U.S. patent application number 14/876634 was filed with the patent office on 2016-01-28 for prodrugs of fluorinated mevalonates to inhibit the mevalonate pathway of streptococcus pneumoniae.
The applicant listed for this patent is Soosung Kang, Thomas S. Leyh, Richard B. Silverman, Mizuki Watanabe. Invention is credited to Soosung Kang, Thomas S. Leyh, Richard B. Silverman, Mizuki Watanabe.
Application Number | 20160024042 14/876634 |
Document ID | / |
Family ID | 49083453 |
Filed Date | 2016-01-28 |
United States Patent
Application |
20160024042 |
Kind Code |
A1 |
Silverman; Richard B. ; et
al. |
January 28, 2016 |
Prodrugs of Fluorinated Mevalonates to Inhibit the Mevalonate
Pathway of Streptococcus pneumoniae
Abstract
Fluorinated prodrug compounds as can be used for selective
streptococcal mevalonate pathway inhibition.
Inventors: |
Silverman; Richard B.;
(Winnetka, IL) ; Watanabe; Mizuki; (Uji, JP)
; Kang; Soosung; (Wilmette, IL) ; Leyh; Thomas
S.; (Katonah, NY) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Silverman; Richard B.
Watanabe; Mizuki
Kang; Soosung
Leyh; Thomas S. |
Winnetka
Uji
Wilmette
Katonah |
IL
IL
NY |
US
JP
US
US |
|
|
Family ID: |
49083453 |
Appl. No.: |
14/876634 |
Filed: |
October 6, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13780929 |
Feb 28, 2013 |
9150534 |
|
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14876634 |
|
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61604261 |
Feb 28, 2012 |
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Current U.S.
Class: |
549/228 ;
549/292; 549/375; 558/276; 560/180; 560/186 |
Current CPC
Class: |
C07D 309/30 20130101;
C07C 69/675 20130101; C07D 319/06 20130101; C07C 235/06
20130101 |
International
Class: |
C07D 319/06 20060101
C07D319/06; C07D 309/30 20060101 C07D309/30; C07C 235/06 20060101
C07C235/06; C07C 69/675 20060101 C07C069/675 |
Goverment Interests
[0002] This invention was made with government support under RO1
AI068989 awarded by the National Institutes of Health. The
government has certain rights in the invention.
Claims
1. A compound of a formula ##STR00073## wherein X is selected from
NH, O and S; R.sub.1 is selected from mono- and
polyfluoro-substituted methyl moieties; and R.sub.2 is selected
from alkyl, mono- and polyfluoro-substituted alkyl, alkanoyloxy,
cycloalkyl, mono- and polyfluoro-substituted cycloalkyl, aryl,
mono- and polyfluoro-substituted aryl, arylalkyl, and mono- and
polyfluoro-substituted arylalkyl moieties; and hydrates and
solvates thereof.
2. The compound of claim 1 wherein X is selected from NH and O; and
R.sub.1 is selected from fluoromethyl and trifluoromethyl
moieties.
3. The compound of claim 2 wherein R.sub.2 is selected from mono-
and polyfluoro-substituted arylalkyl moieties.
4. The compound of claim 3 wherein R.sub.2 is selected from mono-
and difluoro-substituted benzyl moieties.
5. The compound of claim 1 in at least one of human plasma and
contact with an enzyme in a streptococcal mevalonate biosynthetic
pathway.
6. A compound of a formula ##STR00074## wherein X is selected from
NH, O and S; R.sub.1 is selected from mono- and
polyfluoro-substituted methyl moieties; and R.sub.2 is selected
from alkyl, mono- and polyfluoro-substituted alkyl, aryl, mono- and
polyfluoro-substituted aryl, arylalkyl, and mono- and
polyfluoro-substituted arylalkyl moieties; and hydrates and
solvates thereof.
7. The compound of claim 6 wherein X is selected from NH and O; and
R.sub.1 is selected from fluoromethyl and trifluoromethyl
moieties.
8. The compound of claim 7 wherein R.sub.2 is selected from aryl,
mono- and polyfluoroaryl, arylalkyl, mono- and polyfluoroarylalkyl
moieties.
9. The compound of claim 8 wherein R.sub.2 is selected from phenyl,
mono- and polyfluorophenyl, benzyl, monofluorobenzyl and
polyfluorobenzyl moieties.
10. A compound of a formula ##STR00075## wherein X is selected from
NH, O and S; R.sub.1 is selected from mono- and
polyfluoro-substituted methyl moieties; and R.sub.2 is selected
from alkyl, mono- and polyfluoro-substituted alkyl, aryl, mono- and
polyfluoro-substituted aryl, arylalkyl, mono- and
polyfluoro-substituted arylalkyl moieties; and R.sub.3 and R.sub.4
are independently selected from H, alkyl, alkoxyalkyl, aryl and
arylalkyl moieties; and hydrates and solvates thereof.
11. The compound of claim 10 wherein X is selected from NH and O;
and R.sub.1 is independently selected from fluoromethyl and
trifluoromethyl moieties.
12. The compound of claim 11 where R.sub.2 is selected from
arylalkyl, mono- and polyfluoro-substituted arylalkyl moieties.
13. The compound of claim 10 wherein R.sub.3 and R.sub.4 are
independently selected from H and alkyl moieties.
14. The compound of claim 13 wherein at least one of R.sub.3 and
R.sub.4 is methyl.
15. A compound of a formula ##STR00076## wherein X is selected from
NH, O and S; R.sub.1 is selected from mono- and
polyfluoro-substituted methyl moieties; R.sub.2 is selected from
alkyl, mono- and polyfluoro-substituted alkyl, aryl, mono- and
polyfluoro-substituted aryl, arylalkyl, mono- and
polyfluoro-substituted arylalkyl moieties; and R.sub.3 and R.sub.4
are independently selected from H, acetyl, propionyl and
alkoxyalkyl moieties; and hydrates and solvates thereof.
16. The compound of claim 15 wherein X is selected from NH and O;
and R.sub.1 is selected from fluoromethyl and trifluoromethyl
moieties.
17. The compound of claim 16 wherein R.sub.2 is selected from
alkyl, mono- and polyfluoro-substituted alkyl, aryl, mono- and
polyfluoro-substituted aryl, arylalkyl, mono- and
polyfluoro-substituted arylalkyl moieties.
18. The compound of claim 15 wherein X is O; R.sub.2 is selected
from alkyl, aryl and arylalkyl moieties; and R.sub.3 and R.sub.4
are independently selected from H and acetyl moieties.
19. The compound of claim 18 wherein at least one of R.sub.3 and
R.sub.4 is acetyl.
20. A compound of a formula ##STR00077## wherein R.sub.1 is
selected from mono- and polyfluoro-substituted methyl moieties; and
R.sub.2 is selected from acetyl, propionyl and alkoxyalkyl
moieties; and hydrates and solvates thereof.
21. The compound of claim 20 wherein R.sub.1 is selected from
fluoromethyl and trifluoromethyl moieties.
22. The compound of claim 21 wherein R.sub.2 is selected from
acetyl and propionyl moieties.
23. The compound of claim 22 wherein R.sub.2 is acetyl.
Description
[0001] This application is a continuation of and claims priority to
and the benefit of application Ser. No. 13/780,929 filed Feb. 28,
2013 and issued as U.S. Pat. No. 9,150,534 on Oct. 6, 2015, which
claimed priority to and the benefit of application Ser. No.
61/604,261 filed Feb. 28, 2012--each of which is incorporated
herein by reference in its entirety.
BACKGROUND OF THE INVENTION
[0003] Streptococcus pneumoniae is a common bacterium that causes
many types of infection other than pneumonia, including acute
sinusitis, otitis media, and meningitis. Infection by this
bacterium is a significant cause of infant mortality in developing
countries, killing more than 3000 people per day, the majority of
whom are children below the age of five. Unfortunately, the
incidence of strains resistant to penicillin and other
antimicrobial agents has been increasing rapidly worldwide since
the mid-1990s. Thus, developments of new antibiotics are needed to
maintain control of this deadly organism.
[0004] The mevalonate pathway (FIG. 1) is an important cellular
metabolic pathway present in all higher eukaryotes and many
bacteria. Isopentenyl diphosphate, the pathway end-product, is the
five-carbon building block used for the biosynthesis of
isoprenoids, which in turn lead to many biologically active small
molecules, including cholesterol, steroid hormones, and vitamin A.
It was discovered that the mevalonate pathway in S. pneumoniae is
regulated by 5-diphosphomevalonate (DPM). (See, Leyh, T. S., et
al., Biochemistry 2004, 43, 16461.) DPM is a feedback inhibitor of
mevalonate kinase (MVK), the first enzyme in the mevalonate
pathway, binding tightly to an allosteric site of MVK in S.
pneumonia. However, human MVK is not inhibited at DPM
concentrations that essentially completely inhibit the
streptococcal system. It has also been reported that S. pneumoniae
in which the mevalonate pathway is mutated do not survive in mouse
lung or serum.
[0005] On these bases, it appeared that DPM could be a lead
compound for the development of new anti-streptococcal antibiotics
that do not interfere with human metabolism. However, the
highly-charged diphosphate compounds do not penetrate the cell
membrane. Further, phosphatase degradation of the diphosphate also
can occur. For these and other reasons, there remains an ongoing
search in the art for effective inhibitors of the mevalonate
pathway of S. pneumoniae.
SUMMARY OF THE INVENTION
[0006] In light of the foregoing, it is an object of the present
invention to provide various compounds, compositions and/or methods
for their use in the study and/or treatment of Streptococcus
pneumoniae. It will be understood by those skilled in the art that
one or more aspects of this invention can meet certain objectives,
while one or more other aspects can meet certain other objectives.
Each objective may not apply equally, in all its respects, to every
aspect of this invention. As such, the following objects can be
viewed in the alternative with respect to any one aspect of this
invention.
[0007] As can relate to certain other embodiments, it can be an
object of this invention to provide prodrugs of inhibitors of
mevalonate kinase or other enzymes in the biosynthetic pathway of
S. pneumoniae, such prodrug compounds including but not limited to
compounds of the sort described herein.
[0008] As can relate to certain other embodiments, it can be
another object of the present invention to provide such
inhibitory/prodrug compounds, related compositions and/or methods
of use in the study and/or treatment of infectious S.
pneumoniae.
[0009] Other objects, feature, benefits and advantages of this
invention will be apparent from this summary and the following
descriptions of certain embodiments, and will be readily apparent
to those skilled in the art having knowledge of various enzymatic
pathways and mechanistic considerations, together with the design
and synthesis of corresponding inhibitors. Such objects, features,
benefits and advantages will be apparent from above as taken into
conjunction with the accompanying examples, data, figures and all
reasonable inferences to be drawn therefrom, alone or with
consideration of the references incorporated herein.
[0010] In part, the present invention can be directed to a prodrug
of a mevalonate pathway inhibitor compound, such a prodrug compound
as can be selected from compounds of a formula
##STR00001##
wherein X can be selected from NH, O and S; R.sub.1 can be selected
from fluoro-substituted methyl moieties; and R.sub.2 can be
selected from alkyl, fluoro-substituted alkyl, aryl,
fluoro-substituted aryl, arylalkyl and fluoro-substituted arylalkyl
moieties.
[0011] In certain embodiments, X can be NH or O, and/or R.sub.1 can
independently be either a fluoromethyl or trifluoromethyl moiety.
Regardless, in certain such embodiments, R.sub.2 can be an
arylalkyl moiety, optionally mono- or polyfluoro-substituted.
Without limitation, R.sub.2 can be a mono- or di-substituted benzyl
moiety.
[0012] In part, the present invention can also be directed to a
prodrug of a mevalonate pathway inhibitor compound, such a prodrug
compound as can be selected from compounds of a formula
##STR00002##
wherein X can be selected from NH, O and S; R.sub.1 can be selected
from fluoro-substituted methyl moieties; R.sub.2 can be selected
from alkyl, fluoro-substituted alkyl, aryl, fluoro-substituted
aryl, arylalkyl and fluoro-substituted arylalkyl moieties; and
R.sub.3 and R.sub.4 can be independently selected from H, alkyl,
alkoxyalkyl, aryl and arylalkyl moieties.
[0013] In certain embodiments, X can be NH or O, and/or R.sub.1 can
independently be either a fluoromethyl or trifluoromethyl moiety.
Regardless, in certain such embodiments, R.sub.2 can be an
arylalkyl moiety (e.g., benzyl), optionally mono- or
polyfluoro-substituted (e.g., 4-fluoro- or 2,4-difluoro-). Without
limitation, R.sub.3 and R.sub.4 can be independently selected from
H and alkyl moieties. In certain such embodiments, at least one of
R.sub.3 and R.sub.4 can be methyl.
[0014] In part, the present invention can also be directed to a
prodrug of a mevalonate pathway inhibitor compound, such a prodrug
compound as can be selected from compounds of a formula
##STR00003##
wherein X can be selected from NH, O and S; R.sub.1 can be selected
from fluoro-substituted methyl moieties; R.sub.2 can be selected
from alkyl, fluoro-substituted alkyl, aryl, fluoro-substituted
aryl, arylalkyl and fluoro-substituted arylalkyl moieties; and
R.sub.3 and R.sub.4 can be independently selected from H, acetyl,
propionyl and alkoxyalkyl moieties.
[0015] In certain embodiments, X can be NH or O, and R.sub.1 can be
independently either a fluoromethyl or trifluoromethyl moiety.
Regardless, in certain such embodiments, R.sub.2 can be either an
alkyl (e.g., ethyl) or arylalkyl (e.g., benzyl) moiety, each
optionally mono- or polyfluoro-substituted (e.g.,
1,1,1-trifluoroethyl or 1,2,3,4,5-pentafluorobenzyl). Without
limitation as to either R.sub.1 or R.sub.2, R.sub.3 and R.sub.4 can
be independently selected from H and acetyl moieties. In certain
such embodiments, at least one of R.sub.3 and R.sub.4 can be
acetyl.
[0016] In part, the present invention can also be directed to a
prodrug of a mevalonate pathway inhibitor compound, such a prodrug
compound as can be selected from compounds of a formula
##STR00004##
wherein R.sub.1 can be selected from fluoro-substituted methyl
moieties; and R.sub.2 can be selected from acetyl, propionyl and
alkoxyalkyl moieties.
[0017] In certain embodiments, R.sub.1 can be either a fluoromethyl
or trifluoromethyl moiety. Regardless, in certain such embodiments,
R.sub.2 can be either an acetyl or propionyl moiety. Without
limitation, R.sub.2 can be acetyl.
[0018] Generally, the compounds of this invention are without
stereochemical limitation. As illustrated and discussed below, such
compounds and/or their intermediates are available as racemic
mixtures from which isomers can be resolved or are diastereomers,
from which the corresponding enantiomers can be separated.
Accordingly, any stereocenter can be (S) or (R) with respect to any
other stereocenter(s). Further, it will be understood by those
skilled in the art that any one or more the compounds of this
invention can be provided as part of a pharmaceutical composition
comprising a pharmaceutically-acceptable carrier component for use
in conjunction with a treatment method or medicament.
[0019] In part, the present invention can also be directed to a
method of treating Streptococcus pneumoniae. Such a method can
comprise providing a compound of this invention, whether or not
part of a pharmaceutical composition, and administering an
effective amount of such a compound for contact with S. pneumoniae,
mevalonate kinase and/or another enzyme expressed in the
streptococcal mevalonate biosynthetic pathway. In certain such
embodiments, such a compound and/or combination thereof can be
present in an amount at least partially sufficient to bind or
otherwise interact with an enzyme of the mevalonate pathway of such
an organism, inhibit the pathway and/or inactivate the
organism.
BRIEF DESCRIPTION OF THE DRAWINGS
[0020] FIG. 1 illustrates the mevalonate pathway (prior art). The
conversion of mevalonic acid to isopentenyl diphosphate occurs in
three ATP-dependent steps catalyzed by GHMP family kinases: MVK,
mevalonate kinase; PMK, phosphomevalonate kinase; DPM-DC,
dephophomevalonate decarboxylase.
[0021] FIG. 2 provides, without limitation, prodrug compounds of
diphosphomevalomate analogues, in accordance with certain
embodiments of this invention.
[0022] FIG. 3 provides, without limitation, a proposed
decomposition mechanism for certain prodrug compounds in this
invention.
[0023] With reference to FIG. 2, FIG. 4 provides general structures
of non-limiting, representative prodrug compounds, in accordance
with certain embodiments of this invention.
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS
[0024] Illustrating certain non-limiting embodiments of this
invention, various prodrugs of DPM analogs were designed. For
instance, carbonates, acetals, ketals, amides, esters, and lactones
were chosen as promoieties of the carboxyl and hydroxyl
functionalities of mevalonate. Corresponding prodrug compounds,
representative of this invention (see, e.g., FIG. 2), were
synthesized and characterized, as discussed below.
Chemistry of Representative Carbonate Prodrugs.
[0025] A synthetic route to cyclic carbonate 4 is shown in Scheme
1. Commercially available lactone 1 was hydrolyzed under KOH basic
condition, and then produced carboxylic acid 2 underwent
esterification with benzyl bromide to give benzyl ester 3. Although
a portion of ester 3 was converted to mevalonolactone (1) during
column chromatography with silica gel, 3 was isolated as the major
product. Ester 3 was treated with triphosgene to give 4 in good
yields.
##STR00005##
[0026] The other synthetic route to the benzyl ester (3) is shown
in Scheme 2. After the hydroxyl group of 4-hydroxy-2-butanone (5)
was protected with a TBS group, aldol reaction of benzyl acetate
with ketone 6 was performed using LDA as a base to give 7. Desired
product 3 was produced from intermediate 7 by TBS deprotection
using tetrabutylammonium fluoride (TBAF) in THF.
##STR00006##
[0027] As shown in Scheme 3, cyclic carbonate analogues (9-16) were
prepared via Pd/C catalyzed benzyl deprotection followed by
coupling with diverse aryl alcohols (e.g., R=phenyl,
4-fluorophenyl, and 5-indanyl) using EDC as a coupling reagent
(condition A) or with alkyl iodides (e.g., R=cyclohexyl, iPr, and
tBu) using K.sub.2CO.sub.3 as a base (condition B). Carboxylic acid
8 was treated with pivaloyloxymethyl chloride and triethylamine to
give the pivaloyloxymethyl ester (16, Conditions C).
##STR00007##
[0028] Amide analogues of cyclic carbonates were prepared from
carboxylic acid 8 (Scheme 4). A coupling reaction of carboxylic
acid 8 with aniline, benzyl amine, or 4-fluorobenzyl amine using
HBTU as an activation reagent gave the corresponding amides (17-19)
in moderate yields. (If thioalchohols (e.g., Condition A, RSH) are
used in the coupling reaction, the corresponding thioesters
(X.dbd.S) are produced.) Because these amides have an aromatic
ring, we can monitor their stability in human plasma using HPLC
with UV detector. An amide moiety would be much more stable for
hydrolysis than ester moieties; thus this feature could allow
investigation of the stability of the cyclic carbonate moiety in
human plasma. Phenyl amide 17 was made because the N-phenylamide is
chemically less stable than benzyl amide. 4-Fluorobenzyl amide 19
was also thought to be less stable than benzyl amide due to the
electron withdrawing effect of fluorine on the phenyl ring.
##STR00008##
[0029] A synthetic route of 6-fluoromethyl cyclic carbonate
analogues is shown in Scheme 5. Ethyl fluoroacetate (20) was
treated with 1.95 equivalents of magnesium bromide at 0.degree. C.
for 30 min to give diolefin 21. Ozonolysis of crude product 21,
followed by acidic treatment, gave dicarboxylic acid 22. Without
purification of 22, benzylation was conducted to give diester 23 in
more than 50% yield for 4 steps. When 23 was treated with DIBAL-H
(3-4 equiv) at 0.degree. C. in THF, the major product was the diol
(24). Because monoester 24 underwent intramolecular cyclization on
silica gel, a crude mixture of 24 was allowed to react with
triphosgene without further purification to obtain desired cyclic
carbonate 25 in a moderate yield. The benzyl group of 25 was
removed by hydrogenation to give carboxylic acid 26. Esterification
of 26 with iodomethane, iodoethane, 4-fluorobenzyl bromide, and
2,4-difluorobenzyl bromide using sodium bicarbonate as a base were
carried out to give the corresponding esters (27-30),
respectively.
##STR00009##
[0030] 6,6,6-Trifluoromethyl-cyclic carbonate analogues were
synthesized from commercially available
4-ethoxy-1,1,1-trifluoro-3-buten-2-one (Scheme 6). The addition of
benzyl acetate to enone 31 gave 32. The ethylenol was hydrolyzed to
aldehyde under acidic conditions, and then produced aldehyde 33 was
reduced to alcohol 34 by sodium triacetoxyborohydride. When sodium
borohydride was used, the benzyl ester was also reduced to the
hydroxyl group gradually. Cyclic carbonate 35 was obtained by
treatment of triphosgene to the crude product (34). The benzyl
group of 35 was removed by Pd/C catalyzed hydrogenation in ethyl
acetate and hydrogen atmosphere. Carboxylic acid 36 was esterified
with iodomethane, iodoethane, 4-fluorobenzyl bromide, or
2,4-difluorobenzyl bromide.
##STR00010##
Stability of the Cyclic Carbonate Prodrugs in PBS Buffer.
[0031] The stabilities of several cyclic carbonate analogues in PBS
buffer (pH=7.4) were tested first to determine the standard level
of the drug decomposition by media (Table 1). PBS (phosphate
buffered saline) buffer is a standard solution used in plasma
stability tests. The cyclic carbonate analogues (4, 25, 29, 30, 35,
39, and 40) had a UV chromophore, which allowed us to determine the
amount of drug in the culture medium by HPLC analysis. After the
compounds were incubated in PBS buffer at 37.degree. C., the
degradation of the compounds with incubation time was monitored. As
the number of fluorines at the C6-position was increased, the
half-life time (T.sub.1/2) of the molecule was decreased
dramatically (4 vs 25 vs 35). T.sub.1/2 of the C6-methyl analogue
(4) was more than 48 hours and T.sub.1,2 of C6-monofluoromethyl
analogues (25, 29, and 30) were about 25 hours while T.sub.1/2 of
C6-trifluoromethyl analogues (35, 39, and 40) were about 5 hours.
If the substitution of C-6-position (CH.sub.3, CH.sub.2F, or
CF.sub.3) was same, T.sub.1/2 of diverse benzyl esters (benzyl,
4-fluorobenzyl, or 2,4-difluorobenzyl) were not dramatically
changed. These results show that the electron withdrawing effect at
the C6 position affect the stability of the benzyl ester moiety.
These half-lives of benzyl esters and cyclic carbonates in PBS
buffer were sufficient to use these promoieties for the penetration
of the cell membrane.
TABLE-US-00001 TABLE 1 Half-lives of diverse carbonate in PBS
buffer. R.sub.2 R.sub.1 # T.sub.1/2 ##STR00011## Bn Bn 4-F-Bn
2,4-di-F-Bn Bn 4-F-Bn 2,4-di-F-Bn CH.sub.3 CH.sub.2F CF.sub.3 4 25
29 30 35 39 40 >48 h 25 h 26 h 22 h 5 h 6 h 5 h
[0032] Another attempts were carried out to determine whether
desired decompositions occur for the trifluoromethyl analogue (35)
in PBS buffer. While the peak of 35 decreased and the peak of
benzyl alcohol increased in HPLC, the peak of 34 that was resulted
from carbonate moiety decomposition was not observed (FIG. 3). This
result shows that the cyclic carbonate moiety of 35 might be more
stable than the benzyl ester moiety in PBS buffer and compound 34
might be not generated in the process of degradation of 35.
[0033] The stability of cyclic carbonate moiety in carboxylic acid
36 was tested in CD.sub.3OD, D.sub.2O, and PBS buffer. After 36 was
incubated in PBS buffer at 37.degree. C., solvent was removed by
lyophilization, and the residue was monitored by .sup.1H-NMR. As a
result, incubation time-dependent increase of 42 and decrease of
the precursor 36 were observed. The T.sub.1/2 of cyclic carbonate
moiety of 36 in PBS buffer was about 3 h. These stability tests in
PBS buffer show that the cyclic carbonate promoiety can be
converted to the desired linear diol, as expected, and should be
effective prodrugs upon penetration of the cell membrane.
Stability of the Cyclic Carbonate Prodrugs in Human Plasma
[0034] The stabilities of diverse cyclic carbonate prodrugs (4, 9,
10, 11, 17, 18, 19, 25, 29, 30, 39, 40, and 41), which have a
chromosphere for UV detection, in human plasma were tested using
HPLC. After the mixture of test compounds and human plasma were
incubated at 37.degree. C., the degradation of the compounds was
monitored by HPLC (Table 2). There were only slight differences in
the half-lives depending on the ester groups; the half-life of 4
was 4 minutes and that of the other esters (9, 10, 11, 25, 29, 30,
35, 39, 40, and 41) were less than 3 minutes. As the area of the
compounds decreased, the area of produced alcohols increased in
HPLC for all of these compounds. The peaks of the corresponding
diols (3, 24, or 34), which were the compounds that result when
only the carbonate moiety of the analogues was hydrolyzed, were not
observed. The half-lives of the benzyl or phenyl ester moieties may
be shorter than carbonate, under tested conditions for such
compounds to be used effectively as prodrugs.
[0035] The T.sub.1/2 of N-phenyl amide 17 was also below 3 minutes.
As the area of the peak of prodrug 17 decreased, the area of the
peak of aniline increased. The peak of hydrolysis product of the
cyclic carbonate moiety was minor compared with that of the
aniline; therefore, the hydrolysis of N-phenyl amide promoiety was
as fast as the ester groups.
[0036] The T.sub.1/2 of the benzyl amide analogues (18, 19) was 8
minutes. As the area of the peaks of 18 and 19 decreased, the peaks
that result only from hydrolysis of carbonate moiety increased
rather than that of benzyl amine. The peak of benzyl amine was not
shown even further 12-hour incubation. These results show that the
benzyl amide group was stable in human plasma. If the decomposition
of the cyclic carbonate moiety of 4 preceded that of the benzyl
ester moiety, the half-life of 4 should be similar with that of 18.
However, the half-life of 4 was half that of 18. This demonstrates
that the cyclic carbonate moiety of these analogues is more stable
than the benzyl ester moiety, and is consistent with the results of
the stabilities in buffers. Such results suggest that the benzyl
and phenyl ester moieties may not be sufficient, alone, but the
cyclic carbonate moiety may be sufficiently stable in human plasma
for such compounds to be useful as prodrugs.
TABLE-US-00002 TABLE 2 Various half-lives of diverse carbonate in
human plasma. R.sub.1 R.sub.2 # T.sub.1/2 ##STR00012## CH.sub.3
CH.sub.2F Bn Ph 4-F-Ph 5-indanyl Bn 4-F-Bn 2,4-di-F-Bn 4 9 10 11 25
29 30 4 min <3 min <3 min <3 min 2 min 2 min 1 min
CF.sub.3 Bn 35 <1 min 4-F-Bn 39 <1 min 2,4-di-F-Bn 40 <1
min PhEt 41 4 min ##STR00013## CH.sub.3 Ph Bn 4-F-Bn 17 18 19 <3
min 8 min 8 min
Chemistry of Representative Acetal/Ketal Prodrugs.
[0037] Syntheses of acetal/ketal analogues are shown in Scheme 7.
After the ring-opening reaction of 1 or 42 with benzyl amine, the
introduction of the MOM group followed by treatment with
BF.sub.3.Et.sub.2O, gave methylene acetals 45 and 46. The reaction
of 44 and 2,2-dimethoxypropane with a catalytic amount of CSA gave
acetonides 47 and 48. The treatment of 44 with benzaldehyde
dimethoxy acetal and CSA in CH.sub.2Cl.sub.2 gave both
diastereomers 49 and 50, in which the CF.sub.3 and phenyl groups
were anti and syn. These diastereomers were separated by silica gel
chromatography. Compound 44 was converted to the
p-methoxybenzylidene acetals (51 and 52) by treatment with
anisaldehyde dimethyl acetal. These diastereomers were also
separated by silica gel chromatography. In general, the
p-methoxybenzylidene acetal group can undergo acid hydrolysis ten
times faster than the benzylidene acetal group. So, 51 and 52 were
expected to decompose more easily than 49 and 50.
2,4-dimethoxybenzylidene acetal (53) and t-butyl carbonate
functionalized 54 and 55 were also prepared from intermediate
44.
##STR00014##
Stability of the Benzyl Amide, Acetal, Ketal, and t-Butylcarbonate
Promoieties in Human Plasma.
[0038] The stabilities of the benzyl amide analogues with acetal,
ketal, and t-butyl carbonate promoieties (45-55) were tested in
human plasma (Table 3). The half-lives in human plasma for all
acetal, ketal, and t-butyl carbonate compounds were longer than 48
h. The activity of human plasma was confirmed with a compound known
to undergo decomposition in human plasma.
TABLE-US-00003 TABLE 3 Stabilities of the benzyl amide analogues in
human plasma R.sub.1 R.sub.4 R.sub.3 # T.sub.1/2 ##STR00015##
CH.sub.3 CF.sub.3 H CH.sub.3 Ph H CH.sub.3 Ph 4-MeO-Ph H
2,4-diMeO-Ph H CH.sub.3 H H CH.sub.3 H H 4-MeO-Ph H 45 47 49 46 48
50 51 52 53 >48 h >48 h >48 h >48 h >48 h >48 h
>48 h >48 h >48 h ##STR00016## CH.sub.3 CF.sub.3 H H Boc
Boc 54 55 >48 h >48 h
Chemistry of Representative Ester Prodrugs.
[0039] Syntheses of diverse ester analogues are shown in Scheme 8
and Scheme 9. After ring-opening of 1 with various electron
deficient amines, the introduction of the acetyl group gave diverse
acetyl esters (57, 58, 60). The reaction of 56 with excess acetic
anhydride in pyridine gave both mono- and di-acetylated products
(57 and 58). The primary hydroxyl group of 59 was acetylated using
one equivalent of acetic anhydride in pyridine to give 60.
[0040] C-6 fluoromethyl analogous (62, 63, 64, 65) were prepared
from dibenzyl ester 23 (Scheme 9). The dibenyl ester was reduced to
triol using LiBH.sub.4 and then mixed with one equivalent of acetic
anhydride in pyridine to give mono-acetylated intermediate 61. That
intermediate underwent oxidation using PDC and coupled with various
alcohols and an amine to give corresponding products in moderate
yields (62-65). (If thioalchohols are used in the coupling
reaction, the corresponding thioesters (X.dbd.S) are produced.)
##STR00017##
##STR00018##
Stability of the Ester Promoiety in Human Plasma.
[0041] The stabilities of the various amide and ester analogues
(57-65) in human plasma were ascertained (Table 4). The half-life
in human plasma for the amide moieties (56, 59) was greater than 2
h. Therefore, the amide bond of tested compounds is too stable in
human plasma for use as a promoiety. The half-life of the acetyl
groups on the alcohol of 57, 60, and 65 was about 20 minutes,
suggesting that the acetyl group is a promising promoiety for the
alcohols. The half-life of the ester moieties on the carboxylic
acids of 62, 63 and 64 was between 10 and 22 minutes, suggesting
that the ester group is a promising promoiety for the carboxylic
acid.
TABLE-US-00004 TABLE 4 Stabilities tests of the benzyl amide
analogues in human plasma. R.sub.2 R.sub.1 R.sub.3 R.sub.4 #
T.sub.1/2 ##STR00019## ##STR00020## CH.sub.3 CH.sub.3 CH.sub.3 H H
Ac H Ac Ac 56 57 58 >120 min 20 min 12 min ##STR00021##
CH.sub.2F CH.sub.2F H H H Ac 59 60 >120 min 20 min Bn CH.sub.2F
H Ac 65 20 min ##STR00022## ##STR00023## CH.sub.3 H Ac 62 23 min Me
CH.sub.2F H Ac 63 10 min Et CH.sub.2F H Ac 64 11 min
[0042] As discussed above, various studies were undertaken in the
development of prodrugs of diphosphomevalonate, a feedback
inhibitor of mevalonate kinase (MVK). Stability studies of diverse
ester analogues of mevalonate using human plasma and PBS buffer
show that it is converted to mevalonate via hydrolysis mediated by
human plasma or solvent. It is also shown that the rates of
decomposition of the acetates, ethyl esters, and cyclic carbonates
promoieties are not only greater than that of the amides, but also
of cyclic acetals and ketals in the tested analogues. In general,
the ester promoiety and cyclic carbonates are converted to the
desired carboxylic acid and alcohol moieties, respectively, in
human plasma. Although the amides, cyclic ketals, and cyclic
acetals are decomposed relatively slowly in human plasma, this
study shows that the half-lives in human plasma for each functional
group are controllable by modifying the electronic character of the
promoiety. Plasma stability studies of these mevalonate analogues
demonstrated that ester, amide, carbonate, acetal, and ketal
prodrugs of fluorinated mevalonates can be used to enhance membrane
permeability and oral absorption. FIG. 4 shows general and specific
structures of various prodrugs of fluorinated mevalonates, in
accordance with this invention.
[0043] The present invention can also, as would be understood by
those skilled in the art, be extended to or include methods using
or in conjunction with a pharmaceutical composition comprising a
compound of the sort described herein and a physiologically or
otherwise suitable formulation. In certain embodiments, the present
invention includes one or more inhibitory and/or prodrug compounds,
of the sort set forth above, formulated into compositions together
with one or more physiologically tolerable or acceptable diluents,
carriers, adjuvants or vehicles that are collectively referred to
herein as carriers. Compositions suitable for such contact or
administration can comprise physiologically acceptable sterile
aqueous or nonaqueous solutions, dispersions, suspensions or
emulsions. The resulting compositions can be, in conjunction with
the various methods described herein, for administration or contact
with S. pneumonia, mevalonate kinase or another enzyme expressed or
otherwise present in the streptococcal mevalonate pathway. Whether
or not in conjunction with a pharmaceutical composition,
"contacting" means that a mevalonate kinase or other enzyme,
mutation or variation thereof and one or more such
inhibitor/prodrug compounds are brought together for purpose of
binding and/or complexing such a compound to the enzyme. Amounts of
a compound effective for inhibition may be determined empirically,
and making such determinations is within the skill in the art.
Inhibition or otherwise affecting mevalonate/enzyme activity
includes both reduction and/or mitigation, as well as elimination
of kinase activity and/or isopentenyl disphosphate and/or related
intermediate production.
[0044] It is understood by those skilled in the art that dosage
amount will vary with the activity of a particular compound(s),
disease state, route of administration, duration of treatment, and
like factors well-known in the medical and pharmaceutical arts. In
general, a suitable dose will be an amount which is the lowest dose
effective to produce a therapeutic or prophylactic effect. If
desired, an effective dose of such a compound,
pharmaceutically-acceptable salt thereof, or related composition
may be administered in two or more sub-doses, administered
separately over an appropriate period of time.
[0045] Methods of preparing pharmaceutical formulations or
compositions include the step of bringing an inhibitor/prodrug
compound into association with a carrier and, optionally, one or
more additional adjuvants or ingredients. For example, standard
pharmaceutical formulation techniques can be employed, such as
those described in Remington's Pharmaceutical Sciences, Mack
Publishing Company, Easton, Pa.
[0046] Regardless of composition or formulation, those skilled in
the art will recognize various avenues for medicament
administration, together with corresponding factors and parameters
to be considered in rendering such a medicament suitable for
administration. Accordingly, with respect to one or more
non-limiting embodiments, the present invention provides for use of
one or more anti-streptococcal mevalonate pathway inhibitor
compounds for the manufacture of a medicament for therapeutic use
in the treatment of human infectious diseases or the prevention
thereof.
EXAMPLES OF THE INVENTION
[0047] The following non-limiting examples and data illustrate
various aspects and features relating to the compounds and/or
methods of the present invention, including the preparation of
various prodrug compounds, as are available through the synthetic
methodologies described herein. In comparison with the prior art,
the present compounds provide results and data which are
surprising, unexpected and contrary thereto. While the utility of
this invention is illustrated through the use of several compounds
and prodrug moieties which can be incorporated therein, it will be
understood by those skilled in the art that comparable results are
obtainable with various other prodrug compounds and/or moieties, as
are commensurate with the scope of this invention.
Evaluation of Stability in PBS Buffer (pH=7.4).
[0048] The stock solution of the test compound (10 mM in
acetonitrile, 50 .mu.L) was added to PBS buffer (pH=7.4, 450
.mu.L), and the mixture was incubated at 37.degree. C. Aliquots of
the samples were analyzed directly by HPLC (See below). These tests
were conducted two times in each compound.
[0049] The other buffers that were used in the stabilities tests of
34 and 36 are following list; pH=8.2: Tris-HCl (0.1 M), pH=6.4:
sodium dihydrogen phosphate (0.2 M)/sodium phosphate dibasic (0.2
M), pH=5.4: sodium phosphate dibasic (0.2 M)/citric acid (0.1
M).
Evaluation of Stability in Human Plasma.
[0050] Human plasma (100%, 480 .mu.L) was incubated at 37.degree.
C. for 5 min. The stock solution of the test compound (100 mM in
acetonitrile, 20 .mu.L) was added to the human plasma, and the
mixture was incubated at 37.degree. C. Aliquots (80 .mu.L) of the
plasma samples were removed and mixed with an equal volume of
acetonitrile. The mixture was stirred vigorously and centrifuged
(5500 rpm, 5 min). The supernatant was filtered and the filtrate
was analyzed by HPLC or LC-TOF (See below). These tests were
conducted two times in each compound.
HPLC Analysis.
[0051] Analysis was performed on a Phenomenex.RTM. Luna C18 column
(250.times.4.6 mm) eluted with a gradient conditions of
acetonitrile and H.sub.2O. Detection was by UV absorbance at 257 nm
or total count of single ion in Agilent 6210 LC-TOF mass
spectra.
Synthesis and Characterization
Example 1
##STR00024##
[0053] To a solution of (.+-.)-mevalonolanctone 1 (97%, 268 mg,
2.00 mmol) in H.sub.2O (4 mL) was added KOH (.gtoreq.90%, 123 mg,
2.20 mmol) at rt, and the mixture was stirred at 40.degree. C. for
1 h. The pH of the solution was lowered to about pH 7-8 (detected
by pH indicator paper) with aq. HCl (0.1 M). To the mixture were
added benzyl bromide (363 .mu.L, 3.00 mmol), tetrabutylammonium
bromide (967 mg, 3.00 mmol), and THF (8 mL) at rt, and the mixture
was stirred at 50.degree. C. for 4 h. After the mixture was diluted
with AcOEt, the mixture was partitioned between AcOEt and brine.
The organic layer was dried (Na.sub.2SO.sub.4) and evaporated. The
residue was purified by silica gel column chromatography (50% to
75% AcOEt in hexane) to give 3 (329 mg, 69%) as a colorless oil.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.33-7.38 (m, 5H,
aromatic), 5.17 (s, 2H, benzyl-CH.sub.2), 4.04 (br s, 1H, OH), 3.88
(m, 1H, CH.sub.2CH.sub.2O), 3.81 (m, 1H, CH.sub.2CH.sub.2O), 2.90
(br s, 1H, OH), 2.69 (d, 1H, OC(O)CH.sub.2, J=15.5 Hz), 2.52 (d,
1H, OC(O)CH.sub.2, J=15.5 Hz), 1.80 (m, 1H, CH.sub.2CH.sub.2O),
1.74 (m, 1H, CH.sub.2CH.sub.2O), 1.32 (s, 3H, CH.sub.3); .sup.13C
NMR (125 MHz, CDCl.sub.3) .delta. 172.7, 135.3, 128.7, 128.5,
128.4, 72.2, 66.7, 59.4, 45.1, 42.1, 26.9; HRMS (pos. ion ESI) m/z
calcd for (M+Na).sup.+ C.sub.13H.sub.18NaO.sub.4: 261.1103. Found:
261.1105.
Example 2
##STR00025##
[0055] To a solution of 3 (436 mg, 1.83 mmol) in CH.sub.2Cl.sub.2
(15 mL) was added pyridine (224 .mu.L, 2.75 mmol), and the mixture
was stirred at 0.degree. C. for 15 min. A solution of triphosgene
(98%, 665 mg, 2.20 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added to
the mixture, and the resulting mixture was stirred at 0.degree. C.
for 30 min. The reaction was quenched with addition of sat. aq.
NH.sub.4Cl, and extracted with AcOEt. The organic layer was washed
with brine, dried (Na.sub.2SO.sub.4), and evaporated. The residue
was purified by silica gel column chromatography (33% to 50% AcOEt
in hexane) to give 4 (425 mg, 88%) as a light yellow oil.
Example 3
[0056] To a solution of 7 (Example 5, below, 324 mg, 0.919 mmol) in
THF (8 mL) were added tetrabutylammonium fluoride (1.0 M solution
in THF, 1.10 mL, 1.10 mmol) and AcOH (1.0 M solution in THF, 2.20
mL, 2.20 mmol) at 0.degree. C., and the mixture was stirred at
0.degree. C. for 24 h. The mixture was diluted with AcOEt and
washed with sat. aq. NaHCO.sub.3. The organic layer was washed with
brine, dried (Na.sub.2SO.sub.4), and evaporated. The residue was
purified quickly by silica gel column chromatography (50% to 80%
AcOEt in hexane) to give compound 3. After the residue was
dissolved in CH.sub.2Cl.sub.2 (8 mL), pyridine (163 .mu.L, 2.00
mmol) was added to the solution and the mixture was cooled at
0.degree. C. To the mixture was added a solution of triphosgene
(98%, 306 mg, 1.00 mmol) in CH.sub.2Cl.sub.2 (1 mL), and the
resulting mixture was stirred at 0.degree. C. for 30 min. The
reaction was quenched with addition of sat. aq. NH.sub.4Cl, and
extracted with AcOEt. The organic layer was washed with brine,
dried (Na.sub.2SO.sub.4), and evaporated. The residue was purified
by silica gel column chromatography (33% to 50% AcOEt in hexane) to
give 4 (155 mg, 64% for 2 steps) as a light yellow oil, and 7 was
recovered (39 mg, 12%).
[0057] .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.34-7.40 (m, 5H,
aromatic), 5.14 (s, 2H, benzyl-CH.sub.2), 4.42 (m, 2H,
CH.sub.2CH.sub.2O), 2.83 (s, 2H, OC(O)CH.sub.2), 2.36 (m, 1H,
CH.sub.2CH.sub.2O), 2.08 (m, 1H, CH.sub.2CH.sub.2O), 1.57 (s, 3H,
CH.sub.3); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 168.7, 148.5,
135.1, 128.6, 128.5, 128.4, 81.0, 66.9, 64.5, 44.8, 30.4, 25.8;
HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.14H.sub.16NaO.sub.5: 287.0895. Found: 287.0899.
Example 4
##STR00026##
[0059] To a solution of 4-hydroxy-2-butanone (273 .mu.L, 3.00 mmol)
in DMF (20 mL) were added tert-butylchlorodimethylsilane (559 mg,
3.60 mmol) and imidazole (490 mg, 7.20 mmol) at 0.degree. C., and
the mixture was stirred at rt for 12 hr. After the addition of
MeOH, the mixture was diluted with Et.sub.2O and washed with
H.sub.2O (.times.3). The organic layer was washed with brine, dried
(Na.sub.2SO.sub.4), and evaporated. The residue was purified by
silica gel column chromatography (5% AcOEt in hexane) to give 6
(548 mg, 90%) as a colorless liquid. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 3.88 (t, 2H, CH.sub.2CH.sub.2O, J=6.3 Hz), 2.62
(t, 2H, CH.sub.2CH.sub.2O, J=6.3 Hz), 2.19 (s, 3H, CH.sub.3), 0.88
(s, 9H, C(CH.sub.3).sub.3), 0.05 (s, 6H, Si(CH.sub.3).sub.2);
.sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 208.2, 58.8, 46.5, 30.9,
25.8, 18.2, -5.5; HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.10H.sub.22NaO.sub.2Si: 225.1287. Found: 225.1280.
Example 5
##STR00027##
[0061] To a solution of lithium diisopropylamide (1.8 M solution in
heptane/THF/ethyl benzene, 7.93 mL, 14.3 mmol) in THF (105 mL) was
added benzyl acetate (2.04 mL, 14.3 mmol) at -78.degree. C., and
the mixture was stirred at -78.degree. C. for 30 min. A solution of
6 (2.41 g, 11.9 mmol) in THF (5 mL) was added to the mixture via
canule at -78.degree. C. and the resulting mixture was stirred at
-78.degree. C. for 1 h. After addition of sat. aq. NH.sub.4Cl, the
mixture was warmed to room temperature, and evaporated. The residue
was separated between AcOEt and sat. aq. NH.sub.4Cl. The organic
layer was washed with brine, dried (Na.sub.2SO.sub.4), and
evaporated. The residue was purified by silica gel column
chromatography (5% to 8% AcOEt in hexane) to give 7 (3.92 mg, 93%)
as a colorless oil. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
7.31-7.35 (m, 5H, aromatic), 5.17 (d, 1H, benzyl-CH.sub.2, J=7.5
Hz), 5.12 (d, 1H, benzyl-CH.sub.2, J=7.5 Hz), 4.20 (s, 1H, OH),
3.86 (t, 2H, CH.sub.2CH.sub.2O, J=3.6 Hz), 2.66 (d, 1H,
OC(O)CH.sub.2, J=15.6 Hz), 2.60 (d, 1H, OC(O)CH.sub.2, J=15.6 Hz),
1.82 (m, 2H, CH.sub.2CH.sub.2O), 1.30 (s, 3H, CH.sub.3), 0.89 (s,
9H, C(CH.sub.3)), 0.07 (s, 6H, Si(CH.sub.3).sub.2); HRMS (pos. ion
ESI) m/z calcd for (M+Na).sup.+ C.sub.19H.sub.32NaO.sub.4Si:
375.1968. Found: 375.1965.
Example 6
##STR00028##
[0063] To a solution of 4 (340 mg, 1.29 mmol) in AcOEt (15 mL) was
added Pd/C (10%, 33 mg), and the mixture was stirred under H.sub.2
gas at rt for 30 min. The resulting mixture was filtered through
Celite with acetone, and the filtrate was evaporated to give 8 (224
mg, quant.) as a colorless solid. .sup.1H NMR (500 MHz, CD.sub.3OD)
.delta. 4.48 (m, 2H. CH.sub.2CH.sub.2O), 2.82 (d, 1H,
OC(O)CH.sub.2, J=15.5 Hz), 2.78 (d, 1H, OC(O)CH.sub.2, J=15.5 Hz),
2.48 (m, 1H, CH.sub.2CH.sub.2O), 2.12 (m, 1H, CH.sub.2CH.sub.2O),
1.57 (s, 3H, CH.sub.3); .sup.13C NMR (125 MHz, CD.sub.3OD) .delta.
172.5, 151.9, 83.3, 66.2, 45.2, 31.3, 26.1; HRMS (pos. ion ESI) m/z
calcd for (M+Na).sup.+ C.sub.7H.sub.10NaO.sub.5: 197.0426. Found:
197.0426. mp. 83-85.degree. C.
Example 7
##STR00029##
[0065] To a solution of 8 (35 mg, 0.20 mmol) in
CH.sub.3CN/CH.sub.2Cl.sub.2 (1/1, 1.5 mL) were added phenol (85 mg,
0.90 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide
hydrochloride (38 mg, 0.20 mmol), and DMAP (20 mg, 0.16 mmol) at
0.degree. C., and the mixture was stirred at rt for 1 h. The
mixture was diluted with AcOEt and partitioned between AcOEt and
aq. HCl (0.5 M). The organic layer was washed with sat. aq.
NaHCO.sub.3, brine, dried (Na.sub.2SO.sub.4), and evaporated. The
residue was purified by silica gel column chromatography (33% to
50% AcOEt in hexane) to give 9 (27 mg, 54%) as a white solid.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.40 (t, 2H, aromatic,
J=7.9, 8.0 Hz), 7.26 (t, 1H, aromatic, J=8.0 Hz), 7.09 (d, 2H,
aromatic, J=7.9 Hz), 4.49 (t, 2H, CH.sub.2CH.sub.2O, J=5.1 Hz),
3.05 (s, 2H, OC(O)CH.sub.2), 2.47 (m, 1H, CH.sub.2CH.sub.2O), 2.19
(m, 1H, CH.sub.2CH.sub.2O), 1.68 (s, 3H, CH.sub.3); .sup.13C NMR
(125 MHz, CDCl.sub.3) .delta. 167.6, 150.0, 148.4, 129.6, 126.3,
121.3, 81.0, 64.5, 44.9, 30.6, 25.9; HRMS (pos. ion ESI) m/z calcd
for (M+Na).sup.+ C.sub.13H.sub.14NaO.sub.5: 273.0739. Found:
273.0731. mp. 62-62.5.degree. C.
Example 8
##STR00030##
[0067] 10 (35 mg, 65%, white solid) was prepared from 8 (35 mg,
0.20 mmol) as described for the preparation of 9 using
4-fluorophenol instead of phenol. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 7.06-7.08 (m, 4H, aromatic), 4.48 (m, 2H,
CH.sub.2CH.sub.2O), 3.03 (s, 2H, OC(O)CH.sub.2), 2.48 (m, 1H,
CH.sub.2CH.sub.2O), 2.17 (m, 1H, CH.sub.2CH.sub.2O), 1.67 (s, 3H,
CH.sub.3); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 167.6, 160.4
(d), 148.4, 145.8, 122.8, 116.3, 80.9, 64.5, 44.9, 30.6, 25.9; HRMS
(pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.13H.sub.13FNaO.sub.5: 291.0645. Found: 291.0648. mp.
68-69.degree. C.
Example 9
##STR00031##
[0069] 11 (40 mg, 70%, white solid) was prepared from 8 (35 mg,
0.20 mmol) as described for the preparation of 9 using 5-indanol
instead of phenol. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.21
(d, 2H, aromatic, J=8.1 Hz), 6.92 (s, 1H, aromatic), 6.81 (d, 2H,
aromatic, J=8.1 Hz), 4.48 (m, 2H, CH.sub.2CH.sub.2O), 3.03 (s, 2H,
OC(O)CH.sub.2), 2.91 (m, 4H, --CH.sub.2CH.sub.2CH.sub.2--), 2.47
(m, 1H, CH.sub.2CH.sub.2O), 2.19 (m, 1H, CH.sub.2CH.sub.2O), 2.11
(m, 2H, --CH.sub.2CH.sub.2CH.sub.2), 1.68 (s, 3H, CH.sub.3);
.sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 168.0, 148.4, 146.0,
142.3, 124.9, 118.8, 117.3, 81.1, 64.6, 44.9, 32.9, 32.3, 30.6,
25.9, 25.7; HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.16H.sub.18NaO.sub.5: 313.1052. Found: 313.1050. mp.
88-89.degree. C.
Example 10
##STR00032##
[0071] To a solution of 8 (130 mg, 0.746 mmol) in DMF (3 mL) were
added iodomethane (140 .mu.L, 2.24 mmol) and K.sub.2CO.sub.3 (309
mg, 2.24 mmol), and the mixture was stirred at rt for 12 h. The
mixture was diluted with AcOEt and H.sub.2O, and separated. The
organic layer was washed with H.sub.2O (.times.2) and brine, dried
(Na.sub.2SO.sub.4), and evaporated. The residue was purified by
silica gel column chromatography (50% AcOEt in hexane) to give 12
(105 mg, 75%) as a colorless oil. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 4.44 (m, 2H, CH.sub.2CH.sub.2O), 3.71 (s, 3H, OCH.sub.3),
2.80 (s, 2H, OC(O)CH.sub.2), 2.40 (m, 1H, CH.sub.2CH.sub.2O), 2.13
(m, 1H, CH.sub.2CH.sub.2O), 1.58 (s, 3H, CH.sub.3); .sup.13C NMR
(125 MHz, CDCl.sub.3) .delta. 169.4, 148.6, 81.0, 64.6, 52.1, 44.6,
30.5, 25.8; HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.8H.sub.12NaO.sub.5: 211.0582. Found: 211.0590.
Example 11
##STR00033##
[0073] 13 (99 mg, 70%, colorless oil) was prepared from 8 (132 mg,
0.758 mmol) as described for the preparation of 12 using iodoethane
instead of iodomethane. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
4.45 (t, 2H, --CH.sub.2CH.sub.2O--, J=6.6 Hz), 4.17 (q, 2H,
CH.sub.3CH.sub.2O--, J=7.1 Hz), 2.79 (s, 2H. OC(O)CH.sub.2), 2.40
(m, 1H, CH.sub.2CH.sub.2O), 2.12 (m, 1H, CH.sub.2CH.sub.2O), 1.58
(s, 3H, CH.sub.3), 1.28 (t, 3H, CH.sub.3CH.sub.2O--, J=7.1 Hz);
.sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 169.0, 148.7, 81.2,
64.7, 61.3, 45.0, 30.7, 26.0, 14.2; HRMS (pos. ion ESI) m/z calcd
for (M+Na).sup.+ C.sub.9H.sub.14NaO.sub.5: 225.0739. Found:
225.0738.
Example 12
##STR00034##
[0075] 14 (21 mg, 75%, colorless oil) was prepared from 8 (23 mg,
0.13 mmol) as described for the preparation of 12 using
2-iodopropane instead of iodomethane. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 5.03 (m, 1H, CH(CH.sub.3).sub.2), 4.44 (m, 2H,
CH.sub.2CH.sub.2O), 2.75 (s, 2H, OC(O)CH.sub.2), 2.40 (m, 1H,
CH.sub.2CH.sub.2O), 2.11 (m, 1H, CH.sub.2CH.sub.2O), 1.58 (s, 3H,
CH.sub.3), 1.26 (s, 3H, CH(CH.sub.3).sub.2), 1.25 (s, 3H,
CH(CH.sub.3).sub.2); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
168.4, 148.6, 81.1, 68.8, 64.6, 45.2, 30.5, 25.9, 21.7; HRMS (pos.
ion ESI) m/z calcd for (M+Na).sup.+ C.sub.10H.sub.16NaO.sub.5:
239.0895. Found: 239.0895.
Example 13
##STR00035##
[0077] 15 (4.2 mg, 13%, colorless oil) was prepared from 8 (22 mg,
0.13 mmol) as described for the preparation of 12 using
iodocyclohexane instead of iodomethane. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 4.78 (m, 1H, cyclohexyl), 4.44 (m, 2H,
CH.sub.2CH.sub.2O), 2.79 (d, 1H, OC(O)CH.sub.2, J=15.2 Hz), 2.75
(d, 1H, OC(O)CH.sub.2, J=15.2 Hz), 2.40 (m, 1H, CH.sub.2CH.sub.2O),
2.11 (m, 1H, CH.sub.2CH.sub.2O), 1.85 (m, 2H, cyclohexyl), 1.73 (m,
2H, cyclohexyl), 1.58 (s, 3H, CH.sub.3), 1.56 (m, 2H, cyclohexyl),
1.32-1.44 (m, 4H, cyclohexyl); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta. 168.3, 148.6, 81.1, 73.8, 64.6, 45.2, 31.5, 30.5, 26.0,
25.2, 23.7; HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.13H.sub.20NaO.sub.5: 279.1208. Found: 279.1205.
Example 14
##STR00036##
[0079] To a solution of 8 (29 mg, 0.17 mmol) in DMF (1 mL) were
added chloromethyl pivalate (97%, 74 .mu.L, 0.50 mmol),
triethylamine (105 .mu.L, 0.750 mmol) and sodium iodide (2.5 mg,
0.017 mmol), and the mixture was stirred at rt for 12 h. The
mixture was diluted with AcOEt, and partitioned between AcOEt and
aq. HCl (0.5 M). The organic layer was washed with sat. aq.
NaHCO.sub.3, H.sub.2O, and brine, dried (Na.sub.2SO.sub.4), and
evaporated. The residue was purified by silica gel column
chromatography (AcOEt in hexane) to give 16 (18 mg, 38%) as a
colorless oil. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 5.77 (s,
2H. OCH.sub.2O), 4.44 (m, 2H, CH.sub.2CH.sub.2O), 2.84 (s, 2H,
OC(O)CH.sub.2), 2.37 (m, 1H, CH.sub.2CH.sub.2O), 2.12 (m, 1H,
CH.sub.2CH.sub.2O), 1.58 (s, 3H, CH.sub.3), 1.22 (s, 9H,
C(CH.sub.3).sub.3); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
177.1, 167.6, 148.3, 80.7, 79.7, 64.5, 44.6, 38.8, 30.5, 26.8,
25.7; HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.13H.sub.20NaO.sub.7: 311.1107. Found: 311.1109.
Example 15
##STR00037##
[0081] To a solution of 8 (29 mg, 0.17 mmol) in DMF (1 mL) were
added aniline (45 .mu.L, 0.50 mmol), HBTU (64 mg, 0.17 mmol),
N,N-diisopropylethylamine (59 .mu.L, 0.34 mmol) at 0.degree. C.,
and the mixture was stirred at rt for 8 h. The mixture was diluted
with AcOEt and partitioned between AcOEt and aq. HCl (0.5 M). The
organic layer was washed with sat. aq. NaHCO.sub.3, brine, dried
(Na.sub.2SO.sub.4), and evaporated. The residue was purified by
silica gel column chromatography (50%/AcOEt in hexane) to give 17
(23 mg, 55%) as a colorless oil. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 8.03 (br s, 1H, NH), 7.54 (d, 2H, aromatic, J=7.8 Hz), 7.33
(t, 2H, aromatic, J=7.8, 8.2 Hz), 7.13 (t, 1H, aromatic, J=8.2 Hz),
4.51 (m, 2H, CH.sub.2CH.sub.2O), 2.85 (s, 2H, OC(O)CH.sub.2), 2.52
(m, 1H, CH.sub.2CH.sub.2O), 2.20 (m, 1H, CH.sub.2CH.sub.2O), 1.64
(s, 3H, CH.sub.3); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
166.1, 148.9, 137.4, 129.0, 124.7, 120.1, 82.4, 65.0, 48.4, 30.7,
25.6; HRMS (pos. ion ESI) m/z calcd for (M+Na)
C.sub.13H.sub.15NNaO.sub.4: 272.0899. Found: 272.0894.
Example 16
##STR00038##
[0083] 18 (27 mg, 61%, colorless oil) was prepared from 8 (29 mg,
0.17 mmol) as described for the preparation of 17 using benzyl
amine instead of aniline. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
7.26-7.35 (m, 5H, aromatic), 6.47 (br s, 1H, NH), 4.38-4.46 (m, 4H,
CH.sub.2CH.sub.2O and benzyl-CH.sub.2), 2.68 (d, 1H, OC(O)CH.sub.2,
J=14.5 Hz), 2.63 (d, 1H, OC(O)CH.sub.2, J=14.5 Hz), 2.46 (m, 1H,
CH.sub.2CH.sub.2O), 2.09 (m, 1H, CH.sub.2CH.sub.2O), 1.55 (s, 3H,
CH.sub.3); HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.14H.sub.17INNaO.sub.4: 286.1055. Found: 286.1052.
Example 17
##STR00039##
[0085] 19 (19 mg, 38%, colorless oil) was prepared from 8 (29 mg,
0.17 mmol) as described for the preparation of 17 using
4-fluorobenzyl amine instead of aniline. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 7.24-7.27 (m, 2H, aromatic), 6.99-7.02 (m, 2H,
aromatic), 6.70 (br s, 1H, NH), 4.37-4.44 (m, 4H, CH.sub.2CH.sub.2O
and benzyl-CH.sub.2), 2.66 (s, 2H, OC(O)CH.sub.2), 2.45 (m, 1H,
CH.sub.2CH.sub.2O), 2.10 (m, 1H, CH.sub.2CH.sub.2O), 1.54 (s, 3H,
CH.sub.3); HRMS (pos. ion ESI) m/z calcd for
(M+Na)+C.sub.14H.sub.16FNNaO.sub.4: 304.1961. Found: 304.0960.
Example 18
##STR00040##
[0087] To a solution of ethyl fluoroacetate (1.94 mL, 20.0 mmol) in
Et.sub.2O (120 mL) was added allylmagnesium bromide (1.0 M solution
in Et.sub.2O. 39.0 mL, 39.0 mmol) at 0.degree. C., and the mixture
was stirred at 0.degree. C. for 20 min. The reaction was quenched
with addition of sat. aq. NH.sub.4Cl, and separated. The organic
layer was washed with brine, dried (Na.sub.2SO.sub.4), and
evaporated. The residue was used to the next reaction without
further purification. The crude product 35 (2.60 g) was dissolved
in CH.sub.2Cl.sub.2 (40 mL) and cooled to -78.degree. C. Ozone was
bubbled into the solution at -78.degree. C. for 30 min until the
color of the solution turned to light purple. Oxygen was bubbled
into the solution for 20 min to remove ozone, and the solution was
warmed to room temperature. Acetic acid (20 mL) was added to the
solution and then the solvent was reduced in vacuo until the amount
of the solution was a few milliliters. To the residue were added
acetic acid (15 mL), H.sub.2O (15 mL), conc. H.sub.2SO.sub.4 (0.40
mL), and aq. H.sub.2O.sub.2 (30%, 9.0 mL), and the mixture was
stirred under reflux for 4 h. After cooling to room temperature,
the mixture was neutralized with BaCO.sub.3 (1.5 g) and filtered
through Celite with acetone. To the filtrate was added Pd/C (30 mg)
and the mixture was stirred at rt for 8 h to decompose
H.sub.2O.sub.2. The mixture was filtered through Celite with
acetone to remove Pd/C and the filtrate was evaporated. The residue
was co-evaporated with H.sub.2O (.times.2) and toluene (.times.3)
to give the crude of dicarboxylic acid 22 as a brown oil. To a
solution of the crude product 22 in DMF (80 mL) were added benzyl
bromide (98%, 4.85 mL, 40 mmol) and K.sub.2CO.sub.3 (5.53 g, 40
mmol), and the mixture was stirred at rt for 8 h. After filtration
through Celite to remove K.sub.2CO.sub.3, the solvent was
evaporated. The residue was partitioned between AcOEt and aq. HCl
(0.5 M). The organic layer was washed with sat. aq. NaHCO.sub.3 and
brine, dried (Na.sub.2SO.sub.4), and evaporated. The residue was
purified by silica gel column chromatography (10% to 20% AcOEt in
hexane) to give diester 23 (3.87 g, 54% for 4 steps) as a light
yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.32-7.38 (m,
10H, aromatic), 5.14 (s, 4H, benzyl-CH.sub.2.times.2), 4.42 (d, 2H,
CH.sub.2F, J=47 Hz), 4.25 (s, 1H, OH), 2.78 (m, 4H,
C(O)CH.sub.2.times.2); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
171.0, 135.3, 128.6, 128.5, 128.3, 86.4 (d), 71.1, 66.8, 39.9; HRMS
(pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.20H.sub.21FNaO.sub.5: 383.1271. Found: 383.1276.
Example 19
##STR00041##
[0089] To a solution of 23 (280 mg, 0.777 mmol) in THF (8 mL) was
added DIBAL-H (1.0 M solution in THF, 0.932 mL, 0.932 mmol) at 0
OC, and the mixture was stirred at 0.degree. C. for 10 min. DIBAL-H
(1.0 M solution in THF, 1.86 mL, 1.86 mmol) was added to the
mixture at 0.degree. C. and the resulting mixture was stirred at
0.degree. C. for 15 min. The reaction was quenched with addition of
aq. HCl (1.5 M) that was saturated with NaCl, and extracted with
AcOEt. The organic layer was washed with aq. HCl (1.5 M), which was
saturated with NaCl, sat. aq. NaHCO.sub.3, and brine, and then
dried (Na.sub.2SO.sub.4) and evaporated. After the residue was
dissolved in CH.sub.2Cl.sub.2 (60 mL), pyridine (285 .mu.L, 3.50
mmol) was added to the solution and the mixture was cooled at
0.degree. C. To the mixture was added a solution of triphosgene
(98%, 706 mg, 2.33 mmol) in CH.sub.2Cl.sub.2 (4 mL), and the
resulting mixture was stirred at 0 OC for 30 min. The reaction was
quenched with addition of sat. aq. NH.sub.4Cl, and extracted with
AcOEt. The organic layer was washed with brine, dried
(Na.sub.2SO.sub.4), and evaporated. The residue was purified by
silica gel column chromatography (33% to 50% AcOEt in hexane) to
give 25 (81 mg, 37% for 2 steps) as a colorless oil. .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. 7.33-7.40 (m, 5H, aromatic), 5.15 (s,
2H, benzyl-CH.sub.2), 4.62 (dd, 1H, CH.sub.2F, J=10.0, 12.6 Hz),
4.54 (dd, 1H, CH.sub.2F, J=10.0, 12.2 Hz), 4.40 (m, 2H,
CH.sub.2CH.sub.2O), 2.86 (m, 2H, C(O)CH.sub.2), 2.37 (m, 1H,
CH.sub.2CH.sub.2O), 2.30 (m, 1H, CH.sub.2CH.sub.2O); .sup.13C NMR
(125 MHz, CDCl.sub.3) .delta. 168.0, 148.1, 134.9, 128.7, 128.7,
128.5, 85.0 (d), 81.1, 67.2, 64.1, 39.8, 25.9; HRMS (pos. ion ESI)
m/z calcd for (M+Na).sup.+ C.sub.14H.sub.15FNaO.sub.5: 305.0801.
Found: 305.0800.
Example 20
##STR00042##
[0091] 26 (249 mg, 99%, colorless oil) was prepared from 25 (368
mg, 1.30 mmol) as described for the preparation of 8. .sup.1H NMR
(500 MHz, acetone-d.sub.6) .delta. 4.72 (d, 2H, CH.sub.2F, J=48
Hz), 4.48 (m, 2H, CH.sub.2CH.sub.2O), 2.94 (m, 2H, C(O)CH.sub.2),
2.47 (m, 1H, CH.sub.2CH.sub.2O), 2.32 (m, 1H, CH.sub.2CH.sub.2O);
.sup.13C NMR (125 MHz, acetone-d.sub.6) .delta. 170.6, 148.7, 86.4
(d), 82.3, 64.9, 39.4, 26.4; HRMS (pos. ion ESI) m/z calcd for
(M+Na).sup.+ C.sub.7H9FNaO.sub.5: 215.0332. Found: 215.0330.
Example 21
##STR00043##
[0093] 27 (32 mg, 60%/o, white solid) was prepared from 26 (50 mg,
0.26 mmol) as described for the preparation of 12 using NaHCO.sub.3
instead of K.sub.2CO.sub.3. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 4.60 (d, 2H, CH.sub.2F, J=47 Hz), 4.44 (m, 2H,
CH.sub.2CH.sub.2O), 3.73 (s, 3H, OCH.sub.3), 2.84 (m, 2H,
C(O)CH.sub.2), 2.39 (m, 1H, CH.sub.2CH.sub.2O), 2.32 (m, 1H,
CH.sub.2CH.sub.2O); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
168.7, 148.1, 85.1 (d), 81.1, 64.2, 52.4, 39.7, 26.1; HRMS (pos.
ion EST) m/z calcd for (M+Na).sup.+ C.sub.8H.sub.11FNaO.sub.5:
229.0488. Found: 229.0487. mp. 42.0-42.5.degree. C.
Example 22
##STR00044##
[0095] 28 (31 mg, 58%, colorless oil) was prepared from 26 (46 mg,
0.24 mmol) as described for the preparation of 12 using iodoethane
instead of iodomethane. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
4.60 (d, 2H, CHF, J=48 Hz), 4.45 (m, 2H, CH.sub.2CH.sub.2O), 4.18
(q, 2H. CH.sub.3CH.sub.2O, J=7.2 Hz), 2.82 (m, 2H, C(O)CH.sub.2),
2.40 (m, 1H, CH.sub.2CH.sub.2O), 2.32 (m, 1H, CH.sub.2CH.sub.2O),
1.29 (t, 3H, CH.sub.3CH.sub.2O, J=7.2 Hz); .sup.13C NMR (125 MHz,
CDCl.sub.3) .delta. 168.2, 148.2, 85.1 (d), 81.1, 64.2, 39.9, 29.7,
26.0, 14.1; HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.9H.sub.13FNaO.sub.5: 243.0645. Found: 243.0644.
Example 23
##STR00045##
[0097] 29 (41 mg, 77%, colorless oil) was prepared from 26 (34 mg,
0.18 mmol) as described for the preparation of 12 using
4-fluorobenzyl bromide instead of iodomethane. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. 7.33-7.36 (m, 2H, aromatic), 7.05-7.08 (m,
2H, aromatic), 5.12 (s, 2H, benzyl-CH.sub.2), 4.61 (dd, 1H,
CH.sub.2F, J=10.0, 14.0 Hz), 4.52 (dd, 1H, CH.sub.2F, J=10.0, 13.6
Hz), 4.42 (m, 2H, CH.sub.2CH.sub.2O), 2.86 (m, 2H, C(O)CH.sub.2),
2.35 (m, 1H, CH.sub.2CH.sub.2O), 2.28 (m, 1H, CH.sub.2CH.sub.2O);
.sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 168.0, 162.8 (d), 148.0,
130.8, 130.7, 115.7, 85.0 (d), 81.0, 66.5, 64.1, 40.0, 26.1; HRMS
(pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.14H.sub.14F.sub.2NaO.sub.5: 323.0707. Found: 323.0707.
Example 24
##STR00046##
[0099] 30 (36 mg, 72%, colorless oil) was prepared from 26 (30 mg,
0.16 mmol) as described for the preparation of 12 using
2,4-difluorobenzyl bromide instead of iodomethane. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. 7.38 (m, 1H, aromatic), 6.83-6.92 (m, 2H,
aromatic), 5.17 (s, 2H, benzyl-CH.sub.2), 4.62 (dd, 1H, CH.sub.2F,
J=10.0, 13.1 Hz), 4.52 (dd, 1H, CH.sub.2F, J=10.0, 12.8 Hz), 4.42
(m, 2H, CH.sub.2CH.sub.2O), 2.85 (m, 2H, C(O)CH.sub.2), 2.37 (m,
1H, CH.sub.2CH.sub.2O), 2.29 (m, 1H, CH.sub.2CH.sub.2O); .sup.13C
NMR (125 MHz, CDCl.sub.3) .delta. 167.9, 163.4 (d), 161.4 (d),
148.0, 132.2, 118.2, 111.7, 104.2, 85.0 (d), 81.0, 64.2, 60.6,
39.9, 26.0; HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.14H.sub.13F.sub.3NaO.sub.5: 341.0613. Found: 341.0612.
Example 25
##STR00047##
[0101] To a solution of lithium diisopropylamide (1.8 M solution in
heptane/THF/ethyl benzene, 3.05 mL, 5.50 mmol) in THF (55 mL) was
added benzyl acetate (784 .mu.L, 5.50 mmol) at -78.degree. C., and
the mixture was stirred at -78.degree. C. for 30 min. A solution of
4-ethoxy-1,1,1-trifluoro-3-buten-2-one (712 .mu.L, 5.00 mmol) in
THF (5 mL) was added to the mixture via canule at -78.degree. C.
and the resulting mixture was stirred at -78 OC for 30 min. After
the addition of sat. aq. NH.sub.4Cl, the mixture was warmed to room
temperature, and evaporated. The residue was partitioned between
AcOEt and sat. aq. NH.sub.4Cl. The organic layer was washed with
brine, dried (Na.sub.2SO.sub.4), and evaporated. The residue was
purified by silica gel column chromatography (6% AcOEt in hexane)
to give 32 (1.36 g, 86%) as a light yellow oil. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. 7.33-7.40 (m, 5H, aromatic), 6.74 (d, 1H,
CH.dbd.CHOEt, J=12.6 Hz), 5.20 (d, 1H, benzyl-CH.sub.2, J=14.1 Hz),
5.16 (d, 1H, benzyl-CH.sub.2, J=14.1 Hz), 4.76 (s, 1H, OH), 4.71
(d, 1H, CH.dbd.CHOEt, J=12.6 Hz), 3.70 (m, 2H, CH.sub.2CH.sub.3),
2.86 (d, 1H, C(O)CH.sub.2, J=15.6 Hz), 2.71 (d, 1H, C(O)CH.sub.2,
J=15.6 Hz), 1.27 (t, 3H, CH.sub.2CH.sub.3, J=7.0 Hz); .sup.13C NMR
(125 MHz, CDCl.sub.3) .delta. 171.2, 151.8, 134.8, 128.7, 128.5,
128.3, 124.6 (q), 99.4, 73.3 (q), 67.5, 65.5, 38.9, 14.7; HRMS
(pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.15H.sub.17F.sub.3NaO.sub.4: 341.0977. Found: 341.0978.
Example 26
##STR00048##
[0103] To a solution of 32 (334 mg, 1.05 mmol) in acetone (16 mL)
was added aq. HCl (12 M, 4 mL) at 0.degree. C., and the mixture was
stirred vigorously at 0 OC for 8 min. To the mixture was added sat.
aq. NaHCO.sub.3 to neutralize, and extracted with AcOEt. The
organic layer was washed with sat. aq. NaHCO.sub.3 and brine, dried
(Na.sub.2SO.sub.4), and evaporated. The residue was purified by
silica gel column chromatography (20%/o AcOEt in hexane) to give 33
(243 mg, 80%) as a light yellow oil. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 9.85 (s, 1H, CHO), 7.34-7.40 (m, 5H, aromatic),
5.35 (s, 1H, OH), 5.19 (s, 2H, benzyl-CH.sub.2), 2.75-2.93 (m, 4H,
CH.sub.2CHO and C(O)CH.sub.2); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta. 199.0, 170.8, 134.5, 128.8, 128.7, 128.5, 124.8 (q), 73.3
(q), 67.7, 46.1, 36.9; HRMS (pos. ion ESI) m/z calcd for
(M+Na).sup.+ C.sub.13H.sub.13F.sub.3NaO.sub.4: 313.0664. Found:
313.0660.
Example 27
##STR00049##
[0105] To a solution of 33 (320 mg, 1.10 mmol) in benzene (10 mL)
was added sodium triacetoxyborohydride (95%, 701 mg, 3.31 mmol) at
0.degree. C. and the mixture was stirred at rt for 2 h. The mixture
was quenched with addition of sat. aq. NaHCO.sub.3, and extracted
with AcOEt. The organic layer was washed with sat. aq. NaHCO.sub.3,
brine, dried (Na.sub.2SO.sub.4), and evaporated. After the residue
was dissolved in CH.sub.2Cl.sub.2 (10 mL), pyridine (125 .mu.L,
1.54 mmol) was added to the solution and the mixture was cooled at
0.degree. C. To the mixture was added a solution of triphosgene
(98%, 400 mg, 1.32 mmol) in CH.sub.2Cl.sub.2 (2 mL) at 0.degree.
C., and the resulting mixture was stirred at 0.degree. C. for 30
min. The reaction was quenched with addition of sat. aq.
NH.sub.4Cl, and extracted with AcOEt. The organic layer was washed
with brine, dried (Na.sub.2SO.sub.4), and evaporated. The residue
was purified by silica gel column chromatography (33% AcOEt in
hexane) to give 35 (281 mg, 80% for 2 steps) as a colorless solid.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.34-7.41 (m, 5H,
aromatic), 5.18 (s, 2H, benzyl-CH.sub.2), 4.43 (m, 1H,
CH.sub.2CH.sub.2O), 4.37 (m, 1H, CH.sub.2CH.sub.2O), 3.10 (d, 1H,
C(O)CH.sub.2, J=16.5 Hz), 2.87 (d, 1H, C(O)CH.sub.2, J=16.5 Hz),
2.74 (m, 1H, CH.sub.2CH.sub.2O), 2.33 (m, 1H, CH.sub.2CH.sub.2O);
.sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 167.2, 146.8, 134.8,
128.8, 128.7, 128.5, 123.6 (q), 80.2 (q), 67.3, 64.1, 37.9, 24.2;
HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.14H.sub.13F.sub.3NaO.sub.5: 341.0613. Found: 341.0613. mp.
87.5-88.degree. C.
Example 28
##STR00050##
[0107] 36 (176 mg, 98%, colorless oil) was prepared from 35 (250
mg, 0.786 mmol) as described for the preparation of 8. .sup.1H NMR
(500 MHz, D.sub.2O) .delta. 4.61 (m, 2H, CH.sub.2CH.sub.2O), 3.22
(d, 1H, C(O)CH.sub.2, J=16.5 Hz), 3.12 (d, 1H, C(O)CH.sub.2, J=16.5
Hz), 2.76 (m, 1H, CH.sub.2CH.sub.2O), 2.59 (m, 1H,
CH.sub.2CH.sub.2O); .sup.13C NMR (125 MHz, D.sub.2O) .delta. 171.4,
150.4, 123.4 (q), 81.1 (q), 65.3, 36.8, 23.5; HRMS (pos. ion ESI)
m/z calcd for (M+Na).sup.+ C.sub.7H.sub.7F.sub.3NaO.sub.5:
251.0143. Found: 251.0140.
Example 29
##STR00051##
[0109] Ester 37 (36 mg, 76%, colorless oil) was prepared from 36
(45 mg, 0.20 mmol) as described for the preparation of 27. .sup.1H
NMR (500 MHz, CDCl.sub.3) .delta. 4.48 (m, 2H, CH.sub.2CH.sub.2O),
3.76 (s, 3H, OCH.sub.3), 3.07 (d, 1H, C(O)CH.sub.2, J=16.4 Hz),
2.86 (d, 1H, C(O)CH.sub.2, J=16.4 Hz), 2.78 (m, 1H,
CH.sub.2CH.sub.2O), 2.37 (m, 1H, CH.sub.2CH.sub.2O); .sup.13C NMR
(125 MHz, CDCl.sub.3) .delta. 167.7, 146.8, 123.6 (q), 80.1 (q),
64.1, 52.6, 37.5, 24.1; HRMS (pos. ion ESI) m/z calcd for
(M+Na).sup.+ C.sub.8H.sub.9F.sub.3NaO.sub.5: 265.0300. Found:
265.0301.
Example 30
##STR00052##
[0111] Ester 38 (34 mg, 71%, colorless oil) was prepared from 36
(43 mg, 0.19 mmol) as described for the preparation of 27. .sup.1H
NMR (500 MHz, CDCl.sub.3) .delta. 4.47 (m, 2H, CH.sub.2CH.sub.2O),
4.21 (q, 2H, CH.sub.3CH.sub.2O, J=7.1 Hz), 3.05 (d, 1H,
C(O)CH.sub.2, J=16.3 Hz), 2.84 (d, 1H, C(O)CH.sub.2, J=16.3 Hz),
2.78 (m, 1H, CH.sub.2CH.sub.2O), 2.36 (m, 1H, CH.sub.2CH.sub.2O),
1.29 (t, 3H, CH.sub.3CH.sub.2O, J=7.1 Hz); .sup.13C NMR (125 MHz,
CDCl.sub.3) .delta. 167.3, 146.8, 123.6 (q), 80.2 (q), 64.2, 61.9,
37.8, 24.2, 14.0; HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.9H.sub.11F.sub.3NaO.sub.5: 279.0456. Found: 279.0456.
Example 31
##STR00053##
[0113] Ester 39 (48 mg, 88%, white solid) was prepared from 36 (39
mg, 0.17 mmol) as described for the preparation of 27. .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. 7.35 (m, 2H, aromatic), 7.07 (m, 2H,
aromatic), 5.14 (s, 2H, benzyl-CH.sub.2), 4.37-4.47 (m, 2H,
CH.sub.2CH.sub.2O), 3.09 (d, 1H, C(O)CH.sub.2, J=16.4 Hz), 2.86 (d,
1H, C(O)CH.sub.2, J=16.4 Hz), 2.73 (m, 1H, CH.sub.2CH.sub.2O), 2.33
(m, 1H, CH.sub.2CH.sub.2O); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta. 167.1, 162.9 (d), 146.7, 130.8, 130.7, 123.6 (q), 115.6,
80.1 (q), 66.8, 64.0, 37.7, 24.1; HRMS (pos. ion ESI) m/z calcd for
(M+Na).sup.+ C.sub.14H.sub.12F.sub.4NaO.sub.5: 359.0519. Found:
359.0518. mp. 84-85.degree. C.
Example 32
##STR00054##
[0115] Ester 40 (45 mg, 85%, colorless oil) was prepared from 36
(36 mg, 0.16 mmol) as described for the preparation of 27. .sup.1H
NMR (500 MHz, CDCl.sub.3) .delta. 7.38 (m, 1H, aromatic), 6.88 (m,
2H, aromatic), 5.21 (d, 1H, benzyl-CH.sub.2, J=12.2 Hz), 5.17 (d,
1H, benzyl-CH.sub.2, J=12.2 Hz), 4.45 (m, 2H, CH.sub.2CH.sub.2O),
3.09 (d, 1H, C(O)CH.sub.2, J=16.4 Hz), 2.87 (d, 1H, C(O)CH.sub.2,
J=16.4 Hz), 2.75 (m, 1H, CH.sub.2CH.sub.2O), 2.35 (m, 1H,
CH.sub.2CH.sub.2O); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
167.0, 163.4 (d), 161.4 (d), 146.6, 132.3, 123.5 (q), 118.0, 111.6,
104.2, 80.0 (q), 64.1, 60.9, 37.6, 24.1; HRMS (pos. ion ESI) m/z
calcd for (M+Na).sup.+ C.sub.14H.sub.11FNaO.sub.5: 377.0424. Found:
377.0427.
Example 33
##STR00055##
[0117] Ester 41 (54 mg, 75%, colorless oil) was prepared from 36
(50 mg, 0.22 mmol) as described for the preparation of 27. .sup.1H
NMR (500 MHz, CDCl.sub.3) .delta. 7.69-7.25 (m, 5H, aromatic), 4.55
(d, 1H), 4.44 (m, 2H), 3.82 (m, 2H), 3.02 (m, 2H), 2.92 (m, 2H),
2.65 (m, 1H); 13C NMR (125 MHz, CDCl.sub.3) .delta. 165.53, 161.86,
141.62 (q), 131.28, 128.87 (d), 126.83, 124.21, 121.70, 66.01 (d),
61.05, 34.88, 29.96, 21.79; MS (pos. ion ESI) m/z calcd for
(M+H).sup.+ C.sub.14H.sub.11F.sub.5O.sub.5: 333.09. Found:
333.18.
Example 34
##STR00056##
[0119] To a solution of 33 (267 mg, 0.919 mmol) in benzene (10 mL)
was added sodium triacetoxyborohydride (95%, 615 mg, 2.76 mmol) at
0.degree. C. and the mixture was stirred at rt for 2 h. The mixture
was quenched with addition of sat. aq. NaHCO.sub.3, and extracted
with AcOEt. The organic layer was washed with sat. aq. NaHCO.sub.3,
brine, dried (Na.sub.2SO.sub.4), and evaporated. After the residue
was dissolved in CH.sub.2Cl.sub.2 (10 mL), trifluoroacetic acid (1
mL) was added to the solution at 0.degree. C., and the mixture was
stirred at rt for 2 h. The mixture was diluted with AcOEt and
evaporated. The residue was purified by silica gel column
chromatography (33% AcOEt in hexane) to give 42 (135 mg, 80% for 2
steps) as a yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
4.61 (m, 1H, CH.sub.2CH.sub.2O), 4.45 (m, 1H, CH.sub.2CH.sub.2O),
2.86 (d, 1H, CH.sub.2C(O), J=14.7 Hz), 2.84 (br s, 1H, OH), 2.78
(d, 1H, CH.sub.2C(O), J=14.7 Hz), 2.24 (m, 1H, CH.sub.2CH.sub.2O),
2.07 (m, 1H, CH.sub.2CH.sub.2O); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta. 167.5, 124.7 (q), 71.5 (q), 64.4, 36.9, 28.6; HRMS (pos.
ion ESI) m/z calcd for (M+Na).sup.+ C.sub.6H.sub.7F.sub.3NaO.sub.3:
207.0245. Found: 207.0233.
Example 35
##STR00057##
[0121] To a solution of 42 (10 mg, 0.054 mmol) in H.sub.2O (1 mL)
was added KOH (.gtoreq.90%, 3.4 mg, 0.052 mmol) at rt, and the
mixture was stirred at 40.degree. C. for 2 h. The pH of the
solution was lowered to about pH 7-8 (detected by pH indicator
paper) with aq. HCl (0.1 M). The solvent was evaporated and
lyophilized to give 43 (15 mg) as a white powder including KCl.
.sup.1H NMR (500 MHz, D.sub.2O) .delta. 3.79 (m, 2H,
CH.sub.2CH.sub.2O), 2.60 (d, 1H, C(O)CH.sub.2, J=15.5 Hz), 2.52 (d,
1H, C(O)CH.sub.2, J=15.5 Hz), 2.04 (m, 1H, CH.sub.2CH.sub.2O), 1.96
(m, 1H, CH.sub.2CH.sub.2O); .sup.13C NMR (125 MHz, D.sub.2O)
.delta. 178.4, 125.9 (q), 73.2 (q), 56.4, 38.3, 35.9; LRMS (ESI)
m/z=225 (M+Na).sup.+.
Example 36
##STR00058##
[0123] To a solution of (.+-.)-mevalonolanctone (97%, 134 mg, 1.00
mmol) in DMF (1 mL) was added benzyl amine (131 .mu.L, 1.20 mmol)
at rt, and the mixture was stirred at 80.degree. C. for 12 h. After
evaporated, the residue was partitioned between AcOEt and HO0. The
organic layer was washed with brine, dried (Na.sub.2SO.sub.4), and
evaporated. The residue (light yellow oil) was used without further
purification. LRMS (ESI) m/z=260 (M+Na).sup.+.
Example 37
##STR00059##
[0125] To a solution of the crude product 44 (55 mg, .ltoreq.0.23
mmol) in CH.sub.2Cl.sub.2 (2 mL) were added 2,2-dimethoxypropane
(112 .mu.L, 0.91 mmol) and camphor sulfonic acid (4 mg, 0.02 mmol)
at 0 OC and the mixture was stirred at rt for 16 h. After addition
of sat. aq. NaHCO.sub.3, the mixture was extracted with AcOEt. The
organic layer was washed with brine, dried (Na.sub.2SO.sub.4), and
evaporated. The residue was purified by silica gel column
chromatography (50% AcOEt in hexane) to give 47 (17 mg, 27% for 2
steps) as a colorless oil. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 7.27-7.35 (m, 5H, aromatic), 6.97 (br s, 1H, NH), 4.53 (dd,
1H, benzyl-CH.sub.2, J=6.0, 14.8 Hz), 4.39 (dd, 1H,
benzyl-CH.sub.2, J=5.3, 14.8 Hz), 4.02 (m, 1H, CH.sub.2CH.sub.2O),
3.83 (m, 1H, CH.sub.2CH.sub.2O), 2.49 (d, 1H, NHC(O)CH.sub.2,
J=14.4 Hz), 2.44 (d, 1H, NHC(O)CH.sub.2, J=14.4 Hz), 1.91 (m, 1H,
CH.sub.2CH.sub.2O), 1.51 (m, 1H, CH.sub.2CH.sub.2O), 1.41 (s, 6H,
C(CH.sub.3).sub.2), 1.23 (s, 3H, CH.sub.3); .sup.13C NMR (125 MHz,
CDCl.sub.3) .delta. 170.3, 138.4, 128.6, 127.7, 127.4, 98.5, 72.0,
56.5, 51.1, 43.4, 33.3, 29.9, 26.7, 25.7; HRMS (pos. ion ESI) m/z
calcd for (M+Na).sup.+ C.sub.16H.sub.23NNaO.sub.3: 300.1576. Found:
300.1586.
Example 38
##STR00060##
[0127] To a solution of the crude product 44 (54 mg, .ltoreq.0.23
mmol) in CH.sub.2Cl.sub.2 (2 mL) were added benzaldehyde demethyl
acetal (51 .mu.L, 0.34 mmol) and camphor sulfonic acid (4 mg, 0.02
mmol) at 0 OC and the mixture was stirred at rt for 16 h. After
addition of sat. aq. NaHCO.sub.3, the mixture was extracted with
AcOEt. The organic layer was washed with brine, dried
(Na.sub.2SO.sub.4), and evaporated. The residue was purified by
silica gel column chromatography (33-50% AcOEt in hexane) to give
49 (25 mg, 34% for 2 steps) as a white solid. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 7.20-7.32 (m, 10H, aromatic), 6.83 (br s, 1H,
NH), 5.72 (s, 1H, benzyl-CH), 4.47 (dd, 1H, benzyl-CH.sub.2, J=5.8,
14.6 Hz), 4.38 (dd, 1H, benzyl-CH.sub.2, J=5.3, 14.6 Hz), 4.14 (m,
2H, CH.sub.2CH.sub.2O), 2.55 (s, 2H, NHC(O)CH.sub.2), 2.16 (m, 1H,
CH.sub.2CH.sub.2O), 1.53 (s, 3H, CH.sub.3), 1.46 (m, 1H,
CH.sub.2CH.sub.2O); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
169.7, 138.3, 138.1, 128.9, 128.7, 128.4, 127.9, 127.4, 125.8,
95.3, 73.4, 63.2, 50.8, 43.6, 33.6, 20.3; HRMS (pos. ion ESI) m/z
calcd for (M+Na).sup.+ C.sub.20H.sub.23NNaO.sub.3: 348.1576. Found:
348.1569.
Example 39
##STR00061##
[0129] To a solution of the crude product 44 (26 mg, .ltoreq.0.11
mmol) in CH.sub.2Cl.sub.2 were added chloromethyl methyl ether (84
.mu.L, 1.1 mmol), N,N-diisopropylethylamine (383 .mu.L, 2.2 mmol),
and DMAP (1.2 mg) at 0.degree. C. and the mixture was stirred at rt
for 2 h. After dilution with AcOEt, the mixture was partitioned
between AcOEt and 0.5 M aq. HCl. The organic layer was washed with
sat. aq. NaHCO3, brine, dried (Na2SO4), and evaporated. The residue
was dissolved in CH.sub.2Cl2 (17 mL), and BF3.Et2O (30 .mu.L, 0.24
mmol) was added at 0.degree. C. The mixture was stirred at rt for 8
h. After addition of sat. aq. NaHCO3, the mixture was extracted
with AcOEt. The organic layer was washed with brine, dried
(Na2SO4), and evaporated. The residue was purified by silica gel
column chromatography (50% AcOEt in hexane) to give 45 (21 mg, 77%
for 3 steps) as a white solid, 1H NMR (500 MHz, CDCl3) .delta.
7.25-7.35 (m, 5H, aromatic), 6.80 (br s, 1H, NH), 4.92 (d, 1H,
OCH2O, J=6.5 Hz), 4.85 (d, 1H, OCH2O, J=6.5 Hz), 4.48 (d, 2H,
benzyl-CH.sub.2, J=5.5 Hz), 3.95 (m, 1H, CH2CH2O), 3.89 (m, 1H,
CH2CH2O), 2.60 (d, 1H, NHC(O)CH.sub.2, J=14.5 Hz), 2.47 (d, 1H,
NHC(O)CH.sub.2, J=14.5 Hz), 2.02 (m, 1H, CH.sub.2CH.sub.2O), 1.48
(m, 1H, CH.sub.2CH.sub.2O), 1.40 (s, 3H, CH.sub.3); .sup.13C NMR
(125 MHz, CDCl.sub.3) .delta. 169.8, 138.5, 128.6, 127.5, 127.3,
87.8, 72.1, 62.8, 49.0, 43.3, 34.7, 20.9; HRMS (pos. ion ESI) m/z
calcd for (M+Na).sup.+ C.sub.14H.sub.19NNaO.sub.3: 272.1263. Found:
272.1259.
Example 40
##STR00062##
[0131] To a solution of the crude product 44 (24 mg, .ltoreq.0.10
mmol) in acetonitrile (1 mL) were added (Boc).sub.2O (99 mg, 0.45
mmol) and DMAP (1.2 mg, 0.01 mmol) at rt and the mixture was
stirred under reflux conditions for 16 h. After evaporated, the
residue was purified by silica gel column chromatography (50% AcOEt
in hexane) to give 54 (30 mg, 88% for 2 steps) as a colorless oil.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.27-7.36 (m, 5H,
aromatic), 6.23 (br s, 1H, NH), 4.65 (s, 1H, OH), 4.46 (m, 2H,
benzyl-CH.sub.2), 4.22 (m, 2H, CH.sub.2CH.sub.2O), 2.45 (d, 1H,
NHC(O)CH.sub.2, J=14.5 Hz), 2.34 (d, 1H, NHC(O)CH.sub.2, J=14.5
Hz), 1.90 (t, 2H, CH.sub.2CH.sub.2O, J=6.8 Hz), 1.45 (s, 9H,
C(CH.sub.3).sub.3), 1.28 (s, 3H, CH.sub.3); HRMS (pos. ion EST) m/z
calcd for (M+Na).sup.+ C.sub.18H.sub.27NNaO.sub.5: 360.1787. Found:
360.1785.
Example 41
##STR00063##
[0133] To a solution of 41 (120 mg, 0.652 mmol) in DMF was added
benzyl amine (142 .mu.L, 1.30 mmol), and the mixture was stirred at
80.degree. C. for 12 h. After the solvent was evaporated, the
residue was purified by silica gel column chromatography (50-100%
AcOEt in hexane) to give 44 (189 mg, 99%) as a colorless oil:
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.34-7.37 (m, 2H,
aromatic), 7.26-7.32 (m, 3H, aromatic), 6.62 (s, 1H, OH), 6.29 (br
s, 1H, NH), 4.49 (dd, 1H, benzyl-CH.sub.2, J=6.0, 14.5 Hz), 4.43
(dd, 1H, benzyl-CH.sub.2, J=5.5, 14.5 Hz), 3.95 (m, 1H,
CH.sub.2CH.sub.2O), 3.89 (m, 1H, CH.sub.2CH.sub.2O), 2.73 (d, 1H,
NHC(O)CH.sub.2, J=14.7 Hz), 2.51 (m, 1H, OH), 2.49 (d, 1H,
NHC(O)CH.sub.2, J=14.7 Hz), 2.09 (m, 1H, CH.sub.2CH.sub.2O), 1.84
(m, 1H, CH.sub.2CH.sub.2O); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta. 170.6, 137.2, 128.9, 128.7, 127.9, 125.7 (q), 75.0 (q),
58.4, 43.7, 38.0, 35.3; HRMS (pos. ion ESI) m/z calcd for
(M+Na).sup.+ C.sub.13H.sub.16F3NNaO.sub.3: 314.0980. Found:
314.0991.
Example 42
##STR00064##
[0135] 46 (26 mg, 69% for 2 steps, colorless oil) was prepared from
44 (36 mg, 0.12 mmol) as described for the preparation of 45.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.27-7.36 (m, 5H,
aromatic), 6.43 (br s, 1H, NH), 5.06 (d, 1H, OCH.sub.2O, J=6.3 Hz),
4.94 (d, 1H, OCH.sub.2O, J=6.3 Hz), 4.55 (dd, 1H, benzyl-CH.sub.2,
J=6.1, 14.9 Hz), 4.42 (dd, 1H, benzyl-CH.sub.2, J=6.0, 14.9 Hz),
3.99 (m, 2H, CH.sub.2CH.sub.2O), 2.76 (d, 1H, NHC(O)CH.sub.2,
J=14.4 Hz), 2.56 (d, 1H, NHC(O)CH.sub.2, J=14.4 Hz), 2.37 (m, 1H,
CH.sub.2CH.sub.2O), 1.98 (dt, 1H, CH.sub.2CH.sub.2O, J=3.7, 3.7,
14.7 Hz); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 167.4, 137.9,
129.1, 128.7, 128.6, 128.0, 127.5, 125.7 (q), 89.9, 73.4 (q), 62.5,
43.8, 41.2, 25.7; HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.14H.sub.16F.sub.3NNaO.sub.3: 326.0980. Found: 326.0970.
Example 43
##STR00065##
[0137] 55 (50 mg, 68%, colorless oil) were prepared from 44 (55 mg,
0.19 mmol) as described for the preparation of 54. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. 7.26-7.38 (m, 5H, aromatic), 6.55 (s, 1H,
OH), 6.05 (br s, 1H, NH), 4.53 (dd, 1H, benzyl-CH.sub.2, J=6.0,
14.7 Hz), 4.42 (dd, 1H, benzyl-CH.sub.2, J=5.5, 14.7 Hz), 4.29 (m,
2H, CH.sub.2CH.sub.2O), 2.69 (d, 1H, NHC(O)CH.sub.2, J=15.3 Hz),
2.50 (d, 1H, NHC(O)CH.sub.2, J=15.3 Hz), 2.15 (m, 1H,
CH.sub.2CH.sub.2O), 1.99 (m, 1H, CH.sub.2CH.sub.2O), 1.47 (s, 9H,
C(CH.sub.3).sub.3); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
170.8, 153.1, 137.0, 128.9, 127.9, 127.6, 125.6 (q), 82.5, 73.7
(q), 61.8, 43.8, 37.1, 33.6, 27.7; HRMS (pos. ion EST) m/z calcd
for (M+Na).sup.+ C.sub.18H.sub.24F.sub.3NNaO.sub.5: 414.1504.
Found: 414.1500.
Example 44
##STR00066##
[0139] 48 (20 mg, 32%, colorless oil) were prepared from 44 (55 mg,
0.19 mmol) as described for the preparation of 47. HRMS (pos. ion
EST) m/z calcd for (M+Na).sup.+ C.sub.16H.sub.20F.sub.3NNaO.sub.3:
354.1293. Found: 354.1290.
Example 45
##STR00067##
[0141] To a solution of 44 (60 mg, 0.20 mmol) in CH.sub.2Cl.sub.2
(2 mL) were added benzaldehyde dimethyl acetal (151 .mu.L, 1.00
mmol) and camphor sulfonic acid (4.7 mg, 0.02 mmol) at rt and the
mixture was stirred under reflux condition for 12 h. After addition
of sat. aq. NaHCO.sub.3, the mixture was extracted with AcOEt. The
organic layer was washed with brine, dried (Na.sub.2SO.sub.4), and
evaporated. The residue was purified by silica gel column
chromatography (10-25% AcOEt in hexane) to give 50a (25 mg, 33%,
CF.sub.3/Ph=anti) as a colorless oil or 50b (7 mg, 9%.
CF.sub.3/Ph=syn) as a white solid, 50a .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 7.25-7.35 (m, 8H, aromatic), 7.16-7.18 (m, 2H,
aromatic), 6.46 (br s, 1H, NH), 5.92 (s, 1H, benzylidene-CH), 4.41
(d, 2H, benzyl-CH.sub.2, J=5.6 Hz), 4.18 (m, 2H,
CH.sub.2CH.sub.2O), 2.73 (d, 1H, NHC(O)CH.sub.2, J=14.3 Hz), 2.57
(d, 1H, NHC(O)CH.sub.2, J=14.3 Hz), 2.50 (m, 1H,
CH.sub.2CH.sub.2O), 1.97 (m, 1H, CH.sub.2CH.sub.2O); .sup.13C NMR
(125 MHz, CDCl.sub.3) .delta. 167.3, 137.5, 129.4, 128.7, 128.5,
128.0, 127.5, 125.5 (q), 98.0, 74.7 (q), 63.2, 43.9, 42.8, 25.0;
HRMS (pos. ion ESI) m/z calcd for (M+Na)
C.sub.20H.sub.20F.sub.3NNaO.sub.3: 402.1293. Found: 412.1285. 50b
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.27-7.38 (m, 8H,
aromatic), 7.21-7.23 (m, 2H, aromatic), 6.29 (br s, 1H, NH), 5.78
(s, 1H, benzylidene-CH), 4.49 (dd, 1H, benzyl-CH.sub.2, J=5.9, 14.6
Hz), 4.36 (dd, 1H, benzyl-CH.sub.2, J=5.3, 14.6 Hz), 4.27 (m, 1H,
CH.sub.2CH.sub.2O), 4.19 (m, 1H, CH.sub.2CH.sub.2O), 3.01 (d, 1H,
NHC(O)CH.sub.2, J=15.2 Hz), 2.94 (d, 1H, NHC(O)CH.sub.2, J=15.2
Hz), 2.36 (m, 1H, CH.sub.2CH.sub.2O), 2.02 (m, 1H,
CH.sub.2CH.sub.2O); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
167.2, 137.5, 137.0, 129.4, 128.8, 128.4, 127.9, 127.7, 125.9 (q),
96.4, 76.0 (q), 62.5, 44.0, 36.6, 25.5; HRMS (pos. ion ESI) m/z
calcd for (M+Na).sup.+ C.sub.20H.sub.20F.sub.3NNaO.sub.3: 402.1293.
Found: 412.1283.
Example 46
##STR00068##
[0143] 51 (30 mg, 35%, white solid, CF3/PMB=anti) and 52 (22 mg,
25%, colorless oil, CF3/PMB=syn) were prepared from 44 (62 mg, 0.21
mmol) as described for the preparation of 71 using anisaldehyde
dimethylacetal instead of benzaldehyde demethyl acetal: 51 .sup.1H
NMR (500 MHz, CDCl.sub.3) .delta. 7.27-7.31 (m, 3H, aromatic),
7.17-7.20 (m, 4H, aromatic), 6.75 (d, 2H, aromatic, J=8.8 Hz), 6.50
(br s, 1H, NH), 5.86 (s, 1H, benzylidene-CH), 4.42 (d, 2H,
benzyl-CH.sub.2, J=5.6 Hz), 4.15 (m, 2H, CH.sub.2CH.sub.2O), 3.79
(s, 3H, OCH.sub.3), 2.73 (d, 1H, NHC(O)CH.sub.2, J=14.4 Hz), 2.56
(d, 1H, NHC(O)CH.sub.2, J=14.4 Hz), 2.46 (m, 1H,
CH.sub.2CH.sub.2O), 1.96 (m, 1H, CH.sub.2CH.sub.2O); 13C NMR (125
MHz, CDCl.sub.3) .delta. 167.4, 160.2, 137.6, 129.8, 128.7, 128.1,
127.5, 125.5 (q), 113.8, 97.9, 74.7 (q), 63.2, 55.3, 43.9, 42.8,
25.0; HRMS (pos. ion EST) m/z calcd for (M+Na).sup.+
C.sub.21H.sub.22F.sub.3NNaO.sub.4: 432.1399. Found: 432.1408.
[0144] 52 .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.28-7.30 (m,
5H, aromatic), 7.22 (m, 2H, aromatic), 6.84 (d, 2H, aromatic, J=8.8
Hz), 6.30 (br s, 1H, NH), 5.73 (s, 1H, benzylidene-CH), 4.51 (dd,
1H, benzyl-CH.sub.2, J=6.0, 14.6 Hz), 4.35 (dd, 1H,
benzyl-CH.sub.2, J=5.2, 14.6 Hz), 4.25 (m, 1H, CH.sub.2CH.sub.2O),
4.17 (m, 1H, CH.sub.2CH.sub.2O), 3.80 (s, 3H, OCH.sub.3), 2.99 (d,
1H, NHC(O)CH.sub.2, J=15.3 Hz), 2.95 (d, 1H, NHC(O)CH.sub.2, J=15.3
Hz), 2.34 (m, 1H, CH.sub.2CH.sub.2O), 1.98 (m, 1H,
CH.sub.2CH.sub.2O); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
167.2, 160.3, 137.5, 129.4, 128.8, 127.9, 127.6, 124.9 (q), 113.8,
96.3, 76.1 (q), 62.5, 55.3, 44.0, 36.5, 25.5; HRMS (pos. ion ESI)
m/z calcd for (M+Na).sup.+ C.sub.21H.sub.22F.sub.3NNaO.sub.4:
432.1399. Found: 432.1389.
Example 47
##STR00069##
[0146] To a solution of 44 (74 mg, 0.25 mmol) in benzene (3 mL)
were added 2,4-dimethoxybenzaldehyde (98%, 52 mg, 0.31 mmol),
camphor sulfonic acid (4.7 mg, 0.02 mmol), and molecular serves 4A
(powered, 18 mg) at rt and the mixture was stirred under reflux
condition for 36 h. After addition of sat. aq. NaHCO.sub.3, the
mixture was extracted with AcOEt. The organic layer was washed with
brine, dried (Na.sub.2SO.sub.4), and evaporated. The residue was
purified by preparative TLC (50% AcOEt in hexane.times.2) to give
53 (7 mg, 6%) as a colorless oil. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 7.52 (d, 1H, aromatic, J=8.5 Hz), 7.24-7.26 (m, 3H,
aromatic), 7.17-7.19 (m, 2H, aromatic), 6.57 (br s, 1H, NH), 6.52
(d, 1H, aromatic, J=8.5 Hz), 6.32 (s, 1H, aromatic), 6.10 (s, 1H,
benzylidene-CH), 4.44 (d, 2H, benzyl-CH.sub.2, J=5.4 Hz), 4.24 (m,
1H, CH.sub.7CH.sub.2O), 4.14 (m, 1H, CH.sub.2CH.sub.2O), 3.80 (s,
3H, OCH.sub.3), 3.57 (s, 3H, OCH.sub.3), 3.12 (d, 1H,
NHC(O)CH.sub.2, J=15.3 Hz), 2.90 (d, 1H, NHC(O)CH.sub.2, J=15.3
Hz), 2.28 (m, 1H, CH.sub.2CH.sub.2O), 1.88 (m, 1H,
CH.sub.2CH.sub.2O); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta.
167.7, 161.7, 157.3, 137.7, 129.4, 128.6, 128.4, 127.5, 127.3,
124.6 (q), 117.9, 104.9, 98.2, 91.1, 76.1 (q), 62.8, 55.4, 43.9,
36.6, 26.0; HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.22H.sub.24F.sub.3NNaO.sub.5: 462.1504. Found: 462.1503.
Example 48
##STR00070##
[0148] To a solution of 23 (500 mg, 1.39 mmol) in THF (10 mL) was
added LiBH.sub.4 (0.067 g, 3.08 mmol) at 0.degree. C., and the
mixture was stirred at 0.degree. C. for 30 min. The reaction was
quenched with addition of aq. HCl (1.5 M). The solution was
co-evaporated with toluene and the product was extracted from the
dry slurry with THF and the solvent was evaporated. The residue
(250 mg, 1.64 mmol) was dissolved in THF (5 mL) and stirred with
pyridine (0.132 mL, 1.64 mmol) and acetic anhydride (0.24 mL, 2.46
mmol) at room temperature for 8 h. The reaction was quenched with
aq. HCl (1.5 M), and extracted with AcOEt. The organic layer was
washed with aq. HCl (1.5 M), sat. NaHCO.sub.3, and brine, and then
dried (MgSO.sub.4) and evaporated. The residue was purified by
silica gel column chromatography (50% to 75% AcOEt in hexane) to
give 61 (239 mg, 75%) as a colorless oil. .sup.1H NMR (500 MHz,
CDCl.sub.3) 4.40 (dd, 1H, CHHF, J=9.4, 16.5 Hz), 4.29 (m, 3H), 3.94
(m, 2H), 2.06 (s, 3H), 1.97 (m, 2H), 1.85 (m, 2H); .sup.13C NMR
(125 MHz, CDCl.sub.3) .delta. 171.0, 87.3, 85.9, 73.2 (d), 60.3,
59.3, 36.8, 35.6, 21.1. To a solution of 61 (330 mg, 1.54 mmol) in
DMF (5 mL), K.sub.2CO.sub.3 (638 mg, 4.62 mmol) was added to the
mixture followed by the addition of methyl iodide (0.21 mL, 3.08
mmol) and the mixture was stirred for 24 h. The reaction was
quenched with aq. HCl (1.5 M) and extracted with ethyl acetate. The
organic layer was washed with aq. HCl (1.5 M) and brine, dried
(MgSO.sub.4) and evaporated. The residue was purified by silica gel
column chromatography (15% to 25% EtOAc in hexane) to give 63 (105
mg, 31%). .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 4.41 (dd, 1H,
J=9.91 Hz) 4.33-4.24 (m, 3H), 3.92 (s, 1H), 2.72-2.62 (dq, 2H),
2.07 (s, 3H), 1.95 (dt, 2H). .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta. 172.7, 170.9, 87.3, 85.9, 71.2, (d), 59.7, 52.1, 39.5 (d),
35.2, 21.0; HRMS (pos. ion ESI) m/z calcd for (M+Na).sup.+
C.sub.9H.sub.15FO.sub.5: 245.0801. Found: 245.0933.
Example 49
##STR00071##
[0150] To a solution of 62 (300 mg, 1.54 mmol) in DMF (8 mL) was
added pyridinium dichromate at room temperature and the mixture was
stirred at room temperature for 24 h. The reaction was quenched
with aq. HCl (1.5 M) and extracted with AcOEt. The organic layer
was washed with aq. HCl (1.5 M) and brine, then dried (MgSO.sub.4)
and evaporated. After the residue was dissolved in DMF (5 mL),
K.sub.2CO.sub.3 (638 mg, 4.62 mmol) was added to the mixture
followed by the addition of methyl iodide (0.21 mL, 3.08 mmol) and
the mixture was stirred for 24 h. The reaction was quenched with
aq. HCl (1.5 M) and extracted with AcOEt. The organic layer was
washed with aq. HCl (1.5 M) and brine, dried (MgSO.sub.4) and
evaporated. The residue was purified by silica gel column
chromatography (15% to 25% AcOEt in hexane) to give 64 (40 mg, 11%)
as a colorless oil. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 4.37
(dd, 1H), 4.30-4.23 (m, 3H), 4.19 (q, 2H, J=7.19), 3.99 (s, 1H),
2.62 (dq, 2H), 2.04 (s, 3H), 1.93 (m, 2H), 1.28 (t, 3H. J=7.19 Hz);
.sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 172.4, 170.9, 87.3,
85.9, 71.2, 61.2, 59.8, 39.7 (d), 35.2 (d), 21.0, 14.1; HRMS (pos.
ion ESI) m/z calcd for (M+Na).sup.+ C.sub.10H.sub.17FO.sub.5:
259.0957. Found: 259.1080
Example 50
##STR00072##
[0152] To a solution of 43 (R.dbd.CH.sub.2F, 200 mg, 1.35 mmol) in
DMF (5 mL) was added benzylamine (0.18 mL, 1.62 mmol) at room
temperature and the reaction was stirred at 60.degree. C. for 24 h.
The reaction was quenched by aq. HCl (1.5 M) and extracted with
AcOEt. The organic layer was washed with aq. HCl (1.5 M) and brine,
then dried (MgSO.sub.4) and evaporated. After the residue was
dissolved in THF, pyridine (0.11 mL, 1.35 mmol) was added at room
temperature. To the solution was added acetic anhydride (0.25 mL,
2.7 mmol) and the mixture was stirred at room temperature for 24 h.
The reaction was quenched with aq. HCl (1.5 M) and extracted with
AcOEt. The organic layer was washed with aq. HCl (1.5 M) and brine,
then dried (MgSO.sub.4) and evaporated. The residue was purified by
silica gel column chromatography (15% to 25% AcOEt in hexane) to
give 65 as a colorless oil. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 7.24 (m, 5H), 6.03 (s, 1H), 4.98 (s, 1H), 4.43-4.35 (m,
2H), 4.25 (s, 1H), 4.24-4.17 (m, 2H), 4.16 (s, 1H), 2.41 (m, 2H),
1.96 (s, 3H), 1.87-1.84 (m, 2H); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta. 171.5, 170.9, 137.4, 128.9, 127.8, 87.1, 85.7, 71.3, 59.8,
43.6, 40.7, 35.8, 21.0, HRMS (pos. ion ESI) m/z calcd for
(M+Na).sup.+ C.sub.15H.sub.20FNO.sub.4: 320.1274. Found:
320.1268.
* * * * *