U.S. patent application number 12/359490 was filed with the patent office on 2009-06-04 for substituted azetidinones.
This patent application is currently assigned to DAIAMED. Invention is credited to Thomas Bannister, Cassandra Celatka, Nizal S. Chandrakumar, Hongfeng Deng, Zihong Guo, Lei Jin, Tsvetelina Lazarova, Jian Lin, Scott T. Moe, Pamela Nagafuji, Manuel Navia, Amy Ripka, Michael J. Rynkiewicz, Kerry L. Spear, James E. Stickler, Roger Xie.
Application Number | 20090143351 12/359490 |
Document ID | / |
Family ID | 37074064 |
Filed Date | 2009-06-04 |
United States Patent
Application |
20090143351 |
Kind Code |
A1 |
Bannister; Thomas ; et
al. |
June 4, 2009 |
SUBSTITUTED AZETIDINONES
Abstract
Compounds are provided which have the structure ##STR00001##
Wherein A, B, C, D, m, Y, Ra, Rc, Rd, and Rd' are as described
herein, and which are useful as inhibitors of tryptase, thrombin,
trypsin, Factor Xa, Factor VIIa, Factor XIa, and urokinase-type
plasminogen activator and may be employed in preventing and/or
treating asthma, chronic asthma, allergic rhinitis, and thrombotic
disorders.
Inventors: |
Bannister; Thomas; (Palm
Beach Gardens, FL) ; Celatka; Cassandra; (Hull,
MA) ; Chandrakumar; Nizal S.; (North Grafton, MA)
; Deng; Hongfeng; (Acton, MA) ; Guo; Zihong;
(Southbury, CT) ; Jin; Lei; (Wellesley, MA)
; Lazarova; Tsvetelina; (Brookline, MA) ; Lin;
Jian; (Walpole, MA) ; Moe; Scott T.;
(Marlborough, MA) ; Nagafuji; Pamela; (Cambridge,
MA) ; Navia; Manuel; (Lexington, MA) ; Ripka;
Amy; (Winthrop, MA) ; Rynkiewicz; Michael J.;
(Boston, MA) ; Spear; Kerry L.; (Concord, MA)
; Stickler; James E.; (Milton, MA) ; Xie;
Roger; (Southborough, MA) |
Correspondence
Address: |
MORGAN, LEWIS & BOCKIUS LLP (SF)
One Market, Spear Street Tower, Suite 2800
San Francisco
CA
94105
US
|
Assignee: |
DAIAMED
Cambridge
MA
|
Family ID: |
37074064 |
Appl. No.: |
12/359490 |
Filed: |
January 26, 2009 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
11398438 |
Apr 4, 2006 |
7501404 |
|
|
12359490 |
|
|
|
|
60668325 |
Apr 4, 2005 |
|
|
|
Current U.S.
Class: |
514/210.02 ;
540/200; 540/364 |
Current CPC
Class: |
A61P 11/02 20180101;
C07D 401/12 20130101; C07D 405/06 20130101; C07D 205/08 20130101;
A61P 9/00 20180101; A61P 7/02 20180101; A61P 7/00 20180101; C07D
401/04 20130101; C07D 401/06 20130101; C07D 405/14 20130101 |
Class at
Publication: |
514/210.02 ;
540/200; 540/364 |
International
Class: |
A61K 31/397 20060101
A61K031/397; C07D 401/06 20060101 C07D401/06; C07D 205/08 20060101
C07D205/08; A61P 7/00 20060101 A61P007/00; C07D 401/04 20060101
C07D401/04; A61K 31/4427 20060101 A61K031/4427 |
Claims
1. A compound described herein.
2. A pharmaceutical composition comprising an effective amount of a
compound described herein.
3. A method for treating a disease comprising administering a
therapeutically effective amount of a compound described
herein.
4. A method of inhibiting an enzyme in a mammal by administration
of a compound described herein.
5. A method of enhancing thrombolysis or treating thrombosis in a
mammalian species comprising administering an effective amount of
the composition of a compound described herein.
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 11/398,438, filed 4 Apr. 2006, which claims priority of U.S.
Provisional Application Ser. No. 60/668,325, filed 4 Apr. 2005, the
disclosures of all of which are hereby incorporated by reference in
their entirety for all purposes.
BACKGROUND OF THE INVENTION
[0002] Bisacchi in U.S. Pat. No. 6,335,324 explicitly discloses 3
guanidinoalkyl-2-azetidinones which have one of the following two
structures:
##STR00002##
wherein W is an unsubstituted 4-8 membered cycloalkyl ring; Y is
either C.dbd.O or SO.sub.2; Z is either hydrogen or unsubstituted
alkyl; R.sup.H can be any substitutent; and R.sup.2 can be any
substituent.
[0003] Bisacchi in U.S. Pat. Pub. No. 2004/0147502 A1 explicitly
discloses 1-[piperazinecarbonyl]azetidinones of the formula:
##STR00003##
wherein R.sup.3 is any substituent, R.sup.4 is OH, NH.sub.2, alkyl
or heteroalkyl, and R.sup.5 is any substituent.
[0004] Schumacher in U.S. Pat. Pub. No. 2004/0180855 A1 explicitly
discloses methods of treating thrombosis in a mammal comprising
administering a compound of the formula below that is selective for
inhibition of Factor XIa.
##STR00004##
wherein X is COOH, COOR, CONR, unsubstituted alkyl and
unsubstituted arylalkyl, Y is CO or SO.sub.2, Z is H or
unsubstituted alkyl, R.sup.2 is substituted or unsubstituted alkyl,
substituted or unsubstituted aryl, and substituted or unsubstituted
heterocycloalkyl.
[0005] Thrombo-embolic disorders are the largest cause of mortality
(myocardial infarction) and disability (stroke) in the
industrialized world. Arterial thrombosis is initiated by
atherosclerotic plaque rupture, exposure of tissue factor, and
initiation of the coagulation vortex. A number of coagulation
factors are present in the blood as precursors (e.g., Factors
VII-XII), and when the coagulation system is triggered, these
factors undergo a complicated, ordered series of reactions that
ultimately lead to thrombin production. Thrombin is a proteolytic
enzyme that occupies a central position in the coagulation process.
Thrombin catalyzes the conversion of fibrinogen to fibrin, is a key
effector enzyme for blood clotting, and is also pivotal for other
functions. High concentrations of thrombin inhibit fibrinolysis by
activating the Thrombin Activated Fibrinolysis Inhibitor (TAFI),
which can also be activated by modest amounts of thrombin in the
presence of soluble or membrane bound thrombomodulin. TAFIa removes
the C-terminal lysine residues from fibrin, preventing the binding
of t-PA and plasmin and thus, slowing fibrinolysis.
[0006] The complicated coagulation process is initiated by tissue
factor (TF). Tissue factor binds and activates Factor VII (FVII),
which is rapidly converted to activated Factor VIIa (FVIIa) to form
a TF:FVIIa complex. The TF:FVIIa complex activates Factors IX and
X. Factor Xa generates small amounts of thrombin. The small amounts
of thrombin activate Factor V, Factor VIII and platelets,
accelerating thrombin production by Factors IXa and Xa. Activation
of Factor V and FVIII accelerates catalytic activity of FVIIIa:FIXa
and FVa:FXa, resulting in dramatically increased thrombin
production. Another wave of thrombin generation occurs as a result
of thrombin activation of Factor XIa. Factor XI activates more
Factor IX. As the concentration of thrombin increases, more
thrombin is generated, which in turn activates TAFI to then inhibit
fibrinolysis.
[0007] This coagulation process involves an intrinsic pathway and
an extrinsic pathway. In the intrinsic pathway, Factor XII (aka
Hageman Factor) is converted from its inactive form (zymogen) to an
active form, i.e., Factor XIIa. Activated Factor XII enzymatically
activates Factor XI to Factor XIa. Activated Factor XI activates
Factor IXa. Factor IXa then converts Factor X to Factor Xa. FXa
activates prothrombin to thrombin. Thrombin cleaves fibrinogen to
form insoluble fibrin (the clot). In the extrinsic pathway,
addition of thromboplastin (i.e., tissue factor) to plasma
activates Factor VII. This complex, in the presence of calcium ions
and phospholipids, activates Factor X to Factor Xa. Once Factor Xa
is generated, the remainder of the cascade is similar to the
intrinsic pathway. As can be seen, Factor XIa is involved only in
the intrinsic pathway.
[0008] In vitro, the degree to which FXIa contributes to thrombin
generation, platelet activation, and fibrin formation depends on
the concentration of tissue factor. For example, in the absence of
FXI (i.e., in FXIa deficient plasma), plasma stimulated with low
levels of tissue factor (clot formation >10 minutes) showed a
delay in the time required to generate thrombin and form clots. A
FXI deficiency also decreased the amount of thrombin generated and
platelet aggregation in whole blood. However, in blood or plasma
stimulated by higher concentration of tissue factor (clot formation
<5 minutes), a FXI deficiency had no effect on the thrombin
generation or clot formation. Thus, a FXI deficiency will generally
prolong thrombin generation but not in situations where the plasma
is stimulated with high concentrations of tissue factor.
[0009] FXIa, via expanded thrombin generation, also plays a role in
resisting fibrinolysis. Resistance of plasma clots to tPA and
uPA-induced fibrinolysis depends on thrombin concentration
(generated endogenously or added exogenously) in the plasma. The
time required for clot lysis is proportional to the plasma TAFIa
concentrations. However, clot lysis can occur more rapidly, and the
lysis made independent of plasma TAFI concentration, when blocking
antibodies to FXIa are included in the assay.
[0010] Elevated levels of FXIa in the plasma and/or increased
activation of FXIa is associated with various cardiovascular and
other diseases. As an illustration, increased activation of FXIa
occurs in patients with coronary artery disease and is related to
the severity of the disease. Also, Factor IX activation peptide (a
product of FXIa and TF:FVIIa cleavage of FIX) levels have been
found to be significantly higher in patients with acute myocardial
infarction and unstable angina compared with patients with stable
angina. Concentrations of FXIa-.alpha..sub.1AT (FXIa complexed to
the serpin .alpha.1-antitrypsin) were also elevated in patients
with recent myocardial infarction or unstable angina. Patients with
high levels of Factor XI are at risk for deep venous
thrombosis.
[0011] Proteins or peptides that reportedly inhibit Factor XIa are
disclosed in WO 01/27079 to Entremed, Inc. There are advantages in
using small organic compounds, however, in preparing
pharmaceuticals, e.g., small compounds generally have better oral
bioavailability and compatibility in making formulations to aid in
delivery of the drug as compared with large proteins or peptides.
Small organic compounds have been disclosed that reportedly inhibit
coagulation factors besides Factor XIa. For example, compounds
effective in inhibiting Factor Xa are described in U.S. Pat. Nos.
6,344,450 and 6,297,233, and WO 00/47563. Compounds effective in
inhibiting Factors VIIa, Xa, as well as tryptase and urokinase are
described in U.S. Pat. No. 6,335,324. Factor Xa inhibitors are
disclosed in WO 98/57937 to the duPont Merck Pharmaceutical Co.,
and Factor VIIa inhibitors are disclosed in U.S. Pat. No. 6,358,960
to Ono Pharmaceuticals Inc., ("Ono"), and in WO 01/44172 to Axys
Pharm. Inc.
[0012] A possible adverse side effect associated with use of
anti-thrombotic agents for treating cardiovascular diseases
involves the risk of bleeding. For example, heparin is a known
anti-thrombotic agent that has a highly-variable dose-related
response, and its anticoagulant effects must be closely monitored
to avoid a risk of serious bleeding. The erratic anticoagulant
response of heparin is likely due to its propensity to bind
non-specifically to plasma proteins. Aspirin also has been used as
an anti-thrombotic agent but at high doses presents a risk of
gastrointestinal bleeding. Thrombin inhibitors and their drawbacks
are further discussed in WO 96/20689 to duPont Merck Pharmaceutical
Co. Guanidine and beta lactam-containing compounds that are potent
inhibitors of serine proteases including thrombin and tryptase are
described in U.S. Pat. No. 6,335,324, the entire contents of which
is incorporated herein by reference.
SUMMARY OF THE INVENTION
[0013] This invention is directed to the novel .beta.-lactam
compounds of formula I shown below and to a method for the use of
such compounds as inhibitors of various in vivo enzyme systems
including tryptase, thrombin, trypsin, Factor Xa, Factor VIIa,
Factor XIa, and urokinase-type plasminogen activator and their use
in treating and/or preventing asthma and/or allergic rhinitis
and/or thrombotic disorders.
[0014] In a first aspect, the invention provides a compound
according to the following structure:
##STR00005##
In this structure, A is a member selected from CR.sup.1R.sup.2,
NR.sup.1a, O, S and SO.sub.n. The symbol n is an integer selected
from 0 to 2. Each R.sup.1a is a member independently selected from
a negative charge, a salt counterion, H, substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, C(O)R.sup.5,
NR.sup.3R.sup.4, OR.sup.3 and SO.sub.2R.sup.5. R.sup.3 and R.sup.4
are members independently selected from H, substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, C(O)R.sup.6, and
SO.sub.2R.sup.6. R.sup.6 is a member selected from substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl. R.sup.5 is a member
selected from substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl.
R.sup.3 and R.sup.4 can be optionally joined, together with the
atoms to which they are attached, to form a 4-8 membered ring.
R.sup.1 is a member selected from H, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, NR.sup.7R.sup.8, SO.sub.nR.sup.9,
halogen, C(O)R.sup.7, CO.sub.2R.sup.7, C(O)NR.sup.7R.sup.8, and
OR.sup.7. Each R.sup.7 and each R.sup.8 is a member independently
selected from H, substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
C(O)R.sup.10, C(O)OR.sup.10, NR.sup.10R.sup.10a, OR.sup.10,
SO.sub.2R.sup.10 and S(O)R.sup.10 wherein R.sup.10 and R.sup.10a
are members independently selected from H, substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl. R.sup.9 is a member
selected from substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl.
R.sup.7 and R.sup.8 can be optionally joined, together with the
atoms to which they are attached, to form a 4-8 membered ring.
R.sup.2 is a member selected from H, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl and halogen. R.sup.1 and R.sup.2 can
together represent an acyl group. R.sup.1 and R.sup.2 can be
optionally joined together in a 3-8 membered ring. B is a member
independently selected from substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, carboxamido, NR.sup.11, --S--, and --O--.
R.sup.11 is a member selected from H, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, C(O)R.sup.10, C(O)OR.sup.10,
NR.sup.10R.sup.10a, OR.sup.10, SO.sub.2R.sup.10 and S(O)R.sup.10.
The symbol m is an integer selected from 0 to 3. C is a member
selected from a bond, C.dbd.O, SO.sub.2, N.dbd.C, --O-- and
--O--CR.sup.5R.sup.10--. R.sup.5 and R.sup.10 can be optionally
joined, together with the atoms to which they are attached, to form
a 3-8 membered ring. D is a member selected from substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, substituted or unsubstituted amidine,
substituted or unsubstituted amidinohydrazone, substituted or
unsubstituted guanidine, and substituted or unsubstituted amine. A
and B can be optionally joined, together with the atoms to which
they are attached, to form a 3-8 membered ring. A and C can be
optionally joined, together with the atoms to which they are
attached, to form a 3-8 membered ring. A and D can be optionally
joined, together with the atoms to which they are attached, to form
a 3-8 membered ring. B and C can be optionally joined, together
with the atoms to which they are attached, to form a 3-8 membered
ring. B and D can be optionally joined, together with the atoms to
which they are attached, to form a 3-8 membered ring. C and D can
be optionally joined, together with the atoms to which they are
attached, to form a 3-8 membered ring. If m is greater than 2, each
independently selected B can be optionally joined to form a 3-8
membered ring. X is a member selected from S, O, and NR.sup.7. Y is
a member selected from a bond, C=Q, CR.sup.12R.sup.13, and
SO.sub.n. Q is a member selected from S, O, and NR.sup.7. R.sup.12
is a member selected from H, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, and
substituted or unsubstituted heteroaryl, halogen. R.sup.13 is a
member selected from H, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, and
substituted or unsubstituted heteroaryl halogen. R.sup.12 and
R.sup.13 can be optionally joined, together with the atoms to which
they are attached, to form a 4-8 membered ring. R.sub.a is a member
selected from NR.sup.7R.sup.8, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, and
substituted or unsubstituted heteroaryl. R.sup.7 and R.sup.8 can be
optionally joined, together with the atoms to which they are
attached, to form a 4-8 membered ring. R.sub.c is a member selected
from H, substituted or unsubstituted alkyl and halogen. R.sub.d is
a member selected from R.sup.16, (CR.sup.14R.sup.15).sub.pR.sup.16,
C(R.sup.14).dbd.CR.sup.16R.sup.23 and CCR.sup.16. The symbol p is
an integer selected from 0 to 3. R.sup.14 and R.sup.23 are members
independently selected from H, halogen, substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl. R.sup.15 is a member
selected from H, halogen, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl and substituted
or unsubstituted heteroaryl. R.sup.16 is a member selected from
halogen, substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
C(O)R.sup.18, OR.sup.19, --OC(=Q)NR.sup.19R.sup.21,
--NR.sup.19C(=Q)NR.sup.19R.sup.21, --NR.sup.20SO.sub.2R.sup.19,
OSO.sub.2R.sup.19, SO.sub.2R.sup.21, S(O)R.sup.21,
SO.sub.2NR.sup.19R.sup.20, NR.sup.19R.sup.20 and CN. R.sup.18 is a
member selected from substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, OR.sup.21, SR.sup.21, NR.sup.19R.sup.20,
and --NR.sup.19SO.sub.2R.sup.20. R.sup.19 is a member selected from
H, substituted or unsubstituted alkyl, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl and substituted or unsubstituted heteroaryl. R.sup.20 is a
member selected from H, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, C(O)R.sup.22, SO.sub.2R.sup.22. R.sup.22
is a member selected from substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl. R.sup.21 is a member selected from a
negative charge, a salt counterion, H, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, --OR.sup.22 and --SR.sup.22. R.sup.14 and
R.sup.15 can be joined together in a 4-8 membered ring. R.sup.19
and R.sup.21 can be optionally joined, together with the atoms to
which they are attached, to form a 4-8 membered ring. R.sup.14 and
R.sup.23 can be optionally joined, together with the atoms to which
they are attached, to form a 4-8 membered ring. R.sup.19 and
R.sup.20 can be optionally joined, together with the atoms to which
they are attached, to form a 4-8 membered ring. R.sub.d' is a
member selected from H, substituted or unsubstituted alkyl and
halogen. R.sub.c and R.sub.d can be optionally joined, together
with the atoms to which they are attached, to form a 5-8 membered
ring. R.sub.c and R.sub.d' can be optionally joined, together with
the atoms to which they are attached, to form a 5-8 membered
ring.
[0015] In an exemplary embodiment, the compound is subject to one
or more of the following provisos. In an exemplary embodiment, the
structure has the proviso that if
[0016] a) Y is a member selected from C.dbd.O and SO.sub.2; [0017]
ABC, in combination, form unsubstituted alkyl; and [0018] D is
substituted or unsubstituted guanidine; or
[0019] b) Y is a member selected from C.dbd.O and SO.sub.2; [0020]
AB, in combination, form unsubstituted alkyl; and [0021] CD, in
combination, form
[0021] ##STR00006## [0022] wherein the dotted line represents
carbon atoms necessary for the formation of one ring having 4 to 8
atoms; then R.sub.c cannot be a member selected from CH.sub.3,
CH.sub.2CH.sub.3, substituted alkyl, and unsubstituted alkyl; and
R.sub.d and R.sub.d' cannot be members selected from CH.sub.3 and
CH.sub.3; CH.sub.2CH.sub.3 and CH.sub.2CH.sub.3; CH.sub.2CH.sub.3
and CH.sub.3; CH.sub.3 and CH.sub.2CH.sub.3; substituted or
unsubstituted alkyl and substituted or unsubstituted alkyl; H and
substituted or unsubstituted alkyl; substituted or unsubstituted
alkyl and H; H and COOH; H and COOR.sup.1a; H and
CONR.sup.7R.sup.8; unsubstituted alkyl and COOH; unsubstituted
alkyl and COOR.sup.1a; H and unsubstituted phenylalkyl;
unsubstituted alkyl and unsubstituted phenylalkyl.
[0023] In another exemplary embodiment, part (a) of this proviso
is
[0024] a) Y is a member selected from C.dbd.O and SO.sub.2; [0025]
ABC, in combination, form substituted alkyl; and [0026] D is
substituted or unsubstituted guanidine.
[0027] In another exemplary embodiment, R.sub.c of this proviso
cannot be substituted alkyl. In another exemplary embodiment,
R.sub.c of this proviso cannot be unsubstituted alkyl.
[0028] In another exemplary embodiment, R.sub.d and R.sub.d' of
this proviso cannot be members selected from CH.sub.3 and CH.sub.3;
CH.sub.2CH.sub.3 and CH.sub.2CH.sub.3; CH.sub.2CH.sub.3 and
CH.sub.3; CH.sub.3 and CH.sub.2CH.sub.3; H and COOH; H and
COOR.sup.1a; H and CONR.sup.7R.sup.8; unsubstituted alkyl and COOH;
unsubstituted alkyl and COOR.sup.1a; H and unsubstituted
phenylalkyl; unsubstituted alkyl and unsubstituted phenylalkyl.
[0029] In another exemplary embodiment, the structure has the
following proviso: if the compound has the following structure:
##STR00007##
wherein R.sup.30 is a member selected from H, SO.sub.2--R.sup.31,
substituted or unsubstituted alkyl, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl, and substituted or unsubstituted heteroaryl; wherein R.sup.31
is unsubstituted alkyl, then at least one of R.sub.d and R.sub.d'
does not comprise a carbonyl group which is directly attached, or
alpha, to the azetidinone ring. In another exemplary embodiment, at
least one of R.sub.d and R.sub.d' does not comprise a carbonyl
group. In another exemplary embodiment, R.sup.30 does not comprise
a carbon atom.
[0030] In another exemplary embodiment, the structure has the
following proviso: the compound cannot have the following
structure:
##STR00008##
wherein at least one of R.sub.d and R.sub.d' comprise a carbonyl
group which is directly attached, or alpha, to the azetidinone
ring. In another exemplary embodiment, the compound cannot have the
following structure:
##STR00009##
wherein DCB.sub.mA comprises a piperidine ring or a salt thereof.
In another exemplary embodiment, the compound cannot have the
following structure:
##STR00010##
wherein DCB.sub.mA comprises a piperidine ring or a salt
thereof.
[0031] In another exemplary embodiment, the structure has the
following proviso: the compound cannot have the following
structure:
##STR00011##
wherein R.sub.a is NHR.sup.8, and R.sup.8 includes a substituted or
unsubstituted phenyl. In another exemplary embodiment, R.sup.8 is
unsubstituted phenyl. In yet another exemplary embodiment, R.sub.a
is N(H)(unsubstituted phenyl). In another exemplary embodiment, the
structure has the following proviso: the compound cannot have the
following structure:
##STR00012##
[0032] In another exemplary embodiment, the structure has the
following proviso: the compound cannot be a chemical compound which
is explicitly disclosed (ie does not contain any variables) in U.S.
Pat. No. 6,335,324. In another exemplary embodiment, the structure
has the following proviso: the compound cannot be a chemical
compound which is explicitly disclosed (ie does not contain any
variables) in U.S. Pat. Pub. No. 2004/0147502. In another exemplary
embodiment, the structure has the following proviso: the compound
cannot be a chemical compound which is explicitly disclosed (ie
does not contain any variables) in U.S. Pat. Pub. No.
2004/0180855.
[0033] In an exemplary embodiment, D is amidinohydrazone. In
another exemplary embodiment, D is aminopyridine. In an exemplary
embodiment, D is 2-aminopyridine. In an exemplary embodiment, D is
2-aminoaryl. In an exemplary embodiment, D is 2-aminophenyl. In an
exemplary embodiment, A is S.
[0034] In an exemplary embodiment, at least one of said R.sup.1,
R.sup.1a, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7,
R.sup.8, R.sup.9, R.sup.10, R.sup.10a, R.sup.11, R.sup.12,
R.sup.13, R.sup.14, R.sup.15, R.sup.16, R.sup.18, R.sup.19,
R.sup.20, R.sup.21, R.sup.22 and R.sup.23 is a member selected from
substituted or unsubstituted arylalkyl and substituted or
unsubstituted heteroarylakyl. In another exemplary embodiment, D is
a member selected from substituted or unsubstituted guanidine,
substituted or unsubstituted amidinohydrazone, substituted or
unsubstituted pyridine, substituted or unsubstituted
aminopyridine.
[0035] In a second aspect, the invention provides a compound
according to the following structure:
##STR00013##
In this structure, AB.sub.mC, in combination, is a member selected
from substituted or unsubstituted alkyl and substituted or
unsubstituted heteroalkyl. D is a member selected from substituted
or unsubstituted guanidine, substituted or unsubstituted
amidinohydrazone, substituted or unsubstituted pyridine,
substituted or unsubstituted aminopyridine. R.sub.c is a member
selected from H and substituted or unsubstituted alkyl. R.sub.d' is
a member selected from H and substituted or unsubstituted alkyl.
R.sub.d is a member selected from R.sup.16,
(CR.sup.14R.sup.15).sub.pR.sup.16,
C(R.sup.14).dbd.CR.sup.16R.sup.23 and CCR.sup.16. The symbol p is
an integer selected from 0 to 3. R.sup.14 and R.sup.23 are members
independently selected from H, halogen, substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl. R.sup.15 is a member
selected from H, halogen, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl. R.sup.16 is a member selected from
halogen, substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
C(O)R.sup.18, OR.sup.19, --OC(=Q)NR.sup.19R.sup.21,
--NR.sup.19C(=Q)NR.sup.19R.sup.21, --NR.sup.20SO.sub.2R.sup.19,
OSO.sub.2R.sup.19, SO.sub.2R.sup.21, SOR.sup.21,
SO.sub.2NR.sup.19R.sup.20, NR.sup.19R.sup.20, CN. R.sup.18 is a
member selected from substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, OR.sup.21, NR.sup.19R.sup.20, and
--NR.sup.19SO.sub.2R.sup.20. R.sup.19 is a member selected from H,
substituted or unsubstituted alkyl, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl, substituted or unsubstituted heteroaryl. R.sup.20 is a member
selected from H, substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
C(O)R.sup.22, SO.sub.2R.sup.22. R.sup.22 is a member selected from
substituted or unsubstituted alkyl, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl, substituted or unsubstituted heteroaryl. R.sup.21 is a member
selected from substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
substituted or unsubstituted aryl, substituted or unsubstituted
heteroaryl, --OR.sup.22. R.sup.14 and R.sup.15 can be optionally
joined, together with the atoms to which they are attached, to form
a 4-8 membered ring. R.sup.19 and R.sup.21 can be optionally
joined, together with the atoms to which they are attached, to form
a 4-8 membered ring. R.sup.14 and R.sup.23 can be optionally
joined, together with the atoms to which they are attached, to form
a 4-8 membered ring. R.sup.19 and R.sup.20 can be optionally
joined, together with the atoms to which they are attached, to form
a 4-8 membered ring. X is a member selected from S, O, and
NR.sup.35R.sup.35 is a member selected from H, substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, C(O)R.sup.36,
SO.sub.2R.sup.36. R.sup.36 is a member selected from substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl. Y is a member selected
from a bond, C=Q, CR.sup.12R.sup.13 and SO.sub.n. The symbol n is a
member selected from 0 to 2. Q is a member selected from S, O, and
NR.sup.7. R.sup.12 is a member selected from H, halogen,
substituted or unsubstituted alkyl, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl, and substituted or unsubstituted heteroaryl. R.sup.13 is a
member selected from H, halogen, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, and
substituted or unsubstituted heteroaryl. R.sup.12 and R.sup.13 can
be optionally joined, together with the atoms to which they are
attached, to form a 4-8 membered ring. R.sub.a is a member selected
from NR.sup.7R.sup.8, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, and
substituted or unsubstituted heteroaryl. R.sup.7 and R.sup.8 can be
optionally joined, together with the atoms to which they are
attached, to form a 4-8 membered ring.
[0036] In an exemplary embodiment, the compounds of the second
aspect are subject to one or more of the following provisos. In an
exemplary embodiment, the structure has the proviso that when D is
unsubstituted guanidine, AB.sub.mC cannot be unsubstituted alkyl.
In an exemplary embodiment, the structure has the proviso that if
the compound has the following structure:
##STR00014##
in which R.sup.30 is a member selected from H, SO.sub.2--R.sup.31,
substituted or unsubstituted alkyl, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl, and substituted or unsubstituted heteroaryl. R.sup.31 is
unsubstituted alkyl, then at least one of R.sub.d and R.sub.d' does
not comprise a carbonyl group which is directly attached, or alpha,
to the azetidinone ring.
[0037] In an exemplary embodiment, the second aspect has the
proviso that the compound cannot have the following structure:
##STR00015##
wherein DCB.sub.mA comprises a piperidine ring or a salt
thereof.
[0038] In an exemplary embodiment of one of the aspects of the
invention, at least one of said R.sup.1a, R.sup.12, R.sup.13,
R.sup.14, R.sup.15, R.sup.16, R.sup.18, R.sup.19, R.sup.20,
R.sup.21, R.sup.22, R.sup.23, R.sup.35, R.sup.36 and R.sub.a is a
member selected from substituted or unsubstituted arylalkyl and
substituted or unsubstituted heteroarylakyl.
[0039] In an exemplary embodiment of one of the aspects of the
invention, D is a member selected from substituted or unsubstituted
aryl, substituted or unsubstituted heteroaryl, substituted or
unsubstituted amidine, substituted or unsubstituted
amidinohydrazone, substituted or unsubstituted guanidine and
substituted or unsubstituted amine.
[0040] In an exemplary embodiment of one of the aspects of the
invention, D is substituted or unsubstituted guanidine, and
AB.sub.mC, in combination, form a member selected from
D-N.dbd.CR-Z.sup.5, D-O-Z.sup.5, D-SO.sub.2Z.sup.5, --O-D,
D-SO.sub.2Z.sup.5, substituted or unsubstituted alkylene,
substituted or unsubstituted acylene, and substituted or
unsubstituted heteroalkylene, and Z.sup.5 is a member selected from
substituted or unsubstituted alkylene and substituted or
unsubstituted acylene.
[0041] In an exemplary embodiment of one of the aspects of the
invention, D is a member selected from substituted aryl and
substituted or unsubstituted heteroaryl, and AB.sub.mC, in
combination, form a member selected from D-N.dbd.CR.sup.1a-Z.sup.5,
D-O-Z.sup.5, D-SO.sub.2N, D-N.dbd.CR--, --O-D, --SO.sub.2-D,
substituted and substituted alkylene, substituted or unsubstituted
acylene, and substituted or unsubstituted heteroalkylene, and
Z.sup.5 is a member selected from substituted or unsubstituted
alkylene and substituted and unsubstituted acylene.
[0042] In an exemplary embodiment of one of the aspects of the
invention, AB.sub.mCD, in combination, form a member selected
from
##STR00016##
wherein the symbol g is an integer selected from 1 to 4. Each
symbol h is an integer independently selected from 0 to 4.
R.sup.40, R.sup.41 and R.sup.42 are members independently selected
from H, substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, and substituted or unsubstituted
heteroaryl.
[0043] In an exemplary embodiment of one of the aspects of the
invention, R.sub.d is a member selected from
##STR00017##
and CF.sub.3.
[0044] In an exemplary embodiment of one of the aspects of the
invention, --Y--R.sub.a, in combination, form a member selected
from
##STR00018## ##STR00019##
[0045] In an exemplary embodiment of one of the aspects of the
invention, R.sub.c is a member selected from H, methyl and
ethyl.
[0046] In an exemplary embodiment of one of the aspects of the
invention, X is S. In an exemplary embodiment of one of the aspects
of the invention, X is O.
[0047] In an exemplary embodiment of one of the aspects of the
invention, R.sub.a is a member selected from
##STR00020## ##STR00021##
R.sub.d is a member selected from
##STR00022##
##STR00023##
and Y is a member selected from C(O) and S(O).
[0048] In another exemplary embodiment of one of the aspects of the
invention, AB.sub.mCD, in combination, form a member selected
from
##STR00024##
wherein h is an integer selected from 0 to 4.
[0049] In another aspect, the invention provides a compound
according to the following structure:
##STR00025##
wherein
[0050] AB.sub.mCD, in combination, is a member selected from:
##STR00026##
in which the symbol h1 is a member selected from 0 to 4. The symbol
q is a member selected from 0 to 3. The symbol r is a member
selected from 0 to 2. The symbols d and p are members independently
selected from 0 and 1. The symbol Z.sup.1 is member selected from
CR.sup.x13 and N. The symbol Z.sup.4 is a member selected from
NR.sup.7 and S. R.sup.x1, R.sup.x2, R.sup.x9, R.sup.x10 and
R.sup.x13 are members independently selected from H, OR.sup.y1,
S(O).sub.n1R.sup.y1, NR.sup.y1R.sup.y2, halogen, substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
and substituted or unsubstituted heteroaryl. R.sup.x3, R.sup.x4,
R.sup.x5, R.sup.x6, R.sup.x7, R.sup.x8, R.sup.x11 and R.sup.x12 are
members independently selected from a positive charge, a salt
counterion, H, S(O).sub.n1R.sup.y1, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, and
substituted or unsubstituted heteroaryl. The symbol nil is a member
selected from 0 to 2. R.sup.y1 and R.sup.y2 are members
independently selected from a charge (positive or negative), a salt
counterion, H, substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl.
Z.sup.2 and Z.sup.3 are members selected from S(O).sub.n1, O, and
NR.sup.7. The symbol n1 is a member selected from 0 to 2. Each
R.sup.7 is a member selected from a positive charge, salt
counterion, H, substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
C(O)R.sup.10, C(O)OR.sup.10, NR.sup.10R.sup.10a, OR.sup.10,
SO.sub.2R.sup.10 and S(O)R.sup.10. Each R.sup.10 and each R.sup.10a
is a member independently selected from a positive charge, salt
counterion, H, substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, and substituted or unsubstituted heteroaryl.
The symbol X is a member selected from O and S. The symbol Y is a
member selected from C=Q. Q is a member selected from S and O.
R.sub.a is a member selected from R.sup.x14 and
NR.sup.x15R.sup.x16. R.sup.x14 is a member selected from OR.sup.y3,
substituted or unsubstituted alkyl, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl, and substituted or unsubstituted heteroaryl. R.sup.x15 and
R.sup.x16 are members independently selected from H,
SO.sub.2R.sup.y3, NR.sup.y3R.sup.y4, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl and substituted
or unsubstituted heteroaryl. R.sup.y3 and R.sup.y4 are members
independently selected from a charge (either positive or negative),
a salt counterion, H, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl. R.sub.c is a member selected from H,
substituted or unsubstituted alkyl and halogen. R.sub.d is a member
selected from
##STR00027##
The symbol h1 is a member selected from 0 to 4. T is a member
selected from NR.sup.x23 and O. R.sup.x19 is a member selected from
OR.sup.y5, SR.sup.y5, NR.sup.y5R.sup.y6,
NR.sup.x23SO.sub.2R.sup.x22, substituted or unsubstituted alkyl,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, and
substituted or unsubstituted heteroaryl. R.sup.x22 is a member
selected from substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted heterocycloalkyl, substituted or
unsubstituted aryl, and substituted or unsubstituted heteroaryl.
R.sup.x23 is a member selected from H, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, and
substituted or unsubstituted heteroaryl. R.sup.x20 is a member
selected from H, C(O)R.sup.x24, SO.sub.2R.sup.x24, substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
and substituted or unsubstituted heteroaryl. R.sup.x24 is a member
selected from OR.sup.x22, NR.sup.x22R.sup.x23, substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
and substituted or unsubstituted heteroaryl. R.sup.x17 and
R.sup.x18 are members independently selected from H, OR.sup.y5,
S(O).sub.n1R.sup.y5, NR.sup.y5R.sup.y6, halogen, substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
and substituted or unsubstituted heteroaryl. R.sup.y5 and R.sup.y6
are members independently selected from a charge (positive or
negative), a salt counterion, H, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl. R.sub.d' is a member selected from H,
substituted or unsubstituted alkyl and halogen. R.sub.c and R.sub.d
can be optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sub.c and R.sub.d' can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x1 and R.sup.x2 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x2 and R.sup.x3 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x3 and R.sup.x4 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x4 and R.sup.x5 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x4 and R.sup.x6 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x5 and R.sup.x6 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x6 and R.sup.x7 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x4 and R.sup.x8 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x2 and R.sup.x9 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. two R.sup.x10 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x9 and R.sup.x10 can
be optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x12 and R.sup.x11 can
be optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x11 and R.sup.x10 can
be optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.y1 and R.sup.y2 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.y3 and R.sup.y4 can be
optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x15 and R.sup.x16 can
be optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring. R.sup.x22 and R.sup.x23 can
be optionally joined, together with the atoms to which they are
attached, to form a 5-8 membered ring.
[0051] In an exemplary embodiment, the compound is subject to one
or more of the provisos which are described herein. In an exemplary
embodiment, the compound has the proviso that if
[0052] a) Y is C.dbd.O; [0053] ABC, in combination, form
unsubstituted alkyl; and [0054] D is substituted or unsubstituted
guanidine; or
[0055] b) Y is C.dbd.O; [0056] AB, in combination, form
unsubstituted alkyl; and [0057] CD, in combination, form
##STR00028##
[0058] wherein [0059] the dotted line represents carbon atoms
necessary for the formation of one ring having 4 to 8 atoms; then
R.sub.c cannot be a member selected from CH.sub.3 and
CH.sub.2CH.sub.3; and R.sub.d and R.sub.d' cannot be members
selected from CH.sub.3 and CH.sub.3; CH.sub.2CH.sub.3 and
CH.sub.2CH.sub.3; CH.sub.2CH.sub.3 and CH.sub.3; CH.sub.3 and
CH.sub.2CH.sub.3; H and COOH; H and COOR.sup.y5; H and
CONR.sup.10R.sup.10a; unsubstituted alkyl and COOH; unsubstituted
alkyl and COOR.sup.y5; H and unsubstituted phenylalkyl;
unsubstituted alkyl and unsubstituted phenylalkyl. In another
exemplary embodiment, R.sub.c cannot be a member selected from
CH.sub.3, CH.sub.2CH.sub.3, substituted alkyl, and unsubstituted
alkyl; and R.sub.d and R.sub.d' cannot be members selected from
CH.sub.3 and CH.sub.3; CH.sub.2CH.sub.3 and CH.sub.2CH.sub.3;
CH.sub.2CH.sub.3 and CH.sub.3; CH.sub.3 and CH.sub.2CH.sub.3;
substituted or unsubstituted alkyl and substituted or unsubstituted
alkyl; H and substituted or unsubstituted alkyl; substituted or
unsubstituted alkyl and H; H and COOH; H and COOR.sup.y5; H and
CONR.sup.10R.sup.10a; unsubstituted alkyl and COOH; unsubstituted
alkyl and COOR.sup.y5; H and unsubstituted phenylalkyl;
unsubstituted alkyl and unsubstituted phenylalkyl.
[0060] In another exemplary embodiment, part (a) of this proviso
is
[0061] a) Y is a member selected from C.dbd.O and SO.sub.2; [0062]
ABC, in combination, form substituted alkyl; and [0063] D is
substituted or unsubstituted guanidine.
[0064] In another exemplary embodiment, R.sub.c of this proviso
cannot be substituted alkyl. In another exemplary embodiment,
R.sub.c of this proviso cannot be unsubstituted alkyl.
[0065] In another exemplary embodiment, R.sub.d and R.sub.d' of
this proviso cannot be members selected from CH.sub.3 and CH.sub.3;
CH.sub.2CH.sub.3 and CH.sub.2CH.sub.3; CH.sub.2CH.sub.3 and
CH.sub.3; CH.sub.3 and CH.sub.2CH.sub.3; H and COOH; H and
COOR.sup.y5; H and CONR.sup.10R.sup.10a; unsubstituted alkyl and
COOH; unsubstituted alkyl and COOR.sup.y5; H and unsubstituted
phenylalkyl; unsubstituted alkyl and unsubstituted phenylalkyl.
[0066] In another exemplary embodiment, the structure has the
following proviso: if the compound has the following structure:
##STR00029##
wherein R.sup.30 is a member selected from H, SO.sub.2--R.sup.31,
substituted or unsubstituted alkyl, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl, and substituted or unsubstituted heteroaryl; wherein R.sup.31
is unsubstituted alkyl, then at least one of R.sub.d and R.sub.d'
does not comprise a carbonyl group which is directly attached, or
alpha, to the azetidinone ring.
[0067] In another exemplary embodiment, the structure has the
following proviso: the compound cannot have the following
structure:
##STR00030##
wherein at least one of R.sub.d and R.sub.d' comprise a carbonyl
group which is directly attached, or alpha, to the azetidinone
ring. In another exemplary embodiment, the compound cannot have the
following structure:
##STR00031##
wherein DCB.sub.mA comprises a piperidine ring or a salt thereof.
In another exemplary embodiment, the compound cannot have the
following structure:
##STR00032##
wherein DCB.sub.mA comprises a piperidine ring or a salt
thereof.
[0068] In another exemplary embodiment, the structure has the
following proviso: the compound cannot have the following
structure:
##STR00033##
wherein R.sub.a is NHR.sup.8, and R.sup.8 includes a substituted or
unsubstituted phenyl. In another exemplary embodiment, R.sup.8 is
unsubstituted phenyl. In yet another exemplary embodiment, R.sub.a
is N(H)(unsubstituted phenyl). In another exemplary embodiment, the
structure has the following proviso: the compound cannot have the
following structure:
##STR00034##
[0069] In another exemplary embodiment, the structure has the
following proviso: the compound cannot be a chemical compound which
is explicitly disclosed (ie does not contain any variables) in U.S.
Pat. No. 6,335,324. In another exemplary embodiment, the structure
has the following proviso: the compound cannot be a chemical
compound which is explicitly disclosed (ie does not contain any
variables) in U.S. Pat. Pub. No. 2004/0147502. In another exemplary
embodiment, the structure has the following proviso: the compound
cannot be a chemical compound which is explicitly disclosed (ie
does not contain any variables) in U.S. Pat. Pub. No.
2004/0180855.
[0070] In another aspect, the invention provides a method of making
one of the compounds of the invention. General and specific methods
of synthesizing the compounds are provided herein.
[0071] In an exemplary embodiment, the invention provides a
pharmaceutical composition comprising an effective amount of a
compound of the invention including an inner salt or a
pharmaceutically acceptable salt thereof, a hydrolysable ester
thereof, or a solvate thereof and one or more pharmaceutically
acceptable excipients. In an exemplary embodiment, the
pharmaceutical composition is useful for enhancing thrombolysis or
treating thrombosis, asthma, chronic asthma, or allergic
rhinitis.
[0072] In an exemplary embodiment, the invention provides a
pharmaceutical composition useful for treating asthma or allergic
rhinitis comprising an effective amount of a compound of the
invention including an inner salt or a pharmaceutically acceptable
salt thereof, a hydrolysable ester thereof, or a solvate thereof
and one or more pharmaceutically acceptable excipients.
[0073] In an exemplary embodiment, the invention provides a
pharmaceutical composition useful for treating chronic asthma
comprising an effective amount of a compound of the invention
including an inner salt or a pharmaceutically acceptable salt
thereof, a hydrolysable ester thereof, or a solvate thereof and one
or more pharmaceutically acceptable excipients.
[0074] In an exemplary embodiment, the invention provides a method
for treating asthma, chronic asthma or allergic rhinitis in a
mammalian species comprising administering an effective amount of
the composition of the invention.
[0075] In an exemplary embodiment, the invention provides a method
for treating chronic asthma in a mammalian species comprising
administering by inhalation to the bronchioles an effective amount
of the composition of the invention.
[0076] In an exemplary embodiment, the invention provides a method
of inhibiting tryptase in a mammal by administration of a compound
according to the invention. In another exemplary embodiment, the
invention provides a method of inhibiting factor XIa in a mammal by
administration of a compound according to the invention. In yet
another exemplary embodiment, the invention provides a method for
enhancing thrombolysis or inhibiting or preventing thrombosis in a
mammalian species comprising administering an effective amount of
the composition of the invention.
[0077] In an exemplary embodiment, the invention provides a method
of treating thrombosis in a mammal comprising administering to the
mammal a pharmaceutical composition that inhibits thrombosis in the
mammal, wherein the pharmaceutical composition contains a
therapeutically effective amount of a compound according to the
invention that is selective for inhibiting Factor XIa.
[0078] In an exemplary embodiment, the invention provides a method
of treating thrombosis in a mammal comprising administration of a
compound according to the invention to the mammal having sufficient
selectivity and potency for inhibition of Factor XIa, wherein the
administration of the small molecule inhibits thrombosis in the
mammal with no substantial effect on bleeding times in the
mammal.
[0079] In an exemplary embodiment, the invention provides a method
of inhibiting Factor XIa in a mammal by administration of a
compound according to the invention.
BRIEF DESCRIPTION OF THE DRAWINGS
[0080] FIGS. 1A to 1DS show tables that include exemplary compounds
of the invention.
[0081] FIGS. 2A to 2M show tables that include exemplary compounds
of the invention and their IC50 data for Factor XIa (FXIa) and
tryptase. In this figure, (+++) represents an IC50 value between
0.1 nM and 99.9 nM; (++) represents an IC50 value between 100 nM
and 9.99 .mu.M; (+) represents an IC50 value between 10 .mu.M and
99.9 .mu.M; and >(+) represents an IC50 value greater than 99.9
.mu.M.
DETAILED DESCRIPTION OF THE INVENTION AND PREFERRED EMBODIMENTS
I. Definitions and Abbreviations
[0082] AcN-acetonitrile; AcOH acetic acid; Allyl Br allyl bromide;
BH.sub.3 borane; t-Boc tert-butoxycarbonyl; Boc.sub.2O
di-tert-butoxycarbonyl anhydride; BuLi butyl lithium; CAN ceric
ammonium nitrate; CCl.sub.4 carbon tetrachloride; CDI carbonyl
diimimidazole; DCC dicyclohexyl carbodiimide; DCM dichloromethane;
DIEA diisopropylethylamine; DMAP dimethylaminopyridine; DMF
dimethylformamide; EDC
1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide; ESI-MS electrospray
ionization mass spectrometry; EtOAc ethyl acetate; EtOH ethyl
alcohol; HCl hydrochloric acid; HOBt 1-hydroxybenzotriazole; HPLC
high pressure liquid chromatography; LC-MS liquid chromatography
mass spectrometry; LDA lithium diisopropylamide; LiHMDS lithium
hexamethyldisilazide; MeOH methyl alcohol; MS mass spectrometry;
NBS N-bromo succinimide; NMR nuclear magnetic resonance; Pd/C
palladium on carbon; TBAF tetrabutylammonium fluoride; TBDMS
tert-butyldimethylsilyl; TEA triethylamine; TFA trifluoroacetic
acid; THF tetrahydrofuran; TIPS triisopropylsilyl; TLC thin layer
chromatography; TMSCHN.sub.2 trimethylsilyl diazomethane; TMSCl
trimethylsilyl chloride.
[0083] "Reactive functional group," as used herein refers to groups
including, but not limited to, olefins, acetylenes, alcohols,
phenols, ethers, oxides, halides, aldehydes, ketones, carboxylic
acids, esters, amides, cyanates, isocyanates, thiocyanates,
isothiocyanates, amines, hydrazines, hydrazones, hydrazides, diazo,
diazonium, nitro, nitriles, mercaptans, sulfides, disulfides,
sulfoxides, sulfones, sulfonic acids, sulfinic acids, acetals,
ketals, anhydrides, sulfates, sulfenic acids isonitriles, amidines,
imides, imidates, nitrones, hydroxylamines, oximes, hydroxamic
acids thiohydroxamic acids, allenes, ortho esters, sulfites,
enamines, ynamines, ureas, pseudoureas, semicarbazides,
carbodiimides, carbamates, imines, azides, azo compounds, azoxy
compounds, and nitroso compounds. Reactive functional groups also
include those used to prepare bioconjugates, e.g.,
N-hydroxysuccinimide esters, maleimides and the like. Methods to
prepare each of these functional groups are well known in the art
and their application to or modification for a particular purpose
is within the ability of one of skill in the art (see, for example,
Sandler and Karo, eds. ORGANIC FUNCTIONAL GROUP PREPARATIONS,
Academic Press, San Diego, 1989).
[0084] "Non-covalent protein binding groups" are moieties that
interact with an intact or denatured polypeptide in an associative
manner. The interaction may be either reversible or irreversible in
a biological milieu. The incorporation of a "non-covalent protein
binding group" into a chelating agent or complex of the invention
provides the agent or complex with the ability to interact with a
polypeptide in a non-covalent manner. Exemplary non-covalent
interactions include hydrophobic-hydrophobic and electrostatic
interactions. Exemplary "non-covalent protein binding groups"
include anionic groups, e.g., phosphate, thiophosphate,
phosphonate, carboxylate, boronate, sulfate, sulfone, thiosulfate,
and thiosulfonate.
[0085] As used herein, "linking member" refers to a covalent
chemical bond that includes at least one heteroatom. Exemplary
linking members include --C(O)NH--, --C(O)O--, --NH--, --S--,
--O--, and the like.
[0086] The term "targeting group" is intended to mean a moiety that
is: (1) able to actively direct the entity to which it is attached
(e.g., contrast agent) to a target region, e.g., a tumor; or (2) is
preferentially passively absorbed by or entrained within a target
tissue, for example a tumor. The targeting group can be a small
molecule, which is intended to include both non-peptides and
peptides. The targeting group can also be a macromolecule, which
includes, but is not limited to, saccharides, lectins, receptors,
ligand for receptors, proteins such as BSA, antibodies,
poly(ethers), dendrimers, poly(amino acids) and so forth.
[0087] The term "cleavable group" is intended to mean a moiety that
allows for release of the chelate from the rest of the conjugate by
cleaving a bond linking the chelate (or chelate linker arm
construct) to the remainder of the conjugate. Such cleavage is
either chemical in nature, or enzymatically mediated. Exemplary
enzymatically cleavable groups include natural amino acids or
peptide sequences that end with a natural amino acid.
[0088] In addition to enzymatically cleavable sites, it is within
the scope of the present invention to include one or more sites
that are cleaved by the action of an agent other than an enzyme.
Exemplary non-enzymatic cleavage agents include, but are not
limited to, acids, bases, light (e.g., nitrobenzyl derivatives,
phenacyl groups, benzoin esters), and heat. Many cleaveable groups
are known in the art. See, for example, Jung et al., Biochem.
Biophys. Acta, 761: 152-162 (1983); Joshi et al., J. Biol. Chem.,
265: 14518-14525 (1990); Zarling et al., J. Immunol., 124: 913-920
(1980); Bouizar et al., Eur. J. Biochem., 155: 141-147 (1986); Park
et al., J. Biol. Chem., 261: 205-210 (1986); Browning et al., J.
Immunol., 143: 1859-1867 (1989). Moreover a broad range of
cleavable, bifunctional (both homo- and hetero-bifunctional) spacer
arms are commercially available from suppliers such as Pierce.
[0089] The symbol , whether utilized as a bond or displayed
perpendicular to a bond indicates the point at which the displayed
moiety is attached to the remainder of the molecule, solid support,
etc.
[0090] Certain compounds of the present invention can exist in
unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms are equivalent to unsolvated
forms and are encompassed within the scope of the present
invention. Certain compounds of the present invention may exist in
multiple crystalline or amorphous forms. In general, all physical
forms are equivalent for the uses contemplated by the present
invention and are intended to be within the scope of the present
invention.
[0091] Certain compounds of the present invention possess
asymmetric carbon atoms (optical centers) or double bonds; the
racemates, diastereomers, geometric isomers and individual isomers
are encompassed within the scope of the present invention.
[0092] The compounds of the invention may be prepared as a single
isomer (e.g., enantiomer, cis-trans, positional, diastereomer) or
as a mixture of isomers. In a preferred embodiment, the compounds
are prepared as substantially a single isomer. Methods of preparing
substantially isomerically pure compounds are known in the art. For
example, enantiomerically enriched mixtures and pure enantiomeric
compounds can be prepared by using synthetic intermediates that are
enantiomerically pure in combination with reactions that either
leave the stereochemistry at a chiral center unchanged or result in
its complete inversion. Alternatively, the final product or
intermediates along the synthetic route can be resolved into a
single stereoisomer. Techniques for inverting or leaving unchanged
a particular stereocenter, and those for resolving mixtures of
stereoisomers are well known in the art and it is well within the
ability of one of skill in the art to choose an appropriate method
for a particular situation. See, generally, Furniss et al. (eds.),
VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5.sup.TH ED.,
Longman Scientific and Technical Ltd., Essex, 1991, pp. 809-816;
and Heller, Acc. Chem. Res. 23: 128 (1990).
[0093] The compounds of the present invention may also contain
unnatural proportions of atomic isotopes at one or more of the
atoms that constitute such compounds. For example, the compounds
may be radiolabeled with radioactive isotopes, such as for example
tritium (.sup.3H), iodine-125 (.sup.125I) or carbon-14 (.sup.14C).
All isotopic variations of the compounds of the present invention,
whether radioactive or not, are intended to be encompassed within
the scope of the present invention.
[0094] Where substituent groups are specified by their conventional
chemical formulae, written from left to right, they equally
encompass the chemically identical substituents, which would result
from writing the structure from right to left, e.g., --CH.sub.2O--
is intended to also recite --OCH.sub.2--; --NHS(O).sub.2-- is also
intended to represent --S(O).sub.2HN--; etc.
[0095] The term "alkyl," by itself or as part of another
substituent, means, unless otherwise stated, a straight or branched
chain, or cyclic hydrocarbon radical, or combination thereof, which
may be fully saturated, mono- or polyunsaturated and can include
di- and multivalent radicals, having the number of carbon atoms
designated (i.e. C.sub.1-C.sub.10 means one to ten carbons).
Examples of saturated hydrocarbon radicals include, but are not
limited to, groups such as methyl, ethyl, n-propyl, isopropyl,
n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl,
(cyclohexyl)methyl, cyclopropylmethyl, homologs and isomers of, for
example, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. An
unsaturated alkyl group is one having one or more double bonds or
triple bonds. Examples of unsaturated alkyl groups include, but are
not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl,
2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1-
and 3-propynyl, 3-butynyl, and the higher homologs and isomers. The
term "alkyl," unless otherwise noted, is also meant to include
those derivatives of alkyl defined in more detail below, such as
"heteroalkyl." Alkyl groups that are limited to hydrocarbon groups
are termed "homoalkyl".
[0096] The term "alkylene" by itself or as part of another
substituent means a divalent radical derived from an alkane, as
exemplified, but not limited, by
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--, and further includes those
groups described below as "heteroalkylene." Typically, an alkyl (or
alkylene) group will have from 1 to 24 carbon atoms, with those
groups having 10 or fewer carbon atoms being preferred in the
present invention. A "lower alkyl" or "lower alkylene" is a shorter
chain alkyl or alkylene group, generally having eight or fewer
carbon atoms.
[0097] The terms "alkoxy," "alkylamino" and "alkylthio" (or
thioalkoxy) are used in their conventional sense, and refer to
those alkyl groups attached to the remainder of the molecule via an
oxygen atom, an amino group, or a sulfur atom, respectively.
[0098] The term "heteroalkyl," by itself or in combination with
another term, means, unless otherwise stated, a stable straight or
branched chain, or cyclic hydrocarbon radical, or combinations
thereof, consisting of the stated number of carbon atoms and at
least one heteroatom selected from the group consisting of O, N, Si
and S, and wherein the nitrogen and sulfur atoms may optionally be
oxidized and the nitrogen heteroatom may optionally be quaternized.
The heteroatom(s) O, N and S and Si may be placed at any interior
position of the heteroalkyl group or at the position at which the
alkyl group is attached to the remainder of the molecule. Examples
include, but are not limited to, --CH.sub.2--CH.sub.2--O--CH.sub.3,
--CH.sub.2--CH.sub.2--NH--CH.sub.3,
--CH.sub.2--CH.sub.2--N(CH.sub.3)--CH.sub.3,
--CH.sub.2--S--CH.sub.2--CH.sub.3, --CH.sub.2--CH.sub.2,
--S(O)--CH.sub.3, --CH.sub.2--CH.sub.2--S(O).sub.2--CH.sub.3,
--CH.dbd.CH--O--CH.sub.3, --Si(CH.sub.3).sub.3,
--CH.sub.2--CH.dbd.N--OCH.sub.3, and
--CH.dbd.CH--N(CH.sub.3)--CH.sub.3. Up to three heteroatoms may be
consecutive, such as, for example, --CH.sub.2--NH--OCH.sub.3,
--CH.sub.2--O--SO.sub.2--O--CH.sub.3 and
--CH.sub.2--O--Si(CH.sub.3).sub.3. Similarly, the term
"heteroalkylene" by itself or as part of another substituent means
a divalent radical derived from heteroalkyl, as exemplified, but
not limited by, --CH.sub.2--CH.sub.2--S--CH.sub.2--CH.sub.2-- and
--CH.sub.2--S--CH.sub.2--CH.sub.2--NH--CH.sub.2--. For
heteroalkylene groups, heteroatoms can also occupy either or both
of the chain termini (e.g., alkyleneoxy, alkylenedioxy,
alkyleneamino, alkylenediamino, and the like). Still further, for
alkylene and heteroalkylene linking groups, no orientation of the
linking group is implied by the direction in which the formula of
the linking group is written. For example, the formula
--C(O).sub.2R'-- represents both --C(O).sub.2R'-- and
--R'C(O).sub.2--.
[0099] In general, the term "acyl" is also selected from the group
set forth above. As used herein, the term "acyl" refers to groups
attached to, and fulfilling the valence of a carbonyl carbon that
is either directly or indirectly attached to the compounds of the
present invention.
[0100] In general, the term "acylene" refers to an alkylene group
which comprises an acyl group.
[0101] The terms "cycloalkyl" and "heterocycloalkyl", by themselves
or in combination with other terms, represent, unless otherwise
stated, cyclic versions of "alkyl" and "heteroalkyl", respectively.
Additionally, for heterocycloalkyl, a heteroatom can occupy the
position at which the heterocycle is attached to the remainder of
the molecule. Examples of cycloalkyl include, but are not limited
to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl,
cycloheptyl, and the like. Examples of heterocycloalkyl include,
but are not limited to, 1-(1,2,5,6-tetrahydropyridyl),
1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl,
3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl,
tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl,
2-piperazinyl, and the like.
[0102] The terms "halo" or "halogen," by themselves or as part of
another substituent, mean, unless otherwise stated, a fluorine,
chlorine, bromine, or iodine atom. Additionally, terms such as
"haloalkyl," are meant to include monohaloalkyl and polyhaloalkyl.
For example, the term "halo(C.sub.1-C.sub.4)alkyl" is mean to
include, but not be limited to, trifluoromethyl,
2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the
like.
[0103] The term "aryl" means, unless otherwise stated, a
polyunsaturated, aromatic, hydrocarbon substituent that can be a
single ring or multiple rings (preferably from 1 to 3 rings), which
are fused together or linked covalently. The term "heteroaryl"
refers to aryl groups (or rings) that contain from one to four
heteroatoms selected from N, O, and S, wherein the nitrogen and
sulfur atoms are optionally oxidized, and the nitrogen atom(s) are
optionally quaternized. A heteroaryl group can be attached to the
remainder of the molecule through a heteroatom. Non-limiting
examples of aryl and heteroaryl groups include phenyl, 1-naphthyl,
2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl,
3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl,
4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl,
4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,
2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl,
4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl,
2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,
2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl.
Substituents for each of the above noted aryl and heteroaryl ring
systems are selected from the group of acceptable substituents
described below.
[0104] For brevity, the term "aryl" when used in combination with
other terms (e.g., aryloxy, arylthioxy, arylalkyl) includes both
aryl and heteroaryl rings as defined above. Thus, the term
"arylalkyl" is meant to include those radicals in which an aryl
group is attached to an alkyl group (e.g., benzyl, phenethyl,
pyridylmethyl and the like) including those alkyl groups in which a
carbon atom (e.g., a methylene group) has been replaced by, for
example, an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl,
3-(1-naphthyloxy)propyl, and the like).
[0105] Each of the above terms (e.g., "alkyl," "heteroalkyl,"
"aryl" and "heteroaryl") are meant to include both substituted or
unsubstituted forms of the indicated radical. Preferred
substituents for each type of radical are provided below.
[0106] Substituents for the alkyl and heteroalkyl radicals
(including those groups often referred to as alkylene, alkenyl,
heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl,
heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) are
generically referred to as "alkyl group substituents," and they can
be one or more of a variety of groups selected from, but not
limited to: --OR', .dbd.O, .dbd.NR', .dbd.N--OR', --NR'R'', --SR',
-halogen, --SiR'R''R''', --OC(O)R', --C(O)R', --CO.sub.2R',
--CONR'R'', --OC(O)NR'R'', --NR''C(O)R', --NR'--C(O)NR''R''',
--N''C(O).sub.2R', --NR--C(NR'R''R''').dbd.NR'''',
--NR--C(NR'R'').dbd.NR''', --S(O)R', --S(O).sub.2R',
--S(O).sub.2NR'R'', --NRSO.sub.2R', --CN and --NO.sub.2 in a number
ranging from zero to (2m'+1), where m' is the total number of
carbon atoms in such radical. R', R'', R''' and R'''' each
preferably independently refer to hydrogen, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted aryl, e.g.,
aryl substituted with 1-3 halogens, substituted or unsubstituted
alkyl, alkoxy or thioalkoxy groups, or arylalkyl groups. When a
compound of the invention includes more than one R group, for
example, each of the R groups is independently selected as are each
R', R'', R''' and R'''' groups when more than one of these groups
is present. When R' and R'' are attached to the same nitrogen atom,
they can be combined with the nitrogen atom to form a 5-, 6-, or
7-membered ring. For example, --NR'R'' is meant to include, but not
be limited to, 1-pyrrolidinyl and 4-morpholinyl. From the above
discussion of substituents, one of skill in the art will understand
that the term "alkyl" is meant to include groups including carbon
atoms bound to groups other than hydrogen groups, such as haloalkyl
(e.g., --CF.sub.3 and --CH.sub.2CF.sub.3) and acyl (e.g.,
--C(O)CH.sub.3, --C(O)CF.sub.3, --C(O)CH.sub.2OCH.sub.3, and the
like).
[0107] Similar to the substituents described for the alkyl radical,
the aryl substituents and heteroaryl substituents are generally
referred to as "aryl substituents" and "heteroaryl substituents,"
respectively and are varied and selected from, for example:
halogen, --OR', .dbd.O, .dbd.NR', .dbd.N--OR', --NR'R'', --SR',
-halogen, --SiR'R''R''', --OC(O)R', --C(O)R', --CO.sub.2R',
--CONR'R'', --OC(O)NR'R'', --NR''C(O)R', --NR'--C(O)NR''R''',
--NR''C(O).sub.2R', --NR--C(NR'R''R''').dbd.NR'''',
--NR--C(NR'R'').dbd.NR''', --S(O)R', --S(O).sub.2R',
--S(O).sub.2NR'R'', --NRSO.sub.2R', --CN and --NO.sub.2, --R',
--N.sub.3, --CH(Ph).sub.2, fluoro(C.sub.1-C.sub.4)alkoxy, and
fluoro(C.sub.1-C.sub.4)alkyl, in a number ranging from zero to the
total number of open valences on the aromatic ring system; and
where R', R'', R''' and R'''' are preferably independently selected
from hydrogen, substituted or unsubstituted alkyl, substituted or
unsubstituted heteroalkyl, substituted or unsubstituted aryl and
substituted or unsubstituted heteroaryl. When a compound of the
invention includes more than one R group, for example, each of the
R groups is independently selected as are each R', R'', R''' and
R'''' groups when more than one of these groups is present.
[0108] Two of the substituents on adjacent atoms of the aryl or
heteroaryl ring may optionally be replaced with a substituent of
the formula -T-C(O)--(CRR').sub.q--U--, wherein T and U are
independently --NR--, --O--, --CRR'-- or a single bond, and q is an
integer of from 0 to 3. Alternatively, two of the substituents on
adjacent atoms of the aryl or heteroaryl ring may optionally be
replaced with a substituent of the formula
-A-(CH.sub.2).sub.r--B--, wherein A and B are independently
--CRR'--, --O--, --NR--, --S--, --S(O)--, --S(O).sub.2--,
--S(O).sub.2NR'-- or a single bond, and r is an integer of from 1
to 4. One of the single bonds of the new ring so formed may
optionally be replaced with a double bond. Alternatively, two of
the substituents on adjacent atoms of the aryl or heteroaryl ring
may optionally be replaced with a substituent of the formula
--(CRR').sub.s--X--(CR''R''').sub.d--, where s and d are
independently integers of from 0 to 3, and X is --O --, --NR'--,
--S--, --S(O)--, --S(O).sub.2--, or --S(O).sub.2NR'--. The
substituents R', R', R'' and R''' are preferably independently
selected from hydrogen or substituted or unsubstituted
(C.sub.1-C.sub.6)alkyl.
[0109] As used herein, the term "heteroatom" is meant to include
oxygen (O), nitrogen (N), sulfur (S), silicon (Si), phosphorus (P)
and boron (B).
[0110] The symbol "R" is a general abbreviation that represents a
substituent group that is selected from substituted or
unsubstituted alkyl, substituted or unsubstituted heteroalkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted heterocycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl groups.
[0111] The term "amidinohydrazone", as used herein, refers to a
moiety according to the following structure:
##STR00035##
wherein one or more of the hydrogens in this structure can be
replaced with an `R` group.
[0112] "Protecting group," as used herein refers to a portion of a
substrate that is substantially stable under a particular reaction
condition, but which is cleaved from the substrate under a
different reaction condition. A protecting group can also be
selected such that it participates in the direct oxidation of the
aromatic ring component of the compounds of the invention. For
examples of useful protecting groups, see, for example, Greene et
al., PROTECTIVE GROUPS IN ORGANIC SYNTHESIS, John Wiley & Sons,
New York, 1991.
[0113] "Ring" as used herein means a substituted or unsubstituted
cycloalkyl, substituted or unsubstituted heterocycloalkyl,
substituted or unsubstituted aryl, or substituted or unsubstituted
heteroaryl. A ring includes fused ring moieties. The number of
atoms in a ring is typically defined by the number of members in
the ring. For example, a "5- to 8-membered ring" means there are 5
to 8 atoms in the encircling arrangement. The ring optionally
included a heteroatom. Thus, the term "5- to 8-membered ring"
includes, for example pyridinyl and piperidinyl. The term "ring"
further includes a ring system comprising more than one "ring",
wherein each "ring" is independently defined as above.
[0114] The term, "phenylalkyl", as used herein, refers to a moiety
in which the phenyl ring is connected to the rest of the molecule
through an alkyl chain. Examples of phenylalkyl moieties include
phenylmethyl
##STR00036##
and phenylethyl
##STR00037##
Other examples of substituted phenylalkyl is
3-chlorophenylpropyl
##STR00038##
phenyl-2-chloropropyl
##STR00039##
and 3-chlorophenyl-2-chloropropyl
##STR00040##
[0115] The term "pharmaceutically acceptable salts" includes salts
of the active compounds which are prepared with relatively nontoxic
acids or bases, depending on the particular substituents found on
the compounds described herein. When compounds of the present
invention contain relatively acidic functionalities, base addition
salts can be obtained by contacting the neutral form of such
compounds with a sufficient amount of the desired base, either neat
or in a suitable inert solvent. Examples of pharmaceutically
acceptable base addition salts include sodium, potassium, calcium,
ammonium, organic amino, or magnesium salt, or a similar salt. When
compounds of the present invention contain relatively basic
functionalities, acid addition salts can be obtained by contacting
the neutral form of such compounds with a sufficient amount of the
desired acid, either neat or in a suitable inert solvent. Examples
of pharmaceutically acceptable acid addition salts include those
derived from inorganic acids like hydrochloric, hydrobromic,
nitric, carbonic, monohydrogencarbonic, phosphoric,
monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,
monohydrogensulfuric, hydriodic, or phosphorous acids and the like,
as well as the salts derived from relatively nontoxic organic acids
like acetic, trifluoroacetic, propionic, isobutyric, maleic,
malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic,
phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric,
methanesulfonic, and the like. Also included are salts of amino
acids such as arginate and the like, and salts of organic acids
like glucuronic or galactunoric acids and the like (see, for
example, Berge et al., "Pharmaceutical Salts", Journal of
Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds
of the present invention contain both basic and acidic
functionalities that allow the compounds to be converted into
either base or acid addition salts.
[0116] The neutral forms of the compounds are preferably
regenerated by contacting the salt with a base or acid and
isolating the parent compound in the conventional manner. The
parent form of the compound differs from the various salt forms in
certain physical properties, such as solubility in polar solvents,
but otherwise the salts are equivalent to the parent form of the
compound for the purposes of the present invention.
[0117] In addition to salt forms, the present invention provides
compounds, which are in a prodrug form. Prodrugs of the compounds
described herein are those compounds that readily undergo chemical
changes under physiological conditions to provide the compounds of
the present invention. Additionally, prodrugs can be converted to
the compounds of the present invention by chemical or biochemical
methods in an ex vivo environment. For example, prodrugs can be
slowly converted to the compounds of the present invention when
placed in a transdermal patch reservoir with a suitable enzyme or
chemical reagent.
III. Compounds
[0118] Representative compounds of the invention are provided in
FIG. 1 and FIG. 2.
IV. Synthesis and Purification of the Compounds
[0119] The compounds of the invention are synthesized by an
appropriate combination of generally well known synthetic methods.
Techniques useful in synthesizing the compounds of the invention
are both readily apparent and accessible to those of skill in the
relevant art. The discussion below is offered to illustrate certain
of the diverse methods available for use in assembling the
compounds of the invention, it is not intended to define the scope
of reactions or reaction sequences that are useful in preparing the
compounds of the present invention.
IV. a) 2-Guanidinyl 2-oxo ethylene
##STR00041## ##STR00042##
[0121] One method of synthesizing compounds of the invention with
2-guanidinyl-2-oxo-ethylene substituents is shown in Scheme 1. In
this Scheme, 1A can be treated with bromomethyl t-butyl ester 1B
and lithium diisopropyl amide ("LDA") in order to provide 1C. 1C
then undergoes esterification with an alcohol (R.sup.1--OH), EDC,
DMAP in order to provide 1D. 1C can alternatively undergo
esterification under alkylation conditions using a halogenated
compound and sodium carbonate in order to provide 1D. 1D can be
desilylated using a mixture of ammonium fluoride, methanol, and
acetic acid in order to provide 1E. 1E can be treated with an
isocyanate and TEA to produce IF with an urea moiety. 1E can
alternatively be treated with an acyl chloride and TEA to produce
1F with an amide moiety. 1E can alternatively be treated with a
sulfonyl chloride and TEA to produce 1F with a sulfonamide moiety.
1E can alternatively be treated with an aryl boronic acid and
Cu(OAc).sub.2 to produce 1F with an aryl moiety. 1F can be treated
with TFA in order to remove the t-butyl group on 1F to produce 1G.
1G can be treated with guanidine and isobutylchloroformate in order
to produce 1H. 1H can be converted to the acid 1I by treating with
hydrogen and palladium on carbon in ethyl acetate when
R.sup.1=benzyl, or trifluoroacetic acid (TFA) when
R.sup.1=4-methoxybenzyl.
##STR00043## ##STR00044##
[0122] A second method of synthesizing compounds of the invention
with 2-guanidinyl-2-oxo-ethylene substituents is shown in Scheme 2.
In this Scheme, 2A can be treated with trimethylsilylchloride in
THF in order to provide 2B. 2B can then be treated with 2C and an
appropriate metal catalyst in order to provide 2D. 2D can then be
treated with an alcohol (R.sup.1--OH), EDC, DMAP in order to
provide 2E. 2D can alternatively undergo esterification via
alkylation using a halogenated compound and sodium carbonate in
order to provide 2E. 2E can be then treated with a mixture of
ammonium fluoride, methanol, and acetic acid in order to provide
2F. 2F can be treated with an isocyanate and TEA to produce 2G with
a urea moiety. 2F can alternatively be treated with an acyl
chloride and TEA to produce 2G with an amide moiety. 2F can
alternatively be treated with a sulfonyl chloride and TEA to
produce 2G with a sulfonamide moiety. 2F can alternatively be
treated with an aryl boronic acid and Cu(OAc).sub.2 to produce 2G
with a substituted or unsubstituted aryl moiety. A substituted or
unsubstituted heteroaryl moiety may be used in place of the aryl
moiety. 2G can be treated with TFA in order to deprotect the
t-butyl group on 2G and to produce 2H. 2H can then be treated with
guanidine under peptide coupling conditions (e.g. ethyl diisopropyl
carbodiimide (EDC) and hydroxybenzotriazole (HOBt) in order to
product 2I. 2I can be converted to 2J by treating with hydrogen and
palladium on carbon in ethyl acetate when R.sup.1=benzyl, or
trifluoroacetic acid (TFA) when R.sup.1=4-methoxybenzyl.
##STR00045##
[0123] A method of synthesizing azetedinones bearing R.sub.3
substituents containing an acylguanidine moiety is described in
scheme 103. The preparation of 103A is described in scheme 1. This
compound may be coupled via an intermediate mixed anhydride with
guanidine to provide 103B. 103B can be converted to the acid 103C
by treating with hydrogen and palladium on carbon in ethyl acetate
when R.sup.1=benzyl, or trifluoroacetic acid (TFA) when
R.sup.1=4-methoxybenzyl.
##STR00046## ##STR00047##
[0124] A method of synthesizing azetedinones bearing R.sub.3
substituents containing an acylguanidine moiety is described in
scheme 104. Alylation of the dianion of acid 104A using homoallyl
bromide provides 104B. Esterification can provide 104C.
Desilylation affords 104D. 104D can then be treated with an
isocyanate and TEA to produce 104E with a urea functionality. 104D
can alternatively be treated with an acyl chloride and TEA to
produce 104E with an amide functionality. 104D can alternatively be
treated with a sulfonyl chloride and TEA to produce 104E with a
sulfonamide functionality. 104D can alternatively be treated with
an aryl boronic acid and Cu(OAc).sub.2 to produce 104E with an aryl
functionality. Ozonolysis provides aldehyde 104F. Oxidation can
provide 104G. Coupling with guanidine via an active ester provides
104H. 104H can be converted to the acid 1041 by treating with
hydrogen and palladium on carbon in ethyl acetate when
R.sup.1=benzyl, or trifluoroacetic acid (TFA) when
R.sup.1=4-methoxybenzyl.
IV. b) N-guanidinyl 2-iminyl ethylene
##STR00048## ##STR00049##
[0126] One method of synthesizing compounds of the invention with
N-guanidinyl-2-iminyl-ethylene substituents is shown in Scheme 3.
In this Scheme, 3A can be treated with lithium hexamethyldisilazide
(LiHMDS) and 2-propenyl bromide to produce 3B. 3B can be treated
with an alcohol (R.sup.1--OH), EDC, DMAP in order to produce 3C. 3B
can alternatively be treated with a halogenated compound and sodium
carbonate in order to provide 3C. 3C can then be treated with a
mixture of ammonium fluoride, methanol, and acetic acid in order to
provide 3D. 3D can then be treated with an isocyanate and TEA to
produce 3E with an urea moiety. 3D can alternatively be treated
with an acyl chloride and TEA to produce 3E with an amide moiety.
3D can alternatively be treated with a sulfonyl chloride and TEA to
produce 3E with a sulfonamide moiety. 3D can alternatively be
treated with an aryl boronic acid and Cu(OAc).sub.2 to produce 3E
with a substituted or unsubstituted aryl moiety. A substituted or
unsubstituted heteroaryl moiety may be used in place of the aryl
moiety. 3E can be subjected to ozonolysis to produce 3F. 3F can be
treated with substituted or unsubstituted aminoguanidine and a
catalytic amount of an organic acid in order to produce 3G.
Finally, 3G can be converted to 3H by treating with hydrogen and
palladium on carbon in ethyl acetate when R.sup.1=benzyl, or
trifluoroacetic acid (TFA) when R.sup.1=4-methoxybenzyl.
##STR00050## ##STR00051##
[0127] Another method of synthesizing compounds of the invention
with N-guanidinyl-2-iminyl-ethylene substituents is shown in Scheme
4. In this Scheme, 4A can be treated with LDA and allyl bromide to
produce 4B. 4B can be then treated with 4-methoxybenzyl alcohol,
EDC and DMAP in order to produce 4C. 4C can be subjected to
ozonolysis to produce 4D. 4D can be treated with substituted or
unsubstituted aminoguanidine (R.sup.4.dbd.H, alkyl) and a catalytic
amount of an organic acid in order to produce 4E. 4E can then be
treated with (BOC).sub.2O, DIEA and DMAP in order to produce 4F. 4F
can then be treated with a mixture of ammonium fluoride, methanol,
and acetic acid in order to provide 4G. 4G can then be treated with
an isocyanate and TEA to produce 4H with an urea moiety. 4H can
then be treated with TFA in order to produce 4I.
##STR00052## ##STR00053##
[0128] Another method of synthesizing compounds of the invention
with N-guanidinyl-2-iminyl-ethylene substituents is shown in Scheme
5. In this Scheme, 5A can be treated with LiHMDS and 2-propenyl
bromide to produce 5B. 5B can be treated with benzyl alcohol, EDC,
DMAP in order to produce 5C. 5C can be treated with a mixture of
ammonium fluoride and methanol in order to provide 5D. 5D can be
treated with an isocyanate and TEA to produce 5E with an urea
moiety. 5E can be subjected to ozonolysis to produce 5F. 5F can be
subjected to hydrogenolysis in order to produce 5G. 5G can be
treated with substituted or unsubstituted aminoguanidine and a
catalytic amount of an organic acid in order to produce 5H.
##STR00054## ##STR00055##
[0129] Another method of synthesizing compounds of the invention
with N-guanidinyl-2-iminyl-ethylene substituents is shown in Scheme
6. In this Scheme, 6A can be treated with LiHMDS and 2-propenyl
bromide to produce 6B. 6B can be treated with 4-methoxybenzyl
alcohol, EDC, DMAP in order to produce 6C. 6C can be treated with a
mixture of ammonium fluoride, methanol, and acetic acid in order to
provide 6D. 6D can be treated with an isocyanate and TEA to produce
6E with an urea moiety. 6E can be subjected to ozonolysis to
produce 6F. 6F can be treated with substituted or unsubstituted
aminoguanidine and a catalytic amount of an organic acid in order
to produce 6G. 6G can then be treated with TFA in order to produce
6H.
##STR00056## ##STR00057##
[0130] Another method of synthesizing compounds of the invention
with N-guanidinyl-2-iminyl-ethylene substituents is shown in Scheme
7. In this Scheme, 7A can be treated with LiHMDS and 2-propenyl
bromide to produce 7B. 7B can be treated with 4-methoxybenzyl
alcohol, EDC, DMAP in order to produce 7C. 7C can be treated with
ammonium fluoride, methanol, and acetic acid in order to provide
7D. 7D can be treated with an aryl boronic acid and Cu(OAc).sub.2
to produce 7E with a substituted or unsubstituted aryl moiety. A
substituted or unsubstituted heteroaryl moiety may be used in place
of the aryl moiety. 7E can be subjected to ozonolysis to produce
7F. 7F can be treated with substituted or unsubstituted guanidine
and a catalytic amount of an organic acid in order to produce 7G.
7G can be treated with TFA in order to produce 7H.
##STR00058## ##STR00059##
[0131] Another method of synthesizing compounds of the invention
with N-guanidinyl-2-iminyl-ethylene substituents is shown in Scheme
8. In this Scheme, 8A can be treated with LDA and methyl iodide to
produce 8B. 8B can be treated with LiHMDS and 2-propenyl bromide to
produce 8C. 8C can be treated with an alcohol (R.sup.1--OH), EDC,
DMAP in order to produce 8D. 8C can alternatively be treated with a
halogenated compound and sodium carbonate in order to provide 8D.
8D can be treated with ammonium fluoride and methanol in order to
provide 8E. 8E can be treated with an isocyanate and TEA to produce
8F with a urea moiety. 8E can alternatively be treated with an acyl
chloride and TEA to produce 8F with an amide moiety. 8E can
alternatively be treated with a sulfonyl chloride and TEA to
produce 8F with a sulfonamide moiety. 8E can alternatively be
treated with an aryl boronic acid and Cu(OAc).sub.2 to produce 8F
with a substituted or unsubstituted aryl moiety. A substituted or
unsubstituted heteroaryl moiety may be used in place of the aryl
moiety. 8F can be subjected to ozonolysis to produce 8G. 8G can be
treated with substituted or unsubstituted aminoguanidine and a
catalytic amount of an organic acid in order to produce 8H. 8H can
then be deprotected in order to produce 81 by treating with
hydrogen and palladium on carbon in ethyl acetate when
R.sup.1=benzyl, or trifluoroacetic acid (TFA) when
R.sup.1=4-methoxybenzyl.
IV. c) N1-ethylidene-1H-imidazole-1,2-diamine
##STR00060##
[0133] One method of synthesizing compounds of the invention with
N-[2-amino-pyrazinyl]2-iminyl ethylene substituents is shown in
Scheme 9. 9A is a compound that can be produced by the methods
described in Schemes 3-8. 9A can be treated with ClCH.sub.2CHO in
ethanol in order to produce 9B. 9B can be used to produce 9C by
treating with hydrogen and palladium on carbon in ethyl acetate
when R.sup.1=benzyl, or trifluoroacetic acid (TFA) when
R.sup.1=4-methoxybenzyl.
IV. d) 1-(4,5-dihydro-1H-imidazol-2-yl)-2-propylidenehydrazine
##STR00061##
[0135] One method of synthesizing compounds of the invention with
N-[dihydroimidazyl]2-hydrazinylimino ethylene substituents is shown
in Scheme 10. 10A is a compound that can be produced by the methods
described in Schemes 3-8 (See 3F, 5F, 6F, 7F, 8G). 10A can be
treated with 1-(4,5-dihydro-1H-imidazol-2-yl)hydrazine
##STR00062##
in ethanol and acetic acid in order to produce 10B. 10B can be
converted to 10C by treating with hydrogen and palladium on carbon
in ethyl acetate when R.sup.1=benzyl, or trifluoroacetic acid (TFA)
when R.sup.1=4-methoxybenzyl.
IV. e) Heteroarylmethyl
##STR00063## ##STR00064## ##STR00065##
[0137] An alternative method of synthesizing heteroarylmethyl
substituents is shown in Scheme 11. Bis-BOC protection of 11A and
bromination provides 11C. 11C can be used in the LDA-mediated
alkylation of 11E. 11E can in turn prepared from 11D by
LDA-mediated alkylation with iodomethane, where R=Me. The alkylated
product 11F can be converted to 11G using ethanol and a peptide
coupling agent such as pyBOP. 11G can be desilylated to 11H using
ammonium fluoride, converted to urea 11I using and an appropriate
isocyanate and triethylamine, and deprotected with TFA to provide
11J.
##STR00066## ##STR00067## ##STR00068##
[0138] An alternative method of synthesizing heteroarylmethyl
substituents is shown in Scheme 12. Compound 12F was described in
scheme 11 (compound 11F with R.dbd.H). A benzyl ester 12G can be
prepared from 12F using benzyl alcohol and a peptide coupling agent
such as pyBOP. 12G can be desilylated with ammonium fluoride to
provide 12H. Treatment with an appropriate isocyanate can provide
12I. Hydrogenolysis can provide 12J, and BOC removal using TFA can
provide 12K.
##STR00069##
[0139] An alternative method of synthesizing heteroarylmethyl
substituents is shown in Scheme 13. 13D is prepared an analogous
fashion to 12G (with R.dbd.H) in scheme 12. 13D can be treated with
an aryl boronic acid to provide 13E, and the aryl moiety may be
substituted aryl, aryl, or heteroaryl. 13B can treated with TFA to
give acid 13F.
##STR00070##
[0140] A precursor for the synthesis of many heteroarylmethyl
variants can be prepared according to Scheme 14. 14A can be treated
with a mixture of benzyl alcohol, EDC, and di-isopropylethylamine
in DCM to produce 14B. 14B can be treated with excess sodium
borohydride in order to produce 14C. 14A can alternatively be
directed converted to 14C through reduction with BH.sub.3 in
THF.
##STR00071##
[0141] Compound 15A from Scheme 14 can be used to produce 15B by
alkylation with chloromethyl methyl ether. 15B can be readily
converted to 15C as shown in scheme 15, by ammonium fluoride
mediated desilylation, isocyanate acylation, and TFA-mediated BOC
deprotection.
##STR00072##
[0142] 16A from scheme 14 can be used to produce 16B by
base-mediated sulfonylation. 16B can be readily converted to 16C by
3 steps that include ammonium fluoride mediated desilylation,
isocyanate acylation, and BOC deprotection with TFA.
##STR00073##
[0143] The precursor from Scheme 14 (17A) can be used to produce
17B by base-mediated alkylation. 17B can be readily converted to
17C by 3 steps that include ammonium fluoride mediated
desilylation, isocyanate acylation, and BOC deprotection with
TFA.
##STR00074##
[0144] The precursor from Scheme 14 (18A) can be used to produce
18B by base-mediated reaction with a sulfamoyl chloride. 18B can be
readily converted to 18C by 3 steps that include ammonium fluoride
mediated desilylation, isocyanate acylation, and BOC deprotection
with TFA.
##STR00075##
[0145] The precursor from Scheme 14 (19A) can be used to produce
carbamate 19B by base-mediated reaction with an isocyanate. 19B can
be readily converted to 19C by 3 steps that include ammonium
fluoride mediated desilylation, isocyanate acylation, and BOC
deprotection with TFA.
##STR00076##
[0146] One method of synthesizing compounds of the invention with
tetrazole is shown in Scheme 20. LDA-mediated alkylation of 20A can
provide acid 20B. 20B can be converted to the nitrile 20C by
coupling via an acyl halide, reaction with ammonia to form a
primary amide, and finally dehydration. 20C can be desilylated with
ammonium fluoride to form 20D. Treatment of 20D with an isocyanate
provides urea 20E. The nitrile can be used to form tetrazole 20F
using TMS azide and a tin oxide reagent. TFA-mediated BOC
deprotection provides urea 20G.
##STR00077##
[0147] One method of synthesizing compounds of the invention with
tetrazole is shown in Scheme 21. 21A can be treated with
TMSCHN.sub.2, aqueous ammonia and then TFAA in order to produce
21B. 21B can be treated with sodium azide in the presence of LiCl,
NH.sub.4Cl and DMF in order to produce 21C. 21C can also obtained
by reacting 21B with azidotributylstannane. 21C can then be
alkylated with LDA and the Boc-protected bromo-aminopyridine,
protected with tosyl chloride and finally subjected to TBDMS
removal under TBAF conditions to produce 21D. 21D can then be
acylated with an isocyanate and all protecting groups removed with
TFA to produce desired tetazoles 21E.
##STR00078##
[0148] One method of synthesizing compounds of the invention with
trifluoromethyl is shown in Scheme 22. 22A can be hydrogenated to
provide ketone 22B. Ketone 22B can then be subjected to Mitsunobu
conditions followed by elimination to form 22C. 22C can be
transformed by Michael addition of ammonia and protection of the
amine to give 22D. Treatment of 22D with methylmagnesium bromide to
cyclize to the .beta.-lactam followed by methanolysis and TBDMS
protection to give 22E. 22E can then be alkylated to give 22F. 22F
can be treated with TBAF to remove the TBDMS group, acylated and
then all protecting groups removed with TFA to produce desired
CF.sub.3 compounds of the invention 22G.
##STR00079##
[0149] One method of synthesizing compounds of the invention with
geminally substituted methyl groups is shown in Scheme 23. 23A can
be treated with TBDMSC1 and TEA to produce 23B. 23B can be treated
with LDA and then Bis-BOC protected 4-(bromomethyl)pyridin-2-amine
in order to produce 23C. 23C can be treated with ammonium fluoride
in methanol in order to produce 23D. 23D can be treated with NaHMDS
and then an isocyanate in order to produce 23E. 23E can be treated
with TFA in dichloromethane in order to produce 23F.
##STR00080## ##STR00081##
[0150] A method of synthesizing azetedinones bearing R.sub.4
substituents containing sulfonamide, amides, urea, thiourea, and
amino groups is shown in Scheme 102. Compound 102A is described in
scheme 16. 102A can be converted to 102B by a three step sequence
including displacement of the mesylate with azide, desilylation,
and acylation with an appropriate isocyanate. 102B can be converted
to sulfonamide 102C by a sequence involving reduction,
sulfonylation, and BOC deprotection with TFA. Additionally, 102B
can be converted to amide 102D by a sequence involving reduction,
coupling with a carboxylic acid, and BOC deprotection with TFA.
Additionally, 102B can be converted to urea 102E by a sequence
involving reduction, acylation with an isocyanate, and BOC
deprotection with TFA. Additionally, 102B can be converted to amine
102F by a sequence involving reduction, reductive amination with an
aldehyde, and BOC deprotection with TFA.
##STR00082##
[0151] A method of synthesizing azetedinones bearing R.sub.4
substituents bearing a methylene (CH2) group attached to the
azetidinone is shown in scheme 105. Acid 105A (scheme 11) can be
converted to diazoketone 105B via the reaction of its mixed
anhydride with TMS diazomethane. Silver promoted rearrangement
provides 105C. Desilylation can provides 105D. Acylation with an
appropriate isocyanate can provide 105E. TFA deprotection can
provide 105F.
##STR00083##
[0152] A method of synthesizing azetedinones bearing R.sub.4
substituents bearing a methylene (CH.sub.2) group attached to the
azetidinone is shown in scheme 106. Diazoketone 106A (scheme 105)
can undergo silver promoted rearrangement in the presence of an
amine to provide 106B. Desilylation can provides 106C. Acylation
with an appropriate isocyanate can provide 106D. TFA deprotection
can provide 106E.
IV. f) Aryloxy
##STR00084## ##STR00085##
[0154] One method of synthesizing compounds of the invention with
aryloxy substituents is shown in Scheme 24. 24A can be treated with
thionyl chloride and then sodium periodate and ruthenium
trichloride in order to produce 24B. 24B can be treated with sodium
nitrate in sulfuric acid in order to produce 24C. 24C can be
treated with TBMSCl and then triphenylphosphine in water in order
to produce 24D. 24D can be treated with t-BuMgCl and TMSCl in ether
in order to produce 24E. 24E can be treated with an acyl chloride
in order to produce 24F. 24F can then be treated with TBAF and then
triphenylphosphine and isothiocyanate in order to produce 24G. 24G
can be treated with an aryl alcohol (or alternatively a substituted
aryl, heteroaryl, or substituted heteroaryl alcohol) and base in
order to produce 24H. 24H can be subjected to hydrogenation
conditions in order to produce 24I.
IV. g) Heteroaryloxy [pyridinyloxy]
##STR00086##
[0156] N-Heteroaryloxy substituents on azetidinone compounds can be
synthesized as shown in Scheme 25. 25A and 25B can be treated with
4-methoxyaniline under dehydrating conditions to produce an
intermediate imine, which is treated with 2-benzyloxyacetylchloride
to form compound 25C. 25C can be subjected to catalytic
hydrogenation conditions in order to produce 25D. The secondary
alcohol of 25D can be activated with (CF.sub.3SO.sub.2) 20 and
addition of 3-hydroxypyridine can provide 25E. An alternative path
to 23E can be through a Mitsunobu reaction of 25D and
3-hydroxypyridine in the presence of triphenylphosphine and DEAD.
25E can be treated with ceric ammonium nitrate (CAN) in order to
produce 25F. 25F can be treated with various isocyanates in order
to produce 25G.
IV. h) Heteroarylthio
##STR00087## ##STR00088##
[0158] A method of synthesizing heteroarylthio substituents is
shown in Scheme 101. Compound 101A can be treated with sulfide 101B
to provide thioether 101C. 101C can be treated with sulfenyl
chloride 101D to form disulfide 101E. This disulfide can be used to
functionalize the dianion of 101F, forming thioether 101G.
Esterification with a mixture of ethanol, DMAP, and EDC can provide
101H. The TBDMS group can deprotected using a mixture of ammonium
fluoride, acetic acid, and methanol to form 101I. 101I can be
treated with various isocyanates in the presence of triethylamine
to provide 101J. BOC deprotection with TFA can provide 101K.
IV. i) Arylthio
##STR00089##
[0160] A method of synthesizing arylthio substituents is shown in
Scheme 26. Compound 26A can be treated with LDA to form the lithium
dianion. Addition of bis-3-nitrophenyldisulfide provides 26B. The
carboxylate of 26B can be converted to the ethyl ester using
ethanol, DMAP, and EDC. The TBDMS group in the resulting ester 26C
can deprotected using ammonium fluoride in methanol. 26D can be
treated with various isocyanates in the presence of triethylamine
to provide 26E. The nitro group can be reduced using tin (II)
chloride to give the aromatic amine 26F.
IV. j) Heteroarylalkoxy
##STR00090## ##STR00091##
[0162] One method of synthesizing compounds of the invention with
[2-amino-4-azabenzyloxy] substituents is shown in Scheme 27.
Dibenzyltartartic acid 27A can be treated with thionyl chloride and
then sodium periodate in the presence of ruthenium trichloride in
order to produce 27B. 27B can be treated with sodium azide in
sulfuric acid in order to produce 27C. Bis-BOC
protected-2-amino-4-bromomethylpyridine can be treated with 27C in
the presence of a base in order to produce 27D. The azide group of
27D can be reduced with triphenylphosphine (Staudinger reaction) in
the presence of water in order to produce 27E. 27E can be treated
with t-BuMgCl and TMSCl in ether to produce 27F. 27F can be treated
with an isocyanate and TEA, and then subjected to hydrogenation
conditions in order to produce 27G.
##STR00092## ##STR00093##
[0163] A method of synthesizing compounds of the invention with
[2-amino-pyridinyl]methoxy substituents is shown in Scheme 28. 28A
or 28B can be treated with 4-methoxyaniline in ethanol with a
catalytic amount of an organic acid in order to produce 28C. 28C
can be treated with Bis-BOC protected-2-amino-4-bromomethylpyridine
in order to produce 28D. 28D can be treated with t-butylMgCl in
ether in order to produce 28E. Deprotection of the 4-methoxyphenyl
using CAN followed by reaction with various isocyanates, flowed by
BOC removal in TFA/DCM produces 28F.
IV. k) Heteroarylalkylthio
##STR00094##
[0165] Heteroarylthio substituents can be synthesized on
azetidinone compounds as shown in Scheme 29. 29A can be treated
with an equivalent of thiourea, then three equivalents of lithium
hydroxide, and then sodium bicarbonate in order to produce 29B. The
lithium dianion of 29C can be treated with 29B to provide compound
29E. The carboxylic acid of 29D can be protected as the
para-methoxybenzyl ester using para-methoxybenzyl alcohol in the
presences of EDC and DMAP in order to produce 29E. Deprotection of
the TBDMS group of 29E followed by urea formation with isocyanate,
and finally TFA deprotection of the BOC groups can produce 29F.
##STR00095##
[0166] An alternative method of synthesizing heteroarylthio
substituents is shown in Scheme 30. 30A can be treated with 2.4 eq.
of a lithium reagent to provide the dianion of 30A and then 1.2 eq.
of (t-BuS).sub.2 in order to produce 30B. 30B can be treated with
EDC and DMAP in order to produce the benzyl ester 30C. 30C can be
treated with Bis-BOC protected-2-amino-4-bromomethylpyridine and
sodium hydride in DMF in order to produce 30D. 30D can be treated
with ammonium fluoride, then isocyanate and finally TFA in order to
produce 30E.
##STR00096##
[0167] An alternative method of synthesizing heteroarylthio
substituents is shown in Scheme 31. 31A can be condensed with ethyl
glyoxylate, and the resulting imine can be acylated with an
appropriate acid chloride and cyclized to provide 31B.
Hydrogenolysis and treatment with triflic anhydride can provide
31C. Displacement of the triflate with an appropriate mercaptan can
provide thio ether 31D. Removal of the aryl group with CAN,
treatment with an appropriate isocyanate, and deprotection can
provide 31E.
IV. l) Heteroarylalkylamino
##STR00097##
[0169] A method of synthesizing compounds with 4-trifluoromethyl
substituents is shown in Scheme 32. 32A can be treated with
TMS-chloride followed by deprotonation with a Grignard reagent to
form 32B. The resulting beta-lactam 32B can be treated with 2
equiv. of n-butyl lithium to provide the dianion with after
quenching with tosyl azide to give a mixture of 32C and 32D.
Separation of the mixture followed by urea formation and reduction
of the azide can provide 32E. 32E can undergo reductive amination
with an appropriate aldehyde in the presence of a reducing agent
such as NaCNBH.sub.3 to provide, after deprotection of the BOC
groups with TFA, 32F.
##STR00098##
[0170] A method of synthesizing compounds with halomethyl
substituents is shown in Scheme 33. 33B can be prepared by
heterocyclization of 33A with various imines. The resulting
beta-lactam 33B can be hydrogenolyzed to give the intermediate
alcohol with is subsequently activated using
trifluoromethanesulfonic anhydride to give triflate 33C.
Displacement of the trifluomethanesulfonate with S or O
nucleophiles can provide 33D. Deprotection of the
para-methoxyphenyl with CAN, urea formation using various
isocyanates, and deprotection of the BOC groups with TFA can
provide 33E.
IV. m) Thio-.beta.-lactams
##STR00099##
[0172] A method of synthesizing thioazetedinones is shown in scheme
107. Beta lactam 107A can be converted to its thio analog 107B.
Desilylation, acylation with an appropriate isocyanate, and TFA
deprotection can provide 107E. This chemistry may be applied to
other beta lactams 107D of this invention to provide thio analogs
107E.
IV. n) Thiourea
##STR00100##
[0174] A method of synthesizing azetedinone thioureas is shown in
scheme 108. An appropriate ester 108A (see schemes 7 and 11) can be
desilylated to provide 108B. 108B can be converted either directed
to 108D via a thioisocyanate acylation or in a two step procedure
involving intermediate chloride 108C. TFA deprotection can provide
108E.
IV. o) Additional Exemplary Moieties
[0175] In another exemplary embodiment, the compounds of the
invention comprise a moiety that increases the water-solubility of
the parent compound. This moiety can be covalently attached,
directly or indirectly, to the 1, 2, 3 or 4 position on the
azetidinone ring. Exemplary moieties of use for increasing a
compound's water solubility include ethers and polyethers, e.g., a
member selected from ethylene glycol, and ethylene glycol
oligomers, having a molecular weight of from about 60 daltons to
about 10,000 daltons, and more preferably of from about 100 daltons
to about 1,000 daltons.
[0176] Representative polyether-based substituents include, but are
not limited to, the following structures:
##STR00101##
in which b is preferably a number from 1 to 100, inclusive. Other
functionalized polyethers are known to those of skill in the art,
and many are commercially available from, for example, Shearwater
Polymers, Inc. (Alabama).
[0177] In another exemplary embodiment, the compounds of the
invention comprise a moiety that includes a reactive functional
group for conjugating the compound to another molecule or to a
surface. This moiety can be attached, directly or indirectly, to
the 1, 2, 3 or 4 position on the azetidinone ring. The linkers of
use in the compounds of the invention can also include a cleaveable
group. In an exemplary embodiment, the cleaveable group is
interposed between the azetidinone core and a targeting agent or
macromolecular backbone. Representative useful reactive groups are
discussed in greater detail in succeeding sections. Additional
information on useful reactive groups is known to those of skill in
the art. See, for example, Hermanson, BIOCONJUGATE TECHNIQUES,
Academic Press, San Diego, 1996.
Reactive Functional Groups
[0178] As discussed above, the azetidinone core of the compounds of
the invention are optionally tethered to other species by means of
bonds formed between a reactive functional group on the azetidinone
or a linker attached to the azetidinone, and a reactive functional
group of complementary reactivity on the other species. For clarity
of illustration the succeeding discussion focuses on the
conjugation of representative azetidinones of the invention to
polymers, including poly(ethers) and dendrimers, and to targeting
agents useful for translocating the azetidinone-targeting agent
conjugate across a membrane. The focus exemplifies selected
embodiments of the invention from which others are readily inferred
by one of skill in the art. No limitation of the invention is
implied, by focusing the discussion on the representative
embodiments.
[0179] Exemplary azetidinones of the invention bear a reactive
functional group, which is generally located on the azetidinone
ring or on a substituted or unsubstituted alkyl or heteroalkyl
chain attached to the ring, allowing their facile attachment to
another species. A convenient location for the reactive group is
the terminal position of an alkyl or heteroalkyl substituent of the
azetidinone core.
[0180] Reactive groups and classes of reactions useful in
practicing the present invention are generally those that are well
known in the art of bioconjugate chemistry. Currently favored
classes of reactions available with reactive analogues are those
proceeding under relatively mild conditions. These include, but are
not limited to nucleophilic substitutions (e.g., reactions of
amines and alcohols with acyl halides, active esters),
electrophilic substitutions (e.g., enamine reactions) and additions
to carbon-carbon and carbon-heteroatom multiple bonds (e.g.,
Michael reaction, Diels-Alder addition). These and other useful
reactions are discussed in, for example, March, ADVANCED ORGANIC
CHEMISTRY, 3rd Ed., John Wiley & Sons, New York, 1985;
Hermanson, BIOCONJUGATE TECHNIQUES, Academic Press, San Diego,
1996; and Feeney et al., MODIFICATION OF PROTEINS; Advances in
Chemistry Series, Vol. 198, American Chemical Society, Washington,
D.C., 1982.
[0181] Exemplary reaction types include the reaction of carboxyl
groups and various derivatives thereof including, but not limited
to, N-hydroxysuccinimide esters, N-hydroxybenzotriazole esters,
acid halides, acyl imidazoles, thioesters, p-nitrophenyl esters,
alkyl, alkenyl, alkynyl and aromatic esters. Hydroxyl groups can be
converted to esters, ethers, aldehydes, etc. Haloalkyl groups are
converted to new species by reaction with, for example, an amine, a
carboxylate anion, thiol anion, carbanion, or an alkoxide ion.
Dienophile (e.g., maleimide) groups participate in Diels-Alder.
Aldehyde or ketone groups can be converted to imines, hydrazones,
semicarbazones or oximes, or reacted via such mechanisms as
Grignard addition or alkyllithium addition. Sulfonyl halides react
readily with amines, for example, to form sulfonamides. Amine or
sulfhydryl groups are, for example, acylated, alkylated or
oxidized. Alkenes can be converted to an array of new species using
cycloadditions, acylation, Michael addition, etc. Epoxides react
readily with amines and hydroxyl compounds.
[0182] Exemplary combinations of reactive functional groups found
on a ligand of the invention and on a targeting moiety (or polymer
or linker) are set forth in Table 1.
TABLE-US-00001 TABLE 1 Chemical Chemical Functionality 1
Functionality 2 Linkage Hydroxy Carboxy Ester Hydroxy Carbonate
Amine Carbamate SO.sub.3 Sulfate PO.sub.3 Phosphate Carboxy
Acyloxyalkyl Ketone Ketal Aldehyde Acetal Hydroxy Anhydride
Mercapto Mercapto Disulfide Carboxy Acyloxyalkyl Thioether Carboxy
Thioester Carboxy Amino amide Mercapto Thioester Carboxy
Acyloxyalkyl ester Carboxy Acyloxyalkyl amide Amino Acyloxyalkoxy
carbonyl Carboxy Anhydride Carboxy N-acylamide Hydroxy Ester
Hydroxy Hydroxymethyl ketone ester Hydroxy Alkoxycarbonyloxyalkyl
Amino Carboxy Acyloxyalkylamine Carboxy Acyloxyalkylamide Amino
Urea Carboxy Amide Carboxy Acyloxyalkoxycarbonyl Amide N-Mannich
base Carboxy Acyloxyalkyl carbamate Phosphate Hydroxy Phosphate
oxygen ester Amine Phosphoramidate Mercapto Thiophosphate ester
Ketone Carboxy Enol ester Sulfonamide Carboxy Acyloxyalkyl
sulfonamide Ester N-sulfonyl-imidate
[0183] One skilled in the art will readily appreciate that many of
these linkages may be produced in a variety of ways and using a
variety of conditions. For the preparation of esters, see, e.g.,
March supra at 1157; for thioesters, see, March, supra at 362-363,
491, 720-722, 829, 941, and 1172; for carbonates, see, March, supra
at 346-347; for carbanates, see March, supra at 1156-57; for
amides, see, March supra at 1152; for ureas and thioureas, see,
March supra at 1174; for acetals and ketals, see, Greene et al.
supra 178-210 and March supra at 1146; for acyloxyalkyl
derivatives, see, PRODRUGS: TOPICAL AND OCULAR DRUG DELIVERY, K. B.
Sloan, ed., Marcel Dekker, Inc., New York, 1992; for enol esters,
see, March supra at 1160; for N-sulfonylimidates, see, Bundgaard et
al., J. Med. Chem., 31:2066 (1988); for anhydrides, see, March
supra at 355-56, 636-37, 990-91, and 1154; for N-acylamides, see,
March supra at 379; for N-Mannich bases, see, March supra at
800-02, and 828; for hydroxymethyl ketone esters, see, Petracek et
al. Annals NY Acad. Sci., 507:353-54 (1987); for disulfides, see,
March supra at 1160; and for phosphonate esters and
phosphonamidates.
[0184] The reactive functional groups can be chosen such that they
do not participate in, or interfere with, the reactions necessary
to assemble the reactive ligand analogue. Alternatively, a reactive
functional group can be protected from participating in the
reaction by the presence of a protecting group. Those of skill in
the art will understand how to protect a particular functional
group from interfering with a chosen set of reaction conditions.
For examples of useful protecting groups, see Greene et al.,
PROTECTIVE GROUPS IN ORGANIC SYNTHESIS, John Wiley & Sons, New
York, 1991.
[0185] Generally, prior to forming the linkage between the ligand
and the targeting (or other) agent, and optionally, the linker
group, at least one of the chemical functionalities is activated.
One skilled in the art will appreciate that a variety of chemical
functionalities, including hydroxy, amino, and carboxy groups, can
be activated using a variety of standard methods and conditions.
For example, a hydroxyl group of the ligand (or targeting agent)
can be activated through treatment with phosgene to form the
corresponding chloroformate, or p-nitrophenylchloroformate to form
the corresponding carbonate.
[0186] In an exemplary embodiment, the invention makes use of a
targeting agent that includes a carboxyl functionality. Carboxyl
groups may be activated by, for example, conversion to the
corresponding acyl halide or active ester. This reaction may be
performed under a variety of conditions as illustrated in March,
supra pp. 388-89. In an exemplary embodiment, the acyl halide is
prepared through the reaction of the carboxyl-containing group with
oxalyl chloride. The activated agent is combined with a ligand or
ligand-linker arm combination to form a conjugate of the invention.
Those of skill in the art will appreciate that the use of
carboxyl-containing targeting agents is merely illustrative, and
that agents having many other functional groups can be conjugated
to the ligands of the invention.
Targeting Groups
[0187] The compounds of the invention may also be conjugated to an
agent that targets the compound to a specific tissue or region of
disease. The compound of the invention can be targeted for specific
delivery to the cells to be treated by conjugation of the compounds
to a targeting agent. The term "targeting agent" refers to a
species that serves to deliver the compound of the invention to a
specific site. Targeting agents include, for example, molecules
that specifically bind molecules present on a cell surface. Such
targeting agents useful in the invention include anti-cell surface
antigen antibodies; cytokines, including interleukins, factors such
as epidermal growth factor (EGF), and the like, are also specific
targeting agents known to bind cells expressing high levels of
their receptors. Targeting agents include species that are taken up
by cells using either active or passive mechanisms.
[0188] Particularly useful targeting agents for targeting the
compounds of the invention to cells for therapeutic activity
include those ligands that bind antigens or receptors present on
virus-infected cells to be treated. For example, antigens present
on T-cells, such as CD48, can be targeted with antibodies. Antibody
fragments, including single chain fragments, can also be used.
Other such ligand-receptor binding pairs are known in the
scientific literature for targeting anti-viral treatments to target
cells. Methods for producing conjugates of the compounds of the
invention and the targeting moieties are known.
[0189] Membrane translocation polypeptides are another exemplary
targeting agent. Membrane translocation polypeptides" have
amphiphilic or hydrophobic amino acid subsequences that have the
ability to act as membrane-translocating carriers. In one
embodiment, homeodomain proteins have the ability to translocate
across cell membranes. The shortest internalizable peptide of a
homeodomain protein, Antennapedia, was found to be the third helix
of the protein, from amino acid position 43 to 58 (see, e.g.,
Prochiantz, Current Opinion in Neurobiology 6:629-634 (1996)).
Another subsequence, the h (hydrophobic) domain of signal peptides,
was found to have similar cell membrane translocation
characteristics (see, e.g., Lin et al., J. Biol. Chem. 270:
14255-14258 (1995)).
[0190] Examples of peptide sequences include, but are not limited
to: an 11 amino acid peptide of the tat protein of HIV; a 20
residue peptide sequence which corresponds to amino acids 84-103 of
the p16 protein (see Fahraeus et al., Current Biology 6:84 (1996));
the third helix of the 60-amino acid long homeodomain of
Antennapedia (Derossi et al., J. Biol. Chem. 269:10444 (1994)); the
h region of a signal peptide such as the Kaposi fibroblast growth
factor (K-FGF) h region (Lin et al., supra); or the VP22
translocation domain from HSV (Elliot & O'Hare, Cell 88:223-233
(1997)). Other suitable chemical moieties that provide enhanced
cellular uptake may also be chemically linked to the compounds of
the invention.
[0191] Such subsequences can be used to translocate compounds of
the invention across a cell membrane. Compounds of the invention
can be conveniently fused to or derivatized with such sequences.
Typically, the translocation sequence is provided as part of a
fusion protein. Optionally, a linker as described herein can be
used to link the compound of the invention and the translocation
sequence. Any suitable linker can be used, e.g., a peptide linker
or other chemical linkers.
[0192] Toxin molecules also have the ability to transport compounds
across cell membranes. Often, such molecules are composed of at
least two parts (called "binary toxins"): a translocation or
binding domain or polypeptide and a separate toxin domain or
polypeptide. Typically, the translocation domain or polypeptide
binds to a cellular receptor, and then the toxin is transported
into the cell. Several bacterial toxins, including Clostridium
perfringens iota toxin, diphtheria toxin (DT), Pseudomonas exotoxin
A (PE), pertussis toxin (PT), Bacillus anthracis toxin, and
pertussis adenylate cyclase (CYA), have been used in attempts to
deliver peptides to the cell cytosol as internal or amino-terminal
fusions (Arora et al., J. Biol. Chem., 268: 3334-3341 (1993);
Perelle et al., Infect. Immun., 61: 5147-5156 (1993); Stenmark et
al., J. Cell Biol. 113: 1025-1032 (1991); Donnelly et al., PNAS
U.S.A. 90: 3530-3534 (1993); Carbonetti et al., Abstr. Annu. Meet.
Am Soc. Microbiol. 95: 295 (1995); Sebo et al., Infect. Immun.
63:3851-3857 (1995); Klimpel et al., PNAS U.S.A. 89:10277-10281
(1992); and Novak et al., J. Biol. Chem. 267:17186-17193
(1992)).
[0193] Non-covalent protein binding groups are also of use to
target the compounds of the invention to specific regions of the
body and to increase the half-life of the agent through protein
binding.
Macromolecular Conjugates
[0194] In an exemplary embodiment, the invention provides a
macromolecular, i.e., MW>1000 D, conjugate between the
azetidinone core and a macromolecular species. In one embodiment, a
macromolecular conjugate of the invention is formed by covalently
conjugating an azetidinone to a macromolecule via a reactive
functional group. In another embodiment, the macromolecular
conjugate is formed by a non-covalent interaction between a
azetidinone derivative and a macromolecule, e.g., a serum
protein.
[0195] In the following discussion, the invention is described by
reference to specific macromolecules of use for forming conjugates
with the novel azetidinone cores of the invention. Those of skill
in the art will appreciate that the focus of the discussion is for
clarity of illustration and does not limit the scope of the
invention. The invention provides macromolecular conjugates that
include components derived from biomolecules and synthetic
molecules. Exemplary biomolecules include polypeptides (e.g.,
antibodies, enzymes, receptors, antigens, immunogens such as KLH
(keyhole limpet hemocyanin), BSA (bovine serum albumin) and HSA
(human serum albumin); polysaccharides (e.g., starches, inulin,
dextran); lectins, nonpeptide antigens and the like. Exemplary
synthetic polymers include poly(acrylic acid), poly(lysine),
poly(glutamic acid), poly(ethylene imine), etc.
Covalent Conjugation
[0196] Selection of an appropriate reactive functional group on an
azetidinone core of the invention to form a desired macromolecular
species is well within the abilities of one of skill in the art.
Exemplary reactive functional groups of use in forming the covalent
conjugates of the invention are discussed above. It is well within
the abilities of one of skill to select and prepare an azetidinone
core of the invention having an appropriate reactive functional
group of complementary reactivity to a reactive group on its
conjugation partner.
[0197] In one embodiment, the bond formed between reactive
functional groups of the macromolecule and that of the azetidinone
attaches the azetidinone to the macromolecule essentially
irreversibly via a "stable bond" between the components. A "stable
bond", as used herein, is a bond, which maintains its chemical
integrity over a wide range of conditions (e.g., amide, carbamate,
carbon-carbon, ether, etc.). In another embodiment, a "cleaveable
bond" links the macromolecule and the azetidinone. A "cleaveable
bond", as used herein, is a bond that undergoes scission under
selected conditions. Cleaveable bonds include, but are not limited
to, disulfide, imine, carbonate and ester bonds. As discussed in
the preceding sections, the reactive functional group can be
located at one or more positions of the azetidinone.
Polysaccharides
[0198] In an exemplary embodiment, the present invention provides
conjugates between an azetidinone core and saccharides, e.g.,
polysaccharides. In an exemplary embodiment, the invention provides
a conjugate between a azetidinone and inulin. Inulin is a naturally
occurring polysaccharide, which has been previously investigated as
a carrier for diagnostic moieties (Rongved, P. K., J. Carbohydr.
Res. 1991, 214, 315; Corsi, D. M. V. E. et al., Chem. Eur. J. 2001,
7, 64). The structure of inulin can be described as a mixture of
linear .beta.-(2.fwdarw.1)-linked .alpha.-D-fructofuranosyl chains
with a .alpha.-D-glucopyranosyl unit at the terminal end. Inulin is
commercially available in a variety of molecular weights and the
degree of polymerization varies from 10 to 30, resulting in a
molecular weight distribution of 1500 to 5000 Da. The high
hydrophilicity, pH stability, low solution viscosity and
biocompatability of inulin ensure that its conjugates have
favorable pharmacological properties.
Dendrimer-Based Agents
[0199] In another aspect, the present invention provides a
azetidinone as set forth above, which is attached to a dendrimer
via a reactive functional group. Similar to the polymeric group
discussed above, the dendrimer has at least two reactive functional
groups. In one embodiment, one or more formed azetidinone is
attached to the dendrimer. Alternatively, the azetidinone is formed
directly on the dendrimer.
[0200] In an exemplary embodiment, a water-soluble and bio-adapted
polyester (polypropionate) class of dendrimers has been designed to
provide favorable pharmacokinetic properties. See., for example,
Ihre, H. et al., Macromolecules 1998, 31, 4061; Ihre, H. et al., J.
Am. Chem. Soc. 1996, 118, 6388; Anders, H., Ihre, H., Patent
WO/9900440 (Sweden)). In an exemplary embodiment, the termini of
the dendrimers are conjugated to a azetidinone core of the
invention.
Poly(ethylene glycol)-based Agents
[0201] In another exemplary embodiment, the invention provides a
conjugate between a azetidinone core of the invention and
poly(ethylene glycol). Poly(ethylene glycol) (PEG) is used in
biotechnology and biomedical applications. The use of this agent
has been reviewed (POLY(ETHYLENE GLYCOL) CHEMISTRY: BIOTECHNICAL
AND BIOMEDICAL APPLICATIONS, J. M. Harris, Ed., Plenum Press, New
York, 1992). Modification of enzymes (Chiu et al., J. Bioconjugate
Chem., 4: 290-295 (1993)), RGD peptides (Braatz et al.,
Bioconjugate Chem., 4: 262-267 (1993)), liposomes (Zalipsky, S.
Bioconjugate Chem., 4: 296-299 (1993)), and CD4-IgG glycoprotein
(Chamow et al., Bioconjugate Chem., 4: 133-140 (1993)) are some of
the recent advances in the use of polyethylene glycol. Surfaces
treated with PEG have been shown to resist protein deposition and
have improved resistance to thrombogenicity when coated on blood
contacting biomaterials (Merrill, "Poly(ethylene oxide) and Blood
Contact: A Chronicle of One Laboratory," in POLY(ETHYLENE GLYCOL)
CHEMISTRY: BIOTECHNICAL AND BIOMEDICAL APPLICATIONS, Harris, Ed.,
Plenum Press, New York, (1992), pp. 199-220).
[0202] Many routes are available for attaching an azetidinone core
of the invention onto a polymeric or oligomeric species. See, for
example, Dunn, R. L., et al., Eds. POLYMERIC DRUGS AND DRUG
DELIVERY SYSTEMS, ACS Symposium Series Vol. 469, American Chemical
Society, Washington, D.C. 1991; Herren et al., J. Colloid and
Interfacial Science 115: 46-55 (1987); Nashabeh et al., J.
Chromatography 559: 367-383 (1991); Balachandar et al., Langmuir 6:
1621-1627 (1990); and Burns et al., Biomaterials 19: 423-440
(1998).
[0203] Many activated derivatives of poly(ethyleneglycol) are
available commercially and in the literature. It is well within the
abilities of one of skill to choose, and synthesize if necessary,
an appropriate activated PEG derivative with which to prepare a
conjugate useful in the present invention. See, Abuchowski et al.
Cancer Biochem. Biophys., 7: 175-186 (1984); Abuchowski et al., J.
Biol. Chem., 252: 3582-3586 (1977); Jackson et al., Anal. Biochem.,
165: 114-127 (1987); Koide et al., Biochem Biophys. Res. Commun.,
111: 659-667 (1983)), tresylate (Nilsson et al., Methods Enzymol,
104: 56-69 (1984); Delgado et al., Biotechnol. Appl. Biochem., 12:
119-128 (1990)); N-hydroxysuccinimide derived active esters
(Buckmann et al., Makromol. Chem., 182: 1379-1384 (1981); Joppich
et al., Makromol. Chem., 180: 1381-1384 (1979); Abuchowski et al.,
Cancer Biochem. Biophys., 7: 175-186 (1984); Katre et al. Proc.
Natl. Acad. Sci. U.S.A., 84: 1487-1491 (1987); Kitamura et al.,
Cancer Res., 51: 4310-4315 (1991); Boccu et al., Z. Naturforsch.,
38C: 94-99 (1983), carbonates (Zalipsky et al., POLY(ETHYLENE
GLYCOL) CHEMISTRY: BIOTECHNICAL AND BIOMEDICAL APPLICATIONS,
Harris, Ed., Plenum Press, New York, 1992, pp. 347-370; Zalipsky et
al., Biotechnol. Appl. Biochem., 15: 100-114 (1992); Veronese et
al., Appl. Biochem. Biotech., 11: 141-152 (1985)), imidazolyl
formates (Beauchamp et al., Anal. Biochem., 131: 25-33 (1983);
Berger et al., Blood, 71: 1641-1647 (1988)), 4-dithiopyridines
(Woghiren et al., Bioconjugate Chem., 4: 314-318 (1993)),
isocyanates (Byun et al., ASAIO Journal, M649-M-653 (1992)) and
epoxides (U.S. Pat. No. 4,806,595, issued to Noishiki et al,
(1989). Other linking groups include the urethane linkage between
amino groups and activated PEG. See, Veronese, et al., Appl.
Biochem. Biotechnol., 11: 141-152 (1985).
V. Pharmaceutical Compositions and Methods of Treatment
[0204] The pharmaceutical compositions, or pharmaceutical
formulations, of the invention can take a variety of forms adapted
to the chosen route of administration. In general, they include a
compound of the invention and at least one pharmaceutical
excipient. Those skilled in the art will recognize various
synthetic methodologies that may be employed to prepare non-toxic
pharmaceutical formulations incorporating the compounds described
herein. Those skilled in the art will recognize a wide variety of
non-toxic pharmaceutically acceptable solvents that may be used to
prepare solvates of the compounds of the invention, such as water,
ethanol, propylene glycol, mineral oil, vegetable oil and
dimethylsulfoxide (DMSO).
[0205] The compositions of the invention may be administered
orally, topically, parenterally, by inhalation or spray or rectally
in dosage unit formulations containing conventional non-toxic
pharmaceutically acceptable carriers, adjuvants and vehicles. It is
further understood that the best method of administration may be a
combination of methods. Oral administration in the form of a pill,
capsule, elixir, syrup, lozenge, troche, or the like is
particularly preferred. The term parenteral as used herein includes
subcutaneous injections, intradermal, intravascular (e.g.,
intravenous), intramuscular, spinal, intrathecal injection or like
injection or infusion techniques.
[0206] The pharmaceutical formulations containing compounds of the
invention are preferably in a form suitable for oral use, for
example, as tablets, troches, lozenges, aqueous or oily
suspensions, dispersible powders or granules, emulsion, hard or
soft capsules, or syrups or elixirs.
[0207] Compositions intended for oral use may be prepared according
to any method known in the art for the manufacture of
pharmaceutical formulations, and such compositions may contain one
or more agents selected from the group consisting of sweetening
agents, flavoring agents, coloring agents and preserving agents in
order to provide pharmaceutically elegant and palatable
preparations. Tablets may contain the active ingredient in
admixture with non-toxic pharmaceutically acceptable excipients
that are suitable for the manufacture of tablets. These excipients
may be for example, inert diluents, such as calcium carbonate,
sodium carbonate, lactose, calcium phosphate or sodium phosphate;
granulating and disintegrating agents, for example, corn starch, or
alginic acid; binding agents, for example starch, gelatin or
acacia; and lubricating agents, for example magnesium stearate,
stearic acid or talc. The tablets may be uncoated or they may be
coated by known techniques to delay disintegration and absorption
in the gastrointestinal tract and thereby provide a sustained
action over a longer period. For example, a time delay material
such as glyceryl monostearate or glyceryl distearate may be
employed.
[0208] Exemplary compositions for oral administration include
suspensions which may contain, for example, microcrystalline
cellulose for imparting bulk, alginic acid or sodium alginate as a
suspending agent, methylcellulose as a viscosity enhancer, and
sweeteners or flavoring agents such as those known in the art; and
immediate release tablets which may contain, for example,
microcrystalline cellulose, dicalcium phosphate, starch, magnesium
stearate and/or lactose and/or other excipients, binders,
extenders, disintegrants, diluents and lubricants such as those
known in the art. The inventive compounds may be orally delivered
by sublingual and/or buccal administration, e.g., with molded,
compressed, or freeze-dried tablets. Exemplary compositions may
include fast-dissolving diluents such as mannitol, lactose,
sucrose, and/or cyclodextrins. Also included in such formulations
may be high molecular weight excipients such as celluloses (AVICEL)
or polyethylene glycols (PEG); an excipient to aid mucosal adhesion
such as hydroxypropyl cellulose (HPC), hydroxypropyl methyl
cellulose (HPMC), sodium carboxymethyl cellulose (SCMC), and/or
maleic anhydride copolymer (e.g., GANTREZ); and agents to control
release such as polyacrylic copolymer (e.g., CARBOPOL 934).
Lubricants, glidants, flavors, coloring agents and stabilizers may
also be added for ease of fabrication and use.
[0209] Formulations for oral use may also be presented as hard
gelatin capsules wherein the active ingredient is mixed with an
inert solid diluent, for example, calcium carbonate, calcium
phosphate or kaolin, or as soft gelatin capsules wherein the active
ingredient is mixed with water or an oil medium, for example peanut
oil, liquid paraffin or olive oil.
[0210] Aqueous suspensions contain the active materials in
admixture with excipients suitable for the manufacture of aqueous
suspensions. Such excipients are suspending agents, for example
sodium carboxymethylcellulose, methylcellulose,
hydroxypropylmethylcellulose, sodium alginate,
polyvinylpyrrolidone, gum tragacanth and gum acacia; and dispersing
or wetting agents, which may be a naturally-occurring phosphatide,
for example, lecithin, or condensation products of an alkylene
oxide with fatty acids, for example polyoxyethylene stearate, or
condensation products of ethylene oxide with long chain aliphatic
alcohols, for example heptadecaethyleneoxycetanol, or condensation
products of ethylene oxide with partial esters derived from fatty
acids and a hexitol such as polyoxyethylene sorbitol monooleate, or
condensation products of ethylene oxide with partial esters derived
from fatty acids and hexitol anhydrides, for example polyethylene
sorbitan monooleate. The aqueous suspensions may also contain one
or more preservatives, for example ethyl, or n-propyl
p-hydroxybenzoate, one or more coloring agents, one or more
flavoring agents, and one or more sweetening agents, such as
sucrose or saccharin.
[0211] Oily suspensions may be formulated by suspending the active
ingredients in a vegetable oil, for example arachis oil, olive oil,
sesame oil or coconut oil, or in a mineral oil such as liquid
paraffin. The oily suspensions may contain a thickening agent, for
example beeswax, hard paraffin or cetyl alcohol. Sweetening agents
such as those set forth above, and flavoring agents may be added to
provide palatable oral preparations. These compositions may be
preserved by the addition of an anti-oxidant such as ascorbic
acid.
[0212] Dispersible powders and granules suitable for preparation of
an aqueous suspension by the addition of water provide the active
ingredient in admixture with a dispersing or wetting agent,
suspending agent and one or more preservatives. Suitable dispersing
or wetting agents and suspending agents are exemplified by those
already mentioned above. Additional excipients, for example
sweetening, flavoring and coloring agents, may also be present.
[0213] Pharmaceutical formulations of the invention may also be in
the form of oil-in-water emulsions and water-in-oil emulsions. The
oily phase may be a vegetable oil, for example olive oil or arachis
oil, or a mineral oil, for example liquid paraffin or mixtures of
these. Suitable emulsifying agents may be naturally-occurring gums,
for example gum acacia or gum tragacanth; naturally-occurring
phosphatides, for example soy bean, lecithin, and esters or partial
esters derived from fatty acids and hexitol; anhydrides, for
example sorbitan monooleate; and condensation products of the said
partial esters with ethylene oxide, for example polyoxyethylene
sorbitan monooleate. The emulsions may also contain sweetening and
flavoring agents.
[0214] Syrups and elixirs may be formulated with sweetening agents,
for example glycerol, propylene glycol, sorbitol or sucrose. Such
formulations may also contain a demulcent, a preservative, and
flavoring and coloring agents. The pharmaceutical formulations may
be in the form of a sterile injectable aqueous or oleaginous
suspension. This suspension may be formulated according to the
known art using those suitable dispersing or wetting agents and
suspending agents, which have been mentioned above. The sterile
injectable preparation may also be a sterile injectable solution or
suspension in a non-toxic parenterally acceptable diluent or
solvent, for example as a solution in 1,3-butanediol. Among the
acceptable vehicles and solvents that may be employed are water,
Ringer's solution and isotonic sodium chloride solution. In
addition, sterile, fixed oils are conventionally employed as a
solvent or suspending medium. For this purpose any bland fixed oil
may be employed including synthetic mono- or diglycerides. In
addition, fatty acids such as oleic acid find use in the
preparation of injectables.
[0215] The composition of the invention may also be administered in
the form of suppositories, e.g., for rectal administration of the
drug. These compositions can be prepared by mixing the drug with a
suitable non-irritating excipient that is solid at ordinary
temperatures but liquid at the rectal temperature and will
therefore melt in the rectum to release the drug. Such materials
are cocoa butter and polyethylene glycols.
[0216] Alternatively, the compositions can be administered
parenterally in a sterile medium. The drug, depending on the
vehicle and concentration used, can either be suspended or
dissolved in the vehicle. Advantageously, adjuvants such as local
anesthetics, preservatives and buffering agents can be dissolved in
the vehicle.
[0217] For administration to non-human animals, the composition
containing the therapeutic compound may be added to the animal's
feed or drinking water. Also, it will be convenient to formulate
animal feed and drinking water products so that the animal takes in
an appropriate quantity of the compound in its diet. It will
further be convenient to present the compound in a composition as a
premix for addition to the feed or drinking water. The composition
can also added as a food or drink supplement for humans.
[0218] Dosage levels of the order of from about 5 mg to about 250
mg per kilogram of body weight per day and more preferably from
about 25 mg to about 150 mg per kilogram of body weight per day,
are useful in the treatment of the above-indicated conditions. The
amount of active ingredient that may be combined with the carrier
materials to produce a single dosage form will vary depending upon
the condition being treated and the particular mode of
administration. Dosage unit forms will generally contain between
from about 1 mg to about 500 mg of an active ingredient.
[0219] Frequency of dosage may also vary depending on the compound
used and the particular disease treated. However, for treatment of
most disorders, a dosage regimen of 4 times daily or less is
preferred. It will be understood, however, that the specific dose
level for any particular patient will depend upon a variety of
factors including the activity of the specific compound employed,
the age, body weight, general health, sex, diet, time of
administration, route of administration and rate of excretion, drug
combination and the severity of the particular disease undergoing
therapy.
[0220] Preferred compounds of the invention will have desirable
pharmacological properties that include, but are not limited to,
oral bioavailability, low toxicity, low serum protein binding and
desirable in vitro and in vivo half-lives. Penetration of the blood
brain barrier for compounds used to treat CNS disorders is
necessary, while low brain levels of compounds used to treat
peripheral disorders are often preferred.
[0221] Assays may be used to predict these desirable
pharmacological properties. Assays used to predict bioavailability
include transport across human intestinal cell monolayers,
including Caco-2 cell monolayers. Toxicity to cultured hepatocyctes
may be used to predict compound toxicity. Penetration of the blood
brain barrier of a compound in humans may be predicted from the
brain levels of laboratory animals that receive the compound
intravenously.
[0222] Serum protein binding may be predicted from albumin binding
assays. Such assays are described in a review by Oravcova, et al.
(Journal of Chromatography B (1996) volume 677, pages 1-27).
[0223] Compound half-life is inversely proportional to the
frequency of dosage of a compound. In vitro half-lives of compounds
may be predicted from assays of microsomal half-life as described
by Kuhnz and Gieschen (Drug Metabolism and Disposition, (1998)
volume 26, pages 1120-1127).
[0224] The amount of the composition required for use in treatment
will vary not only with the particular compound selected but also
with the route of administration, the nature of the condition being
treated and the age and condition of the patient and will
ultimately be at the discretion of the attendant physician or
clinician.
[0225] Exemplary compositions for nasal aerosol or inhalation
administration include solutions which may contain, for example,
benzyl alcohol or other suitable preservatives, absorption
promoters to enhance absorption and/or bioavailability, and/or
other solubilizing or dispersing agents such as those known in the
art.
[0226] Exemplary compositions for parenteral administration include
injectable solutions or suspensions which may contain, for example,
suitable non-toxic, parenterally acceptable diluents or solvents,
such as mannitol, 1,3-butanediol, water, Ringer's solution, an
isotonic sodium chloride solution, or other suitable dispersing or
wetting and suspending agents, including synthetic mono- or
diglycerides, and fatty acids, including oleic acid.
[0227] Exemplary compositions for rectal administration include
suppositories which may contain, for example, suitable
non-irritating excipients, such as cocoa butter, synthetic
glyceride esters or polyethylene glycols, which are solid at
ordinary temperatures but liquefy and/or dissolve in the rectal
cavity to release the drug.
[0228] The novel compounds of the invention possess tryptase
inhibitory activity. As a result of this tryptase activity, the
compounds of the invention as well as an inner salt thereof, a
pharmaceutically acceptable salt thereof, a hydrolyzable ester
thereof, or a solvate thereof, are useful as antiinflammatory
agents particularly in the treatment of chronic asthma and may also
be useful in treating or preventing allergic rhinitis, inflammatory
bowel disease, psoriasis, conjunctivitis, atopic dermatitis,
rheumatoid arthritis, osteoarthritis, and other chronic
inflammatory joint diseases, or diseases of joint cartilage
destruction. Additionally, these compounds may be useful in
treating or preventing myocardial infarction, stroke, angina and
other consequences of atherosclerotic plaque rupture. Additionally,
these compounds may be useful for treating or preventing diabetic
retinopathy, tumor growth and other consequences of angiogenosis.
Additionally, these compounds may be useful for treating or
preventing fibrotic conditions, for example, fibrosis, scleroderma,
pulmonary fibrosis, liver cirrhosis, myocardial fibrosis,
neurofibromas and hypertrophic scars.
[0229] The compounds of the invention are also inhibitors of Factor
Xa and/or Factor VIIa. As a result, the compounds of the invention
as well as an inner salt or a pharmaceutically acceptable salt
thereof, a hydrolyzable ester thereof, or a solvate thereof may
also be useful in the treatment or prevention of thrombotic events
associated with coronary artery and cerebrovascular disease, venous
or arterial thrombosis, coagulation syndromes, ischemia and angina
(stable and unstable), deep vein thrombosis (DVT), disseminated
intravascular coagulopathy, Kasacach-Merritt syndrome, pulmonary
embolism, myocardial infarction, cerebral infarction, cerebral
thrombosis, transient ischemic attacks, atrial fibrillation,
cerebral embolism, thromboembolic complications of surgery (such as
hip or knee replacement, introduction of artificial heart valves
and endarterectomy) and peripheral arterial occulsion and may also
be useful in treating or preventing myocardial infarction, stroke,
angina and other consequences of atherosclerotic plaque rupture.
The compounds of the invention possessing Factor Xa and/or Factor
VIIa inhibition activity may also be useful as inhibitors of blood
coagulation such as during the preparation, storage and
fractionation of whole blood.
[0230] The compounds of the invention are also inhibitors of
urokinase-type plasminogen activator. As a result, the compounds of
the invention as well as an inner salt or a pharmaceutically
acceptable salt thereof, a hydrolyzable ester thereof, or a solvate
thereof may be useful in the treatment or prevention of restenosis
and aneurysms, in the treatment or prevention of myocardial
infarction, stroke, angina and other consequences of
atherosclerotic plaque rupture, and may also be useful in the
treatment of malignancies, prevention of metastases, prevention of
prothrombotic complications of cancer, and as an adjunct to
chemotherapy.
[0231] The compounds of the invention also possess thrombin and
trypsin inhibitory activity. As a result, the compounds of the
invention as well as an inner salt or a pharmaceutically acceptable
salt thereof, a hydrolyzable ester thereof, or a solvate thereof
may be useful in treating or preventing pancreatitis, in the
treatment or prevention of thrombotic events as described above,
and may also be useful as inhibitors of blood coagulation such as
during the preparation, storage, and fractionation of whole
blood.
[0232] The compounds of the invention are also inhibitors of Factor
XIa. As a result, the compounds of the invention as well as an
inner salt or a pharmaceutically acceptable salt thereof, a
hydrolyzable ester thereof, or a solvate thereof may also be useful
in the treatment or prevention of thrombotic events associated with
coronary artery and cerebrovascular disease, venous or arterial
thrombosis, coagulation syndromes, ischemia and angina (stable and
unstable), deep vein thrombosis (DVT), disseminated intravascular
coagulopathy, Kasacach-Merritt syndrome, pulmonary embolism,
myocardial infarction, cerebral infarction, cerebral thrombosis,
transient ischemic attacks, atrial fibrillation, cerebral embolism,
thromboembolic complications of surgery (such as hip or knee
replacement, introduction of artificial heart valves and
endarterectomy) and peripheral arterial occulsion and may also be
useful in treating or preventing myocardial infarction, stroke,
angina and other consequences of atherosclerotic plaque rupture.
The compounds of the invention possessing Factor XIa inhibition
activity may also be useful as inhibitors of blood coagulation such
as during the preparation, storage and fractionation of whole
blood.
[0233] The compounds of the invention as well as an inner salt or a
pharmaceutically acceptable salt thereof, a hydrolyzable ester
thereof, or a solvate thereof may be administered orally,
topically, rectally or parenterally or may be administered by
inhalation into the bronchioles or nasal passages. The method of
administration will, or course, vary upon the type of disease being
treated. The amount of active compound administered will also vary
according to the method of administration and the disease being
treated. An effective amount will be within the dosage range of
about 0.1 to about 100 mg/kg, preferably about 0.2 to about 50
mg/kg and more preferably about 0.5 to about 25 mg/kg per day in a
single or multiple doses administered at appropriate intervals
throughout the day.
[0234] The pharmaceutical composition used in these therapies can
be in a variety of forms. These include, for example, solid,
semi-solid and liquid dosage forms such as tablets, pills, powders,
liquid solutions or suspensions, liposomes, injectable and
infusible solutions. Such compositions can include pharmaceutically
acceptable excipients, preservatives, stabilizers, and other agents
conventionally employed in the pharmaceutical industry.
[0235] When the compounds of the invention as well as an inner salt
or a pharmaceutically acceptable salt thereof, a hydrolyzable ester
thereof, or a solvate thereof are employed to treat asthma or
allergic rhinitis they will may be formulated as aerosols. The term
"aerosol" includes any gas-borne suspended phase of the active
compound which is capable of being inhaled into the bronchioles or
nasal passage. Aerosol formulations include a gas-borne suspension
of droplets of the active compound as produced in a metered dose
inhaler or nebulizer or in a mist sprayer. Aerosol formulations
also include a dry powder composition suspended in air or other
carrier gas. The solutions of the active compounds of the invention
used to make the aerosol formulation will be in a concentration of
from about 0.1 to about 100 mg/ml, more preferably 0.1 to about 30
mg/ml, and most preferably from about 1 to about 10 mg/ml. The
solution will usually include a pharmaceutically acceptable buffer
such as a phosphate or bicarbonate to give a pH of from about 5 to
9, preferably 6.5 to 7.8, and more preferably 7.0 to 7.6.
Preservatives and other agents can be included according to
conventional pharmaceutical practice.
[0236] Other pharmaceutically active agents can be employed in
combination with the compounds of the invention depending upon the
disease being treated. For example, in the treatment of asthma,
.beta.-adrenergic agonists such as albuterol, terbutaline,
formoterol, fenoterol or prenaline can be included as can
anticholinergics such as ipratropium bromide, anti-inflammatory
cortiocosteroids such as beclomethasone, triamcinolone, flurisolide
or dexamethasone, and anti-inflammatory agents such as cromolyn and
nedocromil.
[0237] In addition to the novel compounds of the invention and the
methods of use for the compounds of the invention, this invention
is also directed to novel intermediates and novel synthetic routes
employed in the preparation of such compounds.
[0238] FXIa inhibition according to the invention represents a more
effective and safer method of inhibiting thrombosis compared to
inhibiting other coagulation serine proteases such as thrombin or
Factor Xa. Administration of a small molecule FXIa inhibitor should
have the effect of inhibiting thrombin generation and clot
formation with no or substantially no effect on bleeding times and
little or no impairment of haemostasis. These results differ
substantially from that of other "direct acting" coagulation
protease inhibitors (e.g. active-site inhibitors of thrombin and
Factor Xa), which demonstrate prolongation of bleeding time and
less separation between antithrombotic efficacy and bleeding time
prolongation. A preferred method according to the invention
comprises administering to a mammal a pharmaceutical composition
containing at least one compound of the invention.
[0239] The methods of the present invention are useful for treating
or preventing those conditions which involve the action of Factor
XIa. Accordingly, the methods of the present invention are useful
in treating consequences of atherosclerotic plaque rupture
including cardiovascular diseases associated with the activation of
the coagulation cascade in thrombotic or thrombophilic states. As
used herein, the terms "treating" or "treatment" encompass
responsive and/or prophylaxis measures, e.g., measures designed to
inhibit or delay the onset of the disease, achieve a full or
partial reduction of the symptoms or disease state, and/or to
alleviate, lessen, or cure the disease or disorder and/or its
symptoms.
[0240] More particularly, the methods of the present invention may
be used to treat acute coronary syndromes such as coronary artery
disease, myocardial infarction, unstable angina (including
crescendo angina), ischemia (e.g., ischemia resulting from vascular
occlusion), and cerebral infarction. The methods of the present
invention further may be useful in treating stroke and related
cerebral vascular diseases (including cerebrovascular accident and
transient ischemic attack); venous thrombosis and thrombo-embolism,
such as deep vein thrombosis (DVT) and pulmonary embolism;
thrombosis associated with atrial fibrillation, ventricular
enlargement, dilated cardiac myopathy, or heart failure; peripheral
arterial disease and intermittent claudication; the formation of
atherosclerotic plaques and transplant atherosclerosis; restenosis
following arterial injury induced endogenously (by rupture of an
atherosclerotic plaque), or exogenously (by invasive cardiological
procedures such as vessel wall injury resulting from angioplasty);
disseminated intravascular coagulopathy, Kasabach-Merritt syndrome,
cerebral thrombosis, cerebral embolism, and disseminated
intravascular coagulopathy.
[0241] Additionally, the methods of the present invention may be
useful in treating thrombo-embolic consequences or complications
associated with surgery (such as hip replacement, endarterectomy,
introduction of artificial heart valves, vascular grafts,
mechanical organs, and implantation or transplantation of organ,
tissue or cells); medications (such as oral contraceptives, hormone
replacement, and heparin, e.g., for treating heparin-induced
thrombocytopenia); and pregnancy or childbirth. The methods of the
present invention may be used to treat thrombosis due to
confinement (i.e. immobilization, hospitalization, bed rest, limb
immobilization, e.g., with immobilizing casts, etc.).
[0242] The methods of the present invention also may be useful in
preventing thrombosis and complications in patients genetically
predisposed to arterial thrombosis or venous thrombosis (including
activated protein C resistance, FVleiden, Prothrombin 20210,
elevated coagulation factors FVII, FVIII, FIX, FX, FXI,
prothrombin, TAFI and fibrinogen), elevated levels of homocystine,
and deficient levels of antithrombin, protein C, and protein S. The
inventive methods may be used for treating heparin-intolerant
patients, including those with congenital and acquired antithrombin
III deficiencies, heparin-induced thrombocytopenia, and those with
high levels of polymorphonuclear granulocyte elastase. The methods
of this invention may be used to treat all forms of
thrombophilia.
[0243] The methods of the present invention may also be used to
maintain blood vessel potency, for example, in patients undergoing
transluminal coronary angioplasty, or in connection with vascular
surgery such as bypass grafting, arterial reconstruction,
atherectomy, vascular grafts, stent patency, and organ, tissue or
cell implantation and transplantation. The inventive methods may be
used to inhibit blood coagulation in connection with the
preparation, storage, fractionation, or use of whole blood. For
example, the inventive methods may be used in maintaining whole and
fractionated blood in the fluid phase such as required for
analytical and biological testing, e.g., for ex vivo platelet and
other cell function studies, bioanalytical procedures, and
quantitation of blood-containing components, or for maintaining
extracorpeal blood circuits, as in dialysis or surgery (e.g.,
coronary artery bypass surgery).
[0244] In addition, the methods of the present invention may be
useful in treating and preventing the prothrombotic complications
of cancer. The methods may be useful in treating tumor growth, as
an adjunct to chemotherapy, for preventing angiogenesis, and for
treating cancer, more particularly, cancer of the lung, prostate,
colon, breast, ovaries, and bone.
[0245] The methods of the present invention also may be used to
treat diabetes mellitus, hypertension, or hypercholesterolemia.
[0246] In carrying out the methods of the present invention, it may
be desired to administer the compounds of the invention (Factor XIa
inhibitors) in combination with each other and one or more other
agents for achieving a therapeutic benefit such as antithrombotic
or anticoagulant agents, anti-hypertensive agents, anti-ischemic
agents, anti-arrhythmic agents, platelet function inhibitors, and
so forth. More particularly, the inventive methods may be carried
out by administering the small molecule Factor XIa inhibitors in
combination with aspirin, clopidogrel, ticlopidine or CS-747,
warfarin, low molecular weight heparins (such as LOVENOX),
GPIIb/GPIIIa blockers, PAI-1 inhibitors such as XR-330 and T-686,
P2Y1 and P2Y12 receptor antagonists; thromboxane receptor
antagonists (such as ifetroban), prostacyclin mimetics, thromboxane
A synthetase inhibitors (such as picotamide), serotonin-2-receptor
antagonists (such as ketanserin); compounds that inhibit other
coagulation factors such as FVII, FVIII, FIX, FX, prothrombin,
TAFI, and fibrinogen, and/or other compounds that inhibit FXI;
fibrinolytics such as TPA, streptokinase, PAI-1 inhibitors, and
inhibitors of .alpha.-2-antiplasmin such as
anti-.alpha.-2-antiplasmin antibody fibrinogen receptor
antagonists, hypolipidemic agents, such as HMG-CoA reductase
inhibitors (e.g., pravastatin, simvastatin, atorvastatin,
fluvastatin, cerivastatin, AZ4522, and itavastatin), and microsomal
triglyceride transport protein inhibitors (such as disclosed in
U.S. Pat. Nos. 5,739,135, 5,712,279 and 5,760,246);
antihypertensive agents such as angiotensin-converting enzyme
inhibitors (e.g., captopril, lisinopril or fosinopril);
angiotensin-II receptor antagonists (e.g., irbesartan, losartan or
valsartan); ACE/NEP inhibitors (e.g., omapatrilat and
gemopatrilat); and/or .beta.-blockers (such as propranolol, nadolol
and carvedilol). The inventive methods may be carried out by
administering the small molecule Factor XIa inhibitors in
combination with anti-arrhythmic agents such as for atrial
fibrillation, for example, amiodarone or dofetilide.
[0247] In carrying out the methods of the present invention, it may
be desired to administer the compounds of the invention (Factor XIa
inhibitors) in combination with agents that increase the levels of
cAMP or cGMP in cells for a therapeutic benefit. For example, the
compounds of the invention may have advantageous effects when used
in combination with phosphodiesterase inhibitors, including PDE1
inhibitors (such as those described in Journal of Medicinal
Chemistry, Vol. 40, pp. 2196-2210 [1997]), PDE2 inhibitors, PDE3
inhibitors (such as revizinone, pimobendan, or olprinone), PDE4
inhibitors (such as rolipram, cilomilast, or piclamilast), PDE7
inhibitors, or other PDE inhibitors such as dipyridamole,
cilostazol, sildenafil, denbutyline, theophylline
(1,2-dimethylxanthine), ARIFLO.TM. (i.e.,
cis-4-cyano-4-[3-(cyclopentyloxy)-4-methoxyphenyl]cyclohexane-1-carboxyli-
c acid), arofyline, roflumilast, C-11294A, CDC-801, BAY-19-8004,
cipamfylline, SCH351591, YM-976, PD-189659, mesiopram,
pumafentrine, CDC-998, IC-485, and KW-4490.
[0248] The inventive methods may be carried out by administering
the compounds of the invention in combination with prothrombolytic
agents, such as tissue plasminogen activator (natural or
recombinant), streptokinase, reteplase, activase, lanoteplase,
urokinase, prourokinase, anisolated streptokinase plasminogen
activator complex (ASPAC), animal salivary gland plasminogen
activators, and the like.
[0249] The inventive methods may be carried out by administering
the compounds of the invention in combination with
.beta.-adrenergic agonists such as albuterol, terbutaline,
formoterol, salmeterol, bitolterol, pilbuterol, or fenoterol;
anticholinergics such as ipratropium bromide; anti-inflammatory
cortiocosteroids such as beclomethasone, triamcinolone, budesonide,
fluticasone, flunisolide or dexamethasone; and anti-inflammatory
agents such as cromolyn, nedocromil, theophylline, zileuton,
zafirlukast, monteleukast and pranleukast.
[0250] The small molecule Factor XIa inhibitors may act
synergistically with one or more of the above agents. Thus, reduced
doses of thrombolytic agent(s) may be used, therefore obtaining the
benefits of administering these compounds while minimizing
potential hemorrhagic and other side effects.
[0251] The effective amount of a small molecule Factor XIa
inhibitor administered according to the present invention may be
determined by one of ordinary skill in the art. The specific dose
level and frequency of dosage for any particular subject may vary
and will depend upon a variety of factors, including the activity
of the specific compound employed, the metabolic stability and
length of action of that compound, the species, age, body weight,
general health, sex and diet of the subject, the mode and time of
administration, rate of excretion, drug combination, and severity
of the particular condition. An exemplary effective amount of
compounds of the invention may be within the dosage range of about
0.1 to about 100 mg/kg, preferably about 0.2 to about 50 mg/kg and
more preferably about 0.5 to about 25 mg/kg (or from about 1 to
about 2500 mg, preferably from about 5 to about 2000 mg) on a
regimen in single or 2 to 4 divided daily doses.
EXAMPLES
[0252] Proton NMR are recorded on Varian AS 300 spectrometer and
chemical shifts are reported as .delta. (ppm) down field from
tetramethylsilane. Mass spectra are determined on Micromass Quattro
II.
Example 1
General method A for the preparation of 3-aminopyridyl beta-lactam
acids
##STR00102## ##STR00103##
[0254] 4-Methyl-2-bis(Boc)-aminopyridine (2): Under argon, in a
1-L, 3-necked flask was placed a solution of 2-aminopicoline (16.23
g, 150.1 mmol, 1.0 equiv), triethylamine (41.8 mL, 30.04 g, 300.2
mmol, 2.0 equiv), dimethylaminopyridine (DMAP) (18.3 g, 150 mmol,
1.0 equiv), and CH.sub.2Cl.sub.2 (300 mL). The mixture was cooled
to 5-7.degree. C. in an ice-water bath with magnetic stirring. A
solution of Boc-anhydride (100 g, 458.2 mmol, 3.1 equiv) in
CH.sub.2Cl.sub.2 (100 mL) was added dropwise over a period of 30
min. The ice bath was removed and the reaction allowed to warm to
room temperature with stirring for 4 h. The reaction mixture was
worked up by washing the CH.sub.2Cl.sub.2 solution with sat.
NH.sub.4Cl (2.times.400 mL) followed by sat. NaHCO.sub.3
(2.times.400 mL), and then passing the solution through silica gel
(350 mL fritted glass funnel) (elution with CH.sub.2Cl.sub.2, 500
mL). Rotary-evaporation of the solvent and tert-butanol at 5 mm Hg,
70.degree. C., 18 h provided the crude product as a
semi-crystalline straw-colored oil. This material was further
purified by silica gel chromatography on a 330 g Combi-flash
pre-packed cartridge, gradient elution with 100% hexanes to 20%
ethyl acetate in hexanes. Fractions containing product were pooled
and evaporated to provide 27.6 g, 60% yield of an oil. The oil was
transformed into a solid by first dissolving it in
CH.sub.2Cl.sub.2, then adding hexane and concentrating in vacuo at
25.degree. C. for 30 min, followed by high vacuum produced a
free-flowing powder 2. TLC: hexanes/EtOAc [2:1] Rf.about.0.6.
[0255] 4-Bromomethyl-2-bis-Boc-aminopyridine (3): To a solution of
4-methyl-2-bis(boc)aminopyridine 2 (26.0 g, 84.4 mmol, 1.0 equiv)
in CCl.sub.4 (421 mL) was added N-bromosuccinimide (15.0 g, 84.4
mmol, 1.0 equiv) and dibenzoylperoxide (97%) (204 mg, 0.84 mmol,
0.01 equiv). The reaction mixture was quickly heated to reflux (hot
plate with assistance from a heat gun) and the reaction was
illuminated with a 500-watt halogen work light and two, 175-watt
incandescent spotlights (Home Depot). The reaction was monitored at
210 nm and when the reaction had reached completion (1 hr at
reflux), the reaction mixture was cooled to room temperature,
filtered through celite/fritted glass to remove much of the
succinimide. The volatiles were removed to give a crude material
which was purified by silica gel chromatography (330 gram
Combiflash silica gel) using gradient elution from 1% acetone in
CH.sub.2Cl.sub.2 to 8% acetone in CH.sub.2Cl.sub.2. Fractions
containing pure product from each of the four chromatography runs
were pooled and evaporated to provide 10 g (31%) of 3 as a yellow
foam.
[0256]
(2S,3R)-1-(tert-butyldimethylsilyl)-3-((2-(di-t-butyloxycarbonylami-
no)pyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid (5):
Commercially available beta-lactam 4 (15.31 mmol, 3.51 g, 1.0 eq.)
was dissolved in 40 mL dry THF at room temperature with magnetic
stirring, in an oven-dried flask with dry stir bar under dry argon.
The solution was cooled to -78.degree. C. and the LDA solution
(2.05 eq., 31.38 mmol, 17.4 mL 1.8M solution from Aldrich) was
added slowly over 5 minutes by syringe to the cooled stirring
solution. The solution was kept at -78.degree. C. for 15 minutes
and warmed to 0.degree. C. with an ice bath. The solution was kept
at 0.degree. C. for 45 minutes, and then cooled to -42.degree. C.
(dry ice-acetonitrile). Bromide 3 (5.34 g, 13.78 mmol, 0.9 eq.) was
dissolved in 20 mL of dry THF and cooled to -42.degree. C. (dry
ice-acetonitrile). The resulting bromide solution was added
dropwise via a dry narrow bore cannula to the enolate solution
using balloon pressure, with both vessels cooled to -42.degree. C.
during the 30 minute addition. A deep dark blue-black color forms
immediately. After the addition was complete, the vial that
contained the bromide solution was rinsed with two 2 mL portions of
dry THF, which was cooled to -42.degree. C. and added to the
enolate solution. The reaction vessel was kept at -42.degree. C.
for 4 hours. A quench solution was prepared by adding 50 g of ice
to 150 mL of 5% aqueous KHSO.sub.4 solution in an Erlenmeyer. The
reaction mixture was poured into the quench solution, and the
vessel was rinsed with 5.times.20 mL aliquots of ethyl acetate,
which were poured into the quench solution. The two layers were
poured into a separatory funnel. The pH of the aqueous layer was
2-3. The organic layer was separated, the organic layer extracted
with 3 portions of ethyl acetate, and the combined organic layers
were washed with brine and concentrated in vacuo. The residue was
dissolved in minimal methylene chloride and put on a 120 g
pre-packed ISCO Combiflash silica column. The column was eluted
with a dual solvent system: solvent A=hexane, solvent B=2% acetic
acid in ethyl acetate. The gradient applied was 0-30% B over 3
minutes, 30% B for 10 minutes, ramped 30 to 50% B over 10 minutes,
50% B for 15 minutes, 50 to 80% B over 10 minutes, 80% for 10
minutes. Product 5 eluted in the 50% B fractions and all pure
fractions containing 5 were concentrated in vacuo to yield 5 (4.76
g, 64%) as a crunchy yellow foam. TLC (3:2 EtOAc/hexanes/1% AcOH)
Rf=0.2.
[0257]
(2S,3R)-3-((2-(di-t-butyloxycarbonylamino)pyridin-4-yl)methyl)-4-ox-
oazetidine-2-carboxylic acid (6): To a solution of pyridyl
beta-lactam acid 5 (0.4 g, 1.0 eq.) in 3.4 mL methanol and 0.12 mL
acetic acid was added 0.5M ammonium fluoride in methanol (1.5 mL,
1.0 eq.). The solution was stirred for 3 hours until disappearance
of all starting material. The reaction was then concentrated in
vacuo and put under high vacuum overnight to yield a yellow solid
which was taken on to the next step without further
purification.
[0258]
(2S,3R)-1-(benzhydrylcarbamoyl)-3-((2-(di-t-butyloxycarbonylamino)p-
yridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid (7): To a
solution of beta-lactam acid 6 (50 mg, 1.0 eq) in dimethylformamide
(1.2 mL) was added diphenylmethylisocyanate (100 uL, 4.45 eq.),
triethylamine (80 uL, 4.8 eq.). The reaction was stirred at room
temperature for 23 hours and then concentrated in vacuo. The crude
material was purified by preparative TLC eluting with 9:1:0.2 ethyl
acetate:methanol:acetic acid to yield 7 (42 mg, 56%) as a yellow
foam.
[0259]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-1-(benzhydrylcarbamoyl)-4-o-
xoazetidine-2-carboxylic acid (8): To solid 7 (42 mg) was added 4N
HCl-dioxane (1.0 mL). The reaction was capped and stirred at room
temperature for 5.5 hrs (or until reaction is complete) and then
triturated with diethyl ether to yield a crude solid which was
further purified via preparative HPLC (C18, acetonitrile/water,
0.1% TFA) to give 8 (10.6 mg, 35%) as a white powder.
[0260]
(2S,3R)-1-(((1S,2S)-2-(benzyloxy)cyclopentyl)carbamoyl)-3-((2-amino-
pyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid (9): Compound
9 was synthesized by general method A using
(1R,2R)-benzyloxycyclopentyl isocyanate (step 3).
[0261]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N1,N2-bis((1S,2S)-2-(benzyl-
oxy)cyclopentyl)-4-oxoazetidine-1,2-dicarboxamide (10): Compound 10
was synthesized by general method A using
(1R,2R)-benzyloxycyclopentyl isocyanate (step 3). Note: This
compound was a byproduct from step 3 which was independently
isolated via the same preparative TLC conditions and deprotected
using the same conditions in step 4 (4N HCl-dioxane).
[0262]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N1,N2-bis(3-benzylphenyl)-4-
-oxoazetidine-1,2-dicarboxamide (11): Compound 11 was synthesized
by general method A using 3-benzylphenylisocyanate (step 3). Note:
This compound was a byproduct from step 3 which was independently
isolated via the same preparative TLC conditions and deprotected
using the same conditions in step 4 (4N HCl-dioxane).
[0263]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-4-oxo-N1,N2-bis(3-phenoxyph-
enyl) azetidine-1,2-dicarboxamide (12): Compound 12 was synthesized
by general method A using 3-phenoxyphenylisocyanate (step 3). Note:
This compound was a byproduct from step 7 which was independently
isolated via the same preparative TLC conditions and deprotected
using the same conditions in step 4 (4N HCl-dioxane).
Example 2
General method B for the preparation of 3-aminopyridyl beta-lactam
methyl esters
##STR00104##
[0265] (2S,3R)-methyl
3-((2-(di-t-butyloxycarbonylamino)pyridin-4-yl)methyl)-4-oxoazetidine-2-c-
arboxylate (13): To a solution of pyridyl beta-lactam acid 5 (0.1
g, 1.0 eq.) in 3 mL methanol and 1.0 mL tetrahydrofuran was added
2.0M trimethylsilyldiazomethane in hexanes (558 .mu.L, 6.0 eq.).
After disappearance of all starting material, the reaction was
concentrated in vacuo. The crude material was purified by reverse
phase HPLC (C18, acetonitrile/water with 0.1% TFA) to yield 13
(45.5 mg, 56%) as a clear oil.
[0266] (2S,3R)-methyl
1-((2-benzylphenyl)carbamoyl)-3-((2-(di-t-butyloxycarbonylamino)
pyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylate (14): To a
solution of beta-lactam ester 13 (11 mg, 1.0 eq) in dichloromethane
(0.75 mL) was added 2'-benzylphenyl isocyanate (10 uL, 2.0 eq.),
triethylamine (11 uL, 3.0 eq.) and a few drops of DMF to solubilize
any undissolved material. The reaction was stirred at room
temperature for 21 hours and then concentrated in vacuo. The crude
material was taken on to the next step without further
purification.
[0267] (2S,3R)-methyl
3-((2-aminopyridin-4-yl)methyl)-1-((2-benzylphenyl)carbamoyl)-4-oxoazetid-
ine-2-carboxylate (15): To crude acylated beta-lactam ester 14 was
added 4N HCl in dioxane (2 mL). The reaction was capped and stirred
for 4 hours and then concentrated in vacuo. The crude material was
purified via reverse phase HPLC (C18, acetonitrile/water with 0.1%
TFA) to yield 15 (3.6 mg) as a white solid.
[0268] (2S,3R)-methyl
1-(((1S,2R)-2-phenylcyclopropyl)carbamoyl)-3-((2-aminopyridin-4-yl)methyl-
)-4-oxoazetidine-2-carboxylate trifluoroacetate (16): Compound 16
was synthesized by general method B using
trans-2-phenyl-cyclopropyl isocyanate (step 3) to yield 16 (3.9 mg)
as a white solid.
[0269] (2S,3R)-methyl
1-(((1R,2R)-2-(benzyloxy)cyclopentyl)carbamoyl)-3-((2-aminopyridin-4-yl)m-
ethyl)-4-oxoazetidine-2-carboxylate trifluoroacetate (17): Compound
17 was synthesized by general method B using
(1S,2S)-2-benzyloxycyclopentyl isocyanate (step 3) to yield 17 (5.0
mg) as a white solid.
Example 3
General method C for the preparation of 3-aminopyridyl beta-lactam
esters and amides
##STR00105##
[0271] (2S,3R)-benzyl
1-(tert-butyldimethylsilyl)-3-((2-(di-t-butoxycarbonylamino)pyridin-4-yl)-
methyl)-4-oxoazetidine-2-carboxylate (18): The beta-lactam acid 5
(136 mg, 0.254 mmol) was dissolved in 4 mL dry dichloromethane at
room temperature. EDC (73 mg, 3 equiv.), benzyl alcohol (33 mg, 32
uL, 1.2 equiv), and catalytic DMAP (3 mg) were added sequentially.
The reaction was stirred overnight under argon, diluted with 20 mL
ethyl acetate and 25 mL water. The water layer was washed with 3
portions of ethyl acetate, and the combined organic layers were
dried over sodium sulfate, filtered, and concentrated in vacuo. The
residue was purified by column chromatography using a gradient of
10 to 30% ethyl acetate in hexane, yielding 18 (72%). MS: 626.3
[M+H].sup.+.
[0272] (2S,3R)-benzyl
3-((2-(di-t-butoxycarbonylamino)pyridin-4-yl)methyl)-4-oxoazetidine-2-car-
boxylate (19): Ester 18 (127 mg, 0.20 mmol) was dissolved in 1 mL
dry methanol at room temperature. 1 mL of a 0.5 M solution of
ammonium fluoride in methanol (0.5 mmol, 2.5 equiv.) was added,
then 60 mg acetic acid (1 mmol, 5 equiv). The reaction was stirred
overnight under argon, diluted with 20 mL ethyl acetate and 20 mL
saturated sodium bicarbonate solution. The water layer was washed
with 3 portions of ethyl acetate, and the combined organic layers
were dried over sodium sulfate, filtered, and concentrated in
vacuo. The residue was purified by column chromatography eluting
with 3:1 methylene chloride/ethyl acetate to yield 19 (72 mg, 69%).
MS: 611.2 [M+H].sup.+.
[0273] (2S,3R)-benzyl
3-((2-(di-t-butoxycarbonylamino)pyridin-4-yl)methyl)-4-oxo-1-(phenylcarba-
moyl)azetidine-2-carboxylate (20): Deprotected lactam 19 (72 mg,
0.14 mmol) was dissolved in 4 mL dry THF at room temperature.
Triethylamine (71 mg, 5 equiv.) was added, then 42 mg (2.5 equiv.)
of the isocyanate. The reaction was stirred overnight under argon
and concentrated in vacuo. The residue was purified by column
chromatography eluting with methylene chloride to yield 20 (75 mg,
85%). MS: 631.3. [M+H].sup.+.
[0274]
(2S,3R)-3-((2-(di-t-butoxycarbonylamino)pyridin-4-yl)methyl)-4-oxo--
1-(phenylcarbamoyl)azetidine-2-carboxylic acid (21): Acylated
lactam ester 20 (56 mg, 0.089 mmol) was dissolved in 2 mL of 1:1
methanol-ethyl acetate in a 20 mL glass vial. 10% palladium on
carbon catalyst (10 mg) was added, and the solution was stirred
under an atmosphere of hydrogen at room temperature overnight under
argon, filtered, and then concentrated in vacuo. The residue was
pure by HPLC/MS and was taken to the next step without
purification. MS: 541.1 [M+H].sup.+.
[0275]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-4-oxo-1-(phenylcarbamoyl)az-
etidine-2-carboxylic acid trifluoroacetate (22): Acylated lactam
acid 21 (from the previous step) was dissolved in 2 mL of 1:1
TFA-methylene chloride in a 20 mL glass vial. The solution was
stirred for 1 hour at room temperature under argon and then
concentrated in vacuo. The residue was purified by preparative HPLC
using a C18 column and a gradient of 20-60% water in acetonitrile.
All fractions containing the desired product were pooled and
concentrated in vacuo. Freeze-drying from 1:1 water:acetonitrile
gave 22 (25 mg, 62% for 2 steps). MS: 341.1 [M+H].sup.+.
[0276]
(2S,3R)-1-(((R)-1-phenylethyl)carbamoyl)-3-((2-aminopyridin-4-yl)me-
thyl)-4-oxoazetidine-2-carboxylic acid trifluoroacetate (23):
Compound 23 was synthesized by general method C using
.alpha.-methylbenzylamine isocyanate (step 3).
[0277]
(2S,3R)-1-(((R)-1-(naphthalen-1-yl)ethyl)carbamoyl)-3-((2-aminopyri-
din-4-yl)methyl)-4-oxoazetidine-2-carboxylic acid trifluoroacetate
(24): Compound 24 was synthesized by general method C using
.alpha.-methylnaphthyl isocyanate (step 3).
[0278] (2S,3R)-ethyl
1-(((R)-1-(naphthalen-1-yl)ethyl)carbamoyl)-3-((2-aminopyridin-4-yl)methy-
l)-4-oxoazetidine-2-carboxylate trifluoroacetate (25): Compound 25
was synthesized by general method C using ethyl alcohol (step 1)
and .alpha.-methylnaphthyl isocyanate (step 3).
[0279] (2S,3R)-ethyl
3-((2-aminopyridin-4-yl)methyl)-4-oxo-1-(phenylcarbamoyl)azetidine-2-carb-
oxylate trifluoroacetate (26): Compound 26 was synthesized by
general method C using ethyl alcohol (step 1) and phenyl isocyanate
(step 3).
[0280] (2S,3R)-benzyl
3-((2-aminopyridin-4-yl)methyl)-4-oxo-1-(phenylcarbamoyl)
azetidine-2-carboxylate trifluoroacetate (27): Compound 27 was
synthesized by general method C using benzyl alcohol (step 1) and
phenyl isocyanate (step 3).
[0281] (2S,3R)-ethyl
1-(((R)-1-phenylethyl)carbamoyl)-3-((2-aminopyridin-4-yl)methyl)-4-oxoaze-
tidine-2-carboxylate trifluoroacetate (28): Compound 28 was
synthesized by general method C using ethyl alcohol (step 1) and
.alpha.-methylbenzylamine isocyanate (step 3).
[0282]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2,N2-diethyl-4-oxo-N1-((R)-
-1-phenylethyl)azetidine-1,2-dicarboxamide trifluoroacetate (29):
Compound 29 was synthesized by general method C using dimethylamine
(step 1) and .alpha.-methylbenzylamine isocyanate (step 3).
[0283]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-ethyl-N2-methyl-4-oxo-N1-
-((R)-1-phenylethyl)azetidine-1,2-dicarboxamide trifluoroacetate
(30): Compound 30 was synthesized by general method C using
N-methyl-N-ethylamine (step 1) and .alpha.-methylbenzylamine
isocyanate (step 3).
[0284]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-2-(morpholine-4-carbonyl)-4-
-oxo-N--((R)-1-phenylethyl)azetidine-1-carboxamide trifluoroacetate
(31): Compound 31 was synthesized by general method C using
morpholine (step 1) and .alpha.-methylbenzylamine isocyanate (step
3).
[0285]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-4-oxo-N1-((R)-1-phenylethyl-
)-N2-(pyridin-3-yl)azetidine-1,2-dicarboxamide trifluoroacetate
(32): Compound 32 was synthesized by general method C using
3-aminopyridine (step 1) and .alpha.-methylbenzylamine isocyanate
(step 3).
[0286] (2S,3R)-benzyl
3-(benzofuran-5-ylmethyl)-4-oxoazetidine-2-carboxylate (33) and
(2S,3R)-3-(benzofuran-5-ylmethyl)-4-oxo-1-(phenylcarbamoyl)azetidine-2-ca-
rboxylic acid (34): Compounds 33 and 34 were synthesized by general
method A (step 1) using 5-(bromomethyl)benzofuran as the
electrophile instead of 4-bromomethyl-2-bis-Boc-aminopyridine and
then the synthesis was completed by following general method C. The
compounds were separated via silica gel chromatography eluting with
10-40% ethyl acetate in hexanes. Yield of 33: 56 mg.
[0287]
(2S,3R)-3-(2-(benzo[d][1,3]dioxol-5-yl)ethyl)-4-oxo-1-(phenylcarbam-
oyl)azetidine-2-carboxylic acid (35): Compound 35 was synthesized
by general method A (step 1) using
5-(2-bromoethyl)benzo[d][1,3]dioxole as the electrophile instead of
4-bromomethyl-2-bis-Boc-aminopyridine. The starting
5-(bromomethyl)benzo[d][1,3]dioxole was made by standard treatment
of benzo[d][1,3]dioxol-5-ylmethanol with carbon tetrabromide and
triphenylphosphine in toluene. The synthesis was then completed by
following general method C.
[0288]
(2S,3R)-3-(benzo[d][1,3]dioxol-5-ylmethyl)-4-oxo-1-(phenylcarbamoyl-
)azetidine-2-carboxylic acid (36): Compound 36 was synthesized by
general method A (step 1) using 5-(bromomethyl)benzo[d][1,3]dioxole
as the electrophile instead of
4-bromomethyl-2-bis-Boc-aminopyridine. The synthesis was then
completed by following general method C.
[0289]
(2S,3R)-3-(3-chlorobenzyl)-4-oxo-1-(phenylcarbamoyl)azetidine-2-car-
boxylic acid (37): Compound 37 was synthesized by general method A
(step 1) by using 1-(bromomethyl)-3-chlorobenzene as the
electrophile instead of 4-bromomethyl-2-bis-Boc-aminopyridine. The
completion of the synthesis then followed general method C using
4-methoxybenzyl alcohol (step 1). Removal of the PMB group was
accomplished by treatment with TFA/DCM at room temperature for one
hour to yield the desired acid 37 (45 mg, 100%).
Example 4
General method D for the preparation of 3-aminopyridyl beta-lactam
alcohols
##STR00106##
[0291]
(2S,3R)-1-(((R)-1-(naphthalen-1-yl)ethyl)carbamoyl)-3-((2-(di-t-but-
oxy carbonylamino)pyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylic
acid (38): Compound 38 was synthesized by general method C using
.alpha.-ethylnaphthyl isocyanate.
[0292]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-2-(hydroxymethyl)-N--((R)-1-
-(naphthalen-1-yl)ethyl)-4-oxoazetidine-1-carboxamide
trifluoroacetate (39) and
((2S,3R)-1-(((R)-1-(naphthalen-1-yl)ethyl)carbamoyl)-3-((2-amino-
pyridin-4-yl)methyl)-4-oxoazetidin-2-yl)methyl
(R)-1-(naphthalen-1-yl)ethylcarbamate trifluoroacetate (40): To a
solution of compound 38 (0.03 g, 0.048 mmol) in 1 ml anhydrous THF
was added 1M borane in THF (0.29 ml, 0.29 mmol) dropwise and the
reaction mixture was stirred at room temperature for 1 h. Then the
reaction was slowly quenched with water, acidified to pH 1 with 1 N
HCl, and then neutralized with sodium bicarbonate solution. The
reaction mixture was extracted twice with ethyl acetate and the
organic extracts were dried over magnesium sulfate and concentrated
in vacuo. The desired product was purified by silica gel
chromatography (12 g pre-packed Si column, hexane/ethyl acetate
eluents) to yield the Boc protected alcohol (0.008 g, 27% yield;
LS/MS M+H 605.1, calc. 605.48). The alcohol (0.008 g, 0.013 mmol)
was treated with 1 ml 1/1 TFA/DCM for 1 h at room temperature.
Afterwards, the reaction mixture was concentrated in vacuo and the
solid washed with ethyl ether (three times) to give the 39 as a TFA
salt (2.2 mg). LS/MS M+H 405.12, calc. 405.5 and 40 as a TFA salt
(2.5 mg).
[0293]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-2-(hydroxymethyl)-4-oxo-N---
((R)-1-phenylethyl)azetidine-1-carboxamide (TFA salt) (41):
Compound 41 was synthesized by general method C using
.alpha.-ethylnaphthyl isocyanate (step 3) and finished following
general method D (2.5 mg, white powder) LC/MS M+H 355.2, calc.
355.17.
Example 5
General method E for the preparation of N-aryl 3-aminopyridyl
beta-lactams
##STR00107##
[0295]
(2S,3R)-4-methoxybenzyl-1-(tert-butyldimethylsilyl)-3-((2-(di-t-but-
oxycarbonylamino)pyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylate
(42): A mixture of 534 mg of crude compound 5 (1.0 mmol), EDC
hydrochloride salt (403 mg, 2.1 mmol), DMAP (24 mg, 0.2 mmol) and
4-methoxybenzyl alcohol (416 mg, 3.0 mmol) in dichloromethane (5
Ml) was stirred at room temperature overnight. The solvent was
removed in vacuo. The residue was dissolved with ethyl acetate,
washed with 5% aqueous KHSO.sub.4, water and brine. The organic
layer was dried over Na.sub.2SO.sub.4, then concentrated to yield a
crude material which was purified by ISCO silica gel column using
0-20% ethyl acetate in hexanes to afford 42 (463 mg, 71%): Anal.
C.sub.34H.sub.49N.sub.3O.sub.8Si, Mol. Wt.: 655.85, Found: ESI-MS:
656.0 (M+H).sup.+.
[0296]
(2S,3R)-4-methoxybenzyl-3-((2-(di-t-butoxycarbonylamino)pyridin-4-y-
l)methyl)-4-oxoazetidine-2-carboxylate (43): A solution of 1.7 Ml
of 0.5 M ammonium fluoride in methanol (0.85 mmol) was added to a
mixture of compound 42 (463 mg, 0.71 mmol), acetic acid (140 Ul,
2.45 mmol) and methanol (7 Ml). The mixture was stirred at room
temperature for 3 h. The solvent was removed and the residue was
taken up in toluene (2.about.3 Ml) to assist removal of AcOH. After
solvent was removed, the residue was taken up in DCM. The resulting
white solids were filtered off. Concentration of the filtrate gave
43 (0.44 g), which was ready for next step reaction without further
purification. Anal. C.sub.28H.sub.35N.sub.3O.sub.8, Mol. Wt.:
541.59, Found: ESI-MS: 541.8 (M+H).sup.+.
[0297] Compounds (44a-d): A mixture of .beta.-lactam 43 (30 mg,
0.055 mmol), aryl boronic acid (0.11 mmol), copper acetate (20 mg,
0.11 mmol), triethylamine (31 Ul, 0.22 mmol) and activated 4 A
molecular sieves (56 mg) in dichloromethane (1.3 Ml) was stirred at
room temperature overnight. The reaction mixture was filtered
through celite and the filtrate was concentrated. Preparative TLC
(20% EtOAc/Hexanes) of the residue gave the desired products 44a-d
in 27-78% yield.
[0298] Compounds (45a-d): The crude esters 44a-d were treated with
30% TFA in DCM (3 Ml). After two hours, LC-MS analyses indicated
completion of the reactions. The solvent was removed and the
residue was purified by preparative HPLC (Vydac reverse phase C-18
column, 22.times.250 mm ID). Mobile phase: A=0.1% TFA in water,
B=0.1% TFA in acetonitrile. The flow rate was 12 Ml/min. The
gradient time was 5% B to 55% B or 10% B to 60% B over 30 min. The
desired products were thus obtained:
[0299]
(2S,3R)-1-(4-((4-fluorophenyl)carbamoyl)phenyl)-3-((2-aminopyridin--
4-yl)methyl)-4-oxoazetidine-2-carboxylic acid (45a): Yield 2.5
mg.
[0300]
(2S,3R)-1-(3-((4-fluorophenyl)carbamoyl)phenyl)-3-((2-aminopyridin--
4-yl)methyl)-4-oxoazetidine-2-carboxylic acid (45b): Yield 3.2
mg.
[0301]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-1-(3-(benzyloxy)phenyl)-4-o-
xoazetidine-2-carboxylic acid (45c): Yield 2.0 mg.
[0302] (2S,3R)-4-methoxybenzyl
3-((2-aminopyridin-4-yl)methyl)-1-(benzofuran-2-yl)-4-oxoazetidine-2-carb-
oxylate (45d): Yield 1.7 mg.
[0303]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-1-(4-(benzyloxy)-2-fluoroph-
enyl)-4-oxoazetidine-2-carboxylic acid (46): Compound 46 was
synthesized by general method E using
4-(benzyloxy)-2-fluorophenylboronic acid.
Example 6
General method F for synthesis of 3,3-alkyl, methyl
beta-lactams
##STR00108##
[0305]
(2S,3R)-1-(tert-butyldimethylsilyl)-3-methyl-4-oxoazetidine-2-carbo-
xylic acid (47): To a solution of
(S)-1-(tert-butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid 4
(2.00 g, 8.73 mmol.) in THF (30 ml) at -78.degree. C. was added LDA
(19 ml, 2.0 eq.). After the solution was stirred for 20 min. at
-78.degree. C., it was warmed to 0.degree. C. for 5 min. The
solution was re-cooled to -78.degree. C. and methyl iodide (3.10 g,
2.5 eq.) in THF (5 ml) was added. The reaction solution was warmed
up to room temperature, quenched with aqueous KHSO.sub.4 solution
(10%, 30 ml), extracted with ethyl acetate (3.times.). The combined
organic layers were washed with brine twice and dried with
MgSO.sub.4. After concentrating in vacuo, 47 was obtained.
H.sup.1NMR (CDCl.sub.3): 0.15, 0.35 (s, 6H), 1.14 (s, 9H), 1.35 (d,
3H), 3.71 (d, 1H), 4.14 (m, 1H). C.sub.11H.sub.21NO.sub.3Si, Mol.
Wt.: 243.37, found: 244 (M+1).
[0306]
(2S,3R)-3-allyl-1-(tert-butyldimethylsilyl)-3-methyl-4-oxoazetidine-
-2-carboxylic acid (48): To a solution of compound 47 (0.200 g,
0.823 mmol) in THF (5 ml) at -78.degree. C. was added a solution of
LDA (1.10 ml, 2.4 eq.). The reaction solution was stirred for 20
min. at that temperature, warmed to -20.degree. C. and allyl
bromide (0.138 Ml, 2.0 eq.) was added slowly. Reaction mixture was
warmed to 0.degree. C. for 30 min. and quenched with KHSO.sub.4 (20
ml, 10%), extracted with ethyl acetate (3.times.). Combined organic
layers were washed with brine twice and dried over MgSO.sub.4.
After concentrating in vacuo, compound 48 was obtained. H.sup.1NMR
(CDCl.sub.3): 0.15, 0.37 (s, 6H), 1.16 (s, 9H), 1.33 (s, 3H), 2.50
(m, 2H), 3.98 (s, 1H), 5.22 (m, 2H), 5.88 (m, 1H).
C.sub.14H.sub.25NO.sub.3Si, Mol. Wt.: 283.44. Found: 284 (M+1).
[0307] (2S,3R)-methyl 3-allyl-3-methyl-4-oxoazetidine-2-carboxylate
(49): TMS diazomethane (1.23 Ml, 3.0 eq. 2 M in THF) was added to a
solution of the acid compound 48 (260 mg, 0.82 mmol) in methanol (5
Ml) at 0.degree. C. and stirred at room temperature overnight to
yield the deprotected product 46. H.sup.1NMR (CDCl.sub.3): 1.30 (s,
3H), 2.50 (m, 2H), 3.80 (s, 3H), 4.08 (s, 3H), 5.30 (m, 2H), 5.85
(m, 1H), 5.92 (s, b, 1H). C.sub.9H.sub.13NO.sub.3, Mol. Wt.: 183.2.
Found: 184 (M+1).
[0308] (2S,3R)-methyl
1-(((R)-1-phenylethyl)carbamoyl)-3-allyl-3-methyl-4-oxoazetidine-2-carbox-
ylate (50): To a solution of 49 (120 mg, 0.66 mmol) in methylene
chloride (15 mL) was added triethylamine (0.27 mL, 3.0 eq.), DMAP
(8.0 mg, 10% mmol) and .alpha.-methyl-benzyl isocyanate (146 mg,
1.5 eq.). The reaction was stirred overnight and concentrated in
vacuo to yield a crude material, which was extracted with ethyl
acetate and aqueous NH.sub.4Cl. Combined organic layers were washed
with brine purified via silica gel chromatography to yield compound
47. H.sup.1NMR (CDCl.sub.3): 1.35 (s, 3H), 1.60 (d, 3H), 2.50 (m,
2H), 3.82 (s, 3H), 4.44 (s, 1H), 5.50 (m, 1H), 5.25 (m, 2H), 5.83
(m, 1H), 6.74 (d, b, 1H), 7.40 (m, 5H).
C.sub.18H.sub.22N.sub.2O.sub.4, Mol. Wt.: 330.38. Found: 331
(M+1).
[0309] (2S,3R)-methyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(2-iminoguanidinoethyl)-3-methyl-4-oxo-
azetidine-2-carboxylate (51): To a solution of 50 (135 mg) in
methylene chloride (5 mL) at -78.degree. C. was bubbled O.sub.3
until a blue color was seen. The reaction was quenched with
dimethylsulfide. Removal of solvents gave the desired aldehyde
product, which was used in the next step without further
purification. To a solution of the freshly prepared aldehyde
compound (118 mg, 0.36 mmol) in ethanol (0.75 mL) was added amino
amidine (HNO.sub.3 salt, 98 mg, 2.0 eq.) and acetic acid (30 uL,
3.0 eq.). The mixture was stirred for 1.5 h and residual after
removal of solvent was purified via reverse-phase HPLC to yield
compound 51 (41 mg). C.sub.18H.sub.24N.sub.6O.sub.4, Mol. Wt.:
388.42. Found: 389 (M+1).
[0310]
(2S,3R)-1-(((R)-1-phenylethyl)carbamoyl)-3-(2-iminoguanidinoethyl)--
3-methyl-4-oxoazetidine-2-carboxylic acid (52): To a solution of
compound 51 (10 mg) in 1:1:1 THF:methanol:water (1.5 mL total) was
added solid LiOH (3.5 eq.). The reaction was stirred at room
temperature for 30 min. LCMS showed formation of the desired acid
and disappearance of the starting methyl ester. The crude material
was purified via reverse-phase HPLC to yield pure 52. C17H22N6O4,
Mol. Wt.: 374.39. Found: 375 (M+1).
[0311] (2S,3R)-ethyl
1-(((R)-1-phenylethyl)carbamoyl)-3-((6-aminopyridin-3-yl)methyl)-3-methyl-
-4-oxoazetidine-2-carboxylate (53) and
(2S,3R)-1-(((R)-1-phenylethyl)carbamoyl)-3-((6-aminopyridin-3-yl)methyl)--
3-methyl-4-oxoazetidine-2-carboxylic acid (54): Compounds 53 and 54
were synthesized by general method F using
4-bromomethyl-2-bis-boc-aminopyridine as the alkylating agent in
step 2 and forming the ethyl ester (step 3) analogous to the ester
synthesis in general method C.
Example 7
General method G for the preparation of N-alkyl amidinohydrazone
beta-lactams
##STR00109## ##STR00110##
[0313]
(2S,3R)-3-Allyl-1-(tert-butyl-dimethyl-silanyl)-4-oxo-azetidine-2-c-
arboxylic acid (57): To a solution of
(S)-1-(tert-butyldimethylsilyl)-4-oxoazetidine-2-carboxylic acid
(4) (2.00 g, 8.73 mmol.) in anhydrous THF (20 mL) at -78.degree. C.
was added 1.0 M solution of LiHMDS (20 ml, 20.1 mmol, 2.3 eq.) in
THF. After the solution was stirred for 30 min. at -78.degree. C.,
it was warmed to 0.degree. C. for 10 min. The solution was recooled
to -78.degree. C. and allyl bromide (1.28 g, 10.5 mmol, 1.2 eq.)
was added slowly. The reaction solution was stirred at -78.degree.
C. for 1 h, then warmed up to room temperature and stirred for
another hour. The reaction mixture was quenched with aqueous 10%
KHSO.sub.4 solution (30 ml) and extracted with ethyl acetate (40
mL.times.3). The combined organic layers were washed with brine
twice and dried over Na.sub.2SO.sub.4. After removal of solvent,
compound 57 was obtained as crystalline solid (2.0 g, 85%); 78%
purity by LCMS, .sup.1H NMR (300 MHz, CDCl.sub.3): 0.12 (s, 3H),
0.30 (s, 3H), 0.95 (s, 9H), 2.55 (m, 2H), 3.41 (m, 1H), 3.80 (d,
J=2.61 Hz, 1H), 5.16 (m, 2H), 5.72 (m, 1H). Anal.
C.sub.13H.sub.23NO.sub.3Si, Mol. Wt.: 269.41, Found: ESI-MS: 270.0
(M+H).sup.+.
[0314]
(2S,3R)-3-Allyl-1-(tert-butyl-dimethyl-silanyl)-4-oxo-azetidine-2-c-
arboxylic acid 4-methoxy-benzyl ester (58): A mixture of 900 mg of
crude compound 57 (3.34 mmol) above, EDC hydrochloride salt (1.34
g, 7.01 mmol), DMAP (83 mg, 0.67 mmol) and 4-methoxybenzyl alcohol
(1.40 g, 10.1 mmol) in dichloromethane (9 mL) was stirred at room
temperature overnight. The solvent was removed in vacuo. The
residue was dissolved with ethyl acetate, washed with 5% aqueous
KHSO.sub.4, water (.times.2) and brine (.times.1). The organic
layer was dried over Na.sub.2SO.sub.4, then concentrated to yield
the crude 58 with LC purity: 60%. Purification by ISCO silica gel
column using 0-20% AcOEt in Hexanes afforded 630 mg of compound 58
(49% yield, 81% purity): .sup.1H NMR (300 MHz, CDCl.sub.3): 0.13
(s, 3H), 0.32 (s, 3H), 0.95 (s, 9H), 2.56 (m, 2H), 3.43 (m, 1H),
3.81 (d, J=2.60 Hz, 1H), 3.82 (s, 3H), 4.67 (s, 2H), 5.17 (m, 2H),
5.72 (m, 1H), 6.94 (d, 2H), 7.34 (d, 2H). Anal.
C.sub.21H.sub.31NO.sub.4Si, Mol. Wt.: 389.56, Found: ESI-MS: 390.0
(M+H).sup.+.
[0315] (2S,3R)-3-Allyl-4-oxo-azetidine-2-carboxylic acid
4-methoxy-benzyl ester (59): A solution of 6.2 mL of 0.5 M ammonium
fluoride in methanol (3.1 mmol) was added to a mixture of compound
58 (1.0 g, 2.57 mmol), acetic acid (530 uL, 9.0 mmol) and methanol
(25 mL). The mixture was stirred at room temperature for 2 h. The
solvent was removed and the residue was taken up in toluene
(2.about.3 mL) to assist removal of AcOH. After solvent was
removed, the residue was taken up in DCM. The resulting white
solids were filtered off. Concentration of the filtrate gave 0.70 g
of the crude 59 with purity: 91.1%, .sup.1H NMR (300 MHz,
CDCl.sub.3): 2.60 (m, 2H), 3.38 (m, 1H), 3.87 (s, 3H), 3.98 (d,
1H), 5.22 (m, 4H), 5.83 (m, 1H), 6.15 (b, s, 1H), 6.90 (d, 2H),
7.35 (d, 2H). Anal. C15H17NO4, Mol. Wt.: 275.3, Found: ESI-MS:
275.8 (M+H).sup.+.
[0316]
(2S,3R)-3-Allyl-4-oxo-1-(1-phenyl-propylcarbamoyl)-azetidine-2-carb-
oxylic acid 4-methoxy-benzyl ester (60): A mixture of compound 59
(41 mg, 0.15 mmol), triethylamine (83 uL, 0.60 mmol),
.alpha.-ethyl-benzyl isocyanate (29 mg, 0.18 mmol) in methylene
chloride (1.2 mL) was stirred overnight. TLC analysis indicated the
completion the reaction. Removal of solvents gave the crude
material, which was purified by preparative TLC plate (15% EtOAc in
hexanes), yielding 38.2 mg of compound 60 (59%): LC-MS purity:
98.1%; .sup.1H NMR (300 MHz, CDCl.sub.3): 0.95 (t, 3H), 1.92 (m,
2H), 2.69 (m, 2H), 3.30 (m, 1H), 3.83 (s, 3H), 4.22 (d, 1H), 4.85
(q, 1H), 5.18 (m, 4H), 5.80 (m, 1H), 6.85 (br, d, 1H), 7.45 (m,
9H). Anal. C25H28N2O5, Mol. Wt.: 436.5, Found: ESI-MS: 437.1
(M+H).sup.+.
[0317]
(3S,4R)-4-Oxo-3-(2-oxo-ethyl)-1-(1-phenyl-propylcarbamoyl)-azetidin-
e-2-carboxylic acid 4-methoxy-benzyl ester (61): Ozone was bubbled
through a solution of 60 (38.1 mg, 0.087 mmol) in dry methylene
chloride (6 mL) at -78.degree. C. until a faint blue color
persisted. The reaction mixture was quenched with dimethylsulfide
(200 uL) at -78.degree. C., then the cooling bath was removed.
After the solvent was concentrated, the residue was taken up in
EtOAc. Removal of solvents gave the desired aldehyde product 61,
which was sufficiently pure for next step without further
purification. Anal. C.sub.24H.sub.26N.sub.2O.sub.6, Mol. Wt.:
338.47, Found: ESI-MS: 439.0 (M+H).sup.+, 461.0 (M+Na).sup.+.
[0318]
(2S,3R)-4-Oxo-3-(3-amindinohydrozone-ethyl)-1-(1-phenyl-propylcarba-
moyl)-azetidine-2-carboxylic acid 4-methoxy-benzyl ester (62): To a
solution of the freshly prepared aldehyde 61 (29 mg, 0.066 mmol) in
ethanol (0.5 mL) was added aminoguanidinium nitrate (18 mg, 0.13
mmol) and acetic acid (11 uL, 0.20 mmol). The mixture was stirred
at room temperature for 2 h. Concentration of solvent gave the
crude desired product 62. Anal. C.sub.25H.sub.30N.sub.6O.sub.5,
Mol. Wt.: 494.54. Found: ESI-MS: 495.0 (M+H).sup.+.
[0319]
(2S,3R)-4-Oxo-3-(3-amindinohydrozone-ethyl)-1-(1-phenyl-propylcarba-
moyl)-azetidine-2-carboxylic acid (63): The crude ester 62 was
treated with a mixture of TFA/DCM (1.5 mL/2 mL). After one and half
hours, LC-MS analysis indicated that no ester was present. The
solvent was removed and the residue was purified by preparative
HPLC (Vydac reverse phase C-18 column, 22.times.250 mm ID). Mobil
phase: A=0.1% TFA in water; B=0.1% TFA in acetonitrile. The flow
rate was 12 mL/min. The gradient time was 5% B to 55% B over 50
min. The peak of interest was eluted at around 27 minutes to give
3.5 mg of the desired product, 63 as a white solid with 95.6%
purity. .sup.1H NMR (300 MHz, CDCl.sub.3): 0.84 (t, 3H), 1.80 (m,
2H), 2.83 (m, 2H), 3.70 (dt, 1H), 4.25 (d, 1H), 4.61 (q, 1H), 6.58
(br s, 1H), 7.15-7.65 (m, 9H), 11.65 (s 1H), 13.10 (br s, 1H);
Anal. C.sub.17H.sub.24N.sub.6O.sub.4, Mol. Wt.: 374.39, Found:
ESI-MS: 375.0 (M+H).sup.+.
[0320] (2S,3R)-cyclobutyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(2-iminoguanidinoethyl)-4-oxoazetidine-
-2-carboxylate (64): Compound 64 was synthesized by general method
G using cyclobutyl alcohol (step 2) and
.alpha.-methylbenzylisocyanate (step 4). Yield: 72.5 mg (48%). Mol.
Wt.: 414.46. Found: ESI-MS: 415.0 (M+H).sup.+.
[0321] (2S,3R)-methyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(2-iminoguanidinoethyl)-4-oxoazetidine-
-2-carboxylate (65): Compound 65 was synthesized by general method
G with step 2 esterification accomplished via TMSCHN.sub.2 in
hexanes and using .alpha.-methylbenzylisocyanate in step 4.
[0322]
(2S,3R)--N2,N2-diethyl-3-(2-iminoguanidinoethyl)-4-oxo-N1-((R)-1-ph-
enylethyl)azetidine-1,2-dicarboxamide (66): Compound 66 was
synthesized by general method G, using diethylamine in step 2 and
.alpha.-methylbenzylisocyanate in step 4, to yield 4.1 mg (35%).
Mol. Wt.: 415.49. Found: ESI-MS: 416.0 (M+H).sup.+.
[0323]
(2S,3R)-3-(2-iminoguanidinoethyl)-4-oxo-1-(biphenylcarbamoyl)azetid-
ine-2-carboxylic acid (67): Compound 67 was synthesized by general
method G using 4-biphenylisocyanate (step 4). Yield: 1.1 mg (6.8%
over 2 steps). Mol. Wt.: 408.41. Found: ESI-MS: 409.0
(M+H).sup.+.
[0324] (2S,3R)-methyl
3-(2-iminoguanidinoethyl)-4-oxo-1-(phenylcarbamoyl)azetidine-2-carboxylat-
e (68): Compound 68 was synthesized by general method G with step 2
esterification accomplished via TMSCHN.sub.2 in hexanes and using
phenylisocyanate in step 4.
[0325]
(2S,3R)-1-(((R)-1-phenylethyl)carbamoyl)-3-(2-iminoguanidinoethyl)--
4-oxoazetidine-2-carboxylic acid (69),
(2S,3R)-1-(((R)-1-phenylethyl)carbamoyl)-3-(2-(R)-1-phenylethyl)carbamoyl-
)-iminoguanidinoethyl)-4-oxoazetidine-2-carboxylic acid (70) and
(2S,3R)-3-(2-(R)-1-phenylethyl)carbamoyl)-iminoguanidinoethyl)-4-oxoazeti-
dine-2-carboxylic acid (71): Compounds 69-71 were synthesized by
general method G using .alpha.-methylbenzylisocyanate (step 4). The
compounds were separated via reverse phase HPLC
(acetonitrile/water) with 0.1% TFA added to the mobile phase.
[0326]
(2S,3R)-3-(2-iminoguanidinoethyl)-4-oxo-1-((4-phenoxyphenyl)carbamo-
yl)azetidine-2-carboxylic acid (72): Compound 72 was synthesized by
general method G using 4-phenoxyphenyl isocyanate (step 4).
[0327]
(2S,3R)-3-(2-iminoguanidinoethyl)-4-oxo-1-(phenylcarbamoyl)azetidin-
e-2-carboxylic acid (73): Compound 73 was synthesized by general
method G using phenyl isocyanate (step 4).
[0328] (2S,3R)-cyclobutyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(2-imino(methylguanidino)ethyl)-4-oxoa-
zetidine-2-carboxylate (74): Compound 74 was synthesized by general
method G using cyclobutyl alcohol (step 2), .alpha.-methylbenzyl
isocyanate (step 4) and N-methylpimagedine (step 6).
[0329] (2S,3R)-benzyl
3-(2-iminoguanidinoethyl)-4-oxo-1-(biphenylcarbamoyl)azetidine-2-carboxyl-
ate (75): Compound 75 was synthesized by general method G using
benzyl alcohol (step 2) and 4-biphenylisocyanate (step 4).
[0330] (2S,3R)-ethyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(2-imino(methylguanidino)ethyl)-4-oxoa-
zetidine-2-carboxylate (76): Compound 76 was synthesized by general
method G using ethyl alcohol (step 2),
.alpha.-methylbenzylisocyanate (step 4) and N-methyl pimagedine
(step 6).
[0331]
(2S,3R)-1-(((R)-1-phenylpropyl)carbamoyl)-3-(2-imino(methylguanidin-
o)ethyl)-4-oxoazetidine-2-carboxylic acid (77): Compound 77 was
synthesized by general method G using .alpha.-ethylbenzylisocyanate
(step 4) and N-methyl pimagedine (step 6).
[0332] (2S,3R)-4-methoxybenzyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(2-iminoguanidino
ethyl)-4-oxoazetidine-2-carboxylate (78): Compound 78 was
synthesized by general method G using .alpha.-methybenzylisocyanate
(step 4).
[0333]
(2S,3R)-1-(((R)-1-phenylpropyl)carbamoyl)-3-((E)-2-(2-(4,5-dihydro--
1H-imidazol-2-amine)imino)ethyl)-4-oxoazetidine-2-carboxylic acid:
Compound 79 was synthesized by general method G using
.alpha.-ethylbenzylisocyanate (step 4) and
1-(4,5-dihydro-1H-imidazol-2-yl)hydrazine in step 6 (2.7 mg,
88%).
Example 8
General method H for the preparation of N-aryl amidinohydrazone
beta-lactams
##STR00111##
[0335] 3-Allyl-4-oxo-1-pyridin-2-yl-azetidine-2-carboxylic acid
4-methoxy-benzyl ester (80): A mixture of .beta.-lactam 59 (60 mg,
0.22 mmol), Pyridin-2-boronic acid (81 mg, 0.66 mmol), copper
acetate (120 mg, 0.66 mmol), triethylamine (0.153 mL, 1.1 mmol) and
activated 4 A molecular sieves (270 mg) in dichloromethane (5 mL)
was stirred at room temperature for 2 days. The reaction mixture
was filtered through celite and the filtrate was concentrated.
Preparative TLC (20% EtOAc/Hexanes) of the residue gave the desired
product 80 (19.2 mg, 24.8%) .sup.1H NMR (300 MHz, CDCl.sub.3): 2.60
(m, 2H), 3.49 (m, 1H), 3.81 (s, 3H), 4.41 (d, J=2.61 Hz, 1H),
5.11-5.21 (m, 4H), 5.82 (m, 1H), 6.85 (d, J=8.7 Hz, 2H), 7.01 (m,
1H), 7.22 (d, J=8.1 Hz, 2H), 7.66-7.72 (m, 2H); 8.19 (d, J=4.7 Hz,
1H) Anal. C.sub.20H.sub.20N.sub.2O.sub.4, Mol. Wt.: 352.38. Found:
ESI-MS: 352.9 (M+H).sup.+.
[0336] 4-Oxo-3-(2-oxo-ethyl)-1-pyridin-2-yl-azetidine-2-carboxylic
acid 4-methoxy-benzyl ester (81): Ozone was bubbled through a
solution of 80 (24.0 mg) in dry methylene chloride (7 mL) at
-78.degree. C. until a faint blue color persisted (about 10 min).
The reaction mixture was quenched with dimethylsulfide (200 uL) at
-78.degree. C., then the cooling bath was removed. After the
solvent was concentrated, the residue was taken up in EtOAc.
Removal of solvents gave the desired aldehyde product 81, which was
sufficiently pure to carry on to the next step without further
purification. Anal. C.sub.19H.sub.18N.sub.2O.sub.5, Mol. Wt.:
354.36. Found: ESI-MS: 354.9 (M+H).sup.+.
[0337]
(2S,3R)-4-methoxybenzyl-3-(2-guanidinoiminoethyl)-4-oxo-1-(pyridin--
2-yl)azetidine-2-carboxylate (82): To a solution of the freshly
prepared aldehyde 81 (32.2 mg, 0.068 mmol) in ethanol (0.85 mL) was
added aminoguanidinium nitrate (19 mg, 0.14 mmol) and acetic acid
(12 uL, 0.20 mmol). The mixture was stirred at room temperature for
3 h. Concentration of solvent gave the crude desired product 82.
Anal. C.sub.20H.sub.30N.sub.6O.sub.4, Mol. Wt.: 410.43. Found:
ESI-MS: 411.0 (M+H).sup.+.
[0338]
(2S,3R)-3-(2-guanidinoiminoethyl)-4-oxo-1-(pyridin-2-yl)azetidine-2-
-carboxylic acid (83): The crude ester 82 was treated with TFA/DCM
(0.75 mL/1 mL). After one and half hours, LC-MS analysis indicated
completion of the reaction. The solvent was removed and the residue
was purified by preparative HPLC (Vydac reverse phase C-18 column,
22.times.250 mm ID). Mobil phase: A=0.1% TFA in water, B=0.1% TFA
in acetonitrile. The flow rate was 12 mL/min. The gradient time was
5% B to 50% B over 45 min. The desired product, 83, was obtained as
a white solid (5.5 mg). Anal. C.sub.19H.sub.24N.sub.6O.sub.4, Mol.
Wt.: 290.28, Found: ESI-MS: 291.0 (M+H).sup.+.
Synthesis of 86 and 87
[0339]
(2R,3R)-1-(isopropyldimethylsilyl)-3-(3-nitrophenylthio)-4-oxoazeti-
dine-2-carboxylic acid (84): Using glassware, starting materials,
and THF that are each rigorously dry is essential for successful
alkylation, as well as keeping the reaction mixture under dry
argon. Commercially available beta-lactam acid 4 (15.31 mmol, 3.51
g) was dissolved in 40 mL dry THF at room temperature with magnetic
stirring, in an oven-dried flask with dry stir bar under dry argon.
The solution was cooled to -78.degree. C. and the LDA solution
(2.05 equivalent relative to acid, 31.38 mmol, 17.4 mL 1.8M
solution from Aldrich) was added slowly over 5 minutes by syringe
to the cooled stirring solution. The solution was kept at
-78.degree. C. for 15 minutes and warmed to 0.degree. C. with an
ice bath. The solution was kept at 0.degree. C. for 45 minutes, and
then cooled to -42.degree. C. (dry ice-acetonitrile). The disulfide
was dissolved in 20 mL of dry THF and cooled to -42.degree. C. (dry
ice-acetonitrile). The disulfide solution was added dropwise via a
dry narrow bore cannula to the enolate solution using balloon
pressure, with both vessels cooled to -42.degree. C. during the 30
minute addition. After the addition was complete, the vial was
rinsed with two 2 mL portions of dry THF, which was cooled to
-42.degree. C. and added to the enolate solution also. The reaction
vessel was kept at -42.degree. C. for 4 hours and then warmed to
0.degree. C. A quench solution was prepared by adding 50 g of ice
to 150 mL of 5% aqueous KHSO.sub.4 solution in an Erlenmeyer. The
reaction mixture was poured into the quench solution, and the
vessel was rinsed with 5.times.20 mL aliquots of ethyl acetate,
which were poured into the quench solution. The two layers were
poured into a separatory funnel. The pH of the aqueous layer was
2-3. The organic layer was separated, the aqueous layer was
extracted with 3 portions of ethyl acetate, and the combined
organic layers were washed with brine and concentrated in vacuo.
The solution was warmed to 0.degree. C. and after 4 hours worked up
in the standard way (KHSO.sub.4 quench). The desired product 84 was
purified by column chromatography on silica gel using 50:50:1
hexane:ethyl acetate: acetic acid as eluent. Product MS: 383.1
[M+H].sup.+.
[0340] (2R,3R)-ethyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(3-nitrophenylthio)-4-oxoazetidine-2-c-
arboxylate (85a) and (2R,3S)-ethyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(3-nitrophenylthio)-4-oxoazetidine-2-c-
arboxylate (85b). The completion of the synthesis of 85a-b was
accomplished by following general method C using ethyl alcohol
(step 1), and .alpha.-methylbenzylisocyanate (step 3).
[0341] (2R,3R)-ethyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(3-aminophenylthio)-4-oxoazetidine-2-c-
arboxylate (86): To a solution of compound 85a (18 mg, 41 umol) in
ethyl acetate (1 mL) was added tin chloride monohydrate (2.5 eq.)
and the solution was heated to 50.degree. C. After 1 h the solution
was concentrated in vacuo. The residue was purified by column
chromatography using a gradient of hexane/ethyl acetate 20-100% as
eluent to yield 86 (12 mg, 72%). Product MS: 414.1 (M+1).
[0342] (2R,3S)-ethyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(3-aminophenylthio)-4-oxoazetidine-2-c-
arboxylate (87): To a solution of 85b (17 mg, 38 umol) in ethyl
acetate (1 mL) at room temperature was added tin chloride
monohydrate (2.5 eq.) and the solution was heated to 50.degree. C.
After 1 h the solution was concentrated in vacuo. The residue was
purified by column chromatography using a gradient of hexane/ethyl
acetate 20-100% as eluent to yield 87 (10 mg, 63%). Product MS:
414.1 (M+1).
Synthesis of 90
[0343] (2S)-ethyl
1-(((R)-1-phenylethyl)carbamothioyl)-3-((2-(tert-butoxycarbonyl)pyridin-4-
-yl)methyl)-4-oxoazetidine-2-carboxylate (89): Compound 88 was
prepared by general method C using ethyl alcohol (step 1). To the
solution of 88 (23 mg) in THF was added LiHMDS (0.14 mL, 1M
solution in THF) at -78.degree. C. The reaction mixture was warmed
up to 0.degree. C. for 5 h and then was quenched with 5% KHSO.sub.4
to pH.about.5. After extraction with ethyl acetate, the organic
layer was washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated in vacuo to yield 89 LC/MS (ESI) m/z 512.8.
[0344] (2S)-ethyl
1-(((R)-1-phenylethyl)carbamothioyl)-3-((2-aminopyridin-4-yl)methyl)-4-ox-
oazetidine-2-carboxylate (90): Compound 89 (crude) was treated with
50% TFA-DCM at 0.degree. C. LCMS was used to monitor the reaction
until it was completed. The reaction mixture was condensed and the
residue was purified by preparative HPLC with 5-70%
acetonitrile-water (with 0.1% TFA) as mobile phase and gradient
time 40 min to afford 90 (9 mg, 26% yield). .sup.1H NMR (DMSO)
.delta. 8.44 (d, 1H), 7.70 (d, 1H), 7.37-7.27 (m, 5H), 7.26 (s,
2H), 6.72 (s, 1H), 6.66 (d, 1H), 5.51 (m, 1H), 4.38 (d, 1H), 4.22
(dd, 2H), 3.40 (d, 1H), 3.14 (m, 2H), 1.62 (d, 3H), 1.22 (t, 3H),
LC/MS (ESI) m/z 413.1.
Synthesis of 93
[0345] (2S)-4-Methoxybenzyl
3-((2-bis(tert-butoxycarbonyl)-aminopyridin-4-yl)methyl)-4-oxoazetidine-2-
-carboxylate (91): Compound 91 was prepared by general method C
using 4-methoxybenzyl alcohol (step 1).
[0346] (2S)-4-Methoxybenzyl
1-(((R)-1-phenylethyl)carbamothioyl)-3-((2-bis(tert-butoxycarbonyl)-amino-
pyridin-4-yl)methyl)-4-oxoazetidine-2-carboxylate (92): Compound 92
was prepared analogously to the synthesis of 89 and the crude
sample was taken on without further purification. LC/MS (ESI) m/z
705.0.
[0347]
(2S)-1-(((R)-1-phenylethyl)carbamothioyl)-3-((2-aminopyridin-4-yl)m-
ethyl)-4-oxoazetidine-2-carboxylic acid (93): Compound 93 was
prepared analogously to the synthesis of 90. The crude sample was
purified by preparative HPLC with 0-70% acetonitrile-water (with
0.1% TFA) as mobile phase and gradient time 40 min to afford 93
(5.6 mg, 16%). HPLC purity, >96%, .sup.1H NMR (DMSO) .delta.9.04
(d, 1H), 8.00 (br, 2H), 7.90 (d, 1H), 7.41-7.27 (m, 5H), 6.90 (m,
2H), 5.44 (m, 1H), 4.33 (d, 1H), 3.69 (m, 1H), 3.2 (m, 2H), 1.53
(d, J=7.0 Hz, 3H), LC/MS (ESI) m/z 385.1.
Synthesis of 96
[0348]
(2S,3R)-3-((2-bis(tert-butoxylcarbonyl)aminopyridin-4-yl)methyl)-N--
(methylsulfonyl)-4-oxoazetidine-2-carboxamide (94): Compound 5 was
prepared by general method A. To a solution of 5 (40 mg, 1 eq.) in
dichloromethane (1 mL) was added EDC-HCl (17 mg, 1.2 eq.), HOBt (12
mg, 1.2 eq.), DIEA (16 uL, 1.2 eq.) and methanesulfonamide (9 mg,
1.2 eq.). The reaction was stirred at room temperature for 12 hrs
and then concentrated in vacuo. The crude material was redissolved
in dichloromethane and washed with saturated NaHCO.sub.3 and water,
dried over sodium sulfate and concentrated in vacuo to give 22 mg
of crude 94 as a tan oil which was taken on to the next step
without further purification.
[0349]
(2S,3R)-3-((2-bis(tert-butoxycarbonyl)aminopyridin-4-yl)methyl)-N2--
(methylsulfonyl)-4-oxo-N1-((R)-1-phenylethyl)azetidine-1,2-dicarboxamide
(95): To a solution of crude 94 in THF (1 mL) was added DIEA (8 uL,
1.5 eq.) and (R)-(+)-.alpha.-methylbenzylisocyanate. The reaction
was stirred for 12 hours at room temperature. Ethyl acetate was
added to the reaction mixture and washed with water. The organic
layer was concentrated in vacuo. The crude product was purified
using a silica gel column and a gradient elution from 10%
EtOAc/hexanes to 100% EtOAc followed by 10% MeOH in DCM to elute
95. Product fractions were concentrated in vacuo to give 20 mg of
compound 95.
[0350]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(methylsulfonyl)-4-oxo-N-
1-((R)-1-phenylethyl)azetidine-1,2-dicarboxamide (96): To a
solution of 95 in 1 mL dichloromethane was added 300 uL of TFA. The
reaction was stirred for 1 hour and then concentrated in vacuo. The
crude material was purified via reverse phase HPLC (C18,
acetonitrile/water with 0.1% TFA) to yield 0.75 mg of 96. MS
[M+H].sup.+=446.0
Synthesis of 101
[0351]
(2S,3R)-3-((2-bis(tert-butoxycarbonyl)aminopyridin-4-yl)methyl)-1-(-
tert-butyldimethylsilyl)-N-(4-fluorophenylsulfonyl)-4-oxoazetidine-2-carbo-
xamide (97): Compound 5 was prepared by general method A. To a
solution of 5 (50.6 mg, 1.0 eq.) in THF (1 mL) was added
carbonyldiimidazole (CDI, 23 mg, 1.5 eq.) and DMAP (17 mg, 1.5
eq.). The reaction was stirred for 1 hour at room temperature. Then
4-fluorobenzenesulfonamide (20 mg, 1.2 eq.) and DBU (15 uL, 1.05
eq.) were added and the capped vial was allowed to stir for 16
hours. The reaction was then concentrated in vacuo to yield crude
97.
[0352]
(2S,3R)-3-((2-bis(tert-butoxycarbonyl)aminopyridin-4-yl)methyl)-N-(-
4-fluorophenylsulfonyl)-4-oxoazetidine-2-carboxamide (98): To a
solution of crude 97 from above in MeOH:AcOH (900 uL: 100 uL) was
added 0.5M NH.sub.4F in MeOH (190 uL, 1.0 eq.). After 2 hours and
20 minutes the reaction was concentrated in vacuo and triturated
with dichloromethane to yield 98 as a yellow foam which was taken
on to the next step without further purification.
[0353]
(2S,3R)-3-((2-bis(tert-butoxycarbonyl)aminopyridin-4-yl)methyl)-N-(-
4-fluorophenylsulfonyl)-N-methyl-4-oxoazetidine-2-carboxamide (99):
To a solution of 98 (18 mg, 1.0 eq.) in MeOH: THF (1 mL: 300 uL)
was added 2.0M TMSCHN.sub.2 in hexanes (266 uL, 17 eq.). After 15
minutes, TLC showed complete conversion of starting material. The
reaction was poured into ethyl acetate and then washed with 0.2N
HCl, 1N NaHCO.sub.3 and brine. The organic layer was then dried
over sodium sulfate and concentrated in vacuo. The product was then
purified via reverse phase HPLC (C18, acetonitrile/water with 0.1%
TFA) to yield 99 (11.9 mg, 65%) as a clear oil.
[0354]
(2S,3R)-3-((2-bis(tert-butoxycarbonyl)aminopyridin-4-yl)methyl)-N2--
(4-fluorophenylsulfonyl)-N2-methyl-4-oxo-N1-((R)-1-phenylethyl)azetidine-1-
,2-dicarboxamide (100): To a solution of 99 in DMF (0.75 mL) was
added (R)-.alpha.-methylbenzylisocyanate (6 uL, 2.0 eq.) and TEA
(8.5 uL, 3.0 eq.). The reaction was stirred under an atmosphere of
argon for 20 hours. The reaction was then concentrated in vacuo and
taken on to the next step without further purification.
[0355]
(2S,3R)-3-((2-aminopyridin-4-yl)methyl)-N2-(4-fluorophenylsulfonyl)-
-N2-methyl-4-oxo-N1-((R)-1-phenylethyl)azetidine-1,2-dicarboxamide
(101): To crude 100 from above was added 4N HCl in dioxane (2 mL).
The reaction was capped, stirred for 4 hours, concentrated in vacuo
and triturated with diethyl ether. The crude product was dissolved
in acetonitrile/methanol, filtered and purified via reverse phase
HPLC (C18, acetonitrile/water with 0.1% TFA) to yield 101 (1.7
mg).
Synthesis of 106
[0356] (2S,3R)-benzyl
3-((2-(bis-N,N-t-butoxycarbonyl)aminopyridin-4-yl)methyl)-1-(tert-butyldi-
methylsilyl)-4-oxoazetidine-2-carboxylate (102): Compound 102 was
synthesized by general method C using .alpha.-methylbenzyl
isocyanate (step 3).
[0357]
(3R,4S)-3-((2-(bis-N,N-t-butoxycarbonyl)aminopyridin-4-yl)methyl)-1-
-(tert-butyldimethylsilyl)-4-(hydroxymethyl)azetidin-2-one (103):
To a solution of compound 102 (0.35 g, 0.56 mmol) in 10 ml
anhydrous methanol was added sodium borohydride (0.128 g, 3.36
mmol) in several batches. The reaction was stirred at room
temperature for 2 hr, then slowly quenched with water and acidified
with 1N HCl. The reaction mixture was extracted with ethyl acetate
(three times), dried over magnesium sulfate and concentrated in
vacuo. The viscous oil was triturated with ether to give the
alcohol 103 as a white solid (0.184 g, 63%). LS/MS M+H 522.3, calc.
522.29.
[0358]
((2S,3R)-3-((2-(bis-N,N-t-butoxycarbonyl)aminopyridin-4-yl)methyl)--
1-(tert-butyldimethylsilyl)-4-oxoazetidin-2-yl)methyl
methanesulfonate (104): To a solution of alcohol 103 (0.047 mmol)
in 2 ml of dichloromethane was added methanesulfonyl chloride
(0.071 mmol) and triethylamine (0.141 mmol) dropwise and the
reaction was stirred at room temperature overnight. Afterwards, it
was concentrated in vacuo, diluted with ethyl acetate and washed
with sodium bicarbonate. The layers were separated and the organic
layer was dried over magnesium sulfate and concentrated in vacuo to
give compound 104 as a yellow oil (85%). LS/MS M+H 600.3, calc.
600.22.
[0359]
((2S,3R)-3-((2-(bis-N,N-t-butoxycarbonyl)-aminopyridin-4-yl)methyl)-
-4-oxoazetidin-2-yl)methyl methanesulfonate (105): To a solution of
compound 104 (0.04 mmol) in 1 ml anhydrous methanol were added
ammonium fluoride (0.048 mmol, 0.5 M in methanol) and acetic acid
(0.144 mmol) and the reaction was stirred at room temperature for 2
h. Afterwards, it was concentrated in vacuo and the residual acetic
acid was azeotroped with toluene. The solid was diluted with
dichloromethane and was filtered through a syringe filter. The
solution was concentrated in vacuo to give compound 105 as a clear
oil (quantitative yield) LS/MS M+H 486.5, calc. 486.13.
[0360]
((2S,3R)-1-(((R)-1-phenylethyl)carbamoyl)-3-((2-aminopyridin-4-yl)m-
ethyl)-4-oxoazetidin-2-yl)methyl methanesulfonate TFA salt (106):
To a solution of compound 105 (0.04 mmol) in 2 ml anhydrous
methylene chloride was added isocyanate (0.06 mmol) and triethyl
amine (0.12 mmol) dropwise and the reaction mixture was stirred at
room temperature for 5 h. Afterwards, the reaction mixture was
concentrated in vacuo, diluted with ethyl acetate and washed with
sodium bicarbonate. The layers were separated and the organic layer
was dried over magnesium sulfate and concentrated in vacuo. Column
chromatography purification (silica, hexane/ethyl acetate) gave the
product in 70% yield. (LS/MS M+H 633.4, calc. 633.2). The product
was treated with 1 ml TFA:dichloromethane for 1 h at room
temperature. Afterwards, it was concentrated in vacuo and washed
with ethyl ether (five times). Compound 106 was obtained as a white
solid (76%). LS/MS M+H 433.3, calc. 433.15.
Synthesis of 112
[0361] (2S,3S)-ethyl
3-(benzyloxy)-1-(4-methoxyphenyl)-4-oxoazetidine-2-carboxylate
(108): To a solution of p-anisidine (107, 5.4 mmol) in 10 ml
anhydrous dichloromethane and 2 ml toluene were added ethyl
glyoxylate (8.0 mmol, 50% solution in toluene) and anhydrous
magnesium sulfate (3.0 g). The reaction mixture was heated at
70.degree. C. for 3 h. Afterwards, the mixture was filtered and
cooled to 0.degree. C. with an ice/water bath. Triethylamine (10.8
mmol) and benzyloxyacetyl chloride (8.1 mmol) were added dropwise
and the reaction mixture was stirred at room temperature for 18 h.
Afterwards, it was concentrated in vacuo and triturated with ethyl
ether and hexane to give compound 108 as a tan solid (69% yield).
LS/MS M+H 356.2, calc. 356.12.
[0362] (2S,3S)-ethyl
1-(4-methoxyphenyl)-4-oxo-3-(trifluoromethylsulfonyloxy)azetidine-2-carbo-
xylate (109): Palladium hydroxide (0.186 mmol, 20% on carbon, wet)
was added to a round bottom flask. The flask was evacuated and
backfilled with hydrogen (three times). Afterwards a solution of
compound 108 (3.72 mmol) in anhydrous methanol (30 ml) was added to
the flask and the reaction was stirred for 18 h at room temperature
under a positive atmosphere of hydrogen. Afterwards, the mixture
was filtered through celite and concentrated in vacuo to give the
free alcohol as a white solid. To a cold (0.degree. C.) solution of
the alcohol (0.377 mmol) in anhydrous dichloromethane (2 ml) were
added triethylamine (1.13 mmol) and trifluoromethanesulfonyl
chloride (0.57 mmol) dropwise and the reaction was stirred at
0.degree. C. for 30 min. Afterwards, it was diluted with ethyl
acetate and washed with sodium bicarbonate. The layers were
separated and the organic layer was dried over magnesium sulfate
and concentrated in vacuo. Compound 109 was obtained as an orange
solid (quant. yield). LS/MS M+H 398.5, calc. 398.3.
[0363] (2R,3R)-ethyl
3-mercapto-1-(4-methoxyphenyl)-4-oxoazetidine-2-carboxylate (110):
To a solution of triisopropylsilane thiol (0.91 mmol) in anhydrous
THF (2 ml) was added sodium hydride (0.68 mmol, 60% in mineral oil)
and the reaction mixture was stirred at room temperature for 15
min. Afterwards, compound 109 (0.45 mmol, dissolved in 2 ml THF)
was added to the reaction and the mixture was stirred at room
temperature for 1 h. Then it was concentrated in vacuo, diluted
with ethyl acetate and washed with sodium bicarbonate. The layers
were separated and the organic layer was dried over magnesium
sulfate and concentrated in vacuo. Column chromatography
purification (12 g pre-packed silica column, hexane/ethyl acetate)
afforded the protected thiol. The thiol (0.126 mmol) was then
diluted with anhydrous methanol (2 ml) and ammonium fluoride (0.15
mmol, 0.5 M in methanol) and acetic acid (0.454 mmol) were added to
the reaction and the mixture was stirred at room temperature for 2
h. Afterwards, it was concentrated in vacuo and the residual acetic
acid was azeotroped with toluene. The solid was diluted with
dichloromethane and was filtered through a syringe filter. The
solution was concentrated in vacuo to give compound 110 as a clear
oil (23% yield for both steps). LS/MS M+H 282.3, calc. 282.1.
[0364] (2R,3R)-ethyl
3-((2-bis(butoxycarbonyl)aminopyridin-4-yl)methylthio)-4-oxoazetidine-2-c-
arboxylate (111): To a solution of compound 110 (0.05 mmol) in
anhydrous THF (2 ml) was added sodium hydride (0.07 mmol, 60% in
mineral oil) and the reaction mixture was stirred at room
temperature for 15 min. Afterwards, bromide A was added (0.08 mmol)
and the reaction was stirred at room temperature for 1 h. Then it
was concentrated in vacuo, diluted with ethyl acetate and washed
with sodium bicarbonate. The layers were separated and the organic
layer was dried over magnesium sulfate and concentrated in vacuo.
Column chromatography purification (4 g pre-packed silica column,
hexane/ethyl acetate) afforded the product as a yellow oil (48%
yield). The product was then diluted with acetonitrile (1 ml) and
water (0.5 ml) and cooled to 0.degree. C. with an ice water bath.
Ceric ammonium nitrate (0.08 mmol) dissolved in water (0.5 ml) was
added and the reaction was stirred at 0.degree. C. for 30 min.
Afterwards, the reaction was concentrated in vacuo and diluted with
ethyl acetate and sodium bicarbonate. The aqueous layer was
extracted with ethyl acetate (three times), dried over magnesium
sulfate and concentrated in vacuo. Compound 111 was obtained was a
clear oil (96% yield). LS/MS M+H 482.3, calc. 482.2.
[0365] (2R,3R)-ethyl
1-(((R)-1-phenylethyl)carbamoyl)-3-((2-aminopyridin-4-yl)methylthio)-4-ox-
oazetidine-2-carboxylate (112): To a solution of compound 111
(0.025 mmol) in 2 ml anhydrous methylene chloride were added
isocyanate B (0.038 mmol) and triethylamine (0.075 mmol) dropwise
and the reaction mixture was stirred at room temperature for 5 h.
Afterwards, the reaction mixture was concentrated in vacuo, diluted
with ethyl acetate and washed with sodium bicarbonate. The layers
were separated and the organic layer was dried over magnesium
sulfate and concentrated in vacuo. The product was treated with 1
ml TFA/dichloromethane for 1 h at room temperature. Afterwards, it
was concentrated in vacuo and washed with ethyl ether. The two
diastereomers were separated on reverse phase HPLC
(acetonitrile/water as eluents). Compound 112 was obtained as a
white solid (0.001 mg). LS/MS M+H 429.62, calc. 429.5
[0366] (2R,3R)-ethyl
3-((2-aminopyridin-4-yl)methylthio)-1-(4-methoxyphenyl)-4-oxoazetidine-2--
carboxylate (113) and (2R,3S)-ethyl
3-((2-aminopyridin-4-yl)methylthio)-1-(4-methoxyphenyl)-4-oxoazetidine-2--
carboxylate (114): Compound 113 and 114 were synthesized by the
same method used to synthesis 112 with the omission of the
NH.sub.4F/AcOH/MeOH deprotection step. The final TFA deprotection
yielded the mixture of the 113 and 114. The isomers were separated
via reverse phase HPLC (acetonitrile/water) with 0.1% TFA added to
the mobile phase.
Synthesis of 119
[0367]
(3R,4S)-3-((2-bis(t-butoxycarbonyl)-aminopyridin-4-yl)methyl)-1-(te-
rt-butyldimethylsilyl)-4-(2-diazoacetyl)azetidin-2-one (115): To a
solution of compound 5 (100 mg, 0.186 mmol) in THF (1 mL) at
-15.degree. C. was added triethylamine (27.8 uL, 0.2 mmol) and
ethyl chloroformate (20 uL, 0.2 mmol). The reaction mixture was
then stirred at -15.degree. C. for 30 minutes and the ppt was then
filtered off under an atmosphere of argon. To the crude anhydride
was then added acetonitrile (1 mL) and 2.0M TMSCHN.sub.2 in hexanes
(0.2 mL, 0.4 mmol). The reaction was stirred at 4.degree. C. for 48
h, at which point LCMS showed complete conversion of starting
material. Diethyl ether (25 mL) was added to the reaction and
washed with 10% NaHSO.sub.4 (25 mL), saturated NaHCO.sub.3 (25 mL)
and brine (25 mL). The crude material was purified by Combiflash
silica gel chromatography (0-3 min: 100% hexanes, 3-10 min: 40%
EtOAc in hexanes) to yield 115 (32 mg). Exact mass. 559.28. Found
ES MS [M+H].sup.+ 559.9.
[0368]
(3R,4S)-3-((2-bis(t-butoxycarbonyl)-aminopyridin-4-yl)methyl)-4-(2--
diazoacetyl)azetidin-2-one (116): To a solution of 115 (50 mg,
0.089 mmol) in methanol (1 mL) at room temperature was added 0.5 M
ammonium fluoride (178 uL, 0.089 mmol) and acetic acid (14 uL,
0.257 mmol). The reaction was stirred for 16 hours and then
purified with Combiflash silica gel chromatography (0-10 min: 100%
ethyl acetate) to yield 116 (39 mg).
[0369]
(2S,3R)-3-((2-bis(t-butoxycarbonyl)-aminopyridin-4-yl)methyl)-2-(2--
diazoacetyl)-4-oxo-N--((R)-1-phenylethyl)azetidine-1-carboxamide
(117): To a solution of 116 (39 mg, 0.089 mmol) in anhydrous DCM (2
mL) was added (R)-(+)-methylbenzylamine (30 mg, 2 eq.) and
triethylamine (36 uL, 3 eq.). The reaction was stirred for 16 hours
and the crude mixture was purified via preparative TLC (70% EtOAc
in hexanes) to yield 117 (43.4 mg).
[0370] Ethyl
2-((2R,3R)-1-(((R)-1-phenylethyl)carbamoyl)-3-((2-bis(t-butoxycarbonyl)-a-
minopyridin-4-yl)methyl)-4-oxoazetidin-2-yl)acetate (118): To a
solution of 117 (10 mg, 0.016 mmol) in ethanol (2 mL) was added
silver benzoate (1 mg, 10% w/w) and the mixture was stirred at room
temperature for 16 hours. The crude material was then purified by
preparative TLC (70% EtOAc in hexanes) to yield 118 (4 mg). Exact
mass. 610.3. Found ES MS [M+H].sup.+ 610.9.
[0371] Ethyl
2-((2R,3R)-1-(((R)-1-phenylethyl)carbamoyl)-3-((2-aminopyridin-4-yl)methy-
l)-4-oxoazetidin-2-yl)acetate (119): Compound 118 was dissolved in
1:1 TFA:DCM (2 mL) and the reaction stirred for 2 hours and then
concentrated in vacuo to yield a crude material which was purified
by reverse phase HPLC (acetonitrile/water with 0.1% TFA) to yield
119 (1.2 mg).
Synthesis of 123
##STR00112##
[0373] 5-Methyl-benzo[d]isoxazol-3-ylamine (121) Acetohydroxamic
acid (1.69 g, 22.5 mmol) in DMF (45 mL) was stirred with
K.sub.2CO.sub.3 and several drops of water at RT for 30 min, the
2-fluoro-5-methylbenzonitrile (120) (1.35 g, 10 mmol) in DMF (5 mL)
was added and the reaction mixture was stirred at RT for 3 days.
The reaction was diluted with water and the mixture was extracted
with ethyl acetate. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated in vacuo to provide 121 (878 mg,
60%). LC-MS (ESI) m/z 149.2.
[0374] Benzo[d]isoxazol-3-bis(N,N-t-butoxycarbonyl)amine (122): To
a solution of 121 (148 mg, 1 mmol) in DCM (2 mL) was added
di-t-butyldicarbonate (546 mg, 2.5 mmol), DIEA (0.348 mL, 2 mmol),
and DMAP (22.2 mg, 1 mmol) at 0.degree. C. The reaction was warmed
up to RT and stirred for 4 hr and then diluted with ethyl acetate
and treated with aqueous ammonium chloride to pH 6. The organic
layer was separated and washed with brine and concentrated in
vacuo. The residue purified by silica gel chromatography using 100%
hexane as the eluent to yield 122 (347 mg, 100%), LC-MS (ESI) m/z
348.8.
[0375]
5-(Bromomethyl)benzo[d]isoxazol-3-(bis(t-butoxycarbonyl))amine
(123): To a solution of 122 (1.37 g, 3.9 mmol) in CCl.sub.4 was
added N-bromosuccinimide (700 mg, 1 eq.) and benzoyl peroxide (54
mg, 0.04 eq.). The mixture was heated to 85.degree. C. for 4 hours
and then filtered after cooling to room temperature. The filtrated
was concentrated in vacuo and purified via silica gel
chromatography eluting with 0-10% EtOAc in hexanes to provide 123
(630 mg).
Synthesis of 129
[0376] (S)-benzyl 4-oxo-1-(phenylcarbamoyl)azetidine-2-carboxylate
(125) A solution of aniline (124) (0.186 mL, 2 mmol) and
triethylamine (0.28 mL, 2 mmol) in dichloromethane (5 mL) was
dropped into a phosgene solution (1.27 mL, 20% in toluene) at
0.degree. C. and stirred for 30 min. Then (S)-benzyl
4-oxoazetidine-2-carboxylate (531 mg, 2.6 mmol) in THF (5 mL) was
added. The resultant reaction mixture was stored in a refrigerator
overnight and then filtered. The filtrate was condensed and the
residue was purified by medium pressure chromatography (silica gel
column) with dichloromethane as the eluent to provide 125 (474 mg,
73%). .sup.1H NMR (CDCl.sub.3) .delta. 8.28 (br, 1H), 7.48-7.29 (m,
9H), 7.14-7.09 (m, 1H), 5.25 (s, 2H), 4.62-4.57 (m, 1H), 3.43-3.35
(m, 1H), 3.12-3.05 (m, 1H), LC-MS (ESI) m/z 324.0.
[0377] (S)-4-oxo-1-(phenylcarbamoyl)azetidine-2-carboxylate acid
(126) Compound 125 was dissolved in a methanol-ethanol solution and
10% Pd--C catalyst was added. The reaction vessel was then
evacuated and flushed with hydrogen. The reaction was stirred under
an atmosphere of hydrogen until all the starting material was
consumed as evidenced by LCMS to yield 126 (quantitative yield).
.sup.1H NMR (MeOD) .delta.7.63-7.46 (m, 2H), 7.34-7.29 (m, 2H),
7.13-7.08 (m, 1H), 4.54-4.51 (m, 1H), 3.48 (dd, J=6.4, 15.8 Hz,
1H), 3.09 (dd, J=2.3, 16.8 Hz, 1H), LC-MS (ESI) m/z 234.9.
[0378]
(2S,3R)-3-((3-di-Boc-aminobenzo[d]isoxazol-5-yl)methyl)-4-oxo-1-(ph-
enylcarbamoyl)azetidine-2-carboxylic acid (128) A solution of 126
(41 mg, 0.174 mmol) in THF was added to 1.0M LiHMDS in THF (0.635
mL) at -78.degree. C. and the reaction mixture was stirred for 30
min and then slowly warmed up to room temperature over 1 h. The
reaction mixture was cooled back to -78.degree. C. followed by
addition of compound 127 (44 mg, 0.103 mmol) dissolved in THF.
After 30 min, the reaction mixture was warmed up to 0.degree. C.
for one hour and then to room temperature for another hour. The
reaction was quenched with aqueous ammonia chloride and extracted
with ethyl acetate. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by preparative TLC to afford 128 (32 mg, 54%), LC/MS (ESI)
m/z 581.0.
[0379]
(2S,3R)-3-((3-aminobenzo[d]isoxazol-5-yl)methyl)-4-oxo-1-(phenylcar-
bamoyl)-azetidine-2-carboxylic acid (129): Compound 128 (32 mg) was
treated with 20% TFA-DCM for 20 min at room temperature. The
reaction was concentrated in vacuo and the residue was purified by
preparative HPLC (Vydac, Protein & Peptide C18 column, 0-60%
H.sub.2O-acetonitrile w/0.1% TFA, gradient change in 50 min) to
afford 129 (7.2 mg, 26%), .sup.1H NMR (DMSO) .delta. 8.95 (s, 1H),
7.77 (s, 1H), 7.55 (d, J=8.3 Hz, 3H), 7.43 (d, J=8.55 Hz, 1H), 7.36
(t, J=7.7 Hz, 2H), 7.13 (t, J=7.4 Hz, 1H), 4.35 (d, J=2.8 Hz, 1H),
3.76 (m, 1H), 3.27 (d, J=7.7 Hz, 2H), LC-MS (ESI) m/z 381.1.
[0380]
(2S,3R)-3-((3-amino-1H-indazol-5-yl)methyl)-4-oxo-1-(phenylcarbamoy-
l)azetidine-2-carboxylic acid (130): Compound 130 was prepared
analogously to 129 using hydrazine instead of acetohydroxamic acid
in the synthesis of the alkylating agent. Yield 2.3 mg. .sup.1H NMR
(DMSO) .delta. 11.60 (br, 1H), 8.90 (s, 1H), 7.63 (s, 1H), 7.51 (d,
J=6.0 Hz, 2H), 7.35-7.20 (m, 5H), 7.09 (t, J=6.9 Hz, 1H), 6.45 (br,
2H), 4.28 (d, J=2.7 Hz, 1H), 3.69 (dt, J=2.7, 7.3 Hz, 1H), 3.17 (d,
J=7.4 Hz, 2H), LC-MS (ESI) m/z 380.1.
Example 9
General method J for the synthesis of 3-substituted-amino
beta-lactams
##STR00113## ##STR00114##
[0382]
(2S,3R)-3-azido-1-(tert-butyldimethylsilyl)-4-oxoazetidine-2-carbox-
ylic acid (131): To LDA (1.8M, 4.99 mL) and THF (10 mL) at
-78.degree. C. was added dropwise
(S)-1-t-butyldimethylsilyl-4-oxo-2-azetidinecarboxylic acid (4, 1.0
g) in THF (15 mL). The resulting solution was warmed to -30.degree.
C. or -10.degree. C. for 30 min. If a slurry formed, additional THF
was added to solubilize the azetidine. After 30 min., the solution
was cooled to -78.degree. C. A pre-cooled solution (-10.degree. C.)
of trisyl azide (1.61 g) in THF (10 mL) was added slowly. The
reaction mixture was stirred for one hour before quenching with
TMSCl (0.82 mL). The cold bath was then removed and the solution
allowed to stir for an additional hour. Saturated sodium
bicarbonate (100 mL) was added and the aqueous layer was extracted
with diethyl ether (2.times.100 mL). The organic extracts were
discarded. The aqueous layer was then brought to pH=7 by slow
addition of 1N HCl and extracted with Et.sub.2O (2.times.100 mL)
and the organic layers were discarded. The aqueous layer was then
brought to pH=3 by slow addition of 1N HCl and extracted with EtOAc
(3.times.100 mL). The combined organic layers were dried over
MgSO.sub.4 and concentrated in vacuo to yield 131 which was taken
on to the next step without further purification.
[0383] (2S,3R)-methyl
3-azido-1-(tert-butyldimethylsilyl)-4-oxoazetidine-2-carboxylate
(132): To a solution of crude 131 (1.58 g) in CH.sub.2Cl.sub.2 (25
mL) was added methanol (0.26 mL, 6.43 mmol). DCC (1.45 g, 7.02
mmol) and catalytic DMAP. The solution was stirred at room
temperature overnight. The solution was then filtered through
Celite and concentrated in vacuo. The crude product was purified by
silica gel column chromatography, eluting with 20% EtOAc in
hexanes, to yield pure 132.
[0384] (2S,3R)-methyl
3-amino-1-(tert-butyldimethylsilyl)-4-oxoazetidine-2-carboxylate
(133): To a solution of 132 (5.85 mmol) in 3:1 methanol:dioxane (25
mL) was added a few drops of TFA and 10% Pd/C (10 wt %). The
reaction was stirred under atmospheric hydrogen for one hour and
then filtered through Celite. The solvent was removed and the
residue triturated with diethyl ether to afford 133, which was used
in the next step without further purification.
[0385] (2S,3R)-methyl
1-(tert-butyldimethylsilyl)-3-((2-(di-(t-butoxycarbonyl)-amino)pyridin-4--
yl)methylamino)-4-oxoazetidine-2-carboxylate (134): To a solution
of 133 and 3 in acetonitrile was added potassium carbonate and
catalytic NaI. The resultant solution was heated at reflux until
complete by TLC (.about.1-2 h). The solution was then cooled and
the solvent removed in vacuo. The residue was taken up in ethyl
acetate and the organic layer was washed with water (2.times.X mL)
and brine (x mL.). The organic layer was then dried over
MgSO.sub.4, and concentrated in vacuo. The product was then
purified by silica gel chromatography, eluting with 50-100% ethyl
acetate in hexanes, to yield pure 134.
[0386] (2S,3R)-methyl
3-((2-(di-(t-butoxycarbonyl)-amino)pyridin-4-yl)methylamino)-4-oxoazetidi-
ne-2-carboxylate (135): Compound 135 was synthesized following the
procedure outlined in steps 2 of General method C.
[0387] (2S,3R)-methyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(1-((2-(di-(t-butoxycarbonyl)-amino)py-
ridin-4-yl)methyl)-3-((R)-1-phenylethyl)ureido)-4-oxoazetidine-2-carboxyla-
te (136): Compound 136 was synthesized following the procedure
outlined in steps 3 of General method C.
[0388] (2S,3R)-methyl
1-(((R)-1-phenylethyl)carbamoyl)-3-(1-((2-aminopyridin-4-yl)methyl)-3-((R-
)-1-phenylethyl)ureido)-4-oxoazetidine-2-carboxylate (137), TFA
salt: Compound 137 was synthesized following the procedure outlined
in steps 5 of General method C.
Example 10
General method K for the preparation of geminal dimethyl
beta-lactams
##STR00115##
[0390]
1-(Tert-butyldimethylsilyl)-3-(3-chloropropyl)-4,4-dimethylazetidin-
-2-one (138): 4,4-dimethylazetidin-2-one was prepared as described
in J. Medicinal Chemistry 1994, 37, 897-906 and then as described
in U.S. Pat. No. 6,335,324.
[0391]
3-((2-(N,N-bis-tert-butoxycarbonyl)-aminopyridin-4-yl)methyl)-1-(te-
rt-butyldimethylsilyl)-4,4-dimethylazetidin-2-one (139): The
alkylation procedure from U.S. Pat. No. 6,335,324, p. 22 except a)
3,4-bromomethyl-2-bis-Boc-aminopyridine, was used as the alkylating
agent instead of 3-chloropropyl iodide and b) the reaction was
allowed to warm up to room temperature after the addition of the
3.
[0392]
3-((2-(N,N-bis-tert-butoxycarbonyl)-aminopyridin-4-yl)methyl)-4,4-d-
imethylazetidin-2-one (140): To 139 (31 mg) was added 0.5M
NH.sub.4F in methanol (143 uL) and acetic acid (11 uL) and stirred
until disappearance of the starting material. The reaction was
concentrated in vacuo and re-dissolved in DCM. After 2 hr, the
precipitate was filtered and the filtrate purified via preparative
TLC (50% ethyl acetate in hexanes) to yield 140 (20 mg, 84%).
[0393]
3-((2-(N,N-bis-tert-butoxycarbonyl)-aminopyridin-4-yl)methyl)-2,2-d-
imethyl-4-oxo-N--((R)-1-phenylethyl)azetidine-1-carboxamide (141):
To a solution of 140 (0.03 mmol) in anhydrous THF (1 mL) was cooled
to -78.degree. C. and then 1.0 M LiHMDS (1.2 eq) was added slowly
via syringe. After stirring for 30 minutes at -78.degree. C.,
.alpha.-methylbenzyl isocyanate (1.2 eq.) was added via syringe.
The reaction was then allowed to warm up to 0.degree. C. over 30
minutes (or until LCMS showed no starting material). The reaction
was then quenched with saturated ammonium chloride, extracted with
ethyl acetate, and the organic layer was dried over sodium sulfate.
The reaction was concentrated in vacuo and taken on without further
purification.
[0394]
3-((2-aminopyridin-4-yl)methyl)-2,2-dimethyl-4-oxo-N--((R)-1-phenyl-
ethyl)azetidine-1-carboxamide (142): A solution of crude 141 in DCM
(1 mL) was cooled to 0.degree. C. and then TFA (200 uL) was added.
After 4 hr, the reaction was concentrated in vacuo and the crude
material purified via reverse phase HPLC (acetonitrile/water) with
0.1% TFA to yield 142 (13.3 mg) as the TFA salt.
Example 11
General method L for the preparation of 3-propylguanidine
beta-lactams
##STR00116## ##STR00117##
[0396] 4(2S,3R)-3-(3-chloropropyl)-4-oxoazetidine-2-carboxylic acid
(145): Commercially available beta-lactam acid 144 (1 g, 1.0 eq.)
was dissolved in anhydrous THF (15 mL) and the solution was cooled
to -78.degree. C. under an atmosphere of argon. 1.8M LDA in
heptane/THF/ethylbenzene (5.9 mL, 2.4 eq.) was added slowly to the
cooled solution via syringe. The reaction was stirred for 20
minutes at -78.degree. C. and then 1-chloro-3-iodopropane (0.6 mL,
1.2 eq.) was slowly added to the cooled solution. The reaction was
stirred a further hour at -78.degree. C., warmed to room
temperature and poured into 1:1 cold 1N HCl: brine (75 mL). The
aqueous phase was extracted with ethyl acetate (2.times.75 mL) and
the organic layers were combined and washed with saturated
NaHCO.sub.3 (2.times.25 mL). The basic layer was then washed again
with ethyl acetate (75 mL). The basic aqueous layer was then
acidified with 1N HCl-brine and extracted with ethyl acetate
(2.times.50 mL) and the organic layers were combined and dried over
sodium sulfate. Concentration in vacuo yielded 145 (0.88 g) as a
dark yellow oil.
[0397] (2S,3R)-3-(3-azidopropyl)-4-oxoazetidine-2-carboxylic acid
(146): To a solution of 145 (1.0 g, 1.0 eq.) in anhydrous DMF (5
mL) was added tetrabutylammonium iodide (60 mg, 0.05 eq.) and
tetrabutylammonium azide (1.1 g, 1.2 eq.). The reaction was stirred
at room temperature for 72 hours and the poured into 1N HCl-brine
and extracted with ethyl acetate (2.times.20 mL). Then the organic
layers were extracted with saturated NaHCO.sub.3 (3 15 mL). The
combined basic aqueous layers were then acidified with 1N-HCl:brine
and then the acidic solution was extracted with ethyl acetate
(3.times.20 mL). These last organic layers were combined, dried
over magnesium sulfate and concentrated in vacuo to yield 146 (0.59
g) as a brown oil.
[0398] (2S,3R)-3-(3-aminopropyl)-4-oxoazetidine-2-carboxylic acid
(147): To a solution of 146 (0.59 g, 1.0 eq.) in DMF (5 mL) and
acetic acid (1 mL) was added 10% Pd/C (300 mg). The flask was
evacuated and flushed with hydrogen. The reaction was then stirred
at room temperature under an atmosphere of hydrogen for 16 hours.
The reaction was then filtered through Celite and crude amine 147
was immediately used in the next step as a solution in DMF.
[0399]
(2S,3R,E)-3-(3-(2,3-bis(3-phenylpropanoyl)guanidino)propyl)-4-oxoaz-
etidine-2-carboxylic acid (148): To the DMF solution of amine 147
(330 mg of amine) was added
N,N'bis(benzyloxycarbonyl)-1H-pyrazole-1-carboxamide (718 mg, 1.0
eq.) and DIEA (1.5 mL). The reaction was stirred at room
temperature for 6 hours and then poured into 1N HCl:brine and
extracted with ethyl acetate (2.times.30 mL). The organic layers
were combined and extracted with saturated NaHCO.sub.3 (2.times.25
mL). The basic aqueous layers were then acidified with 1N HCl:brine
and extracted with ethyl acetate (2.times.25 mL). These last
organic layers were dried over sodium sulfate and concentration in
vacuo yielded a crude material (200 mg) as a colorless oil. The
crude material was purified via silica gel chromatography eluting
with a gradient elution from ethyl acetate to 1% acetic acid in
EtOAc to give 148 (74 mg).
[0400] (2S,3R,E)-benzyl
3-(3-(2,3-bis(3-phenylpropanoyl)guanidino)propyl)-4-oxoazetidine-2-carbox-
ylate (149): To a solution of 148 (74 mg, 1.0 eq.) in DMF (1.5 mL)
was added benzyl alcohol (19 uL, 1.2 eq.), EDC-HCl (35 mg, 1.2
eq.), HOBt (24 mg, 1.2 eq.) and DIEA (60 .mu.L, 2.4 eq.). The
reaction was stirred at room temperature for 16 hours and then
diluted with ethyl acetate and washed with 1N HCl. The aqueous
layer was then washed with ethyl acetate (2.times.10 mL). The
combined organics were then washed with saturated NaHCO.sub.3
(2.times.10 mL), water (10 mL), dried over magnesium sulfate and
concentrated in vacuo to give the crude product (55 mg) as a
colorless oil. The product was purified by silica gel
chromatography using a gradient elution from 20% EtOAc in hexanes
to 50% EtOAc in hexanes to yield 149 (50 mg).
[0401] (2S,3R,E)-benzyl
3-(3-(2,3-bis(3-phenylpropanoyl)guanidino)propyl)-4-oxo-1-(phenylcarbamoy-
l)azetidine-2-carboxylate (150): To a solution of 149 (50 mg, 1.0
eq.) in DCM (2 mL) was added phenylisocyanate (29 .mu.L, 3.0 eq.),
triethylamine (12 .mu.L, 1 eq.) and DMAP (1 mg). The reaction was
stirred for 18 hours at room temperature and then diluted with
dichloromethane and washed with 1N HCl and water. The organic layer
was then concentrated in vacuo and purified via silica gel
chromatography eluting with 15% EtOAc in hexanes to give 150 (10
mg).
[0402]
(2S,3R)-3-(3-guanidinopropyl)-4-oxo-1-(phenylcarbamoyl)azetidine-2--
carboxylic acid (151): To a solution of 150 (10 mg, 1.0 eq.) in THF
(3 mL) and 1N HCl (0.5 mL) was added 10% Pd/C (20 mg). The reaction
flask was then evacuated and flushed with hydrogen and stirred
under an atmosphere of hydrogen for 18 hours. The reaction was then
filtered through Celite and concentrated in vacuo. The crude
material was purified via HPLC (C18, acetonitrile/water, 0.1% TFA)
to yield 151 (0.5 mg).
[0403] (2S,3R)-Ethyl
3-(3-guanidinopropyl)-4-oxo-1-(phenylcarbamoyl)azetidine-2-carboxylate
(152): Compound 152 was synthesized by general method L using ethyl
alcohol (step 5) and phenyl isocyanate (step 6) to yield 152.
[0404]
(2S,3R)-1-((4-Methoxybenzyl)carbamoyl)-3-(3-guanidinopropyl)-4-oxoa-
zetidine-2-carboxylic acid (153): Compound 153 was synthesized by
general method L using 4-methoxylbenzylisocyanate (step 6) to yield
153 (6.8 mg).
[0405]
(2S,3R)-1-((4-Methoxyphenethyl)carbamoyl)-3-(3-guanidinopropyl)-4-o-
xoazetidine-2-carboxylic acid (154): Compound 154 was synthesized
by general method L using 4-methoxyphenethylisocyanate (step 6) to
yield 154 (23 mg).
[0406]
(2S,3R)-1-(((R)-1-(Naphthalen-1-yl)ethyl)carbamoyl)-3-(3-guanidinop-
ropyl)-4-oxoazetidine-2-carboxylic acid (155): Compound 155 was
synthesized by general method L using (R)-1-naphthylethylisocyanate
(step 6) to yield 155 (31.1 mg, 66%).
[0407]
(2S,3R)-1-((3-Ethylphenyl)carbamoyl)-3-(3-guanidinopropyl)-4-oxoaze-
tidine-2-carboxylic acid (156): Compound 156 was synthesized by
general method L using 4-ethylphenylisocyanate (step 6) to yield
156 (25.1 mg, 61%).
[0408]
(2S,3R)-1-(((S)-1-Phenylethyl)carbamoyl)-3-(3-guanidinopropyl)-4-ox-
oazetidine-2-carboxylic acid (157): Compound 157 was synthesized by
general method L using (S)-1-methylbenzylisocyanate (step 6) to
yield 157.
[0409]
(2S,3R)-1-(((S)-1-(naphthalen-1-yl)ethyl)carbamoyl)-3-(3-guanidinop-
ropyl)-4-oxoazetidine-2-carboxylic acid (158): Compound 158 was
synthesized by general method L using (S)-1-naphthylethylisocyanate
(step 6) to yield 158 (25.3 mg, 77%).
[0410]
(2S,3R)-1-((4-ethylphenyl)carbamoyl)-3-(3-guanidinopropyl)-4-oxoaze-
tidine-2-carboxylic acid (159): Compound 159 was synthesized by
general method L using 4-ethylphenylisocyanate (step 6) to yield
159 (24.8 mg, 63%).
[0411]
(2S,3R)-1-(((R)-1-phenylethyl)carbamoyl)-3-(3-guanidinopropyl)-4-ox-
oazetidine-2-carboxylic acid (160): Compound 160 was synthesized by
general method L using (R)-1-methylbenzylisocyanate (step 6) to
yield 160.
* * * * *