.100 g, 0.207 mmol) in hexamethyldisilazide (0.227 mL,
1.04 mmol) under nitrogen at 23.degree. C. was treated with
trimethylsilylchloride (one drop, 0.13 uL, 0.001 mmol). The mixture
was heated at 140.degree. C. for 2 hours; upon heating the reaction
became a clear brown solution. The mixture was cooled to room
temperature and held under high vacuum for 1 hour producing a brown
glass-like material. Separately, a suspension of C13 (0.103 g,
0.207 mmol) in dichloromethane (0.1 mL) under nitrogen at 0.degree.
C. was treated with chlorosulfonylisocyanate (0.019 mL, 0.207 mmol)
and stirred until the mixture became homogeneous (approximately 5
minutes). The solution was stirred for 1.5 hours at 0.degree. C.
The silylated triazolinone (0.207 mmol) was treated with DCM (0.2
mL), the brown solution was cooled to -40.degree. C. and stirred
under nitrogen. The prepared sulfamoyl chloride solution (0.1 mL,
0.207 mmol) was then transferred via syringe to the complex
prepared from C42 and the mixture stirred at -40.degree. C. for 30
minutes, warmed to room temperature over 1 hour and stirred for 2
hours at room temperature. The mixture was quenched by the addition
of methanol (0.5 mL), the solvent was removed in vacuo and the
crude material purified by column chromatography (silica-gel, 5%
methanol in dichloromethane) to give 0.050 g (22%) of C43. LCMS m/z
1085.1 (M+1). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 1.34-1.48
(m, 24H), 2.06-2.10 (m, 1H), 2.25-2.28 (m, 1H), 3.40-3.44 (m, 1H),
3.55 (s, 1H), 4.92 (br. s, 1H), 5.20-5.63 (m, 4H), 5.56-5.63 (m,
1H), 7.25 (d, J=4.9 Hz, 1H), 7.28-7.50 (m, 10H), 7.58 (s, 1H), 8.33
(s, 1H), 9.01 (br. s, 2H).
[0162] B. Preparation of
2-({[(1Z)-1-(2-amino-1,3-thiazol-4-yl)-2-({(3S)-1-[({4-[(1,5-dimethyl-1H--
pyrazol-3-yl)methyl]-3-(5-hydroxy-4-oxo-1,4-dihydropyrid
in-2-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl}sulfonyl)carbamoyl]-2-ox-
oazetidin-3-yl}amino)-2-oxoethylidene]amino}oxy)-2-methylpropanoic
acid (8). Compound C43 was deprotected and HPLC purified in an
analogous manner to that described for compound 1 in Example 1
affording 0.015 g (9%) of compound 8 as a pink solid. LCMS m/z
748.9 (M+1). .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 1.41 (s,
3H), 1.41 (s, 3H), 2.39 (s, 1H), 3.55 (s, 3H), 3.67 (m, 1H), 3.91
(dd, J=6.3, 6.3 Hz, 1H), 5.05 (m, 1H), 5.58 (s, 2H), 6.73 (s, 1H),
7.26 (s, 1H), 7.34 (br s, 1H), 7.80 (s, 1H), 9.10 (d, J=8.0 Hz,
1H), 11.85 (s, 1H).
[0163] Additional Examples 9-30 are shown below in Table A and were
prepared analogously to the Examples above using either cyclization
method 1 (described in Example 1, step 3B) or cyclization method 2
(described in Example 4, step 1 B), in combination with either
coupling method 1 (described in Example 1, step 6A) or coupling
method 2 (described in Example 4, step 1B).
TABLE-US-00001 TABLE A .sup.1H NMR 400 MHz, DMSO-d.sub.6
Cyclization (unless otherwise Method, indicated); Ex. Coupling
Observed MS Ion No. Method Structure IUPAC Name (m/z) 9 1.1
##STR00056## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-{[(3S)-1-({[3- (5-hydroxy-4-oxo-1,4-
dihydropyridin-2-yl)-5-oxo-4- propyl-4,5-dihydro-1H-1,2,4-
triazol-1- yl]sulfonyl}carbamoyl)-2- oxoazetidin-3-yl]amino}-2-
oxoethylidene]amino}oxy)-2- methylpropanoic acid 0.76 (t, J = 7.5
Hz, 3H), 1.42 (br s, 6H), 1.51 (m, 2H), 3.37 (dd, J = 6.2, 3.3 Hz,
1H), 3.69 (dd, J = 6.6, 6.6 Hz, 1H), 3.94 (t, J = 7.5 Hz, 2H), 4.91
(m, 1H), 6.81 (s, 1H), 7.34 (s, 1H), 8.01 (s, 1H), 9.06 (d, J = 7.9
Hz, 1H) 10 1.1 ##STR00057## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-{[(3S)-1-({[4- (2,3-dihydroxypropyl)-3-(5-
hydroxy-4-oxo-1,4- dihydropyridin-2-yl)-5-oxo-
4,5-dihydro-1H-1,2,4-triazol- 1-yl]sulfonyl}carbamoyl)-2-
oxoazetidin-3-yl]amino}-2- oxoethylidene]amino}oxy)-2-
methylpropanoic acid 1.43 (s, 6H), 3.28 (m, 2H), 3.38 (m, 1H), 3.65
(m, 1H), 3.70 (dd, J = 6.2, 6.2 Hz, 1H), 3.93 (m, 2H), 4.91 (m,
1H), 6.82 (s, 1H), 7.38 (s, 1H), 8.02 (s, 1H), 9.06 (d, J = 7.9 Hz,
1H); 715.5 (M + 1) 11 1.1 ##STR00058## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-{[(3S)-1-({[3- (5-hydroxy-4-oxo-1,4-
dihydropyridin-2-yl)-5-oxo-4- (2-pyrrolidin-1-ylethyl)-4,5-
dihydro-1H-1,2,4-triazol-1- yl]sulfonyl}carbamoyl)-2-
oxoazetidin-3-yl]amino}-2- oxoethylidene]amino}oxy)-2-
methylpropanoic acid Selected peaks; 1.38 (s, 3H), 1.39 (s, 3H),
1.78 (m, 2H), 1.89 (m, 2H), 3.15-3.36 (m, assumed 7H, obscured by
water peak), 3.58 (m, 2H), 4.13 (m, 1H), 5.68 (br s, 1H), 6.06 (br
s, 1H), 6.86 (s, 1H), 7.30 (br s, 2H), 8.85 (br s, 1H); 736.1 (M -
1) 12 1.1 ##STR00059## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-({(3S)-1-[({4- [2-(diethylamino)ethyl]-3-(5-
hydroxy-4-oxo-1,4- dihydropyridin-2-yl)-5-oxo-
4,5-dihydro-1H-1,2,4-triazol- 1-yl}sulfonyl)carbamoyl]-2-
oxoazetidin-3-yl} amino)-2- oxoethylidene]amino}oxy)-2-
methylpropanoic acid (MeOD) Selected peaks: 1.27-1.35 (m, assumed
6H), 1.55 (s, 3H), 1.56 (s, 3H), 3.39 (m, 4H), 3.59 (dd, J = 6.6.
6.6 Hz, 1H), 3.66 (t, J = 5.4 Hz, 2H), 3.92(dd, J = 6.6, 6.6 Hz,
1H), 4.51 (t, J = 5.4 Hz, 2H), 5.34 (m, 1H), 6.89 (s, 1H), 7.50 (s,
1H), 8.05 (s, 1H); 740.5 (M + 1) 13 2.1 ##STR00060##
2-({[(1Z)-1-(2-amino-1,3- thiazol-4-yl)-2-{[(3S)-1-({[3-
(5-hydroxy-4-oxo-1,4- dihydropyridin-2-yl)-5-oxo-4-
(2,2,2-trifluoroethyl)-4,5- dihydro-1H-1,2,4-triazol-1-
yl]sulfonyl}carbamoyl)-2- oxoazetidin-3-yl]amino}-2-
oxoethylidene]amino}oxy)-2- methylpropanoic acid 1.41 (s, 3H), 1.41
(s, 3H), 3.36 (dd, J = 6.2, 3.2 Hz, 1H), 3.69 (dd, J = 6.2, 6.2 Hz,
1H), 4.91 (m, 1H), 5.11 (m, 2H), 6.76 (s, 1H), 7.37 (s, 1H), 8.00
(s, 1H), 9.01 (d, J = 8.3 Hz, 1H); 723.1 (M + 1) 14 2.2
##STR00061## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-{[(3S)-1-({[4- (3,3-dimethylbutyl)-3-(5-
hydroxy-4-oxo-1,4- dihydropyridin-2-yl)-5-oxo-
4,5-dihydro-1H-1,2,4-triazol- 1-yl]sulfonyl}carbamoyl)-2-
oxoazetidin-3-yl]amino}-2- oxoethylidene]amino}oxy)-2-
methylpropanoic acid 0.89 (s, 9H), 1.40 (m, 2H), 1.43 (s, 3H), 1.43
(s, 3H), 3.37 (dd, J = 6.4, 3.1 Hz, 1H), 3.70 (dd, J = 6.4, 6.4 Hz,
1H), 4.01 (m, 2H), 4.92 (m, 1H), 6.80 (s, 1H), 7.99 (s, 1H), 9.05
(br d, J = 8.8 Hz, 1H); 725.2 (M + 1) 15 1.2 ##STR00062##
2-({[(1Z)-1-(2-amino-1,3- thiazol-4-yl)-2-{[(3S)-1-({[3-
(5-hydroxy-4-oxo-1,4- dihydropyridin-2-yl)-4-(2-
methoxyethyl)-5-oxo-4,5- dihydro-1H-1,2,4-triazol-1-
yl]sulfonyl}carbamoyl)-2- oxoazetidin-3-yl]amino}-2-
oxoethylidene]amino}oxy)-2- methylpropanoic acid, disodium salt
1.42 (s, 3H), 1.50 (s, 3H), 3.16 (s, 3H), 3.32 (HOD, obscures
region), 3.48 (m, 2H), 3.78 (dd, J = 6.3, 6.3 Hz, 1H), 4.18 (m,
2H), 5.14 (m, 1H), 6.79 (s, 1H), 7.19 (br s, 2H), 7.40 (s, 1H),
7.88 (s, 1H); 699.2 (M + 1) 16 1.2 ##STR00063##
2-({[(1Z)-1-(2-amino-1,3- thiazol-4-yl)-2-({(3S)-1-[({3-
(5-hydroxy-4-oxo-1,4- dihydropyridin-2-yl)-5-oxo-4-
[(2S)-tetrahydrofuran-2- ylmethyl]-4,5-dihydro-1H- 1,2,4-triazol-1-
yl}sulfonyl)carbamoyl]-2- oxoazetidin-3-yl}amino)-2-
oxoethylidene]amino}oxy)-2- methylpropanoic acid Selected peaks:
1.42 (br s, 6H), 3.38 (m, 1 H), 3.48 (t, J = 5.1, 2 H), 3.70 (dd, J
= 6.2, 6.2, 2H), 3.97 (m, 2H), 4.13 (m, 1 H), 4.92 (m, 1 H), 6.82
(s, 1 H), 7.35 (s, 1 H), 8.01 (s, 1H), 9.04 (d, J = 8.0 Hz, 1H);
725.2 (M + 1) 17 1.2 ##STR00064## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-{[(3S)-1-({[3- (5-hydroxy-4-oxo-1,4-
dihydropyridin-2-yl)-4-(3- hydroxypropyl)-5-oxo-4,5-
dihydro-1H-1,2,4-triazol-1- yl]sulfonyl}carbamoyl)-2-
oxoazetidin-3-yl]amino}-2- oxoethylidene]amino}oxy)-2-
methylpropanoic acid (500 MHz) 1.44 (s, 3H), 1.44 (s, 3H), 1.67 (m,
2H), 3.35 (t, J = 6.3 Hz, 2H), 3.38 (dd, J = 6.5, 3.3 Hz, 1H), 3.70
(dd, J = 6.4, 6.4 Hz, 1H), 4.01 (m, 2H), 4.92 (m, 1H), 6.84 (s,
1H), 7.35 (s, 1H), 8.01 (s, 1H), 9.07 (d, J = 8.5 Hz, 1H); 699.0 (M
+ 1) 18 1.2 ##STR00065## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-{[(3S)-1-({[4- (2-hydroxyethyl)-3-(5-
hydroxy-4-oxo-1,4- dihydropyridin-2-yl)-5-oxo-
4,5-dihydro-1H-1,2,4-triazol- 1-yl]sulfonyl}carbamoyl)-2-
oxoazetidin-3-yl]amino}-2- oxoethylidene]amino}oxy)-2-
methylpropanoic acid (500 MHz) 1.40(s, 3H), 1.41 (s, 3H), 3.29-3.38
(m, assumed 4H, obscured by water peak), 3.47 (m, 1H), 4.08 (dd, J
= 6.3, 6.3, 1H), 4.89 (m, 1 H), 6.54 (s, 1H), 6.71 (s, 1H), 7.31
(m, 2H), 8.00 (s, 1H), 9.06 (m, 1H); 685.0 (M + 1) 19 1.2
##STR00066## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-{[(3S)-1-({(3- (5-hydroxy-4-oxo-1,4-
dihydropyridin-2-yl)-5-oxo-4- (tetrahydro-2H-pyran-4-
ylmethyl)-4,5-dihydro-1H- 1,2,4-triazol-1-
yl]sulfonyl}carbamoyl)-2- oxoazetidin-3-yl]amino}-2-
oxoethylidene]amino}oxy)-2- methylpropanoic acid 1.11 (m, 2H), 1.39
(m, 2H), 1.42 (s, 3H), 1.43 (s, 3H), 1.84 (m, 1H), 3.15 (m, 2H),
3.36 (dd, J = 6.4, 3.5 Hz, 1H), 3.68 (dd, J = 6.2, 6.2 Hz, 1H),
3.75 (br d, J = 10 Hz, 2H), 3.97 (d, J = 6.8 Hz, 2H), 4.91 (m, 1H),
6.79 (s, 1H), 7.36 (s, 1H), 8.01 (s, 1H), 9.02 (d, J = 8.4 Hz, 1H);
739.2 (M + 1) 20 1.2 ##STR00067## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-({(3S)-1-[({3- (5-hydroxy-4-oxo-1,4-
dihydropyridin-2-yl)-5-oxo-4- [3-(2-oxopyrrolidin-1-
yl)propyl]-4,5-dihydro-1H- 1,2,4-triazol-1-
yl}sulfonyl)carbamoyl]-2- oxoazetidin-3-yl}amino)-2-
oxoethylidene]amino}oxy)-2- methylpropanoic acid 1.44 (s, 6H), 1.74
(m, 2H), 1.91 (m, 2H), 2.19 (t, J = 8.1 Hz, 2H), 3.17 (t, J = 6.9
Hz, 2H), 3.31 (t, J = 6.9 Hz, 2H), 3.37 (dd, J = 6.6, 3.3 Hz, 1H),
3.70 (dd, J = 6.2, 6.2 Hz, 1H), 3.94 (m, 2H), 4.92 (m, 1H), 6.84
(s, 1H), 7.36 (s, 1H), 8.01 (s, 1H), 9.05 (d, J = 8.8 Hz, 1H);
766.2 (M + 1) 21 1.2 ##STR00068## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-({(3S)-1-[({3- (5-hydroxy-4-oxo-1,4-
dihydropyridin-2-yl)-5-oxo-4- [2-(2-oxoimidazolidin-1-
yl)ethyl]-4,5-dihydro-1H- 1,2,4-triazol-1-
yl}sulfonyl)carbamoyl]-2- oxoazetidin-3-yl}amino)-2-
oxoethylidene]amino}oxy)-2- methylpropanoic acid Product peaks:
1.41 (s, 3H), 1.41 (s, 3H), 3.29-3.56 (m, assumed 4H, obscured by
water peak), 3.77 (m, 1H), 3.82 (m, 2H), 4.15 (t, J = 6.6, 2H),
5.00 (m, 1H), 6.72 (s, 1H), 7.32 (s, 1H), 8.01 (s, 1H), 8.99 (d, J
= 8.2 Hz, 1H); 753.2 (M + 1) 22 1.2 ##STR00069##
2-({[(1Z)-1-(2-amino-1,3- thiazol-4-yl)-2-{[(3S)-1-({[4-
(2-ethoxyethyl)-3-(5-hydroxy- 4-oxo-1,4-dihydropyridin-2-
yl)-5-oxo-4,5-dihydro-1H- 1,2,4-triazol-1-
yl]sulfonyl}carbamoyl)-2- oxoazetidin-3-yl]amino}-2-
oxoethylidene]amino}oxy)-2- methylpropanoic acid 0.93 (t, J = 6.9
Hz, 3H), 1.44 (s, 6H), 3.31 (q, J = 6.9 Hz, 2H), 3.37 (m, 1H), 3.46
(t, J = 5.7 Hz, 2H), 3.70 (dd, J = 5.9, 5.9 Hz, 1H), 4.14 (m, 2H),
4.92 (m, 1H), 6.82 (s, 1H), 7.36 (s, 1H), 8.01 (s, 1H), 9.04 (d, J
= 8.6 Hz, 1H); 713.2 (M + 1) 23 1.2 ##STR00070##
2-({[(1Z)-1-(2-amino-1,3- thiazol-4-yl)-2-({(3S)-1-[({3-
(5-hydroxy-4-oxo-1,4- dihydropyridin-2-yl)-5-oxo-4-
[(2R)-tetrahydrofuran-2- ylmethyl]-4,5-dihydro-1H- 1,2,4-triazol-1-
yl}sulfonyl)carbamoyl]-2- oxoazetidin-3-yl}amino)-2-
oxoethylidene]amino}oxy)-2- methylpropanoic acid Selected peaks:
1.42 (br s, 6H), 3.36 (m, 1 H), 3.48 (t, J = 5.3 Hz, 2 H), 3.70
(dd, J = 6.2, 6.2 Hz, 1H), 3.97 (m, 2H), 4.14(m, 1 H), 4.91 (m, 1
H), 6.78 (br s, 1 H), 7.34 (s, 1 H), 8.01 (s, 1H), 9.02 (d, J = 7.8
Hz, 1H) 24 2.2 ##STR00071## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-{[(3S)-1-({[4- (2-hydroxy-2-methylpropyl)-
3-(5-hydroxy-4-oxo-1,4- dihydropyridin-2-yl)-5-oxo-
4,5-dihydro-1H-1,2,4-triazol- 1-yl]sulfonyl}carbamoyl)-2-
oxoazetidin-3-yl]amino}-2- oxoethylidene]amino}oxy)-2-
methylpropanoic acid 1.00 (s, 6H), 1.42 (s, 6H), 3.37 (m, 1H), 3.69
(dd, J = 6.2, 6.2 Hz, 1H), 3.96 (s, 2H), 4.91 (m, 1H), 6.75 (s,
1H), 7.31 (s, 1H), 7.98 (s, 1H), 8.98 (d, J = 8.6 Hz, 1H); 713.2 (M
+ 1) 25 1.2 ##STR00072## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-{[(3S)-1-({[4- (3-ethoxy-2-hydroxypropyl)-
3-(5-hydroxy-4-oxo-1,4- dihydropyridin-2-yl)-5-oxo-
4,5-dihydro-1H-1,2,4-triazol- 1-yl]sulfonyl}carbamoyl)-2-
oxoazetidin-3-yl]amino}-2- oxoethylidene]amino}oxy)-2-
methylpropanoic acid 1.05 (t, J = 6.9 Hz, 3H), 1.43 (s, 3H), 1.43
(s, 3H), 3.25 (d, J = 5.3 Hz, 2H), 3.36 (q, J = 6.9 Hz, 2H), 3.39
(m, 1H), 3.70 (dd, J = 6.2, 6.2 Hz, 1H), 3.79 (m, 1H), 3.97 (m,
2H), 4.91 (m, 1H), 6.80 (s, 1H), 7.36 (s, 1H), 8.00 (s, 1H), 9.03
(d, J = 8.4 Hz, 1H); 743.2 (M + 1) 26 1.2 ##STR00073##
2-({[(1Z)-1-(2-amino-1,3- thiazol-4-yl)-2-({(3S)-1-[({4-
[(1R)-2-hydroxy-1- methylethyl]-3-(5-hydroxy-4-
oxo-1,4-dihydropyridin-2-yl)- 5-oxo-4,5-dihydro-1H-1,2,4-
triazol-1- yl}sulfonyl)carbamoyl]-2- oxoazetidin-3-yl}amino)-2-
oxoethylidene]amino}oxy)-2- methylpropanoic acid (500 MHz) 1.35 (d,
J = 6.8 Hz, 3H), 1.43 (s, 3H), 1.44 (s, 3H), 3.39 (dd, J = 6.3, 3.2
Hz, 1H), 3.50 (dd, J = 10.9, 5.5 Hz, 1H), 3.71 (dd, J = 6.3. 6.3
Hz, 1H), 3.82 (dd, J = 10.9, 8.9 Hz, 1H), 4.65 (m, 1H), 4.92 (m,
1H), 6.83 (s, 1H), 7.32 (s, 1H), 8.04 (s, 1H), 9.07 (d, J = 8.3 Hz,
1H); 699.0 (M + 1) 27 1.2 ##STR00074## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-({(3S)-1-[({3- (5-hydroxy-4-oxo-1,4-
dihydropyridin-2-yl)-4-[(5- methylisoxazol-3-yl)methyl]-
5-oxo-4,5-dihydro-1H-1,2,4- triazol-1- yl}sulfonyl)carbamoyl]-2-
oxoazetidin-3-yl}amino)-2- oxoethylidene]amino}oxy)-2-
methylpropanoic acid 1.42 (s, 6H), 2.30 (s, 3H), 3.38 (m, 1H),
3.42-3.74(m, assume 1 H, obscured by water peak), 4.92 (m, 1H),
5.35 (s, 2H), 6.05 (s, 1H), 6.78 (s, 1H), 7.35 (s, 1H), 7.93 (s,
1H), 9.01 (d, J = 8.0 Hz, 1H); 736.1 (M + 1) 28 2.2 ##STR00075##
2-({[(1Z)-1-(2-amino-1,3- thiazol-4-yl)-2-{[(3S)-1-({[4-
(3-hydroxy-2,2- dimethylpropyl)-3-(5-hydroxy-
4-oxo-1,4-dihydropyridin-2- yl)-5-oxo-4,5-dihydro-1H-
1,2,4-triazol-1- yl]sulfonyl}carbamoyl)-2-
oxoazetidin-3-yl]amino}-2- oxoethylidene]amino}oxy)-2-
methylpropanoic acid 0.58 (s, 6H), 1.43 (s, 3H), 1.44 (s, 3H), 2.97
(s, 2H), 3.38 (dd, J = 6.3, 3.2 Hz, 1H), 3.70 (dd, J = 6.3, 6.3 Hz,
1H), 4.05 (s, 2H), 4.92 (m, 1H), 6.83 (s, 1H), 7.28 (s, 1H), 8.02
(s, 1H), 9.05 (d, J = 8.4 Hz, 1H); 726.9 (M + 1) 29 1.1
##STR00076## 2-({[(1Z)-1-(2-amino-1,3-
thiazol-4-yl)-2-({(3S)-1-[({3- (5-hydroxy-4-oxo-1,4-
dihydropyridin-2-yl)-4-[2-(3- methylphenyl)ethyl]-5-oxo-
4,5-dihydro-1H-1,2,4-triazol- 1-yl}sulfonyl)carbamoyl]-2-
oxoazetidin-3-yl}amino)-2- oxoethylidene]amino}oxy)-2-
methylpropanoic acid Selected peaks: 1.40 (s, 9H), 2.74 (t, J = 7.5
Hz, 2H), 4.13 (t, J = 8.1 Hz, 2H), 4.86- 4.92 (m, 1H), 6.78- 6.95
(m,3H), 7.07 (t, J = 7.8 Hz, 1H), 7.12 (s, 1H), 8.01 (s, 1H), 9.05
(d, J = 7.8 Hz, 1H); 759.2 (M + 1) 30 2.2 ##STR00077##
2-({[(1Z)-1-(2-amino-1,3- thiazol-4-yl)-2-{[(3S)-1-({[3-
(5-hydroxy-4-oxo-1,4- dihydropyridin-2-yl)-4-(2-
methoxy-1-methylethyl)-5- oxo-4,5-dihydro-1H-1,2,4- triazol-1-
yl]sulfonyl}carbamoyl)-2- oxoazetidin-3-yl]amino}-2-
oxoethylidene]amino}oxy)-2- methylpropanoic acid 1.36 (d, J = 6.5
Hz, 3H), 1.44 (d, J = 3.0 Hz, 6H), 3.38 (dd, J = 6.1 Hz, 3.0 Hz,
1H), 3.41 (dd, J = 10.0 Hz, 5.6 Hz, 1H), 3.71 (t, J = 6.5 Hz, 2H),
3.86 (t, J = 10.0 Hz, 2H), 4.90- 4.95 (m, 1H), 5.03 (br. s, 1H),
6.83 (s, 1H), 7.29 (s, 1H), 8.03 (s, 1H), 9.08 (d, J = 8.7 Hz, 1H);
713.2 (M + 1)
Biological Properties
[0164] In some embodiments, compounds of the invention exhibit a
targeted and effective activity against bacteria. Compounds of the
invention can therefore be used, e.g., for treating and/or
preventing a variety of diseases caused by pathogenic bacteria in
human beings and animals.
[0165] Table 1 below shows in vitro MIC data for specified strains
of Pseudomonas aeruginosa, Klebsiella pneumonia, and Acinetobacter
baumanii. Culture collection strain 1045-06 is resistant to several
classes of known antimicrobial agents including carbapenems,
aminoglycosides and fluoroquinolones, while strains 1000-02 and
3167 are resistant to cephalosporins.
[0166] Strain PA0200 is a derivative of laboratory strain PAO1 that
lacks a functional MexAB-oprM efflux pump. The compounds listed are
highly active against all three of these screening strains
demonstrating their broad activity against gram-negative bacterial
pathogens.
TABLE-US-00002 TABLE 1 MIC of Examples 1-30 MIC MIC MIC MIC
Pseudomonas Pseudomonas Klebsiella Acinetobacter aeruginosa
aeruginosa pneumoniae baumanii PA0200 MexAB- Ex. 1045-06 1000-02
3167 oprM KO No. (mg/mL) (mg/mL) (mg/mL) (mg/mL) 1 N.T. N.T. N.T.
<0.0625 2 N.T. N.T. N.T. 0.188.sup.1 3 0.5 0.5 0.5 N.T. 4 0.5 1
1 N.T. 5 0.25 0.5 0.5 N.T. 6 0.25 0.5 1 N.T. 7 0.25 0.25 1 N.T. 8
32 8 >64.0 N.T. 9 N.T. N.T. N.T. <0.0625 10 N.T. N.T. N.T.
0.25.sup.1 11 N.T. N.T. N.T. 16 12 2 2 16 N.T. 13 0.25 0.25 16 N.T.
14 0.5 0.06 2 N.T. 15 0.5 0.5 4 N.T. 16 0.5 0.5 2 N.T. 17 0.25 0.25
1 N.T. 18 2 1 8 N.T. 19 0.5 0.25 2 N.T. 20 0.5 0.25 2 N.T. 21 4 32
>64.0 N.T. 22 0.5 0.5 2 N.T. 23 0.5 0.25 1 N.T. 24 0.5 0.25 2
N.T. 25 0.5 0.5 4 N.T. 26 0.5 0.125 2 N.T. 27 1 1 2 N.T. 28 2 0.5 4
N.T. 29 2 0.5 >64 N.T. 30 0.5 8 1 N.T. N.T. = Not Tested
.sup.1Value represents average 2 MIC determinations
[0167] Table 2 below shows several compounds of the invention
compared to cefipime (a cephalosporin antibiotic indicated to treat
bacterial infections caused from Pseudomonas aeruginosa), imipenem
(a carbapenem antibiotic used to treat infections caused by P.
aeruginosa) and Comparative Example A (example 23 in EP 0281289,
published Sep. 7, 1988).
TABLE-US-00003 TABLE 2 In vitro and In Vivo Comparison Against P.
aeruginosa ##STR00078## RTI PD.sub.50 vs. Pa 1091-05 Pa (mg/kg)
1091-05 (95% MIC confidence Pa Compound Structure (R.sup.1) (mg/mL)
interval) MIC90.sup.2 Cefepime -- 2 22 64 Imipenem -- 0.5 1.04
>64 Comarative --CH.sub.3 0.5 >150.sup.1 1 Example A Example
15 ##STR00079## 0.5 >70.8 1 Example 6 ##STR00080## 0.125 32.7
(23.7-42.0) 1 Example 7 ##STR00081## 0.125 15.7 (8.45-22.96) 1
Example 3 ##STR00082## 0.125 20.6 (8.7-32.52) 1 Example 5
##STR00083## 0.125 18.6 (8.94-28.29) 1 Example 4 ##STR00084## 0.125
25.0 (24.8-25.2) 1 .sup.1Data are from a previous experiment
.sup.291 clinical isolate
[0168] Table 2 shows the results for compounds of the invention
which were evaluated for efficacy in the murine respiratory tract
infection model against P. aeruginosa 1091-05. For this model,
C3H/HeN mice were immunosuppressed with cyclophosphamide given
orally at 150 mg/kg and 100 mg/kg on days--4 and --1 relative to
challenge, respectively. Mice were anesthetized with isoflurane (5%
in oxygen) and the bacterial inoculum was given to each mouse via
intranasal instillation in a 40 .mu.L volume (-2.8.times.10.sup.3
cfu per mouse). Mice were dosed with compound administered via
subcutaneous injection beginning at four hours post-challenge, and
continuing for two days of BID therapy. Lethalities were followed
over ten days and the 50% protective doses (PD.sub.50s) were
determined. include interpretative comment on The known monocarbam
prototype Comparative Example A (example 23b in EP 0281289,
published Sep. 7, 1988) typically has a PD.sub.50 of 100 mg/kg in
this model. However, the exemplified monocarbams of the present
invention were evaluated in this model and many demonstrated better
efficacy than Comparative Example A, for example, Example 4 (25.0
mg/kg), Example 6 (32.7 mg/kg), Example 5 (18.6 mg/kg), Example 3
(20.6 mg/kg), and Example 7 (15.7 mg/kg).
[0169] PD.sub.50 is a measure of the ability of a compound to
protect mice from a lethal infection. Hence, a lower value in this
study is indicative of improved efficacy. Since the 95% confidence
intervals (the range that predicts where the actual value will lie
with 95% confidence) calculated for the compounds Example 3,
Example 6, Example 4, Example 7 and Example 5 do not overlap with
the PD.sub.50 value determined for Comparative Example A, it can be
concluded that these compounds are significantly more efficacious
relative to Comparative Example A. This result was unexpected given
the similar MICs against the pathogen used (P. aeruginosa 1091-05).
Importantly, performance in these pre-clinical in vivo models is
predictive of outcomes of clinical efficacy against these types of
infections.
* * * * *