U.S. patent application number 09/973493 was filed with the patent office on 2002-09-05 for polypyrrolinone based inhibitors of matrix metalloproteases.
Invention is credited to Hirschmann, Ralph F., Nittoli, Thomas, Smith, Amos B. III, Sprengeler, Paul.
Application Number | 20020123635 09/973493 |
Document ID | / |
Family ID | 26931612 |
Filed Date | 2002-09-05 |
United States Patent
Application |
20020123635 |
Kind Code |
A1 |
Smith, Amos B. III ; et
al. |
September 5, 2002 |
Polypyrrolinone based inhibitors of matrix metalloproteases
Abstract
A compound of the formula 1 wherein R1, R2, R3, R4, R5, R6 are
as defined herein, useful for the inhibition of inhibition of
matrix metalloproteases (MMPs)and for treating conditions mediated
by elevated levels of MMPs such as osteoarthritis, rheumatoid
arthritis, septic arthritis, periodontal disease, gingivitis, solid
tumor growth and tumor invasion by secondary metastasis, corneal
ulceration, dermal ulceration, epidermolysis bullosa, neural
degeneration, multiple sclerosis and surgical wound healing.
Inventors: |
Smith, Amos B. III; (Merion,
PA) ; Hirschmann, Ralph F.; (Blue Bell, PA) ;
Nittoli, Thomas; (West Caldwell, NJ) ; Sprengeler,
Paul; (El Granada, CA) |
Correspondence
Address: |
MATHEWS, COLLINS, SHEPHERD & GOULD, P.A.
100 THANET CIRCLE, SUITE 306
PRINCETON
NJ
08540-3674
US
|
Family ID: |
26931612 |
Appl. No.: |
09/973493 |
Filed: |
October 9, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60238375 |
Oct 6, 2000 |
|
|
|
Current U.S.
Class: |
548/314.7 ;
548/465; 548/519 |
Current CPC
Class: |
C07D 207/36
20130101 |
Class at
Publication: |
548/314.7 ;
548/465; 548/519 |
International
Class: |
C07D 43/14; C07D
403/02 |
Goverment Interests
[0002] This invention was made with Government support under Grant
No. AI-42010 awarded by the National Institutes of Health through
the National Institute of Allergy and Infectious Diseases. The
government has certain rights in this invention.
Claims
We claim:
1. A compound having formula (1): 8wherein: R.sup.1 is --NHOH or
--OH; R.sup.2 is hydrogen, C.sub.1-6 alkyl; R.sup.3 and R.sup.4 are
selected independently from a group consisting of the side chains
of the naturally occurring .alpha.-amino acids, C.sub.1-6 alkyl,
(CH.sub.2).sub.nAr wherein the aryl group is optionally substituted
with up to two groups independently selected from the group
consisting of phenyl, hydroxy, C.sub.1-4 alkoxy, phenoxy,
--O(CH.sub.2).sub.mOH, C.sub.1-4 thioalkyl, halogens, nitro, cyano,
C.sub.1-4 alkylsulfonyl, and C.sub.1-4 alkylsulfinyl wherein m is 1
or 2 and n is 0 to 3; R.sup.5 is hydrogen, C.sub.1-6 alkyl; R.sup.6
is OR.sup.7, NR.sup.7R.sup.8 wherein R.sup.7 and R.sup.8 taken
independently are selected from a group consisting of hydrogen,
C.sub.1-4 alkyl, C.sub.1-4 branched alkyl, alkyl aryl and benzyl;
and, a stereoisomer, enantiomer, diastereomer, hydrate or
pharmaceutically acceptable salt thereof.
2. A compound according to claim 1 wherein: R.sup.1 is --NHOH or
--OH; R.sup.2 is H or CH.sub.3; R.sup.3 is n-C.sub.1-6 alkyl,
s-C.sub.4H.sub.9, i-C.sub.4H.sub.9, (CH.sub.2).sub.nAr,
(CH.sub.2).sub.n-p-C.sub.6H.sub.4OM- e,
(CH.sub.2).sub.np-C.sub.6H.sub.4--C.sub.6H.sub.5 or
(CH.sub.2).sub.n-p-C.sub.6H.sub.4OC.sub.6H.sub.5 and n is 0 to 2;
R.sup.4 is hydrogen, Me, i-C.sub.4H.sub.9 or n-C.sub.4H.sub.9;
R.sup.5 is hydrogen or Me; R.sup.6 is OR.sup.7 or NR.sup.7R.sup.8
wherein R.sup.4 and R.sup.8 are selected independently from the
group consisting of hydrogen, Me, Et or CH.sub.2Ph.
3. A compound according to claim 1 wherein: R.sup.1 is --NHOH or
--OH; R.sup.2 is H or CH.sub.3; R.sup.3 and R.sup.4 are selected
independently from the group consisting of the side chains of
naturally occurring .alpha.-amino acids; R.sup.5 is hydrogen,
C.sub.1-6 alkyl; R.sup.6 is OR.sup.7 or NR.sup.7R.sup.8 wherein
R.sup.7 and R.sup.8 are selected independently from the group
consisting of hydrogen, Me, Et or CH.sub.2Ph.
4. A compound according to claim 1 wherein: R.sup.1 is --NHOH or
--OH; R.sup.2is H or CH.sub.3; R.sup.3 and R.sup.4 are selected
independently from the group consisting of the side chains of
naturally occurring hydrophobic .alpha.-amino acids; R.sup.5 is
hydrogen, C.sub.1-6 alkyl; R.sup.6 is OR.sup.7 or NR.sup.7R.sup.8
wherein R.sup.7 and R.sup.8 are selected independently from the
group consisting of hydrogen, Me, Et or CH.sub.2Ph.
5. A compound according to claim 1 wherein: R.sup.1 is NHOH;
R.sup.2 is H; R.sup.3 and R.sup.4 are i-C.sub.3H.sub.7;
i-C.sub.4H.sub.9, s-C.sub.4H.sub.9, CH.sub.2C.sub.6H.sub.5,
CH.sub.2C.sub.6H.sub.4-p-OMe, (3-indolinyl)methyl; R.sup.5 is
hydrogen, CH.sub.3; R.sup.6 is OR.sup.7; R.sup.7 is OMe or OEt.
6. A method of inhibiting pathological changes mediated by elevated
levels of matrix metalloproteases in mammals comprising
administration to a mammal in need thereof a therapeutically
effective amount of a matrix metalloprotease inhibiting compound
according to claim 1.
7. A method of treating an inflammatory disorder comprising
administration to a mammal in need thereof a therapeutically
effective amount of a matrix metalloprotease inhibiting compound
according to claim 1
8. A method according to claim 6 wherein the condition treated is
of treating is osteoarthritis, rheumatoid arthritis, septic
arthritis, periodontal disease, gingivitis, solid tumor growth and
tumor invasion by secondary metastasis, corneal ulceration, dermal
ulceration, epidermolysis bullosa, neural degeneration, multiple
sclerosis and surgical wound healing.
9. A pharmaceutically composition comprising a pharmaceutical
carrier and a therapeutically effective amount of a matrix
metalloprotease inhibiting compound according to claim 1.
Description
CROSS-REFERENCE TO PRIOR APPLICATIONS
[0001] This application claims the benefit of the following
provisional application: U.S. Ser. No. 60/238,735 filed Oct. 6,
2000.
SUMMARY OF THE INVENTION
[0003] Most tissues exist in a highly regulated dynamic equilibrium
wherein new tissue is formed and existing tissue is degraded and
eliminated. The degradation of the extracellular matrix (ECM),
including connective tissue and basement membranes, is effected by
the metalloproteinases which are released from connective tissue
and invading inflammatory cells. There are at least for distinct
groups of the more than 20 matrix metalloproteinases (MMP) which
have been identified (Birkedal-Hansen, H. J. Oral Pathol. 1988
17:445; Birkedal-Hansen, H. Curr. Opin. Cell Biol. 1995 7:728;
Emonard, H.; Grimaud, J. A. Cell. Mol. Biol. 1990 36:131; Murphy,
G.; Docherty, A. J. P. Am. J. Respir. Cell Mol. Biol. 1992 7:120;
Baramova, E.; Foidart, J. Cell Biol. Int. 1995 19:239; Borkakoti,
N. Prog. Biophys. Mol. Biol. 1998 70:73; Johnson, L. L., Dyer, R.,
Hupe, D. J. Curr. Opin. Chem. Biol. 1998 2:466; Shapiro, S. D.;
Senior, R. M. Am. J. Respir. Cell Mol. Biol. 1999 20:1100): the
collagenases (interstitial collagenase, MMP-1; PMN collagenase,
MMP-8, collagenase-3, MMP-13), the gelatinases (gelatinase A,
MMP-2, 72 kDa-gelatinase, Type IV collagenase; gelatinase B, MMP-9,
92 kDa-gelatinase, Type IV collagenase) the stromelysins
(Proteoglycanase, MMP-3, stromelysin-1, transin; stromelysin-2,
MMP-10; stromelysin 3, MMP-11) and the membrane type matrix
metalloproteinases (MT-1, MMP-14; MT-2, MMP-15; MT-3, MMP-16 and
MT-4, MMP-17). Excessive unregulated activity of these enzymes can
result in undesirable tissue destruction and their activity is
regulated at the transcription level, by controlled activation of
the latent proenzyme and, after translation, by intracellular
specific inhibitory factors such as TIMP ("Tissue Inhibitors of
MetalloProteinase") or by more general proteinase inhibitors such
as .alpha.2-macroglobulins.
[0004] Inhibitors of MMPs also have been found to inhibit the
release of the pleiotropic proinflammatory cytokine, tumor necrosis
factor alpha which has be associated with the pathogenesis of
numerous inflammatory, autoimmune, and neoplastic diseases. The
protease, TNF.alpha.-Converting Enzyme (TACE), catalyzes the
release of TNF.alpha. from a membrane bound precursor protein.
[0005] The MMPs are a family of related proteolytic enzymes. They
are zinc-binding metalloproteases linked by structural homology and
by proteolytic activity against various components of the ECM while
exhibiting divergent substrate specificity and activities. Calcium
is generally required for maximum activity. They are distinguished
from other metalloproteases by their susceptibility to activation
of the zymogen by thiol-modifying reagents, mercurial compounds,
N-ethylmaleimide and oxidized glutathione, by their inhibition by a
group of endogenous substances known collectively as TIMPs, and by
the presence of a consensus sequence in their propeptide forms. (H.
Nagase, "Matrix Metalloproteinases," chapter 7, pp153-204 in Zinc
Metalloproteases in Health and Disease" N. M. Hooper (ed.), Taylor
and Francis, London (1996)).
[0006] Many pathological conditions are associated with the rapid
unregulated breakdown of extracellular matrix tissue by MMPs. Some
of these conditions include rheumatoid arthritis, osteoarthritis,
septic arthritis, corneal, epidermal or gastric ulceration;
periodontal disease, proteinuria, coronary thrombosis associated
with atherosclerotic plaque rupture and bone disease. The process
of tumor metastasis and angiogenesis also appears to be dependent
on MMP activity. Since the cycle of tissue damage and response is
associated with a worsening of the disease state, limiting
MMP-induce tissue damage due to elevated levels of the proteinases
with specific inhibitors of these proteases is a generally useful
therapeutic approach to many of these debilitating diseases (for a
general review see R C Wahl, et al. Ann. Rep, Med. Chem. 1990
25:175-184; Zask, A.; Levin, J. I.; Killar, L. M., Skotnicki, J. S.
Curr. Pharm. Des. 1996, 2, 624).
[0007] It is an object of the present invention to provide novel
selective, small molecule inhibitors of matrix metalloproteinases
which can be used to modulate the progression of the underlying
diseases and to treat diseases associated with excessive
MMP-induced tissue damage.
BRIEF DESCRIPTION OF THE DRAWINGS
[0008] Specific embodiments of the invention have been chosen for
the purpose of illustration and description but are not intended in
any way to restrict the scope of the invention. These embodiments
are shown in the accompanying drawings wherein:
[0009] FIG. 1 depicts the preparation
2-(trimethylsilyl)ethoxymethyl protected aldehyde 13.
[0010] FIG. 2 depicts the intramolecular cyclization of a
metalloenamine to prepare the bis-pyrrolidone 16.
[0011] FIG. 3 depicts the functional group manipulation required to
convert the bis-pyrrolinone to the ester 18c.
[0012] FIG. 4 depicts the conversion of the ester 18c into a
hydroxamic acid 1a.
[0013] FIG. 5 depicts an alternate protection strategy utilizing
and alloc protecting group in place of the CBZ group and the use of
O-trityl hydroxylamine to prepare the hydroxamic acid 1b.
DETAILED DESCRIPTION OF THE INVENTION
[0014] The invention encompasses novel polypyrrolidone compounds of
formula 1 which are useful inhibitors of matrix metalloproteinases
associated with inflammatory neoplastic and degenerative diseases
and/or inhibitors of TNF.alpha. activity associated with
inflammatory, autoimmune and neoplastic diseases.
[0015] Novel compounds of the present invention are of general
formula 1: 2
[0016] wherein:
[0017] R.sup.1 is --NHOH or --OH;
[0018] R.sup.2 is hydrogen, C.sub.1-6 alkyl;
[0019] R.sup.3 and R4 are selected independently from a group
consisting of the side chains of naturally occurring .alpha.-amino
acids, C.sub.1-6 alkyl, (CH.sub.2).sub.nAr wherein the aryl group
is optionally substituted with up to two groups independently
selected from the group consisting of phenyl, hydroxy, C.sub.1-4
alkoxy, phenoxy, --O(CH.sub.2).sub.mOH, C.sub.1-4 thioalkyl,
halogens, nitro, cyano, C.sub.1-4 alkylsulfonyl, and C.sub.1-4
alkylsulfinyl wherein m is 1 or 2 and n is 0 to 3;
[0020] R.sup.5 is hydrogen, C.sub.1-6 alkyl;
[0021] R.sup.6 is OR.sup.7, NR.sup.7R.sup.8 wherein R.sup.7 and
R.sup.8 taken independently are selected from a group consisting of
hydrogen, C.sub.1-4 alkyl, C.sub.1-4 branched alkyl, alkyl aryl and
benzyl; and,
[0022] a stereoisomer, enantiomer, diastereomer, hydrate or
pharmaceutically acceptable salt thereof.
[0023] Another embodiment of the invention is a compound of formula
1 wherein:
[0024] R.sup.1 is --NHOH or --OH
[0025] R.sup.2 is H or CH.sub.3;
[0026] R.sup.3 is n-C.sub.1-6 alkyl, s-C.sub.4H.sub.9,
i-C.sub.4H.sub.9, (CH.sub.2).sub.nAr,
(CH.sub.2).sub.n-p-C.sub.6H.sub.4OMe,
(CH.sub.2).sub.n-p-C.sub.6H.sub.4-C.sub.6H.sub.5 or
(CH.sub.2).sub.n-p-C.sub.6H.sub.4OC.sub.6H.sub.5 and n is 0 to
2;
[0027] R.sup.4 is hydrogen, Me, i-C.sub.4H.sub.9 or
n-C.sub.4H.sub.9;
[0028] R.sup.5 is hydrogen, Me;
[0029] R.sup.6 is OR.sup.7 or NR.sup.7R.sup.8 wherein R.sup.7 and
R.sup.8 are selected independently from the group consisting of
hydrogen, Me, Et or CH.sub.2Ph.
[0030] Yet another embodiment is a compound according to formula 1
wherein:
[0031] R.sup.1 is --NHOH or --OH;
[0032] R.sup.2 is H or CH.sub.3;
[0033] R.sup.3 and R.sup.4 are selected independently from the
group consisting of the side chains of naturally occurring
.alpha.-amino acids;
[0034] R.sup.5 is hydrogen, C.sub.1-6 alkyl;
[0035] R.sup.6 is OR.sup.7 or NR.sup.7R.sup.8 wherein R.sup.7 and
R.sup.8 are selected independently from the group consisting of
hydrogen, Me, Et or CH.sub.2Ph.
[0036] A further embodiment is a compound according to formula 1
wherein:
[0037] R.sup.1 is --NHOH or --OH;
[0038] R.sup.2 is H or CH.sub.3;
[0039] R.sup.3 and R.sup.4 are selected independently from the
group consisting of the side chains of naturally occurring
hydrophobic .alpha.-amino acids;
[0040] R.sup.5 is hydrogen, C.sub.1-6 alkyl;
[0041] R.sup.6 is OR.sup.7 or NR.sup.7R.sup.8 wherein R.sup.7 and
R.sup.8 are selected independently from the group consisting of
hydrogen, Me, Et or CH.sub.2Ph.
[0042] An alternate embodiment is a compound according to formula 1
wherein:
[0043] R .sup.1 is --NHOH
[0044] R.sup.2 is H;
[0045] R.sup.3 and R.sup.4 are i-C.sub.3H.sub.7; i-C.sub.4H.sub.9,
s-C.sub.4H.sub.9, CH.sub.2C.sub.6H.sub.5,
CH.sub.2C.sub.6H.sub.4-p-OMe, (3-indolinyl)methyl;
[0046] R.sup.5 is hydrogen, CH.sub.3;
[0047] R.sup.6 is OR.sup.7; and,
[0048] R.sup.7 is Me or Et;
[0049] Yet another embodiment is a method of inhibiting
pathological changes mediated by elevated levels of matrix
metalloproteases in mammals comprising administration to a mammal
in need thereof a therapeutically effective amount of a matrix
metalloprotease inhibiting compound according to formula 1.
[0050] Still another embodiment of the present invention is a
method of treating an inflammatory disorder comprising
administration to a mammal in need thereof a therapeutically
effective amount of a matrix metalloprotease inhibiting compound
according to formula 1.
[0051] Yet another embodiment is a method for treating a condition
mediated by elevated MMP levels with a compound according to
formula 1 wherein the condition treated is of treating is
osteoarthritis, rheumatoid arthritis, septic arthritis, periodontal
disease, gingivitis, solid tumor growth and tumor invasion by
secondary metastasis, corneal ulceration, dermal ulceration,
epidermolysis bullosa, neural degeneration, multiple sclerosis and
surgical wound healing.
[0052] Another embodiment of the current invention is a
pharmaceutically composition comprising a pharmaceutical carrier
and a therapeutically effective amount of a matrix metalloprotease
inhibiting compound according to formula 1.
[0053] The phrase "a" or "an" entity as used herein refers to one
or more of that entity; for example, a compound refers to one or
more compounds or at least one compound. As such, the terms "a" (or
"an"), "one or more", and "at least one" can be used
interchangeably herein.
[0054] The terms "comprising", "including" and "having" are used
interchangeably. Furthermore, a compound "selected from the group
consisting of" refers to one or more of the compounds in the list
that follows, including mixtures (i.e., combinations) of two or
more of the compounds.
[0055] The compounds of this invention may contain one or more
asymmetric centers and may thus give rise to optical isomers and
diastereomers. Carbons marked with an asterisk in formula 1 are
asymmetric or can be asymmetric when R.sup.2 and/or R.sup.5
substituent is other than hydrogen. Furthermore, the substituents
R.sup.2-R.sup.6 can contain additional asymmetric carbon atoms. The
present invention includes all such optical isomers and
diastereomers; as well as the racemic and resolved,
enantiomerically pure R and S stereoisomers; as well as other
mixtures of the R and S stereoisomers and pharmaceutically
acceptable salts thereof. It is recognized that one optical isomer,
including diastereomer and enantiomer, or stereoisomer may have
favorable properties over the other. Thus when disclosing and
claiming the invention, when one racemic mixture is disclosed, it
is clearly contemplated that both optical isomers, including
diastereomers and enantiomers, or stereoisomers substantially free
of the other are disclosed and claimed as well.
[0056] For the purpose of the present invention, the carbon content
of various hydrocarbon containing moieties is indicated by a prefix
designating the minimum and maximum number of carbon atoms in the
moiety, i.e., the prefix C.sub.i-j defines the number of carbon
atoms present from the integer "i" to the integer "i",
inclusive.
[0057] As used herein, the term "alkyl" refers to a straight or
branched chain alkyl moiety having from one to six carbon atoms,
including for example, methyl, ethyl, propyl, iso-propyl, butyl,
tert-butyl, pentyl, hexyl and the like.
[0058] As used herein, the term "alkoxy" refers to a straight chain
or branched chain alkoxy group containing a maximum of six carbon
atoms, such as methoxy, ethoxy, propoxy, iso-propoxy, butoxy,
tert-butoxy and the like.
[0059] As used herein, the term "aryl" means an optionally
substituted phenyl or naphthyl group with the substituent(s) being
selected, for example, from halogen, trifluoromethyl, C.sub.1-6
alkyl, C.sub.1-6 alkoxy, phenyl and the like.
[0060] The term "aralkyl" as used herein refers to a optionally
substituted alkylenephenyl group, wherein alkyl is lower alkyl and
preferably from 1 to 3 carbon atoms, and aryl is as previously
defined.
[0061] As used herein, the term "halogen" means bromine, chlorine,
fluorine, and iodine.
[0062] As used herein, the term "naturally occurring amino acids"
means the L-isomers of the naturally occurring amino acids. The
naturally occurring amino acids are glycine, alanine, valine,
leucine, isoleucine, serine, methionine, threonine, phenylalanine,
tyrosine, tryptophan, cysteine, proline, histidine, aspartic acid,
asparagine, glutamic acid, glutamine, .gamma.-carboxyglutamic acid,
arginine, ornithine and lysine.
[0063] As used here, the term "side chain of an .alpha.-amino acid"
refers to the substituents on the alpha carbon of the natural amino
acids and include: hydrogen, methyl, i-propyl, i-bu, s-bu,
--CH.sub.2OH, --CH(OH)CH.sub.3, --CH.sub.2SH,
--CH.sub.2CH.sub.2SMe, --(CH.sub.2).sub.pCOR wherein R is --OH or
--NH.sub.2 and p is 1 or 2, --(CH.sub.2).sub.q--NH.sub.2 where q is
3 or 4, --(CH.sub.2).sub.3--NHC(.- dbd.NH)NH.sub.2,
--CH.sub.2C.sub.6H.sub.5, --CH.sub.2-p-C.sub.6H.sub.4--OH- ,
(3-indolinyl)methylene, (4-imidazolyl)methylene.
[0064] As used herein, the term "hydrophobic amino acid" means any
amino acid having an uncharged, nonpolar side chain that is
relatively insoluble in water. Examples of naturally occurring
hydrophobic amino acids are alanine, leucine, isoleucine, valine,
phenylalanine, tryptophan, tyrosine and methionine.
[0065] Those skilled in the art will recognize that certain
reactions are best carried out when other potentially reactive
functionality on the molecule is masked or protected, thus avoiding
undesirable side reactions and/or increasing the yield of the
reaction. The terms "protected amino" and "protected carboxy" mean
amino and carboxy groups which are protected in a manner familiar
to those skilled in the art. Examples of these protecting group
moieties may be found in T. W. Greene, P. G. M. Wuts "Protective
Groups in Organic Synthesis", 2nd Edition, 1991, Wiley & Sons,
New York. Reactive side chain functionalities on amino acid
starting materials are preferably protected. The need and choice of
protecting groups for a particular reaction is known to those
skilled in the art and depends on the nature of the functional
group to be protected (hydroxy, amino, carboxy, etc.), the
structure and stability of the molecule of which the substituent is
part and the reaction conditions. For example, an amino group can
be protected by a benzyloxycarbonyl, tert-butoxycarbonyl, acetyl or
like groups, or in the form of a phthalimido or like group.
[0066] Pharmaceutically acceptable salts can be formed from organic
and inorganic acids, for example, acetic, propionic, lactic,
citric, tartaric, succinic, fumaric, maleic, malonic, mandelic,
malic, phthalic, hydrochloric, hydrobromic, phosphoric, nitric,
sulfuric, methanesulfonic, naphthalenesulfonic, benzenesulfonic,
toluenesulfonic, camphorsulfonic, and similarly known acceptable
acids when a compound of this invention contains a basic moiety.
Salts may also be formed from organic and inorganic bases,
preferably alkali metal salts, for example, sodium, lithium, or
potassium, when a compound of this invention contains an acidic
moiety.
1 The following abbreviations have been used in this application:
Bn benzyl cbz benzyloxycarbonyl CI chemical ionization EDCl
1-ethyl-3(3'-dimethylaminopropyl)carbodiimide hydrochloride ESI
electrospray ionization HOBt 1-hydroxybenzotriazole hydrate KHMDS
potassium hexamethyldisilazane MMP Matrix Metalloprotease SEM
2-(trimethylsilyl)ethoxymethyl SEMCI 2-(trimethylsilyl)ethoxymethyl
chloride TFA trifluoroacetic acid THF tetrahydrofuran TIMP Tissue
Inhibitors of Metalloproteinase Ts p-toluenesulfonyl
[0067] The role of the MMPs in a variety of serious and
debilitating diseases has prompted attempts to identify potent and
selective inhibitors of individual members of this class of
proteases. Proteases have been excellent model systems for rational
drug design studies and these efforts have provided numerous
approaches which have been adapted to these enzymes. Naturally
occurring polypeptides which are substrates or inhibitors of this
class of proteases provide a natural starting point for chemical
design and modification. The 2.2 .ANG. resolution X-ray structure
of potent peptidyl MMP inhibitor Ro-31-4724 (Johnson, W. H., et al.
J. Enzyme Inhib. 1987 2:1), (3) co-crystallized with MMP-1
(Borkakoti, N., et al. Struct. Biol. 1994 1:106) provides useful
insight from which non-peptide analogs can be envisioned. The
well-established difficulties associated with the use and
administration of labile peptides or large polypeptides as
therapeutic agents, especially for long term treatment of chronic
diseases, have made the identification of stable, easily
administered peptide mimics highly desireable. Some of these
classes of compounds now include, thiols (4) phosphorus
(phosphonamides, phosphonates and phosphinates) (5) hydroxamates
(6) and carboxylates (7) peptide derivatives. (R. C. Wahl and R. P.
Dunlap, Biochemistry and Inhibition of Collagenase and Stromelysin,
Ann. Rep. Med. Chem. (1990) 25:177-184; J. B. Summers and S. K.
Davidson, Matrix Metalloproteinases and Cancer, Ann. Rep. Med.
Chem. (1998) 33:131-140; R. Babine and S. Bender, Chem. Rev. (1997)
97:1359).
2 (3) 3 4 X R.sup.1/R.sup.2 R.sup.3 (4) CHSH amino acid aryl
sidechains alkyl (5) PO.sub.2 alkyl alkoxy 5 X R.sup.1/R.sup.2 (6)
NHOH amino acid (7) CO.sub.2H sidechains
[0068] Despite the continuing efforts to identify specific and
potent inhibitors of these potentially pathological mediators,
there remains a need to identify MMP inhibitors, particularly low
molecular weight inhibitors, with sufficient selectivity, potency
and bioavailability to be useful clinical candidates.
Peptidomimetics, non-peptides that mimic the structure of
endogenous peptides, are stable to physiologic conditions and are
bioavailable after oral administration. Although a variety of
scaffolds have been identified which mimic secondary conformations
of proteins and polypeptides, the enormous variety of conformations
found in nature affords a continuing need to identify useful
templates to mimic polypeptides. 6
[0069] The 3,5,5-trisubstituted pyrrolin-4-one ring system, (8) has
proven to be a versatile template for the design of peptidomimetics
and polypyrrolinones (9) have been shown to be effective surrogates
for polypeptides. Depending on their structure, polypyrrolinones,
which are stable to both strong acid and proteases, can adopt
diverse conformations including those analogous to .beta.-strands
(Smith, A. B., III et al., J. Am. Chem. Soc. 1992, 114, 10672;
Smith, A. B., Ell et al., J. Am. Chem. Soc. 1994, 116, 9947),
.beta.-turns and helices (Smith, A. B., III et al., Bioorg. Med.
Chem. 1999, 9). The .beta.-strand structural motif was successfully
utilized in the design and synthesis of several potent,
bioavailable inhibitors of the HIV-1 aspartic acid protease which
exhibited improved membrane transport properties relative to their
peptidal counterparts. (Smith, A. B., III et al., J. Med. Chem.
1994, 37, 215; Smith, A. B., III, et al., J. Am. Chem. Soc. 1995,
117, 11113; Smith, A. B., III et al., J. Med. Chem. 1997, 40, 2440;
Thompson, W. J., et al., J. Med. Chem. 1992, 35, 1685.) The
improved transport was attributed to the presence of intramolecular
hydrogen bonds between adjacent pyrrolinone rings (NH and CO),
which led to a reduction in desolvation energy upon membrane
transport (Hirschmann, R., et al. In New Perspectives in Drug
Design; Dean, P. M., Jolles, G., Newton, C. G., Eds.; Academic:
London, 1995; pp 1-14.). A bis-pyrrolinone was successfully used in
the construction of a pyrrolinone-peptide hybrid ligand, which
bound the Class II MHC protein HLA-DR1 in an extended
.beta.-strand-like conformation with similar potency to the native
peptide. (Smith, A. B., III, et al. J. Am. Chem. Soc. 1998, 120,
12704; Smith, A. B., III; et al. J. Am. Chem. Soc. 1999, 121,
9286.) All references cited in this application are hereby
incorporated into this application in their entirety.
[0070] The 3,5-linked (nitrogen displaced) pyrrolinone scaffold
directly substitutes on a per residue basis for R-amino acids
(except proline and glycine). (Smith, A. B. III, et al. Biopolymers
(Peptide Science) 1995 37:29) Importantly, this structural motif
derived from the D-amino acids maintains both the spacial
orientation of the amino acid side chains and the capacity to form
intermolecular hydrogen bonds with the receptor or enzyme. The
advantage of nonpeptidyl peptidomimetics, in general, is their
ability to resist degradation by proteases and to possess
additional favorable pharmacokinetic properties as a result of
reduced solvation.
[0071] Compounds of the present invention are available from a
protocol exploiting the intramolecular cyclization of a
metalloenamine derived from an (.alpha.-amino acid derivative.
Those skilled in the art will recognize that the nature and order
of the synthetic steps presented may be varied for the purpose of
optimizing the formation of the compounds of the invention.
[0072] The synthesis of aldehyde 13 (FIG. 1) began with Evans
alkylation of (S)-propionyloxazolidinone (+)-10 with prenyl bromide
to furnish the oxazolidinone (+)-11 in 64% yield (>98% ee).
Reduction with lithium borohydride in wet THF (Penning; T. D. et
al. Synth. Comm. 1990 20:307) followed by protection of the
hydroxyl with 2-(trimethylsilyl)ethoxymethy- l chloride (SEM-Cl)
led to the SEM ether, which was subjected to ozonolysis to furnish
aldehyde (+)-13; the overall yield from (+)-10 was 39%.
[0073] To construct monopyrrolinone (+)-15, amino ester (-)-14a was
condensed with aldehyde (+)-13 (FIG. 2); dehydration and subsequent
base-promoted pyrrolinone ring formation with KHMDS furnished
(+)-15 in 93% yield (two steps). Hydrolysis of the dimethylacetal
with TsOH at 40.degree. C. led to the corresponding aldehyde in
nearly quantitative yield. A second pyrrolinone ring was
constructed using amino ester (-)-14a which led to bis-pyrrolinone
(-)-16a. The efficiency of our iterative pyrrolinone construction
protocol was clearly demonstrated by the 77% overall yield of
(-)-16a from (+)-13. Hydrolysis of the dimethylacetal in (-)-16a
was next achieved with 1 N HCl at 40.degree. C. Unfortunately,
oxidation of the derived aldehyde to the corresponding carboxylic
acid by under a variety of different conditions (e.g., Jones,
sodium chlorite, PCC, etc.) proceeded only in low yield. Careful
examination of the product mixture revealed that the pyrrolinone
rings were not stable to the oxidation conditions. To circumvent
this problem, (-)-16a was protected as the bis-Cbz derivative
(+)-17c (FIG. 3); although this operation led to a less reactive
pyrrolinone ring, the acetal proved resilient to hydrolysis. The
problem was solved by first removing the SEM group. Treatment of
(+)-17c with TsOH and methanol at 40.degree. C. furnished alcohol
(+)-18a. A two-step oxidation with Dess-Martin periodinane (Dess,
D. B. and Martin, J. C. J. Org. Chem. 1983 48:4155; Dess, D. B. and
Martin, J. C. J. Am. Chem. Soc. 1991 113:7277; Ireland and R. E.;
Liu, L. J. Org. Chem. 1993 58:2899) and then with sodium chlorite
produced the acid 18b in 81% yield. Esterification followed by
removal of the acetal (TsOH in wet THF at 40.degree. C.) furnished
the intermediate bis-pyrrolinone aldehyde; immediate oxidation with
sodium chlorite led to (+)-18c. Completion of the synthesis was
achieved via coupling (+)-19b with O-benzyl hydroxylamine (EDCI and
HOBt), followed by hydrogenolysis with Pd/BaSO.sub.4 (Nikam, S. S.
Tetrahedron Lett. 1995 36:197-200). The overall yield of (-)-1a for
the two steps was 51%. The corresponding carboxylic acid 19d was
prepared by hydrogenolysis of 19b.
[0074] Those skilled in the art will appreciate that a variety of
protecting groups and reagents can be employed. The hydroxamic acid
1b was prepared by a similar scheme (FIG. 5). The base catalyzed
intramolecular cyclization was carried out with 14b which yielded
the bis-pyrrolidone 16b. In this case the allyloxycarbonyl (Alloc)
protecting group was used in place of the cbz group and 16b was
protected as the bis-alloc derivative 19a which was converted to
19b. The alloc protecting group was removed with
tetrakis-triphenylphosphine palladium(0) and dimedone. In this case
the hydroxamic acid was introduced as the O-trityl derivative 20b
which was deprotected with TFA to yield 1b.
[0075] Compounds of the present invention were assayed for
metalloprotease activity by a published procedure (see, for
example, J. Duan, WO/0059285). The bis-pyrrolinone (-)-1a exhibited
inhibitory activity against gelatinase (MMP-2), matrilysin (MMP-7),
and the membrane type 2 matrix metalloprotease (MMP-15) with
K.sub.i values of 2.9, 6.4 and 6.8 5 .mu.M, respectively. The
bis-pyrrolinone carboxylic acid (-)-19d, on the other hand, failed
to inhibit the ten proteases assayed, a result presumably of the
shorter overall chain length compared to (-)-1a and/or the known
reduced affinity of the carboxylate for zinc(II) compared to the
hydroxamate functionality. (Whittaker, M. et al. Chem. Rev. 1999
99:2735; Borkakoti, N. et al. Struct. Biol. 1994 1:106)
[0076] Compounds of this invention may be administered neat or with
a pharmaceutical carrier to a patient in need thereof. The
pharmaceutical carrier may be solid or liquid.
[0077] Applicable solid carriers can include one or more substances
which may also act as flavoring agents, lubricants, solubilizers,
suspending agents, fillers, glidants, compression aids, binders or
tablet-disintegrating agents or an encapsulating material. In
powders, the carrier is a finely divided solid which is in
admixture with the finely divided active ingredient. In tablets,
the active ingredient is mixed with a carrier having the necessary
compression properties in suitable proportions and compacted in the
shape and size desired. The powders and tablets preferably contain
up to 99% of the active ingredient Suitable solid carriers include,
for example, calcium phosphate, magnesium stearate, talc, sugars,
lactose, dextrin, starch, gelatin, cellulose, methyl cellulose,
sodium carboxymethyl cellulose, polyvinylpyrrolidine, low melting
waxes and ion exchange resins.
[0078] Liquid carriers may be used in preparing solutions,
suspensions, emulsions, syrups and elixirs. The active ingredient
of this invention can be dissolved or suspended in a
pharmaceutically acceptable liquid carrier such as water, an
organic solvent, a mixture of both or pharmaceutically acceptable
oils or fat. The liquid carrier can contain other suitable
pharmaceutical additives such a solubilizers, emulsifiers, buffers,
preservatives, sweeteners, flavoring agents, suspending agents,
thickening agents, colors, viscosity regulators, stabilizers or
osmo-regulators. Suitable examples of liquid carriers for oral and
parenteral administration include water (particularly containing
additives as above, e.g., cellulose derivatives, preferable sodium
carboxymethyl cellulose solution), alcohols (including monohydric
alcohols and polyhydric alcohols, e.g., glycols) and their
derivatives, and oils (e.g., fractionated coconut oil and arachis
oil). For parenteral administration the carrier can also be an oily
ester such as ethyl oleate and isopropyl myristate. Sterile liquid
carriers are used in sterile liquid form compositions for
parenteral administration.
[0079] Liquid pharmaceutical compositions which are sterile
solutions or suspensions can be utilized by, for example,
intramuscular, intraperitoneal or subcutaneous injection. Sterile
solutions can also be administered intravenously. Oral
administration may be either liquid or solid composition form.
[0080] The compounds of this invention may be administered rectally
in the form of a conventional suppository. For administration by
intranasal or intrabronchial inhalation or insufflation, the
compounds of this invention may be formulated into an aqueous or
partially aqueous solution, which can then be utilized in the form
of an aerosol. The compounds of this invention may also be
administered transdermally through the use of a transdermal patch
containing the active compound and a carrier that is inert to the
active compound, is non-toxic to the skin, and allows delivery of
the agent for systemic absorption into the blood stream via the
skin. The carrier may take any number of forms such as creams and
ointments, pastes, gels, and occlusive devices. The creams and
ointments may be viscous liquid or semi-solid emulsions of either
the oil in water or water in oil type. Pastes comprised of
absorptive powders dispersed in petroleum or hydrophilic petroleum
containing the active ingredient may also be suitable. A variety of
occlusive devices may be used to release the active ingredient into
the blood stream such as a semipermeable membrane covering a
reservoir containing the active ingredient with or without a
carrier, or a matrix containing the active ingredient. Other
occlusive devices are known in the literature.
[0081] The dosage to be used in the treatment of a specific patient
suffering a MMP dependent condition must be subjectively determined
by the attending physician. 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, rate of excretion,
drug combination and the severity of the particular disease
undergoing therapy. Treatment will generally be initiated with
small dosages less than the optimum dose of the compound.
Thereafter the dosage is increased until the optimum effect under
the circumstances is reached. Precise dosages for oral, parenteral,
nasal, or intrabronchial administration will be determined by the
administering physician based on experience with the individual
subject treated and standard medical principles.
[0082] General Experimental Procedures
[0083] All reactions were carried out in oven-dried or flame-dried
glassware under an argon atmosphere, unless otherwise noted. All
solvents were reagent or high performance liquid chromatography
grade. Tetrahydrofuran (THF) was freshly distilled from
sodium/benzophenone under argon prior to use unless otherwise
noted. Triethylamine and diisopropylethylamine were distilled from
calcium hydride and stored over potassium hydroxide. Anhydrous
dimethylformamide was purchased from Aldrich and used without
purification. n-Butyllithium was purchased from Aldrich and
standardized by titration with sec-butyl alcohol. All reactions
were magnetically stirred and monitored by thin layer
chromatography using 0.25 mm E. Merck pre-coated silica gel plates.
Flash column chromatography was performed with the indicated
solvents using silica gel-60 (particle size 0.040-0.062 mm)
supplied by E. Merck. Yields refer to chromatographically and
spectroscopically pure compounds, unless otherwise stated. The IR
and NMR spectra were obtained for CHCl.sub.3 and CDCl.sub.3
solutions respectively unless otherwise noted. Infrared spectra
were recorded with a Perkin-Elmer 1600 series FTIR spectrometer.
Proton and carbon-13 NMR spectra were recorded on a Bruker AM-500
spectrometer and obtained at 305 K. Chemical shifts are reported
relative to chloroform (.delta.7.26 for proton and .delta.77.0 for
carbon-13). Optical rotations were obtained with a Perkin-Elmer
model 341 polarimeter in the solvent indicated. High-resolution
mass spectra were obtained at the University of Pennsylvania Mass
Spectrometry Service Center on a Micromass (UK) AutoSpec
spectrometer in electrospray or chemical ionization mode.
EXAMPLE 1
Oxazolidinone (+)-11
[0084] To a solution of (S)-propionyl-oxazolidinone (10; 9.34 mmol)
in THF (125 mL) at -78.degree. C. was added 1.0 M NaHMDS in THF (41
mL, 41 mmol) over 1 h. The resulting solution was stirred for 15
min and then freshly distilled prenyl bromide (11.83 mL, 103 mmol)
was added dropwise via syringe over 30 min. The clear yellow
solution was stirred for 15 min at -78.degree. C. and then warmed
to 0.degree. C. and stirred 45 min, where upon the solute became
cloudy. The solution was then poured into 10 % aqueous NaHSO4 (100
mL). The resulting biphasic mixture was extracted with EtOAc
(2.times.100 mL) and the organic phase washed with saturated
NaHCO.sub.3 and brine (100 mL each), dried over MgSO.sub.4 and
concentrated in vacuo. The resulting yellow oil was purified by
flash chromatography using ethyl acetate-hexanes (1:5) as the
eluant to afford the alkylated oxazolidinone (6.6 g, 64% yield,
>98% ee) as a clear colorless oil:
[a].sup.23.sub.D+40.90.degree. (c 1.70, CHCl.sub.3); IR (neat,
film) 3380(w), 2973(s), 2917(s), 1770(s), 1694(s), 1289(s),
1212(s), 1100(s), 1055(s), 1016(s) cm.sup.-1; .sup.1H NMR (500 MHz,
CDCl.sub.3) 07.28 (m, 5 H), 5.19 (t, J=7.4 Hz, 1H),4.70 (m, 1H),
4.17 (m, 2H), 3.82 (sx, J=6.7 Hz, 1H), 3.25 (dd, J=3.3 Hz, 1H),
2.74 J=9.3 Hz, 1H), 2.47 (m, 1H), 2.21 (m, 1H),1.72 (s, 3H), 1.66
(s, 3H), 1.18 (d, J=7.1 Hz, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta.176.88, 152.99, 135.32, 133.77, 129.30, 128.80, 127.18,
121.04, 65.82, 55.10, 37.83, 37.79, 32.28, 25.70, 17.76, 16.32;
high resolution mass spectrum (CI, CH.sub.4) m/z 302.1746
[(M+H)].sup.+, calcd. for C.sub.18H.sub.24NO.sub.3 302.1756.
EXAMPLE 2
Alcohol (+)-12a
[0085] To a solution of (+)-11 (13.9 g, 46 mmol) in Et.sub.2O (700
mL) at 0.degree. C. was added H.sub.2O (2.65 g, 147 mmol) and 2.0 M
LiBH.sub.4 in THF (25 mL, 50 mmol) dropwise over 30 min. The
resulting solution was stirred for 1 h and then warmed to room
temperature. The reaction was quenched with saturated NaHCO.sub.3
(200 mL). The resulting biphasic mixture was extracted with
Et.sub.2O (3.times.125 mL), dried over NaSO.sub.4, and concentrated
in vacuo. The resulting yellow oil was purified by flash
chromatography using Et.sub.2O-hexanes (1:1) as the eluant to
afford 12 (5.1 g, 86% yield) as a volatile clear colorless oil:
[.alpha.].sup.23.sub.D+4.3.degree. (c 1.40, CH.sub.2Cl.sub.2); IR
(CHCl.sub.3) 3626(m), 3450(b), 3009(s), 2965(s), 2929(s), 2877(s),
1672(w), 1377(s), 1028(s) cm.sup.-1; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta.5.16 (m, 1H), 3.52 (m, J=6.0 Hz, 1H), 3.44 (dd,
J=6.3, 6.0 Hz, 1H), 2.07 (m, 1H), 1.88 (m, 1H), 1.71 (s, 3H), 1.69
(m, 1H), 1.62 (s, 3H), 1.37 (s, 1H). 0.92 (d, J=6.7 Hz, 3H);
.sup.13C NMR (125 MHz, CDCl.sub.3) .delta.132.47, 122.56, 67.96.
36.47, 31.80, 25.70, 17.66, 16.48; high resolution mass spectrum
(CI, CH.sub.4) m/z 128.1201 (M).sup.+, calcd. for C.sub.8H.sub.16O
128.1201.
EXAMPLE 3
SEM Ether 12b
[0086] To a solution of (+)-12a (4.81 g, 38 mmol) in
dichloromethane (18 mL) at 0.degree. C. was added (i-Pr).sub.2NEt
(33.10 mL, 190 mmol) dropwise over 15 min. To the resulting
solution was added SEM-Cl (20.18 mL, 114 mmol) dropwise over 15
min. and the resulting solution was stirred for 2 h. The solution
was then poured into 10% aqueous NaHSO.sub.4 (100 mL). The
resulting biphasic mixture was extracted with Et.sub.2O
(3.times.100 mL), dried over MgSO.sub.4 and concentrated in vacuo.
The resulting orange oil was purified by flash chromatography using
ethyl acetate-hexanes (1:5) as the eluant to afford the
corresponding prenyl SEM ether (9.81 g, 99% yield) as a clear
colorless oil: [.alpha.].sup.23.sub.D-1.9.degree. (c 2.59,
CHCl.sub.3); IR (neat film) 2953(s), 2921(s), 1248(s), 1109(s),
1058(s), 1038(s), 859(s), 835(s) cm.sup.-1; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta.5.12 (m, 1H), 4.65 (s, 2H), 3.61 (m, 2H), 3.40
(dd, J=3.6, 6.0 Hz, 1H), 3.30 (m, J=6.7 Hz, 1H), 2.07 (m, 1H), 1.85
(m, 1H), 1.74 (sx, J=6.7 Hz, 1H), 1.69 (s, 3H), 1.59 (s, 3H), 0.93
(m, 2H), 0.91 (d, J=6.7 Hz, 3H), 0.01 (s, 9H); .sup.13C NMR (125
MHz, CDCl.sub.3) .delta.132.38, 122.56, 94.94, 72.86, 64.82, 34.27,
31.98, 25.76, 18.11, 17.74, 16.96, -1.45; high resolution mass
spectrum (CI, NH3) m/z 276.2360 [(M+NH.sub.4)].sup.+, calcd. for
C.sub.14H.sub.30O.sub.2Si.NH.sub.4 276.2359. Anal. Calcd. for
C.sub.14H.sub.30O.sub.2Si: C, 65.06; H,11.70. Found: C, 65.26;
H,11.95.
EXAMPLE 4
Aldehyde (+)-13
[0087] Ozone was bubbled through a solution of (-)-prenyl SEM ether
12b (6.6 g, 26 mmol) in dichloromethane (200 mL) at -78.degree. C.
until a pale blue color persisted. At -78.degree. C., Ph.sub.3P
(6.69 g, 26 mmol) was then added and the reaction mixture allowed
to stir and warm to room temperature overnight. The resulting clear
oil was purified by flash chromatography using EtOAc-hexanes (1:5
then 3:10) as the eluant to afford the aldehyde 13 (4.2 g, 71%
yield) as a clear colorless oil: [.alpha.].sup.23.sub.D+6.6.degree.
(c 1.07, CHCl.sub.3); 1R (neat film) 2954(m), 1732(s), 1713(s),
1416(m), 1250(m), 1057(s), 859(s), 835(s) cm.sup.-1; .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta.9.69 (t, J=2.0 Hz, 1H), 4.56 (m, 2H),
3.53 (m, 2H), 3.40 (m, 1H), 3.31 (m, J=7.1 Hz, 1H), 3.26 (dd,
J=7.5, 7.1 Hz, 1H), 1.00 (m, 5H), 0.86 (m, 2H),-0.05 (s, 9H);
.sup.13C NMR (125MHz, CDCl.sub.3) .delta.202.07, 94.86, 72.08,
65.08, 38.11, 29.00, 18.10, 17.08, -1.45; high resolution mass
spectrum (CI, NH.sub.3) m/z 250.1828 [(M+NH.sub.4)].sup.+, calcd
for C.sub.11H.sub.24O.sub.3Si.NH.sub.4 250.1838.
EXAMPLE 5
Monopyrrolinone (+)-15
[0088] To a solution of (-)-7 (2.0 g, 8.6 mmol) in toluene (60 mL)
was added to (+)-6 (2.4 g, 8.6 mmol). The solution was stirred for
15 min; then the solution was concentrated in vacuo and the residue
azeotropically dehydrated with additional toluene (5.times.60 mL).
To a solution of the residue in THF (80 mL) was added 0.5 M KHMDS
In toluene (43 mL, 21 mmol) rapidly via syringe. The resulting
yellow-orange solution was stirred for 20 min and then 10% aqueous
NaHSO.sub.4 (100 mL) was added and diluted with EtOAc (100 mL). The
resulting biphasic mixture was extracted with EtOAc (2.times.100
mL) and washed with saturated NaHCO.sub.3 and brine (100 mL each).
The resultant yellow solution was dried over MgSO.sub.4 and
concentrated in vacuo. The residue was purified by flash
chromatography using methanol-dichloromethane (1:19) as the eluant
to afford the monopyrrolinone (3.3 g, 93% yield) as a yellow oil:
[a].sup.23.sub.D+38.5.degree. (c 1.07, CHCl.sub.3); IR (CHCl.sub.3)
3425(m), 3008(s), 2956(s), 2873(s), 2836(m), 1710(m), 1661(s),
1583(s), 1465(s), 1422(s), 1368(s), 1250(s), 1121(s), 1057(s),
861(s), 837(s) cm.sup.-1; 1H NMR (500 MHz, CDCl.sub.3) .delta.7.77
(d, J=3.7 Hz, 1H), 5.61 (s, 1H), 4.65 (s, 2H), 4.47 (dd, J=4.1, 4.5
Hz, 1H), 3.61 (m, 3H), 3.42 (m, J=6.7 Hz, 1H), 3.35 (s, 3H), 3.28
(s, 3H), 2.81 (sx, J=6.7 Hz, 1H), 1.91 (dd, J=4.1 Hz, 1H), 1.66 (m,
2H), 1.52 (m, 2H), 1.16 (d, J=7.1 Hz 3H), 0.93 (m, 2H), 0.85 (d,
J=6.3 Hz, 3H), 0.81 (d, J=6.7 Hz, 3H), 0.02 (s, 9H); .sup.13C NMR
(125 MHz, CDCl.sub.3) .delta.203.80, 160.54, 115.81, 102.30, 94.96,
71.76, 67.99, 64392, 54.00, 53.14, 43.66, 40.29, 28.56, 24.33,
24.26, 24.13, 18.06, 16.85, -1.452; high resolution mass spectrum
(ESI) m/z 438.2667 [(M+Na)].sup.+, calcd. for
C.sub.21H.sub.41NO.sub.5SiNa 438.2652.
EXAMPLE 6
Monopyrrolinone Aldehyde
[0089] 7
[0090] To a solution of (+)-15 (2.1 g, 5 mmol) in a 3:1 mixture of
THF and water (70 mL) was added p-TsOH hydrate (943 mg, 5 mmol).
The solution was heated at 50.degree. C. for 4 h and was then
cooled to room temperature and diluted with EtOAc (400 mL) and
saturated NaHCO.sub.3 (300 mL). The resulting biphasic mixture was
extracted with EtOAc (2.times.200 mL) and washed with brine (200
mL). The yellow solution was then dried over MgSO.sub.4 and
concentrated in vacuo. The residue was purified by flash
chromatography using methanol-dichloromethane (3:47) as the eluant
to afford the monopyrrolinone aldehyde (1.82 g, 99% yield) as a
yellow oil: [a].sup.23.sub.D-43.8.degree. (c 0.95, CHCl.sub.3); IR
(CHCl.sub.3) 3447(m), 3008(s), 2957(s), 2928(s), 2873(s), 1722(s),
1661(s), 1582(s), 1452(m), 1426(m), 1368(m), 1250(s), 1212(s),
1058(s), 1029(s), 861(s), 837(s) cm.sup.-1; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta.9.60 (d, J=2.6 Hz, 1H), 7.81 (d, J =3.7 Hz, 1H),
5.59 (s, 1H), 4.64 (s,2H), 3.61 (m, 3H), 3.45 (m, J=6.7 Hz, 1H),
2.81 (m, 1H), 2.76 (m, J=2.6 Hz, 1H), 2.56 (d, J=16.8 Hz, 1H). 1.73
(m, J=5.6 Hz, 1H),1.66 (m, J=6.7 Hz, 1H),1.58 (m, 1H), 1.16 (d,
J=6.7 Hz, 3H), 0.93 (m, 2H), 0.88 (d,J=6.7 Hz, 3H), 0.81 (d, J=6.7
Hz, 3H), 0.02 (s, 9H); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta.202.44, 200.10, 160.51, 115.13. 94.88, 71.45, 67.41, 64.92,
50.34, 44.42, 28.56, 24.26, 18.02, 16.82, -1.48; high resolution
mass spectrum (ESI) m/z 392.2224 [(M+Na)].sup.+, calcd for
C.sub.19H.sub.35NO.sub.4SiNa 392.2233.
EXAMPLE 7
bis-pyrrolinone (-)-16a
[0091] To a solution of (-)-14a (1.0 g, 4.5 mmol in toluene (60 mL)
was added to (-)-monopyrrolinone aldehyde (1.8 g, 5 mmol). The
solution was stirred for 15 min to allow formation of the imine
after which the solution was concentrated in vacuo and the residue
azeotropically dried with additional toluene (5.times.30 mL). To a
solution of the residue in THF (40 ml) was added 0.5 M KHMDS in
toluene (45 mL, 22 mmol) rapidly via syringe. The resulting
yellow-orange solution was stirred for 45 min, and then 10 %
aqueous NaHSO.sub.4 (100 mL) was added and diluted with EtOAc (100
mL). The resulting biphasic mixture was extracted with EtOAc
(2.times.100 mL) and the combined organic phase was washed with
saturated NaHCO.sub.3 and brine (100 mL each). The yellow solution
was dried over MgSO.sub.4 and concentrated in vacuo. The residue
was purified by flash chromatography using ethyl acetate as the
eluant to afford the bis-pyrrolinone (2.3 g, 84% yield) as a yellow
oil: [a].sup.23.sub.D-97.2.degree. (c 1.21, CHCl.sub.3; IR (neat,
film) 3272(s), 2953(s), 1708(s), 1643(s), 1556(s), 1468(s),
1248(s), 1188(s), 1122(s), 1057(s), 860(s), 836(s) cm.sup.-1;
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta.8.22 (d, J=4.1 Hz, 1H),
7.80 (d, J=3.4 Hz, 1H), 7.03 (d, J=3.4 Hz, 1H), 5.99 (d, J=3.7 Hz,
1H), 4.65 (s, 2H), 4.48 (q, J=3.7, 4.1 Hz, 1H), 3.60 (m, 3H), 3.44
(d, J=7.1 Hz, 1H), 3.42 (d, J=7.4 Hz, 1H), 3.35 (s, 3H), 3.01 (s,
3H), 2.80 (m, 1H), 1.93 (dd, J=3.7, 4.1 Hz, 1H),1.86 (dd, J=4.1,
4.5 Hz, 1H), 1.72 (dd, J=7.5 7.4 Hz, 1H), 1.61 (m, 2H), 1.38 (hp,
J=6.7 Hz, 1H), 1.16 (m, 1 H), 1.12 (d, J=7.1 Hz, 3H), 1.02 (d,
J=6.7 Hz, 1 H), 0.94 (m, 2H), 0.87 (d, J=6.3 Hz, 3H), 0.85 (d,
J=6.3 Hz, 3H), 0.77 (d, J=6.7 Hz, 3H), 0.69 (d, J=6.3 Hz, 3H), 0.01
(s, 9H); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.203.02, 202.95,
161.48, 160.89, 114.82, 110.28, 102.00, 94.93, 71.80, 68.72, 67.78,
64.88, 53.91, 53.23, 47.62, 43.72, 39.88, 28.45, 24.79, 24.47,
24.29, 24.05, 23.72, 18.06, 16.92, -1.44; high resolution mass
spectrum (ESI) m/z 575.3475 [(M+Na)].sup.+, calcd for
C.sub.29H.sub.52N.sub.2O.sub.6SiNa 575.3492.
EXAMPLE 8
bis-CBZ Protected bis-pyrrolinone (+)-17c
[0092] To a solution of (-)-17a (2.1 g, 3.7 mmol) in THF (40 mL) at
a -78.degree. C. was added 1.0 M NaHMDS in THF (11 mL, 11 mmol)
dropwise over 30 min. The resulting yellow solution was stirred for
5 min and then benzyl chloroformate (1.6 mL, 11 mmol) was added
dropwise via syringe over 30 min. The yellow solution was stirred
for 15 min at -78.degree. C. and was warmed to room temperature.
The solution was then poured into 10% aqueous NaHSO.sub.4 (300 mL).
The resulting biphasic mixture was extracted with EtOAc
(2.times.200 mL) and the organic phase washed with saturated
NaHCO.sub.3 and brine (200 mL each), dried over MgSO.sub.4 and
concentrated in vacuo. The resulting yellow oil was purified by
flash chromatography using EtOAc-hexanes (3:7) as the eluant to
afford the bis-benzyloxycarbonyl-bis-pyrrolinone (2.5 g, 80%,
yield) as a yellow oil: [.alpha.].sup.23.sub.D+47.90.degree. (c
1.07, CHCl.sub.3); IR (neat, film) 2955(m), 1698(s), 1614(s),
1402(s), 1278(s), 1057(s), 860(m), 836(m) cm.sup.-1; .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta.8.65 (br s), 8.41 (m), 8.25 (s), 8.35
(m), 5.24 (m), 4.60 (s), 4.08 (m), 3.57 (m), 3.50 (d, J=6.3 Hz),
3.48 {d, J=6.7 Hz), 3.23 (br s), 3.13 (br s), 3.04 (m), 2.81 (m),
2.43 (m), 2.11 (m), 1.84 (m), 1.55 (m), 1.42 (br s), 1.34 (br s),
1.17 (m), 0.90 (m), 0.79 (m), 0.70 {m), 0.57 (m), -0.01 (br s);
.sup.13C NMR (125 MHz, CDCl.sub.3) .delta. (complex spectrum due to
rotomers); high resolution mass spectrum (ESI) m/z 843.4225
[(M+Na)].sup.+, calcd for C.sub.45H.sub.64N.sub.2O.sub.10SiNa
843.4228.
EXAMPLE 9
Alcohol (+)-18a
[0093] To a solution of (+)-17c (640 mg, 0.78 mmol) in a 1:1
mixture of THF and methanol (60 mL} was added p-TsOH (445 mg, 2.3
mmol). The solution was heated at 40.degree. C. for 2.5 h and was
then cooled to room temperature and diluted with Et.sub.2O and
saturated NaHCO.sub.3 (200 mL each). The resulting biphasic mixture
was extracted with Et.sub.2O (2.times.100 mL) and washed with brine
(100 mL). The yellow solution was then dried over MgSO.sub.4 and
concentrated in vacuo. The residue was purified by flash
chromatography using EtOAc-hexanes (1:1) as the eluant to afford
the bis-benzyloxycarbonyl-bis-pyrrolinone alcohol (500 mg, 93%
yield) as a yellow oil: [.alpha.].sup.23.sub.D+22.0.degree. (c
1.22, CHCl.sub.3); IR (CHCl.sub.3) 3520(b), 2956(m), 1722(m),
1693(m), 1682(m), 1606(m), 1402(m), 1203(m), 1148(m), 1062 (m)
cm.sup.-1; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.9.13 (s), 8.44
(m), 8.24 (m),7.35 (m), 5.27 (m), 5.06 (m), 4.07 (m), 3.93 (m),
3.73 (m), 3.52 (m), 3.44 (m), 3.18 (br 5), 3.11 (br s), 3.04 (br
s), 2.96 (br s), 2.92 (br s), 2.67 (m), 2.50 (m), 2.16 (m}, 1.90
(m), 1.60 (m), 1.41 (m), 1.23 (m), 1.15 (d, J=7.1 Hz), 0.82 (m),
0.74 (m), 0.66 (m); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
(complex spectrum due to rotomers}: high resolution mass spectum
(ESI) m/z 713.3416 [(M+Na)].sup.+, calcd for
C.sub.39H.sub.50N.sub.2O.sub.9Na 713.3414.
EXAMPLE 10
bis-pyrrolinone acid 18b
[0094] To a solution of (+)-18a (426 mg, 0.62 mmol) in
dichloromethane (8 mL) was added the Dess-Martin periodinane (928
mg, 2.5 mmol). The heterogeneous mixture was stirred under air for
1.5 h. To the mixture was added saturated NaHCO.sub.3 (30 mL),
Na.sub.2S.sub.2O.sub.3 (30 mL), and Et.sub.2O (40 mL). The mixture
was stirred until the Et.sub.2O layer was clear (ca. 30 min). The
resulting biphasic mixture was extracted with Et.sub.2O (3.times.40
mL) and the organic phase washed with saturated NaHCO.sub.3 and
brine (40 mL each), dried over MgSO.sub.4 and concentrated in
vacuo. To a solution of the above residue in t-BuOH (20 mL) was
added 2-methyl-2-butene (1.21 mL, 2.4 mmol), premixed NaClO.sub.2
(164 mg, 1.8 mmol) and Na.sub.2H.sub.2PO.sub.4 (142 mg, 0.9 mmol)
in water (4 mL). The solution was stirred for 2 h and then 10%
aqueous NaHSO.sub.4 (80 mL) and Et.sub.2O (80 mL) was added. The
resulting biphasic mixture was extracted with Et.sub.2O (3.times.80
mL) and dried over NaSO.sub.4 and concentrated in vacuo. The
resulting clear oil was purified by flash chromatography using
EtOAc-HOAc-hexanes (49:1:50) as the eluant to afford the
bis-benzyloxycarbonyl-bis-pyrrolinone acid (350 mg, 81% yield, 2
steps) as a clear oil: [a].sup.23.sub.D+67.40.degree. (c 2.78,
CHCl.sub.3); IR (CHCl.sub.3) 2956(m), 1698(m), 1605(m), 1402(m),
1360(m), 1200(m), 1147(m), 1091(m), 1058(m) cm.sup.-1; .sup.1H
NMR(500 MHz, CDCl.sub.3) .delta.8.69 (brs), 8.58 (brs), 8.43 (brs),
7.35 (m), 5.27 (m), 5.14 (m), 4.09 (m), 3.54 (m), 3.23 (br s), 3.13
(br s), 3.04 (br s), 2.97 (br s), 2.46 (m), 2.23 (m), 2.09 (m),
1.87 (m), 1.56 (br s), 1.42 (m), 1.24 (m), 0.79 (m), 0.70 (br s),
0.59 (m); .sup.13C NMR (125 MHz, CDCI3) .delta. (complex spectrum
due to rotomers); high resolution mass spectrum (ESI) m/z 727.3224
[(M+Na)].sup.+, calcd. for C.sub.39H.sub.48N.sub.2O.sub.10Na
727.3207.
EXAMPLE 11
[0095] Ester (+)-18c
[0096] To a solution of (+)-bis-pyrrolinone acid (64 mg, 0.1 mmol)
in DMF (3 mL) was added diisopropylcarbodiimide (34 mg, 0.3 mmol),
1-hydroxybenzotriazol (37 mg, 0.3 mmol), and DMAP (ca. 1 mg). After
5 min, absolute ethanol (0.02 mL, 0.3 mmol) was added and the
solution stirred for 7 h. The solution was diluted water and
Et.sub.2O (40 mL each), separated, and the Et.sub.2O phase washed
with brine (50 mL). The solution was dried over MgSO.sub.4 and
concentrated in vacuo. The residue was purified by flash
chromatography using EtOAc-hexanes (4:6) as the eluant to afford
the bis-benzyloxycarbonyl-bis-pyrrolinone ethyl ester (39 mg, 58%
yield) as a light yellow oil: [.alpha.].sup.23.sub.D+89.9.deg- ree.
(c 2.00, CHCl.sub.3); IR (CHCl.sub.3) 3019(m), 3012(m), 2961(m),
1726(s), 1608(m), 1404(s), 1224(s), 1060(s) cm.sup.-1; .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta.8.67 (br s), 8.54 (br s), 8.42 (br s),
7.38 (m), 5.26 (m), 4.12 (m, 4H), 4.07 (br s), 3.51 (q, J=7.1 Hz,
1H), 3.45 (m), 3.24 (br s), 3.14 (br s), 3.05 (br s), 2.98 (br s),
2.45 (m), 2.10 (m), 1.86 (m), 1.56 (m), 1.41 (m), 1.26 (br s), 1.21
(t, J=7.1 Hz), 0.80 (m), 0.72 (br s), 0.59 (m); .sup.13C NMR (125
MHz, CDCl.sub.3) .delta. (complex spectrum due to rotomers); high
resolution mass spectrum (ESI) m/z 755.3535 [(M+Na)].sup.+, calcd
for C.sub.41H.sub.52N.sub.2O.sub.10Na 755.3520.
EXAMPLE 12
Acid (+)-19b
[0097] To a solution of (+)-18c (70 mg, 0.1 mmol) in wet THF (3 mL)
was added p-TsOH (184 mg, 1 mmol). The solution was heated at
40.degree. C. for 3 h and was then cooled to room temperature and
diluted with Et.sub.2O (20 mL) and saturated NaHCO.sub.3 (30 mL).
The resulting biphasic mixture was extracted with Et.sub.2O
(3.times.20 mL) and the combine Et.sub.2O phases washed with brine
(20 mL). The solution was dried over MgSO.sub.4 and concentrated in
vacuo. To a solution of the above residue in t-BuOH (3.5 mL) was
added 2-methyl-2-butene (0.19 mL, 0.38 mmol), premixed NaClO.sub.2
(26 mg, 0.3 mmol) and Na.sub.2H.sub.2PO.sub.4 (22 mg, 0.14 mmol) in
water (0.7 mL). The solution was chilled to 0.degree. C. and
stirred for 1.5 h. To the mixture was added 10% aqueous NaHSO.sub.4
(50 mL) and Et.sub.2O (50 mL). The resulting biphasic mixture was
extracted with Et.sub.2O (3.times.40 mL), dried over NaSO.sub.4 and
concentrated in vacuo. The resulting clear oil was purified by
flash chromatography using EtOAc-HOAc-hexanes (49:1:50) as the
eluant to afford the bis-benzyloxycarbonyl-bis-pyrrolino- ne acid
(40 mg, 59% yield for the 2 steps) as a light yellow oil:
[.alpha.].sup.23.sub.D+68.2 (c 1.54, CHCl.sub.3); IR (CHCl.sub.3)
3019(m), 2962(w), 1727(s), 1609(w), 1405(s), 1212(s), 1091(w),
1061(w) cm.sup.-1; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.8.46
(m), 7.36 (m), 5.22 (m), 4.12 (m,), 3.54 (m), 3.20 (br s), 2.74
(m), 2.49 (br s), 2.20 (m), 2.04 (m), 1.79 (brs), 1.58 {brs), 1.40
(m), 1.26 (brs), 1.21 (t, J=7.1 Hz), 0.78 (m), 0.71 {m), 0.69 {m),
0.63 (m); .sup.13C NMR (125 MHz, CDCl.sub.3) 6 (complex spectrum
due to rotomers); high resolution mass spectrum (ESI) m/z 725.3046
[(M+Na)].sup.+, calcd for C.sub.39H.sub.46N.sub.20.sub.10Na
725.3050.
EXAMPLE 13
bis-benzyloxycarbonyl-bis-pyrrolinone O-benzylhydroxyl amide
19c
[0098] To a solution of (+)-19b (25 mg, 0.04 mmol) in
dichloromethane (2 mL) at 0.degree. C. was added EDCl.HCI (10 mg,
0.054 mmol) and 1-hydroxybenzotriazole (7 mg, 0.054 mmol) and
stirred for 30 min. The solution was warmed to room temperature for
1 h then cooled to 0.degree. C. To the solution was added
O-benzylhydroxylamine.HCl (17 mg, 0.108 mmol) and
di-isopropylethyl-amine (0.04 mL, 0.252 mmol) and the resulting
solution was stirred for 4 h. The solution was diluted with water
and Et.sub.2O (30 mL each) ant the Et.sub.2O phase washed with
saturated NaHCO.sub.3 and brine (20 mL each), dried over MgSO.sub.4
and concentrated in vacuo. The residue was purified by flash
chromatography using methanol-dichloromethane (3:47) as the eluant
to afford the bis-benzyloxycarbonyl-pyrrolinone
O-benzylhydroxyl-amide (26 mg, 90% yield) as a light yellow oil:
[.alpha.].sup.23.sub.D+72.6.degree. (c 0.34, CHCl.sub.3); IR
(CHCl.sub.3) 3018(w), 3010(w), 2962(w), 1726(s), 1607(w), 1404(s),
1210(s), 1091(w), 1061(w) cm.sup.-1; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta.8.41 (m, 3H), 7.34 (m), 5.28 (m), 5.08 (m), 4.81
(m), 4.12 (q, J=7.1 Hz, 2H), 3.52 (m, 1H), 2.86 (br s), 2.48 (br
s), 2.16 (br s), 2.00 (brs), 1.88 (m), 1.59 (m), 1.39 (m), 1.26
(brs), 1.21 (t, J=7.1 Hz, 3H), 0.88 (m), 0.78 (m), 0.69 (m), 0.62
(m); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta. (complex spectrum
due to rotomers); high resolution mass spectrum (ESI) m/z 830.3664
[(M+Na)].sup.+, calcd for C.sub.46H.sub.53N.sub.3O.sub.10Na
830.3629.
EXAMPLE 14
Bispyrrolinone Hydroxamic Acid (-)-1a
[0099] To a solution of (+)-bis-benzyloxycarbonyl-bis-pyrrolinone
O-benzylhydroxylamide (26 mg, 0.032 mmol) in ethanol (6 mL) was
added 5% Pd/BaSO.sub.4 (26 mg) and mixture was stirred under a
hydrogen atmosphere (hydrogen filled balloon) for 17 h. The
heterogeneous mixture was filtered through a 0.45 .mu.m filter disc
syringe 1/4 filled with CELITE and then concentrated in vacuo. The
resulting residue was purified by flash chromatography using
isopropyl alcohol-hexanes (3:7) as the eluant to afford the
hydroxamic acid (8 mg, 57% yield) as a light yellow film:
[.alpha.].sup.23.sub.D-172.5.degree. (c 0.80, CHCl.sub.3); IR
(CHCl.sub.3) 3243(w), 3021(w),2959(w), 2929 (w), 2872(w), 1724(w),
1650(m), 1576(m), 1446(w), 1174(w) cm.sup.-1; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. (concentration dependent spectrum) 10.10 (s br,
1H), 8.31 (s, 1H), 8.01 (d, J=3.7 Hz, 1H), 7.51 (s, 1H), 7.27 (s,
1H), 4.12 (q, J=6.9, 7.4 Hz, 2H), 3.49 (q, J=6.9, 7.4 Hz, 1H), 2.54
(d, J=13.9 Hz, 1H), 2.29 (d, J=14.3 Hz, 1H), 1.94 (m, 1H), 1.63 (m,
5H), 1.40 (s,1H), 1.32 (d, J=7.4 Hz, 3H), 1.23 (t, J=6.9 Hz, 3H),
0.86 (d, J=6.0 Hz, 3H), 0.81 (d, J=6.5 Hz, 3H), 0.78 (d, J=6.5 Hz,
3H), 0.66 (d, J=6.5 Hz, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta.202.39, 201.62, 174.95, 167.01, 162.57, 162.04, 111.49,
108.98, 68.93, 68.09, 60.71, 46.45, 43.51, 40.45, 33.79, 24.58,
24.49, 24.29, 24.15, 23.78, 23.48, 17.44, 14.14; high resolution
mass spectrum (ESI) m/z 472.2432 [(M+Na)].sup.+ calcd. for
C.sub.23H.sub.35N.sub.3O.sub.6Na 472.2424.
EXAMPLE 15
bis-pyrrolinone carboxylic acid (-)-19d
[0100] To a solution of (+)-19b (11 mg, 0.016 mmol) in ethanol (3
mL) was added 5% Pd/BaSO.sub.4 (10 mg) and the mixture stirred
under a hydrogen atmosphere (hydrogen filled balloon) for 2 h. The
heterogeneous mixture was filtered through a 0.45 .mu.m filter disc
syringe 1/4 filled with CELITE and concentrated in vacuo. The
resulting residue was purified by flash chromatography using acetic
acid-methanol-dichloromethane (1:10:90) as the eluant to afford the
bis-pyrrolinone carboxylic acid (4 mg. 58 % yield) as a light
yellow film: [.alpha.].sup.23.sub.D-235.1.degree. (c 0.70
CHCl.sub.3); IR (CHCl3) 3436(m), 3026(m), 3018(m), 2958(s),
2932(s), 2872(m), 1723(s), 1648(s), 1576(s), 1448(s), 1368(m),
1168(s) cm.sup.-1; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
(concentration dependent spectrum) 8.30 (d, J=3.3 Hz, 1H), 7.98 (d,
J=2.9 Hz, 1H), 7.12 (s, 1H), 6.72 (s, 1H), 4.12 (q, J=7.5, 7.0 Hz,
2H), 3.50 (q, J=7.1 Hz, 1H), 2.69 (d, J=16.4 Hz, 1H). 2.41 (d,
J=16.8 Hz,. 1H), 1.95 (m, 1H), 1.82 (dd, J=14.1, 4.8 Hz, 1H), 1.63
(m, 4H), 1.42 (m, 1H), 1.31 (d, J=7.1 Hz, 3H), 1.23 (t, J=7.1 Hz,
3H), 0.87 (d, J=6.3 Hz, 3H), 0.82 (d, J=6.3 Hz, 3H), 0.80 (d, J=6.7
Hz, 3H), 0.67 (d, J=6.7 Hz, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta.202.48, 201.62, 174.84, 173.38, 162.04, 161.69, 111.68,
108.89, 68.12, 67.96, 60.72, 46.67, 43.31, 41.23, 33.68, 24.62,
24.42, 24.27, 24.23, 23.79, 23.38, 17.53, 14.14; high resolution
mass spectrum (ESI) m/z 457.2317 [(M+Na)].sup.+, calcd. for
C.sub.23H.sub.34N.sub.2O.sub.6Na 457.2315.
[0101] Thus, while there have been described what are presently
believed to be the preferred embodiments of the invention, those
skilled in the art will realize that other and further embodiments
can be made without departing from the spirit of the invention, and
thus it is intended to include all such further modifications and
changes come within the true scope of the claims as set forth
herein.
* * * * *